October 17-21 2015 Baltimore Maryland

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ASRM 2015
SCIENTIFIC ABSTRACTS to be presented at the 71st Annual Meeting
of the American Society for Reproductive Medicine, October 17-21,
2015, Baltimore, Maryland.
(e2) ORAL SESSION
(e106) POSTER SESSION
(e362) AUTHOR INDEX
(e378) TOPIC INDEX
(e380) AUTHOR AND SPOUSE/PARTNER DISCLOSURES INDEX
October 17-21, 2015
Baltimore, Maryland
These abstracts of research studies, printed as submitted by the authors, are presented
in the ASRM 2015 meeting sessions and are published in the order of their
presentation. Abstracts of plenary lectures, symposia and interactive sessions are
not included.
Copyright ª2015 American Society for Reproductive Medicine,
1209 Montgomery Highway, Birmingham, Alabama 35216-2809

The first six papers are candidates for the ASRM Scientific Program
Prize Paper Awards. Six additional candidates will be presented during
the Prize Paper Candidates’ session on Tuesday.
SCIENTIFIC PROGRAM PRIZE PAPER SESSION 1
O-1 Monday, October 19, 2015 11:15 AM
DIETARY PROTEIN INTAKE AND REPRODUCTIVE HORMONES
AND OVULATION: THE BIOCYCLE STUDY. S. L. Mumford, a
A. Alohali, b J. Wactawski-Wende. c a NICHD, NIH, Rockville, MD; b George
Mason University, Arlington, VA; c University at Buffalo, Buffalo, NY.
OBJECTIVE: Protein intake has been associated with changes in steroidogenesis
in women with polycystic ovary syndrome, likely through reducing
hyperinsulinemia. However, the associations among premenopausal women
without a history of infertility or polycystic ovary syndrome are not well
characterized. The objective of this study was to evaluate the association between
protein intake and reproductive hormone concentrations and ovulation
in a healthy population.
DESIGN: Prospective cohort study of 259 healthy premenopausal women
followed for up to two menstrual cycles.
MATERIALS AND METHODS: Estradiol, progesterone, luteinizing hormone,
follicle-stimulating hormone, and testosterone were measured up to
eight times per cycle for up to two cycles, with visits scheduled using fertility
monitors to coincide with menstrual cycle phases. Percent energy from total
protein, animal protein, and vegetable protein were assessed by 24-hour
recall up to four times per cycle. Linear mixed models and generalized linear
mixed models were used to evaluate the association between protein intake
and reproductive hormone levels, and with ovulatory status (peak progesterone
%5ng/mL with no LH peak on the mid or late luteal phase visit), respectively.
All models were adjusted for total energy intake, age, body mass
index, race, fat intake, and physical activity.
RESULTS: Dietary protein consumption, specifically animal protein, was
found to be inversely associated with testosterone concentrations. In particular,
the highest tertile of percent energy from total protein intake was associated
with lower testosterone concentrations (beta -0.05, 95% CI -0.01,
-0.005) compared to the lowest tertile of intake. The highest tertile of percent
energy from animal protein intake was also associated with lower testosterone
(beta -0.02, 95% CI -0.04, -0.0001). No associations were observed
between protein intake and other hormones, including estradiol, progesterone,
luteinizing hormone, or follicle stimulating hormone levels, nor was
there an association between protein intake and ovulation.
CONCLUSIONS: These findings suggest that a diet high in protein, particularly
animal protein, is significantly associated with reduced testosterone
levels among healthy women. These results highlight the importance of
diet on reproductive function and the potential role of protein intake in
androgen synthesis.
Supported by: Intramural Research Program, DIPHR, NICHD, NIH.
O-2 Monday, October 19, 2015 11:30 AM
ROCKING THE DOGMA OF SEMINAL ROUND CELLS. Q. V. Neri,
T. Cozzubbo, Z. Rosenwaks, G. D. Palermo. Reproductive Medicine, Weill
Cornell Medical College, New York, NY.
OBJECTIVE: To revisit the origin and significance of the sporadic presence
of round cells in the ejaculates of men screened for male infertility.
DESIGN: In a prospective fashion, a total of 4,051 men undergoing male
infertility screening in a period of 24 months were included in the study. RC
cells were characterized for WBC components versus exfoliated germ cells
by testing for multiple markers of ploidy as well as protamine assays. Cases
displaying R2 million RC were screened for bacteria. The effect of RC on
clinical outcome was assessed in specimens used for ART.
MATERIALS AND METHODS: Raw specimens containing RC were
processed by peroxidase and other leukocyte assays, specific stains for protamines
were used to identify spermiogenic stage, aneuploidy (9 chromosome
FISH) assessment was carried out, and the presence of various Sertoli-cell
cytoplasmic remnants was analyzed to identify and characterize immature
germ cells (IGC).
RESULTS: A total of 4,810 ejaculated samples were processed for semen
analysis. The average age of the men involved was 39.2 7yrs. Semen samples
had a mean concentration of 40.7 31 million, motility of 42.6 35%,
and morphology of 2.3 2%. Round cells were identified in 261 of the specimens
evaluated, representing a proportion of 5.4%. Men presenting with
round cell had comparable age but lower sperm concentration and
morphology than the control (P < 0.0001). Aneuploidy was detected in 91
specimens, of which 30.8% (28/91) presented with round cells. The aneuploidy
rate of 4.3%, remarkably higher than the control (2.3%; P < 0.001).
Sperm aneuploidy rate positively correlate with the amount of RC (P ¼
0.0001).In 44 men, 17 of them in 18 cycles had up to 1.9 million RC without
affecting fertilization and clinical pregnancy rates when compared to control
(n¼365 cycles). In 27 men with 33 ICSI cycles with over R2 million round
cells, the fertilization rate trended lower and the miscarriage rate was significantly
increased (P ¼ 0.05).The absence of any correlation between RC and
bacteriological growth as well as the results of marker testing indicates that
seminal RC are mostly immature germ cells that stain for vimentin and
inhibin B. Moreover, their modest protamine content and their haploid status
confirm that they are post-meiotic. Sequential observation in the same man
showed that the RC episode was followed by an amelioration of the semen
parameters and that the presence of RC corresponds to flu season peaks.
CONCLUSIONS: Seminal round cell presence is not a marker of infectiousness
but rather a transient indicator of spermatogenic insult that possibly
occurs in most men following a mild and transient ailment such as the flu.
Supported by: WCMC.
O-3 Monday, October 19, 2015 11:45 AM
THE SUPEROVULATED ENVIRONMENT, INDEPENDENT OF
EMBRYO VITRIFICATION, RESULTS IN LOW BIRTHWEIGHT
FOLLOWING EMBRYO TRANSFER IN A MOUSE
MODEL. R. S. Weinerman, T. S. Ord, C. Coutifaris, M. A. Mainigi. Division
of Reproductive Endocrinology and Infertility, University of Pennsylvania,
Philadelphia, PA.
OBJECTIVE: Babies born following fresh embryo transfer are of lower
birthweight than babies born following frozen embryo transfer. The objective
of this study was to determine, in a mouse model, if this difference is due to
the superovulated (SO) peri-implantation environment or embryo vitrification
(VIT).
DESIGN: Laboratory research.
MATERIALS AND METHODS: Female CF1 mice were superovulated
with gonadotropins and mated to male mice heterozygous for green-fluorescent
protein (GFP). 2-pronuclear embryos were collected and cultured to the
blastocyst stage and a subset of embryos were vitrified. For each transfer
experiment, 10 blastocysts, 5 fresh and 5 vitrified/thawed, were transferred
into a host mouse, using GFP to tag the embryos as fresh or frozen. Transfers
were performed into pseudopregnant females created through either natural
mating to vasectomized males (n¼30) or superovulation followed by mating
to vasectomized males (n¼45). This resulted in 4 experimental groups: 1)
Natural environment, fresh embryos (Nat-Fresh) 2) Natural environment,
frozen embryos (Nat-VIT) 3) Superovulated environment, fresh embryos
(SO-Fresh) 4) Superovulated environment, frozen embryos (SO-VIT). Pregnant
mice were sacrificed near term (E18.5) for assessment of fetal and
placental weights and GFP status. An a-priori power calculation determined
that 17 fetuses per group would provide 80% power to detect a 15% difference
in fetal weight with an alpha of 0.05.
RESULTS: There was no difference in litter size between natural (n¼15
litters) and SO (n¼13 litters) hosts. Although there was no difference in
placental weight, there was a highly significant difference (p

O-4 Monday, October 19, 2015 12:00 PM
THE SYNCHRONIZATION OF THE LEIOMYOMA
EXTRACELLULAR MATRIX SIGNALING PATHWAYS OF
SURGICAL SPECIMENS IN RESPONSE TO ULIPRISTAL
ACETATE. M. Malik, a J. Cox, b J. Britten, a A. Patel, c L. K. Nieman, d
W. H. Catherino. a a Obstetrics and Gynecology, Uniformed Services University
of the Health Sciences, Bethesda, MD; b Program in Reproductive and
Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child
Health and Human Development, Bethesda, MD; c Henry Jackson Foundation,
Bethesda, MD; d Reproductive Biology and Medicine Branch, Eunice
Kennedy Shriver National Institute of Child Health and Human Development,
Bethesda, MD.
OBJECTIVE: To elucidate the pathways that may be involved in the action
of Ulipristal acetate, a selective progesterone receptor modulator, on leiomyoma
extracellular matrix (ECM) production and breakdown.
DESIGN: Uterine leiomyomas are characterized by increased stiffness as a
result of excessive and disordered ECM which contributes to the total bulk of
the tumor. Ulipristal acetate reduced leiomyoma size in several randomized
placebo-controlled studies. We have previously demonstrated that Ulipristal
acetate reduces the total ECM that forms the bulk of the tumor in over 60% of
patients studied. Clinical tissue was sent for RNASeq analysis to identify
pathways that may be involved in leiomyoma ECM breakdown. In-vitro analysis
was carried out to analyze the effect of the medroxyprogesterone acetate
(MPA) and Ulipristal acetate.
MATERIALS AND METHODS: RNA isolated from clinically collected
fibroid and myometrial tissue from patients treated with Ulipristal acetate
in a placebo-controlled, randomized trial, underwent RNASeq analysis
(Beckman Coulter Genomics Inc). To further evaluate the impact of Ulipristal
acetate, 3-dimensional leiomyoma cultures were treated with MPA, Ulipristal
acetate, and combinations. The expressions of ECM genes as well
as Wnt/b-catenin pathway genes were analyzed. Proteins were analyzed using
Western Blot.
RESULTS: Both TGFb3 and TGFb1 transcripts were reduced in tumors of
women treated with Ulipristal acetate. The reduction was also observed at
protein levels. Multiple components of the TGFb signaling pathway demonstrated
an overall reduction. Reduced transcript and protein expressions were
observed in matrix metalloproteinases (MMPs), such as MMP-9 (2.38-fold)
and MMP-13 (1.97-fold). Decreased WNT5a protein indicated an involvement
of the WNT/b-catenin pathway that was Supported by changes in
expression of Frizzled gene transcripts such as FZD6. In addition, there
was increased expression of aquaporins and other osmotic stress regulators
such as NFAT5, which are known to be regulated by WNT pathway.
Leiomyoma cells in 3D culture, in response to MPA, demonstrated a
3.23+/-0.15-fold increase in b-catenin expression. Ulipristal acetate alone
and in combination with MPA decreased b-catenin expression below that
of untreated leiomyoma cells.
CONCLUSIONS: TGFb signaling pathway in association with both
canonical and non-canonical WNT pathway may participate in Ulipristal
acetate clinical activity by reducing the total amount of ECM proteins produced,
and by decreasing osmotic stress. Ulipristal acetate may also reduce
the progesterone-mediated fibrosis.
Supported by: This research was Supported by Intramural grant from Uniformed
Services University of the Health Sciences, QP85GF13 and
RO85298815. The research was also Supported, in part, by the intramural
research Program in Reproductive and Adult Endocrinology, NIH; NIH
R21 and EMD Serono.
O-5 Monday, October 19, 2015 12:15 PM
ELECTIVE SINGLE EMBRYO TRANSFERS (ESET) AS COMPARED
TO COMPREHENSIVE CHROMOSOME SCREENING
(CCS). A. R. Anderson, D. Taylor, K. C. Chiles, U. Balthazar,
A. S. Browne. Reproductive Medicine Associates of Texas, San Antonio, TX.
OBJECTIVE: To investigate the association with eSET with optimal embryo
development and CCS testing for embryo selection.
DESIGN: A prospective cohort study.
MATERIALS AND METHODS: CCS aneuploidy testing was performed
on 297 embryo transfers. Alternately, 172 eSET blastocyst transfers were
completed based on at least two high quality day 5 embryos available for
transfer where one is transferred and at least one selected for cryopreservation
on day 5. In the CCS subgroup, 5 to 7 cells were biopsied from day 5 and day 6
blastocysts in a HEPES buffered Medium. Biopsied cells were placed in
separate PCR tubes and analyzed for 24 chromosomes at an outside reference
laboratory. All embryos in the CCS subgroup were subjected to vitrification
for a delayed embryo transfer in a subsequent frozen embryo cycle.
RESULTS: There was no significant difference in ongoing pregnancy,
average age, embryos transferred, or miscarriage rates when eSET criterion
or CCS was utilized for embryo selection. In the eSET group there were 172
transfers with 126 (73%) positive pregnancies, 25 (20%) losses, and 101
(59%) ongoing pregnancies. From 297 CCS embryo transfers there were
209 (70%) positive pregnancies, 47 (22%) losses, and 162 (54%) ongoing
pregnancies respectively with no significant difference between these two
subgroups. However, there was a significant (P

expression of AMH, FSHR, Inhibina and Inhibinb in growing follicles in
group 3 versus group 2.Tracking studies demonstrated the human MSCs
evenly infiltrating and repopulating growing follicles in treated ovaries.
Finally, breeding data showed significant increase in both the number of
pregnancies and total number of pups per litter in group 3 compared to group
2(P¼ 0.02).
CONCLUSIONS: Our study shows that intra-ovarian injected human
BMSCs were able to restore ovarian hormone production, reactivate folliculogenesis
in chemotherapy-damaged ovaries and reverse infertility in this
preclinical model. This approach carries high promise to women with chemotherapy-induced,
and potentially other types of, premature ovarian failure.
MENOPAUSE
O-7 Monday, October 19, 2015 11:15 AM
DRUG METABOLIZING ENZME POLYMORPHISMS ARE
ASSOCIATEDWITH CHEMOTHERAPY RELATEDAMENORRHEA
IN YOUNG BREAST CANCER SURVIVORS. L. M. Charo, a
M. V. Homer, a L. Natarajan, a C. Haunschild, b A. DeMichele, c I. Su. a
a UC San Diego, San Diego, CA;
b Stanford University, Stanford, CA;
c University of Pennsylvania, Philadelphia, PA.
OBJECTIVE: To test if candidate single nucleotide polymorphisms
(SNPs) in enzymes involved in cyclophosphamide activation or detoxification
are associated with time to ovarian failure after chemotherapy in young
breast cancer survivors.
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: Premenopausal breast cancer survivors
(n ¼ 116) with Stages 0 to III disease and planned cyclophosphamide-based
chemotherapy were enrolled at diagnosis from three academic breast programs
and followed longitudinally for menstrual pattern. Participants were
genotyped for SNPs in genes involved in cyclophosphamide activation
(CYP3A4 [rs1067910] and CYP2C19 [rs42244285]) and detoxification
(GSTP1 [rs1695] and GSTA1 [rs4715332]). The primary endpoint was
time to chemotherapy-related amenorrhea (CRA), defined as >12 months
of amenorrhea after the end of chemotherapy. Using the time-to-event
method, the association between SNPs and CRA were assessed using Cox
proportional hazard regression models. A priori sample size calculations estimated
80% power to detect relative risks of 1.7-2.6.
RESULTS: The cohort had a median age of 39.7 years (range 20.8-46.1) at
end of chemotherapy and median follow up of 594.5 days (range 23-2119).
28% experienced CRA. Survivors with at least one major allele of GSTA1
had significantly lower hazards of developing CRA compared to survivors
who were homozygous for the minor allele (HR 0.22 [95% CI 0.61-0.91],
p¼0.04). Survivors with at least one major allele of CYP2C19 had significantly
higher hazards of developing CRA compared to survivors who were
homozygous for the minor allele (HR 4.56 [95% CI 1.54-13.57], p¼0.01).
CYP3A4 and GSTP1 SNPs were not related to CRA. Increased age was
also associated with CRA. In separate multivariable models adjusting for
age and BMI, GSTA1 remained significantly associated with CRA (HR
0.23 [95% CI 0.06-0.96], p¼0.04) while CYP2C19 was attenuated (HR
2.75 [0.89-8.49], p¼0.08).
CONCLUSIONS: In younger breast cancer patients undergoing cyclophosphamide-based
chemotherapy, the presence of one or more major alleles
of GSTA1 was found to have lower risk of developing CRA. Inter-individual
variation in enzymes involved in chemotherapy metabolism is related to posttreatment
ovarian function.
Supported by: MRSG-08-110-01-CCE, HD058799, T32 HD007203.
O-8 Monday, October 19, 2015 11:30 AM
PRESCRIBING OF COMPOUNDED AND COMMERCIALLY
AVAILABLE MENOPAUSAL HORMONE THERAPY BY
OBSTETRICIAN-GYNECOLOGISTS AND FAMILY MEDICINE
PHYSICIANS. J. P. Dubaut, F. Dong, B. L. Tjaden, D. A. Grainger,
J. Duong, L. L. Tatpati. University of Kansas School of Medicine -
Wichita, Wichita, KS.
OBJECTIVE: Compounded bioidentical hormone use has risen in the
United States (1). The American College of Obstetricians and Gynecologists
(ACOG) released a committee opinion on this topic, including guidelines for
its members (2). We explored factors that may influence prescribing practices
of compounded and bioidentical menopausal hormonal therapy (MHT) and
level of agreement with relevant ACOG statements among obstetrician-gynecologists
(OB/GYNs) and family medicine physicians (FPs).
DESIGN: Cross-sectional physician survey.
MATERIALS AND METHODS: After Institutional Review Board
approval, investigators created an online anonymous survey. The survey
link was emailed to Kansas OB/GYN and FP physicians. The initial and
reminder emails were sent 4 weeks apart; the survey was available for 3
months. Survey constructs included: demographics, MHT knowledge,
MHT prescribing practices, and opinions on statements published in the
ACOG Committee Opinion 532. Chi-square analyses were conducted to
identify associations between specialties, practices and opinions on ACOG
statements.
RESULTS: Overall response rate was 11.1% (150 of 1349). The response
rate of OB/GYNs (27%) exceeded that of FPs (7.2%). Of 150 respondents,
53.3% were FPs. The majority of respondents identified as female (64%),
were under 50 years old (57.4%), and worked in cities with populations
over 100,000 (70.5%). In the past year, 84.5% prescribed conventional
MHT, 83% prescribed commercial bioidentical MHT, and 58.9% prescribed
compounded bioidentical MHT. OB/GYNs prescribed more than
FPs in each category; the difference in prescribing commercial bioidentical
MHT was statistically significant (p¼0.03). Hormone levels were monitored
in at least some patients by 40% of physicians. When asked whether
compounded MHT was regulated by the Food and Drug Administration
(FDA), 76.7% answered ‘‘no’’, 4% answered ‘‘yes’’, and 19.3% were
‘‘not sure’’ or declined to answer. Most respondents stated efficacy, risks,
tolerability, cost, patient preference, and experience of previous patients
were important factors influencing their MHT prescribing practices. FDA
regulation was not important to 15.3%, while customization was important
to 62% of physicians. The majority of respondents agreed with 10 ACOG
statements regarding MHT (range 53-97%), but at least 15% showed
disagreement with 7 of 10 statements. More OB/GYNs than FPs agreed
‘‘saliva levels are not biologically meaningful’’ (p

RESULTS: Ethnic distribution did not differ in each reproductive category:
39 premenopausal South Asians and 34 Europeans; 10 perimenopausal
South Asians and 14 Europeans; and 42 postmenoapusal South Asians and 39
Europeans. Body fatness variables increased with reproductive ageing to an
almost similar degree in both ethnic groups (p for trend greater than 0.05 for
all associations) without ethnicity modifying the gradient of the associations
between menopausal state and body fatness (p for interaction with ethnicity
greater than 0.05 for all associations). Moderate to vigorous physical activity
and VO2max decreased in a similar fashion across the reproductive stages in
both ethnic groups whereas energy intake remained unchanged. Body
fatness, physical activity and fitness did not differ among the ethnic groups
for each reproductive stage either. Metabolic biomarkers (insulin, total
cholesterol, triglycerides and blood pressure) deteriorated with reproductive
ageing in both ethnic groups in a similar degree. Notably, HbA1c levels
increased to a much greater degree with reproductive ageing in the South
Asians than in the Europeans (p¼0.02 for interaction with ethnicity).
CONCLUSIONS: The increase in HbA1c levels in healthy women
without overt T2DM during menopausal transition was much greater in the
South Asians than in their White counterparts and this discrepancy was not
explained by a greater deterioration in body composition or physical activity
variables along with reproductive ageing.
O-10 Monday, October 19, 2015 12:00 PM
ASSOCIATION BETWEEN POLYCYSTIC OVARY SYNDROME
AND HOT FLASH PRESENTATION DURING THE
MENOPAUSE TRANSITION. O. Yin, a H. A. Zacur, b J. A. Flaws, c
M. S. Christianson. a a Johns Hopkins University School of Medicine,
Lutherville, MD;
b Reproductive Endocrinologist, Lutherville, MD;
c University of Illinois, Urbana, IL.
OBJECTIVE: While polycystic ovary syndrome (PCOS) is the most common
endocrinopathy in reproductive-age women, most research has focused
on young women and the impact of PCOS on the menopause transition remains
poorly understood. This study aims to determine the influence of
PCOS on hot flash presentation in midlife women.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Subjects were recruited from an ongoing
cohort study involving 748 midlife women aged 45-54 from an urban
metropolitan area. Subjects completed detailed questionnaires that included
hot flash symptom and demographic information. Between June 2014 and
March 2015, 656 patients were contacted by telephone. Those who agreed to
participate were screened for history of PCOS using the Rotterdam criteria.
Chi square analysis and Wilcoxon rank sum test were used as needed to compare
subjects with a history of PCOS with other midlife women. Multivariate logistic
regression was performed to identify factors associated with hot flashes at
midlife; odds ratios (OR) with 95% confidence intervals (CI) were calculated.
RESULTS: A total of 453 women (69%) responded to the telephone interview
and 9.3% (n¼42) met diagnostic criteria for PCOS. The remaining 411
were included as controls. Mean PCOS subject age was 48.0 and body mass
index (BMI) was 27.3. The majority of subjects were Caucasian (73%) with a
smaller proportion African American (21%) and other ethnicities (2%).
PCOS and control groups were not statistically different with respect to
age, BMI, race, income, smoking, drinking, physical activity, number of periods
in the last year, and testosterone, progesterone, or estradiol levels.
Multivariate logistic regression analysis demonstrated that PCOS was not
associated with increased odds of hot flash prevalence, frequency, duration,
or severity. Smoking was the only variable that demonstrated an increased association
with experiencing hot flashes (OR 2.0, 95% CI 1.05-3.98).
CONCLUSIONS: A history of PCOS was not associated with increased
hot flashes during the menopause transition in this study. These data suggest
that women with PCOS have similar hot flash presentations in midlife as
compared to the general population. Additional research should continue
to investigate the health and quality of life implications associated with a
history of PCOS in the aging population.
Supported by: 2R01AG018400 - 05A2.
O-11 Monday, October 19, 2015 12:15 PM
ORAL TIBOLONE (2.5 MG) VERSUS TRANSDERMAL ESTRA-
DIOL GEL (0.06%, 2.5 GM) - EFFECTS ON SERUM CALCIUM
AND 25-HYDROXY VITAMIN D3 LEVELS AFTER SURGICAL
MENOPAUSE. S. M. Bhattacharya a A. Jha. b a Obstetrics and Gynecology,
KPC Medical College, Kolkata, India; b Research Associate, West Virginia,
WV.
OBJECTIVE: To compare the effects of oral Tibolone (2.5 mg) and Transdermal
Estradiol gel (0.06%; 2.5g) on serum Calcium and 25-hydroxy
Vitamin D3 levels in surgically postmenopausal women after 6 months of
treatment.
DESIGN: Open label randomized controlled study.
MATERIALS AND METHODS: 144 women (40-52 years of age) with
surgical menopause (duration 3-18 months and done for benign gynecological
causes and having distressing menopausal symptoms) with preset inclusion
and exclusion criteria were randomized in 1:1 ratio, between
01.03 2013 and 30.06.2014 into 2 groups. Prior ethical approval and
informed written consents (from all participants) were obtained. Group A
received Tibolone (2.5mg, daily) orally and Group B received Transdermal
Estradiol gel (0.06%; 2.5 g, daily). The primary outcomes were comparison
of the absolute changes in serum Calcium and 25- hydroxy Vitamin D3
levels following 6 months of treatment between the two groups. Body
mass index (BMI) and blood pressure (systolic, SBP: diastolic, DBP)
were recorded. Serum calcium and 25- hydroxyl vitamin D3 levels were
measured. All parameters were studied at baseline and after 6 months of
treatment. Sample size was calculated based on a pilot study where 6
months treatment with Tibolone increased vitamin D3 level by 4.91
ng/ml (SD ¼ 6.54) and Transdermal estradiol gel increased vitamin D3
level by 2.08 ng/ml (SD ¼ 3.99). It was calculated that a sample size of
58 patients per group would have 80% power and 95% confidence level
with 2-sided test of significance to detect this mean difference between
the two groups. 6 cases in the Group A and 9 cases in the Group B were
lost to follow up after 6 months of treatment.
RESULTS: Intent-to-treat analysis showed that after 6 months of treatment,
effects of the two treatment interventions were identical in the studied
population. In both intervention arms, patients recorded increase in serum
Calcium and Vitamin D3 levels, but the mean differences were not statistically
significant.
CONCLUSIONS: There were no differences in therapeutic effects of oral
Tibolone and Transdermal Estradiol gel on BMI, blood pressure, serum Calcium
and Vitamin D3 levels in surgically postmenopausal women after 6
months of treatment. Effects of the changes in serum Calcium and Vitamin
D3 levels, by either mode of treatment, on bone remodelling in menopausal
women would need more studies.
Support: None.CTRI registration no. - CTRI/2013/02/003341.
O-12 Monday, October 19, 2015 12:30 PM
LONG TERM HORMONE REPLACEMENT THERAPY (HT)
DOES NOT AFFECT POST-MENOPAUSAL TOTAL BODY
COMPOSITION. A. H. Bayer, a K. N. Goldman, b R. Mauricio, a
M. J. Nachtigall, b F. Naftolin, b L. E. Nachtigall. b a NYU School of Medicine,
New York, NY; b Department of Obstetrics & Gynecology, NYU School of
Medicine, New York, NY.
OBJECTIVE: The impact of HT on menopause-related changes in body
composition is not resolved. We sought to evaluate differences in total
body composition in post-menopausal women who had taken HT for an
average of 14 years.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Post-menopausal women (40-100 years)
on HT for a minimum of 6 years in a university-affiliated menopause clinic
underwent annual Dual Energy X-ray Absorptiometry (DXA) from August
2004 to October 2014. Annual DXA scans from untreated post-menopausal
women seen in the same clinic during the same time period were evaluated as
controls. Exclusion criteria included the diagnosis of primary ovarian insufficiency,
DXA data unavailable or incomplete, and DXA scan not performed
while on HT (for HT group). Primary outcomes were percent (%) total body
fat and % total lean body mass. Secondary outcomes included body mass index
(BMI) and relevant co-morbidities. Data were analyzed using Student’s
t-test and Fisher’s exact test where appropriate (p

differences in medical co-morbidities in women on HT compared to controls,
including but not limited to osteoporosis, diabetes, hypertension, hyperlipidemia,
coronary artery disease, fibrocystic breast disease, endometrial polyps,
colonic polyps, endometrial cancer, ovarian cancer, mammogram BIRADS
(breast imaging-reporting data system) 3, 4, or 5, or breast cancer.
CONCLUSIONS: Evaluation by DXA of post-menopausal women
receiving HT for up to 25 years demonstrates that long-term HT has no significant
impact on body composition. It is notable that there was no increased
prevalence of medical co-morbidities between the treated and control groups.
These findings may inform the risk-benefit ratio when considering long-term
HT for post-menopausal women.
Supported by: Pfizer Corporation.
REPRODUCTIVE ENDOCRINOLOGY: RESEARCH 1
O-13 Monday, October 19, 2015 11:15 AM
ELEVATED SERUM ANTI-MULLERIAN HORMONE (AMH)
STALLS OVARIAN FOLLICLE DEVELOPMENT BY
DOWNREGULATING FSH- AND LH-RECEPTORS AND
INHIBIN-B PRODUCTION. L. Detti, a L. J. Williams, a
S. E. Osborne, a N. M. Fletcher, b G. M. Saed. b a Obstetrics and Gynecology,
University of Tennessee, Memphis, Memphis, TN; b Wayne State University,Detroit,MI.
OBJECTIVE: In granulosa cell cultures AMH inhibits the FSH-dependent
follicular growth and the cyclic selection for dominance [1-3]. Women with
polycystic ovary syndrome (PCOS) have high serum AMH levels from
increased production [4] which are correlated to the follicle number and
AMH levels [5]. We tested the hypothesis that administration of recombinant
AMH to ovarian cortex fragments would inhibit follicular development by
downregulating hormone receptors’ expression.
DESIGN: Pilot experimental study with ovarian cortex obtained from 3
patients.
MATERIALS AND METHODS: Immediately after explant the ovarian
cortex specimens were divided into 5 equal fragments. One fragment was
flash-frozen (untreated) and four were incubated for 48 hours 37 Cina
pH-adjusted gamete buffer media with increasing AMH concentrations of
0-5-25-50 ng/ml. After incubation, all specimens were rinsed and flashfrozen
for PCR analyses, which were executed in triplicates. We utilized
real time RT-PCR to determine mRNA levels for FSH-R, LH-R and
Inhibin-B in ovarian cortex tissue. We performed ANOVA with Tukey post
hoc tests to evaluate changes in mRNA levels among the five different fragments.
A p

were collected. We used microanalytical assays to measure the levels of ATP
and citrate in single oocytes. To evaluate spindle and chromosome alignment,
mice were injected with PMSG and hCG, sacrificed and ovulated meiosis II
(MII) oocytes were collected. Mature oocytes were stained with tubulin and
DAPI. Imaging was performed on a Leica confocal microscope and analysis
performed blindly with ImageJ software. ANOVA, students t-tests, and chisquare
analysis were used in the statistical analysis as appropriate.
RESULTS: HF mice weighed significantly more than mice on the control
diet (28g vs 21.7g, p

Supported by: Expanding the Boundaries Research Grant, Brigham &
Women’s Hospital, Harvard Medical School NIH Grant RO1 HD053112,
R21 HD061259.
O-18 Monday, October 19, 2015 12:30 PM
SELECTIVE PROGESTERONE RECEPTOR MODULATOR
(SPRM), CDB-4142 INHIBITS DECIDUALIZATION OF HUMAN
AND MOUSE ENDOMETRIAL STROMAL CELLS AND PREVENTS
EMBRYO IMPLANTATION IN THE MOUSE
UTERUS. S. Kuokkanen, a L. Zhu, b B. McAvey, c J. Pollard. d a Obstetrics
& Gynecology, Albert Einstein College of Medicine, Bronx, NY; b Department
of Molecular Biology, Albert Einstein College of Medicine, Bronx,
NY; c Obstetrics & Gynecology, Icahn School of Medicine, Mt. Sinai and
RMA of NY, New York, NY; d College of Medicine and Veterinary Medicine,
University of Edinburgh, Edinburg, United Kingdom.
OBJECTIVE: SPRMs with selective effects on hormone responsive tissues
hold promise for long-term medical management of fibroids and endometriosis.
Telapristone acetate (CDB-4124) is a derivative form of ulipristal
acetate, a reliable emergency contraceptive (EllaÒ). Here, we examined the
impact of CDB-4124 on in vivo decidualization and embryo implantation in
the mouse uterus and in vitro decidualization of human endometrial stromal
cells (hESC).
DESIGN: Controlled laboratory study.
MATERIALS AND METHODS: Endometrial tissue was collected by biopsy
from healthy volunteers. After isolation, stromal cells were seeded in 6-
well plates in DMEM-F12 with 2% charcoal/dextran-treated FBS. The decidualization
media was supplemented with 1uM progesterone (P), 30 nM
estradiol (E) and CDB or 0.1% EtoH vehicle control. hESC decidualization
was monitored by morphology and prolactin (PRL) and IGFBP1 mRNA by
qrtPCR. Decidualization of the mouse uterus was induced in ovariectomized,
E/P or E/P/CDB treated mice by intraluminal injection of peanut oil or PBS
(control) and decidual response was assessed by uterine horn weight and
morphology. For implantation study, copulation of wild type CD1 mice
was monitored with daily vaginal plugs and CDB was administered on pregnancy
d 3-6. The mice were euthanized on pregnancy d7 and implantation
sites were quantified.
RESULTS: After 9 days of incubation, hESC in the decid. media transformed
from fibroblast-like cells to round decidual cells and produced
increased levels of the decidualization markers compared to the control cells
(PRL FC¼41, p¼0.0003; IGFBP1 FC¼76, p¼0.01). hESC decidualized in
the decid. media with CDB 0.1uM (PRL FC¼8.5, p¼0.0006; IGFBP1
FC¼10, p¼0.03). In contrast, hESC cultured in the decid. media with
CDB at 1, 3 or 9 uM remained spindle-shaped and PRL and IGFBP1
mRNAwere at the level of the control cells. CDB inhibited decidual response
in the mouse uterus and the weight ratio of the oil stimulated to the control
mouse horn was approximately 3 in E/P treated mice (0.1095 mg 0.034
mg vs. 0.034 mg 0.0072, p¼0.01), but only 1.03 in E/P/CDB treated
mice (0.042 mg 0.011 mg vs. 0.041 mg 0.011, p¼0.25). Implantation
sites were absent in the CDB treated uteri after copulation compare to an
average of 12 sites in the control mice.
CONCLUSIONS: CDB-4124 acts as progesterone receptor antagonist in
human and mouse endometrial stromal cells, completely inhibiting stromal
decidualization and inhibiting embryo implantation in the mouse uterus.
These findings describe a novel mechanism as to how SPRM can be efficacious
in treating sex-steroid dependent conditions and, as post-coital contraceptive,
delay endometrial maturation and extend contraceptive efficacy
beyond the time of ovulation.
Supported by: U54 HD058155, ABOG/AAOGF Bridge Funding (S.K).
MALE REPRODUCTION AND UROLOGY:
TRAVELING SCHOLARS
O-19 Monday, October 19, 2015 11:15 AM
COMPARISON OF THREE METHODS OF PENILE VIBRATORY
STIMULATION (PVS) IN MEN WITH SPINAL CORD INJURY
(SCI). W. Chong, a E. Ibrahim, b T. Aballa, b C. Lynne, a N. L. Brackett. b
a University of Miami/Jackson Memorial Hospital, Miami, FL; b The Miami
Project to Cure Paralysis, Miami, FL.
OBJECTIVE: PVS is recommended as the first line of therapy for semen
retrieval in anejaculatory men with SCI. This study compared ejaculatory
success rates and patient preference for three methods of PVS within the
same group of men with SCI.
DESIGN: Prospective, three-way crossover design.
MATERIALS AND METHODS: Subjects were 15 men with SCI whose
level of injury was T10 or rostral. Each subject received the following three
methods of PVS, with an interval of 2-4 weeks between each method.
Method 1 (M1): applying one FertiCare PersonalÔ (Multicept, Denmark)
device to the dorsum or frenulum of the glans penis; Method 2 (M2): ‘‘sandwiching’’
the glans penis between two FertiCare devices; Method 3 (M3):
sandwiching the glans penis between the two vibrating surfaces of the Viberect
X3Ô (Reflexonic, Frederick, MD) device. To control for sequencing effects,
5 subjects received PVS in the following sequence: M1, M2, M3; 5
received the sequence M2, M3, M1; and 5 received the sequence M3, M2,
M1. For trials with M1 and M2, FertiCare device(s) were set at 2.5 mm
amplitude and 100 Hz. For trials with M3, stimulation parameters of the Viberect
X3 were preset by the manufacturer and were not adjustable. For all
methods, PVS was delivered in 2 minute increments with inspection of the
penile skin between increments. PVS was stopped if ejaculation occurred
or after 10 minutes of PVS with no ejaculation. Following each PVS trial,
subjects were asked to rate their experience on a questionnaire with scaled
responses.
RESULTS: Please see Table 1. Ejaculation success rates were high for
each method, however, ejaculation latency was significantly longer with
M3 compared with M1 or M2. When analyzing subject responses to survey
questions, there were no significant differences in ratings of M1 compared to
M2. In contrast, M3 was rated lower than M1 and M2 for all survey questions.
These differences were significant for survey questions 1, 2 and 4. Semen
collection was more problematic with M3 versus M1 or M2 due to the configuration
of the Viberect X3 device, which hampered proximity of the specimen
cup to the urethral meatus.
CONCLUSIONS: Based on these findings, our recommended algorithm is
to attempt PVS with one FertiCare device. If that fails, use two FertiCare devices.
Although the Viberect X3 was preferred less by patients, it is a lower
cost alternative that may be suitable for home use by some patients.
Table 1
Success rate (% of men
ejaculating)
Ejaculation latency in
seconds (mean SEM)
Survey Questions:
(Values represent
means SEM)
1. How much did this
method meet your
expectations?
0 ¼ Did not meet
expectations
100 ¼ Met expectations
2. How comfortable
did you feel during
stimulation?
0 ¼ Not comfortable
100 ¼ Very comfortable
3. How comfortable do
you feel about using
this method at home
either by yourself or
with a partner?
0 ¼ Not comfortable
100 ¼ Very comfortable
4. Would you recommend
this method to other
men with spinal
cord injury?
0 ¼ Would not recommend
100 ¼ Would recommend
One FertiCare
Device (M1)
Two Ferticare
Devices (M2)
87 100 87
Viberect X3
Device (M3)
29.6 5.0 32.2 4.4 56.8 5.0 a
74.9 6.1 77.2 6.1 52.8 5.9 b
82.2 5.7 82.2 5.7 64.3 5.5 c
79.1 6.5 71.5 6.5 67.5 6.2
91.7 7.0 83.6 7.0 68.4 6.7 d
SEM ¼ standard error of the mean
Unless indicated by a superscripted letter, comparisons between groups
were not significant.
a Significantly different from M1 (p ¼ 0.0006) and M2 (p ¼ 0.001)
b Significantly different from M1 (p ¼ 0.01) and M2 (p ¼ 0.03)
c Significantly different from M1 (p ¼ 0.03) and M2 (p ¼ 0.03)
d Significantly different from M1 (p ¼ 0.02)
e8 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

O-20 Monday, October 19, 2015 11:30 AM
REDEFINING AND CLARIFYING THE RELATIONSHIP
BETWEEN TOTAL MOTILE SPERM COUNTS (TMSC) AND
INTRAUTERINE INSEMINATION (IUI) PREGNANCY
RATES. R. S. Rubin, a K. S. Richter, b F. Naeemi, b S. Shipley, b
P. R. Shin, c J. E. O’Brien. b a Urology, Medstar Georgetown University
Hospital, Washington, DC; b Shady Grove Fertility Reproductive Science
Center, Rockville, MD; c Shady Grove Fertility Center, Washington, DC.
OBJECTIVE: The value of post-wash TMSC for predicting IUI outcomes
is not well defined. Limitations of previous reports include small sample
sizes and attempts to identify clinically meaningful thresholds as opposed
to gradual trends across a continuum of TMSC. To clarify the relationship between
post-wash TMSC and IUI outcomes we evaluated a large single institution
sample.
DESIGN: Retrospective review
MATERIALS AND METHODS: All stimulated clomiphene citrate, letrozole,
and/or injectable gonadotropin IUI cycles performed at a single institution
from 2004 to 2014 were reviewed, excluding double insemination.
Generalized estimating equations (GEE) analysis was used to account for
multiple cycles by individual patients and to adjust for age, BMI, infertility
diagnoses, stimulation protocol, and pre-insemination endometrial thickness,
serum estradiol, and numbers of follicles R14 mm.
RESULTS: 47,553 insemination cycles were available to evaluate the relationship
between TMSC and clinical pregnancy (defined as ultrasound
confirmation of an intrauterine gestational sac). Pregnancy rates were highest
with a clear threshold noted at R9 million TMSC. Pregnancy rates declined
gradually as TMSC decreased.
Complete data for the adjusted GEE analysis were available for 40,655 cycles.
Adjusted GEE analysis among cycles with R9 million TMSC
confirmed that TMSC in this range was unrelated to pregnancy (p¼0.47).
Conversely, TMSC was highly predictive of pregnancy (Wald c 2 ¼120) in
adjusted GEE analysis among cycles with

(not gender specific). Among all cancer centers, only 60% include information
on fertility preservation specifically directed toward men, such as sperm
cryopreservation. Survivorship information on family building after cancer
was available on 32% of cancer center web sites. State population density
had no significant effect on whether a web site included risks of treatment
on fertility (p¼0.90) or information on fertility preservation (p¼0.29).
CONCLUSIONS: Forty percent of NCI designated cancer center web sites
do not discuss options for male fertility preservation, and over one-third
make no mention of the ramifications of cancer treatment on male fertility.
Given the increasing recognition of the importance of oncofertility in cancer
survivorship, more education should be available about options for fertility
preservation, particularly among men.
References: [1] There are 68 NCI designated cancer centers, of which 61
engage in clinical activity. Cleveland Clinic Taussig Cancer Institute was
included as a separate data point although it is also a member of the Case
Comprehensive Cancer Center, for final n¼62.
O-23 Monday, October 19, 2015 12:15 PM
BLOCKADE OF PHOSPHATIDYLSERINE ON MURINE SPERM
INHIBITS IN VITRO FERTILIZATION. L. Smith-Harrison, a
K. Wheeler, b C. Barberry, a W. Xu, b R. Smith, c J. Lysiak. b a Urology,
University of Virginia, Charlottesville, VA; b University of Virginia, Charlottesville,
VA; c UVA Urology, Charlottesville, VA.
OBJECTIVE: Phosphatidylserine (PtdSer) expression is not only a marker
for apoptotic cells but also a ligand for engulfment receptors essential for the
recognition and efficient removal of these dead cells. The engulfment of the
dead/apoptotic cells by phagocytes or neighboring cells is actually the final
step of apoptotic cell death. Our lab has recently shown that efficient clearance
of apoptotic germ cells is essential for normal spermatogenesis. Studies
in the boar, ram, mouse, rat, and human have found mature sperm positive for
the expression of PtdSer. PtdSer positive sperm were found to be functional
and may even positively correlate with embryo formation in humans. In this
current study we test the novel hypothesis that egg-sperm interactions during
fertilization may hijack the machinery normally used for engulfing apoptotic
cells, with the sperm mimicking an apoptotic cell and the oocyte a phagocyte.
DESIGN: In vitro fertilization (IVF) experiments were performed using
oocytes from super-ovulated mice and sperm isolated from the murine cauda
epididymis.
MATERIALS AND METHODS: Expression of PtdSer on sperm was performed
with fluorescenctly tagged Annexin V (AnV). Blockade of PtdSer on
sperm was accomplished with unlabeled AnV.
RESULTS: The percentage of fertilized embryos after PtdSer blockade
with AnV was 57.6% versus 87.2% in controls (p < 0.001). Sperm motility
was not affected by AnV. Additionally, immunofluorescence identified
PtdSer on the midpiece and acrosome regions of sperm from the cauda but
not caput epididymis.
CONCLUSIONS: The exact mechanisms involved in sperm entry into the
oocyte remain elusive. These results suggest that PtdSer on sperm play an
important role in fertilization and supports our hypothesis that ligands and
receptors involved in apoptotic cell engulfment may be used during sperm
oocyte interactions. Whether oocytes have receptors for PtdSer, or whether
PtdSer interacts with cumulus cells or the zona pellucida remains to be elucidated.
The results of this study will have potential impacts on in vitro fertilization
and contraceptive technologies.
O-24 Monday, October 19, 2015 12:30 PM
PROTEOMIC PATHWAY IN SEMINAL PLASMA OF MEN WITH
SPINAL CORD INJURY (SCI) BEFORE AND AFTER ORAL
ADMINISTRATION OF PROBENECID. M. Camargo, a E. Ibrahim, b
T. C. Aballa, b V. Carvalho, c K. Cardozo, c C. M. Lynne, b,d R. Bertolla, a
N. Brackett. b,d a Department of Surgery, Division of Urology, Human Reproduction
Section, Sao Paulo Federal University, Sao Paulo, Brazil; b Miami
Project To Cure Paralysis, University of Miami, Miami, FL; c Fleury Group,
Sao Paulo, Brazil; d Urology, University of Miami, Miller School of Medicine,
Miami, FL.
OBJECTIVE: Previous results demonstrate that oral administration of
probenecid increases sperm motility in men with SCI (J Urol 193(4S)
e344-e345). Our objective was to evaluate the proteome of seminal plasma
in SCI patients before and after treatment with oral probenecid in order to
demonstrate changes in associated pathways.
DESIGN: Prospective Study.
MATERIALS AND METHODS: This study included 10 men with SCI
who ejaculated regularly by penile vibratory stimulation or electroejaculation.
Probenecid tablets (Watson Pharma Inc., Corona, CA) were administered
in two phases. In Phase 1, subjects received 250 mg orally twice a
day for 1 week. In Phase 2, subjects who completed Phase 1 with no complications
or side effects were administered 500 mg orally twice a day for 3
weeks. Semen analyses were performed at two time points: Pre-treatment
(Pre-Rx group, 1-2 days before Phase 1) and Post-treatment (Post-Rx group,
within 4 weeks after completion of Phase 2). Seminal plasma proteomics was
performed by a label-free quantitative approach, in which 50 mg of total proteins
were pooled, digested into tryptic peptides and analyzed by liquid chromatography
followed by tandem mass spectrometry (LC-MS/MS). Each
sample was run in triplicate. Significant proteins (Student’s t-test) were
used for functional enrichment analysis performed using the Cytoscape platform.
RESULTS: In total, 806 proteins were identified, of which 695 were not
significantly changed in both groups. Thirteen proteins were down-regulated
and 65 other proteins were exclusive to the Pre-Rx group. Five proteins were
up-regulated and 28 other proteins were exclusive to the Post-Rx group. The
Pre-Rx group expressed increased immunologic functions, such as antigen
processing and preservation of peptide antigen via MHC class I. Catabolic
activities such as amino acid degradation, lysosome organization, as well
as the pentose-phosphate shunt oxidative functions were also observed in
the Pre-Rx group. The Post-Rx group expressed enriched energy production
pathways (glycine-tRNA ligase activity) as well as 4-hydroxyproline catabolic
processes.
CONCLUSIONS: Oxidative and immune functions, as well as catabolic
processes were enhanced in patients with SCI. Treatment with oral probenecid
enhanced the energy-production pathways that play an important role
in the biological process of improving sperm motility in men with SCI.
Supported by: Craig H. Neilsen Foundation #224598.
MALE REPRODUCTION AND UROLOGY: CLINICAL 1
O-25 Monday, October 19, 2015 11:15 AM
SEMINAL VESICLE SPERM ASPIRATION FROM WOUNDED
WARRIORS: A CASE SERIES. M. W. Healy, a B. Yauger, a
A. N. James, b R. Dean. c a Department of Obstetrics and Gynecology, Division
of Reproductive Endocrinology and Infertility, Walter Reed National
Military Medical Center, Bethesda, MD; b ART Institute of Washington,
Inc, Bethesda, MD; c Department of Urology, Division of Andrology, Walter
Reed National Military Medical Center, Bethesda, MD.
OBJECTIVE: There has been an increase in blast injuries from dismounted
Improvised Explosive Devices (IEDs) encountered on the battlefield
(i.e. ambulatory soldiers not within the protection of a vehicle).
Associated injuries involve the lower extremities, pelvis, and perineum.
Thus, options to retrieve sperm may be limited due to type and extent of
injury. An alternative technique to the standard Testicular Sperm Extraction
or Microsurgical Epididymal Sperm Aspiration is seminal vesicle sperm
aspiration (SVSA), described in cases of ejaculatory duct obstructions or primary
anorgasmy. Given the type of pelvic injuries seen in these wounded
warriors, we assessed SVSA as an option to retrieve sperm with the goal
of cryopreservation for future use in In Vitro Fertilization (IVF) with intracytoplasmic
sperm injection (ICSI).
DESIGN: Retrospective case series.
MATERIALS AND METHODS: Wounded warriors who underwent
SVSA at Walter Reed National Military Medical Center between 2012-
2014 were included. Patient age, type and date of injury, date of SVSA, specimen
fluid analysis, post-thaw analysis, fertilization rates during IVF/ICSI,
pregnancy rates, and live birth outcomes were evaluated.
RESULTS: Six patients who presented with lower extremity, pelvic, and
perineal injuries resulting from dismounted IEDs underwent SVSA within
5-12 days of the initial injury. Sperm retrieved were analyzed (volume:
0.4mL to 1.8mL, concentration: 40K to 2,200K, motility: 0% to 5%). Sperm
was washed and cryopreserved. In two cases, IVF/ICSI cycles were performed
using the frozen samples. Sperm retrieval for these cases occurred
5 and 9 days after the initial injuries. In one couple, 13 mature oocytes underwent
ICSI with morphologically normal sperm that responded to the touch
test with a fertilization rate of 38%. One grade V embryo was transferred
on day 4 with a negative pregnancy test. The second couple underwent
two IVF/ICSI cycles. In the first cycle, 9 mature oocytes underwent ICSI
with 4 sperm that responded to Pentoxifylline and 5 sperm that responded
e10 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

to touch test notable for a 44% fertilization rate. On day 5, one expanded blastocyst,
B/B, was transferred with a negative pregnancy test. In the second cycle,
17 mature oocytes underwent ICSI with sperm that responded to
Pentoxifylline, with a 47% fertilization rate. Two B/B expanded blastocysts
were cryopreserved due to concern for ovarian hyperstimulation in the female
partner. Among all three cycles, fertilization rate was 43.5%.
CONCLUSIONS: SVSA is a reasonable option to retrieve sperm in
wounded warriors or trauma patients with pelvic or perineal injuries.
Supported by: This research was Supported, in part, by the Program in
Reproductive and Adult Endocrinology, NICHD, NIH.
O-26 Monday, October 19, 2015 11:30 AM
EVALUATION OF REPRODUCTIVE SYSTEM ANATOMY AND
GONADAL FUNCTION IN PATIENTS WITH PRUNE-BELLY
SYNDROME. M. Cocuzza, a B. C. Tiseo, a R. Park, a G. P. Padovani, a
R. H. Baroni, b A. Tavares, a F. T. Denes, a M. Srougi. a a Division of Urology,
Hospital das Clinicas, University of Sao Paulo Medical School, Sao Paulo,
Brazil; b Radiology Institute, Hospital das Clinicas, University of Sao Paulo
Medical School, Sao Paulo, Brazil.
OBJECTIVE: To report the first series of Prune-belly syndrome (PBS) patients
that were submitted to surgical treatment during early childhood
focusing in the evaluation of reproductive system anatomy and gonadal function.
To date, PBS patients were diagnosed as infertile and fertility was often
given lowest priority. Their infertility is multifactorial and is probably related
to the undescended testes, prostatic hypoplasia and retrograde ejaculation.
There are no documented cases of unassisted paternity; few successful pregnancies
have been reported through assisted reproduction, but all using
epididymal or testicular sperm extraction.
DESIGN: Series of case analysis.
MATERIALS AND METHODS: We accessed 30 patients with PBS from
our service that were submitted to any surgical procedure during their
childhood and now are at least 14 years old. Patient records were accessed
to identify age at orchidopexy. All patients were submitted to pelvic MRI in
order to evaluate anatomical findings of reproductive system, including
prostate size, characteristics of seminal vesicles and vas deferentia. Serum
levels of LH, FSH, testosterone and also creatinine were evaluated. Sperm
analysis was conducted and analysis of the urine after masturbation when
needed.
RESULTS: We contacted 18 patients. Of those, 15 had reliable data from
patient records including physical examination and hormone profile. The
average of age of this group at evaluation is 18.2 years. The average age at
orchidopexy was 17 months and an average follow-up was 17.4 years. All
had normal physical development and stable anatomy and function of the reconstructed
urinary tract with an average creatinine of 1.64 mg/dL. Fourteen
patients had both testes in scrotum, and the testicular volume varied from
2.1cc to 9.4cc, averaging 6.9cc. Eight patients collected semen with a sperm
count of 5.07 million/mL, whereas motile sperm was found in 62.5%
including three in the ejaculate and two in urine after masturbation. Average
hormones levels were LH: 5.3 mg/dL, FSH: 6.9 mg/dL, total testosterone:
531.2 mg/dL and free testosterone: 450.6 mg/dL. MRI revealed prostates hypoplastic
in 55.6% and absent in 22.3%, while 55.6% had absence of at least
one of the seminal vesicles. There was no vasal abnormality.
CONCLUSIONS: Our data enlightens findings in patients with PBS that
were not described yet. A high prevalence of hypoplastic or absent prostate
and seminal vesicle abnormalities was observed in our patients; those findings
may represent their main cause of infertility. Probably, early orchidopexy
increases testicular function salvage preserving their fertility
potential patients leading to the finding of motile sperm in the ejaculate or
in urine after masturbation. The next step is to provide a better understanding
of their fertility potential improving quality of life.
O-27 Monday, October 19, 2015 11:45 AM
THE EFFECTS OF TRANSVERSE AND LONGITUDINAL
INCISION OF TUNICA ALBUGINEA IN MICRODISSECTION
TESTICULAR SPERM EXTRACTION. T. Ishikawa, K. Yamaguchi,
R. Nishiyama, Y. Takaya, K. Kitaya, H. Matsubayashi. Reproduction Clinic
Osaka, Osaka, Japan.
OBJECTIVE: The optimal technique of sperm extraction would be
minimally invasive and avoid destruction of testicular function without
compromising the chance of retrieval adequate numbers of spermatozoa to
perform ICSI. In general, the tunica albuginea should be opened on an equatorial
plane because antimesenteric incision increases the risk of testicular
devascularization and may adversely affect access to the seminiferous tubules.
To demonstrate the differences of sperm retrieval rate (SRR) and postoperative
complication between transverse or longitudinal incision of tunica
albuginea in microdissection testicular sperm extraction (micro TESE).
DESIGN: A retrospective study.
MATERIALS AND METHODS: A total of 1080 patients (including 970
46XY males with NOA and 110 Klinefelter syndrome (KS) patients) underwent
micro TESE were subjected to sperm retrieval procedures. All operations
were performed by one surgeon (TI). Karyotyping test was
performed on a sample of blood to all patients. SRR and postoperative
complication were analyzed in the 960 patients with transverse incision
(group T) and the 120 patients with longitudinal incision (group L). Ninety
and 20 non-mosaic KS cases each were included in group T and group L,
respectively.
RESULTS: Testicular sperm were successfully retrieved by micro-TESE
in 415 of 960 (43.2%) NOA (including 47 of 90 KS: 52.2%) and 56 of 120
(46.7%) NOA (including 10 of 20 KS: 50%) patients with group T and group
L incision in micro TESE, respectively. There was no significant difference
of sperm retrieval rate with either approach. No patients had postoperative
complications such as major hematoma or leakage of seminiferous tubules
through this series. The hospital stay was all the same in both groups
(a-day-surgery). For the 46XY males with NOA patients, after micro-
TESE, the testosterone level did not drop significantly in both group T and
group L. In addition, there were no significant differences in the levels of
testosterone between group T and group L for the 46XY males with NOA
patients. Of the KS patients who underwent micro TESE, the mean serum
testosterone level showed an average decline of 30-35% from baseline
when assessed at 1, 3, and 6 months after micro-TESE, but no significant differences
were also shown in the levels of testosterone between group T and
group L.
CONCLUSIONS: There were no significant differences between transverse
or longitudinal incision for SRR and postoperative complication in
micro TESE. We should take care of the hypogonadism in KS patients after
even microdissection procedure by either transverse or longitudinal incision.
O-28 Monday, October 19, 2015 12:00 PM
COMPARISON OF SEMEN QUALITY BETWEEN UNIVERSITY
AND PRIVATE CLINIC LABORATORIES. O. Khan, a C. F. Jensen, b
J. Sonksen, b M. Fode, b T. Shah, c D. A. Ohl. a a Urology, University of Michigan,
Ann Arbor, MI;
b Urology, Herlev University Hospital, Herlev,
Denmark; c Obstetrics and Gynecology, University of Michigan, Ann Arbor,
MI.
OBJECTIVE: Obtaining an ejaculate and performing a semen analysis
(SA) is an essential first step in evaluating the infertile man. Based on the
SA the physician determines the nature of further evaluation and treatment.
Multiple treatment options are available and range from simple counseling to
the more complicated and expensive assisted reproduction techniques
(ART). The objective of this study was to investigate inter-laboratory variation
in semen quality between private ART laboratories and university-based
ART laboratories respectively.
DESIGN: Clinical retrospective study.
MATERIALS AND METHODS: IRB approval was obtained to retrospectively
evaluate patients who had undergone a SA at both the University
of Michigan Center For Reproductive Medicine and private practice IVF
clinics. When more than one SA was available from both clinics the SA
with the highest total motile sperm was selected for analysis. Major semen
parameters from both SA’s were compared using Wilcoxon signed-rank
test.
RESULTS: A total of 20 men aged 35 6 (mean SD) years were
included in the study. Table 1 shows comparisons of the major semen parameters
obtained at the two laboratories. Morphology was reported significantly
lower at the private clinics.
CONCLUSIONS: In this small series, morphology was significantly lower
in semen analyses performed at private IVF clinics. Since sperm morphology
under 5% is commonly used to recommend IVF with ISCI, underestimation
of sperm morphology at private clinics may lead to over-utilization of high
level assisted reproductive techniques at these sites.
FERTILITY & STERILITY Ò
e11

TABLE 1.
University Based ART Laboratory
Private ART Laboratory
25th percentile Median 75th percentile 25th percentile Median 75th percentile
p-value
Volume (mL)(n¼19) 1.7 3.3 4.1 1.75 3.2 4.5 0.43
Motility (%)(n¼19) 40 40 50 16.9 56 69.2 0.07
Concentration (million/mL)(n¼20) 0.45 10 44 0.89 7 26 0.30
Total Motile Sperm (N/ejaculate)(n¼19) 0.3 8 54 0.35 9 31 0.44
Morphology (%)(n¼9) 6.5 7 10 2 3 6 0.01
O-29 Monday, October 19, 2015 12:15 PM
CLOMID HAS GOTA BRIGHT SIDE AND A DARK SIDE. WHAT DO
WE REALLY KNOWAFTER ALLTHESE YEARS? EVIDENCE FOR
TOXICITY. R. Wiehle G. K. Fontenot. Research and Development,
Repros Therapeutics Inc, The Woodlands, TX.
OBJECTIVE: Clomid has been approved in women with PCOS/ovulation
induction. In men it is used off-label to increase testosterone or to relieve
symptoms of androgen abuse. Clomid is a mixture of two isomers: zuclomiphene
and enclomiphene. The former is a weak estrogen agonist and the latter
is a strong estrogen antagonist. Repros Therapeutics is developing enclomiphene
for elevating endogenous testosterone in men with secondary hypogonadism.
DESIGN: We have administered each isomer of clomid to mice, both
chronically and acutely, in an attempt to determine the relative differences.
MATERIALS AND METHODS: Enclomiphene and zuclomiphene were
separated from clomiphene as pure mono-isomers. Mice were administered
each isomer chronically (90 days) then tissues were examined histologic.
In a mouse ADME, pigmented mice were acutely administered each of the
14C-labelled isomers. Tissue and fluid distribution were followed up to 45
days.
RESULTS: In animals chronically administered each isomer, zuclomiphene
had pernicious effects on the male reproductive tract (testes, epididymis,
and seminal vesicles) and the kidney. Significant reduction in size of
testes with testicular degeneration was seen, including Leydig cell loss
with absence of sperm in the seminiferous tubules and reduction in size of
the epididymis, seminal vesicles and kidneys. In the ADME study, for 47 tissues
assessed, 97.5% of the 14C-enclomiphene seen at 4 hours was lost by 24
hours. In contrast, only 40.8% of the 14C-zuclomiphene seen at 4 hours was
lost. Zuclomiphene was retained selectively. Among those that accumulated
zuclomiphene that seen in the blood were the pigmented portions of the eye,
the brain, adrenal gland, the kidneys and the testes.
CONCLUSIONS: We infer that zuclomiphene accumulates in excess over
enclomiphene. Human studies have suggested this as well. We concede that
the antagonist effects of enclomiphene can overwhelm effects of zuclomiphene
when used chronically, however the extreme high build-up of the
agonist isomer may have lasting effects. These results justify the case for a
monoisomeric preparation and the development of Enclomiphene citrate,
for clinical use in men with secondary hypogonadism to increase testosterone.
It is interesting to speculate whether clomiphene citrate would have
been granted FDA approval given the differences between its constitutive
isomers.
Supported by: Repros Therapeutics Inc.
O-30 Monday, October 19, 2015 12:30 PM
THE EFFECT OF CLOMIPHENE CITRATE IN THE TREATMENT
OF SUBFERTILE MALES WITH BODY MASS INDEX (BMI) ‡ 25
KG/M 2 . B. Patel, a T. Shah, a D. Shin. b a Urology, Rutgers New Jersey
Medical School, Newark, NJ; b Urology, Hackensack University Medical
Center, Hackensack, NJ.
OBJECTIVE: Elevated BMI has been shown to have a negative impact on
male fertility. Clomiphene citrate (CC), a selective estrogen receptor modulator,
is often used in the empiric treatment of subfertile males to increase
testosterone levels and improve spermatogenesis. The objective of this study
was to assess the effect of CC in the treatment of subfertile males with
elevated BMI R 25 kg/m2.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Fifty-six subfertile males with BMI
R 25 kg/m2 were treated with CC between 2009 and 2014. Semen analysis
was conducted on all 56 patients at baseline and at minimum, 3 months
follow-up. Fifty of the 56 patients had baseline and 5 month follow-up measurements
of follicle-stimulating hormone (FSH), luteinizing hormone (LH),
total testosterone (TT), bioavailable testosterone (BT) and estradiol (E)
levels. Pregnancy status was obtained when possible. Paired t-test was
used to compare baseline and follow-up hormonal profiles and semen analyses.
RESULTS: Significant increase in sperm concentration, from 19.0 2.7
M to 28.2 4.0 M (p

MATERIALS AND METHODS: Patients with normal ovarian reserve
% 42 years of age were recruited. Following routine IVF care, trophectoderm
biopsy was performed. Embryos were subsequently selected for transfer per
routine. No NGS CCS analysis was done prior to transfer. A novel targeted
amplification method of NGS based CCS which does not require whole
genome amplification was then performed. The outcome for each transferred
embryo was compared to the NGS screening result to determine the
predictive value of that result. In the case of a two embryo transfer, DNA
fingerprinting was utilized to ensure that the embryo responsible for the pregnancy
was identified correctly. Implantation rates were compared between
embryos designated euploid, aneuploid, and for the population as a whole.
RESULTS: 117 patients had 187 blastocysts transferred. Of the 41 embryos
assigned an aneuploid karyotype, none sustained implantation yielding
a predictive value of an aneuploid result of 100%. 97 of 146 embryos designated
as euploid implanted and progressed to delivery yielding a predictive
value for a euploid result of 66.0%. No embryo designated as euploid subsequently
developed into an aneuploid gestation. Euploid embryos had a higher
sustained implantation rate than the population as a whole (50.8%,
p

OBJECTIVE: The objective of this study was twofold; first we wanted to
determine the number of blastocysts needed at a certain age to produce at
least one euploid blastocyst (>95%) through PGS using whole comprehensive
chromosome screening. Secondly, we evaluated the chance of obtaining
a euploid embryo in the next cycle after obtaining all aneuploid embryos.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: 10,852 cycles of PGS were performed
using blastocyst biopsy and analyzed by array Comparative Genomic
Hybridization (aCGH). A total of 58,798 embryos were analyzed. IVF
cycles were performed at multiple fertility centers and biopsies sent to a
reference laboratory for analysis. Some poor prognosis patients accumulated
embryos from several cycles and then performed PGS (‘‘embryo
banking’’).
RESULTS: There was no correlation between cohort size of blastocyst
analyzed and euploidy rates, but there were significant differences between
euploidy rates and maternal age (p< 0.001). Euploidy rates 65%, 56%,
46%, 33%, 19% and 13% for egg donors were observed for ages 42 years
old, respectively.The cumulative number of blastocyst needed to produce
at least one euploid blastocyst with 95% or higher chance was calculated
from single cycles or embryo banking cycles from the same patient and is
shown in the below table.
% of patients with normal blastocysts
#of
embryos
egg
donors
42 years old, respectively, produced
no euploid embryos in the first cycle. Of those with no euploid embryos
in the first cycle, 38% (41-42 years old) and 25% (> 42 yeas old)
of those that produced 17 embryos produced euploid embryos in successive
cycles.
CONCLUSIONS: This is the largest study so far on the relationship between
aneuploidy and maternal age and cohort size. In women, 35 and
older more than 50% of embryos are chromosomally abnormal, with those
41 and older needing 18 or more embryos to secure one euploid one. However,
a cycle with no euploid embryos does not preclude finding euploid
ones in the next cycle(s) provided those cycles are not far apart and
enough embryos are generated. This data will help couples assess their
odds at producing a viable pregnancy and how many cycles and embryos
it might involve.
O-35 Monday, October 19, 2015 12:15 PM
41-42
years
>42 years
1-3 83% 80% 71% 57% 36% 22%
4-6 97% 95% 92% 82% 59% 43%
7-10 99% 98% 96% 89% 74% 50%
10-17 100% 99% 99% 97% 88% banked 64% banked
>17 100% 100% 100% 99% 97% banked 87% banked
WHY DO ARRAY-CGH (ACGH) EUPLOID EMBRYOS MISCARRY?
REANALYSIS BY NGS REVEALS UNDETECTED ABNORMALITIES
WHICH WOULD HAVE PREVENTED 56% OF THE
MISCARRIAGES. J. Grifo, a P. Colls, b L. Ribustello, b T. Escudero, b
E. Liu, b S. Munne. b a NYU Langone Medical Center, NY, NY; b Reprogenetics,
Livingston, NJ.
OBJECTIVE: aCGH, qPCR and SNP arrays have been extensively used
for preimplantation genetic screening (PGS) but a more robust and sensitive
technique, Next Generation Sequencing (NGS) can detect mosaicism and
polyploidy more effectively. The objective of this study was to determine
if miscarriages occurring after PGS were due to abnormalities not detected
by aCGH.
DESIGN: Retrospective Analysis with prospective sample re-analysis by
aCGH and NGS
MATERIALS AND METHODS: A total of 43 miscarriages were reported
upon follow up of spontaneous pregnancy loss resulting from
2442 cycles with euploid blastocysts obtained from PGS by blastocyst biopsy
and array CGH. Karyotype analysis of products of conception (POC)
in 18 samples revealed aneuploidy (full aneuploidy, mosaic aneuploidy
and partial aneuploidy) which was inconsistent with the aCGH analysis
from the blastocyst biopsy and went undetected prior to embryo transfer.
In the remaining 25 samples no POC data was available. Saved amplified
DNA samples from these 43 blastocyst biopsies previously diagnosed as
‘‘euploid’’ by aCGH were re-analyzed by aCGH yielding the same result.
A third aliquot of the same amplified DNA was then analyzed by NGS to
determine if abnormalities not detectable by aCGH were present.
RESULTS: The DNA from the embryos replaced that resulted in pregnancy
and miscarriage were reanalyzed by NGS and the results are shown
in the table below:
NGS Reanalysis of TE Biopsy Specimen
Normal
Triploid
Mosaic
Whole
Aneuploidy
Mosaic
Partial
Aneuploidy
Full
Trisomy
No
Diagnosis
POC Not 11 (44%) 2 (8%) 3 (12%) 7 (28%) 0 2 (8%)
Analyzed
N¼25
POC 6 (33%) 0 9 (50%) 1 (6%) 0 2 (11%)
Aneuploid
N¼18
Total
N¼43
17 (40%) 2 (5%) 12 (28%) 8 (19%) 0 4 (9%)
CONCLUSIONS: NGS is known to be able to detect mosaicism as well as
triploidy, which is also well-known cause of spontaneous pregnancy loss.
Whole and partial mosaicism could also play a role in spontaneous pregnancy
loss depending on the chromosomes involved as well as the percentage of
abnormal cells in the embryo. Of 16 embryos diagnosed euploid by aCGH
that resulted in an aneuploid loss 10 (62%) were diagnosed as abnormal by
NGS (2 not analyzable). Similarly, of the 23 embryos diagnosed euploid
by aCGH that resulted in loss but not diagnosed by POC analysis, 12
(52%) were abnormal by NGS (2 not analyzable). The use of NGS would
have avoided 56%(22/39) of the pregnancy losses resulting from aCGH
tested embryos. Likely, mosaicism and triploidy accounted for most of these
losses and NGS is more sensitive at detecting them. These data support the
notion that NGS is the most powerful technique for PGS analysis in predicting
euploid outcome but will not predict all euploid losses. It reduces the
miscarriage rate by more than 50% over aCGH and will significantly improve
outcomes.
O-36 Monday, October 19, 2015 12:30 PM
GENETIC CARRIER SCREENING IN AN EGG DONOR
PROGRAM. A. Quinteiro Retamar, a C. M. Borghi, a G. Fiszbajn, a
S. Papier, a S. Munne, b J. Hamer, a C. Alvarez Sedo. a a CEGYR (Reproductive
Medicine and Genetics), Buenos Aires, Argentina; b Reprogenetics, Livingston,
NJ.
OBJECTIVE: With the emergence of genomics platforms and the possibility
of studying multiple diseases in only one study and the lack of local updated
data, raised the necessity to establish the prevalence of genetic carriers
(250 diseases) in our egg donor population, in order to improve our egg donation
program.
DESIGN: Retrospective prevalence study.
MATERIALS AND METHODS: Three hundred two oocyte donors were
included (21-33 y/o), from December 2013-March 2015. All donor signed an
informed consent to participate in this study. Donors were included in this
study after fulfilling the egg donation program inclusion criteria (antral follicle
count >16, negative serology, psychological counseling, genetic counseling,
and normal karyotype). All patients were evaluated by a clinical
geneticist, in order to detect relevant family history, before and after the
study.A blood sample was taken, and DNA was extracted in our molecular
biology laboratory. Dried DNA was sent to Recombine Laboratory (Livingston,
NJ, USA). A genomics platform was used to detect more than 1700 mutations
that correspond to 250 autosomal recessive diseases by microarray
SNPs array technique.
RESULTS: Considering the 250 tested autosomal recessive diseases, our
donors have mutations for 35 (14%), 74.3% (26) of the latter have a high
impact over life expectancy and quality of life, and 25.7% (9) have a moderate
impact.Among the high impact diseases, 61.54% (16) may be subject to
medical treatment, but the 38.46% (10) do not. Cystic Fibrosis (1:19), the
nonsyndromic hearing loss and deafness: GJB2-Related (1:50) and Biotinidase
Deficiency (1:50) were the most prevalent high impact diseases
with medical treatment. Spinal Muscular Atrophy: SMN1 Linked (1:23)
was the most prevalent disease in the group without treatment.Moderate
e14 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

impact diseases were the least present. Within this group, 44.4%(4) have not
treatment and 55.6%(5) have a possible treatment. The 21-Hydroxylase-
Deficient Congenital Nonclassical Adrenal Hyperplasia (1:9), Familial Mediterranean
Fever (1:100) and Pseudocholinesterase Deficiency (1:50) were
the most prevalent detected diseases.
CONCLUSIONS: It is clear that the screening of genetic diseases
for oocyte donors is part of the necessary studies to reduce the risk of children
born from this kind of treatment. We can infer that the genetic screening of
recessive mutations in people who donate their gametes will further reduce
the risk of certain genetic diseases transmission.
OUTCOME PREDICTORS - CLINICAL: ART 1
O-37 Monday, October 19, 2015 11:15 AM
MATERNAL PREGNANCY AND BIRTH COMPLICATIONS BY
FERTILITY STATUS: THE MASSACHUSETTS OUTCOMES STUDY
OF ASSISTED REPRODUCTIVE TECHNOLOGIES. B. Luke, a
D. Gopal, b J. E. Stern, c E. Declercq, b L. Hoang, b M. Kotelchuck, d
H. Diop. e a Obstetrics, Gynecology, and Reproductive Biology, Michigan
State University, East Lansing, MI; b Community Health Sciences, Boston
University, Boston, MA; c Geisel School of Medicine at Dartmouth, Lebanon,
NH; d MGH Center for Child & Adolscent Health Research and Policy, Mass-
General Hospital for Children, Boston, MA; e Bureau of Family Health and
Nutrition, Massachusetts Department of Public Health, Boston, MA.
OBJECTIVE: To evaluate the effect of maternal fertility status on the risk
of pregnancy and birth complications.
DESIGN: Longitudinal cohort study, linking cycles from the SART
CORS, hospital discharge, and vital records from 2004-2010 in Massachusetts.
MATERIALS AND METHODS: The study included three fertility
groups: women without ART or other infertility treatment (fertile); women
with indicators of subfertility but no ART treatment (subfertile), and women
with ART treatment. The risks of seven adverse outcomes were modeled using
logistic regression, adjusted for parental ages, race/ethnicity, education,
payor status, maternal pre-existing conditions (diabetes and chronic hypertension),
and plurality, and reported as adjusted odds ratios (AORs) and
95% confidence intervals.
Pregnancy Outcomes and Multiple Pregnancy Rate based on Age and Number
of Embryos Transferred
Compliant
(1 ET)

O-39 Monday, October 19, 2015 11:45 AM
ELECTIVE SINGLE EMBRYO TRANSFER (ESET) IS ASSOCIATED
WITH NON-LOW BIRTHWEIGHT TERM SINGLETON
OUTCOMES: AN ANALYSIS OF 263,375 CYCLES. A. K. Styer, a
W. Vitek, b M. S. Christianson, c V. Baker, d A. Armstrong, e N. Santoro, f
B. Luke, g A. J. Polotsky. f a Massachusetts General Hospital/Harvard Medical
School, Boston, MA; b University of Rochester School of Medicine, Rochester,
NY; c Johns Hopkins University School of Medicine, Lutherville, MD;
d Stanford University, Palo Alto, CA; e NICHD, Bethesda, MD; f University
of Colorado, Aurora, CO; g Research, Ann Arbor, MI.
OBJECTIVE: Multiple gestation and its concomitant risk of prematurity is
the major complication of ART. The adoption of ‘‘good perinatal outcome’’
as a more relevant measure of term live birth with a normal birth weight
neonate has been recently advocated. The objective of this study is to evaluate
the likelihood of non-low birth weight (NLBW) term singleton live birth
outcomes with single (SET) and double embryo transfer (DET) in the United
States.
DESIGN: Historical cohort study from the Society for Assisted Reproductive
Technology Clinic Online Reporting System between 2004 and 2012.
MATERIALS AND METHODS: Fresh autologous IVF cycles among
women ages 18-37 years using partner’s semen with either SET or DET
were assessed and categorized into groups with or without cryopreserved
(cryo) supernumerary embryos. SET (cryo) was designated as eSET. SET
(no cryo) were non elective SET due to limited embryo cohort. Gestational
carrier, preimplantation genetic screening or diagnosis, and research cycles
were excluded. Logistic regression (adjusted for age, race/ethnicity, infertility
diagnosis, year of cycle, number of oocytes retrieved, day of transfer)
was used to determine the likelihood of NLBW term singleton live birth
outcome (birthweight R2,500 g and gestation R259 days). Results are reported
as adjusted odds ratios (AORs) and 95% confidence intervals.
RESULTS: 263,375 cycles were analyzed. Compared to eSET, the likelihood
of clinical pregnancy and live birth was significantly increased for DET
(cryo). The odds of NLBW term singleton live birth was significantly lower
in all groups compared to eSET [Table].
Pregnancy Outcomes Following SET and DET
Group
SET(cryo)
[eSET]
n¼20,996
DET(cryo)
n¼117,091
SET(no cryo)
n¼21,917
DET(no cryo)
n¼103,371
Clinical Pregnancy Live Birth NLBW Term Singleton
% AOR (95% CI) % AOR (95% CI) % AOR (95% CI)
57.6 Reference 49.3 Reference 39.1 Reference
62.5 1.38 (1.34, 1.42) 55.0 1.40 (1.36, 1.44) 25.8 0.55 (0.53, 0.57)
26.6 0.39 (0.37, 0.41) 21.7 0.41 (0.39, 0.43) 17.3 0.36 (0.34, 0.38)
47.8 0.86 (0.83, 0.89) 40.9 0.89 (0.86, 0.91) 23.0 0.48 (0.46, 0.49)
CONCLUSIONS: Elective SET increases the likelihood of NLBW by 45-
52% as compared to DET. The use of NLBW term singleton live birth as a
measure of ART success may be useful for the development of clinical strategies
which optimize good perinatal birth outcomes.
Supported by: Clinical Research Scientist Training Program/NICHD,
R25HD075737.
O-40 Monday, October 19, 2015 12:00 PM
THE IMPACT OF INSURANCE MANDATES ON MULTIPLE BIRTH
RATES FOLLOWING IN VITRO FERTILIZATION. M. P. Provost, a
J. S. Yeh, a S. M. Thomas, b W. W. Hurd, a J. L. Eaton. a a Division of Reproductive
Endocrinology & Infertility, Duke University Medical Center,
Durham, NC; b Department of Biostatistics & Bioinformatics, Duke University
Medical Center, Durham, NC.
OBJECTIVE: To examine the relationship between state-mandated insurance
coverage for in vitro fertilization (IVF) and multiple birth rate.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: We utilized the Society for Assisted
Reproductive Technologies-Clinical Outcomes Reporting System (SART-
CORS) database to identify fresh, autologous IVF cycles performed between
2007 and 2011 in women aged 20-42 years. Only first IVF cycles performed
in each woman’s state of residence were included in the analysis. Cycles were
excluded if the indication for IVF was ‘‘non-infertile’’ or ’’preimplantation
genetic diagnosis,’’ if they were performed in a state with only one reporting
clinic, or if embryo transfer occurred on days other than 3 or 5. Among the 40
states with more than one clinic, 6 have legislation requiring insurance
coverage for at least one IVF cycle and were designated ‘‘mandated:’’ CT,
HI, IL, MA, MD, and NJ. The remaining 34 states were designated ‘‘nonmandated.’’
Student’s t-test and the X 2 test were used to analyze continuous
and categorical variables, respectively. P < 0.00019 was considered statistically
significant after Bonferroni adjustment for multiple comparisons. Logistic
regression was performed to model IVF outcomes while controlling
for potential confounders.
RESULTS: Of the 173,968 cycles included in the analysis, 45,011 (26%)
were performed in mandated states and 128,957 (74%) were performed in
non-mandated states. The multiple birth rate per live birth was significantly
lower in mandated vs. non-mandated states (29% vs. 33%, P < 0.00001).
This association remained significant after adjusting for potential confounders
(OR 0.87, 95% CI 0.83-0.91). After stratification by SARTage category
and day of transfer, the relationship between mandate status and
multiple birth rate remained statistically significant only in women

OBJECTIVE: ASRM and SART regularly update guidelines on the
maximum number of embryos to transfer per cycle. Providers/patients, however,
often elect to transfer more than the recommended number of embryos
which can lead to an unnecessary increase in the number of multiple pregnancies.
The objective of this study was to determine the prevalence of cleavage
transfer cycles that do not conform to current recommended embryo transfer
(ET) limits and assess the impact of noncompliance on multiple pregnancy
rate (MPR) in first IVF cycles with a favorable prognosis.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: 30,567 first fresh autologous IVF cycles
in women under 43 undergoing cleavage stage embryo transfer (ET) from the
2011-2012 SART registry were stratified into cohorts based on ASRM
defined age bins. Cycles were classified as compliant (C) versus noncompliant
(NC) based on their adherence to the published 2009/2013 guidelines for
first IVF cycles with a favorable prognosis. Main outcomes measured were
percentage of C and NC cycles in each age group as well as the MPR (R2
fetal heart beats on ultrasound) in each of these groups. Secondary outcomes
included clinical pregnancy rate (CPR), live birth rate (LBR), and singletons
(1 fetal heart beat on ultrasound). Data were analyzed using two-sided
Welch’s t-test for each age category and the Benjamini-Hochberg procedure
was used to control the false discovery rate.
RESULTS: The percentage of NC cycles ranged from 2 to 27.4% in
different age groups. Compared to C cycles, noncompliance resulted in
higher MPR in every age group, but statistical significance was reached
only in the two youngest age bins (p

CONCLUSIONS: There is no significant difference of epigenetic profiling
between PBs and corresponding female nuclear genome in the ooplasm at
specific stage from mouse to human, suggesting PB1 and PB2 could be the
optimum candidate of oocyte genome replacement for preventing the transmission
of inherited mitochondrial diseases.
References:
1. Wakayama T, Hayashi Y, Ogura A. Participation of the female pronucleus
derived from the second polar body in full embryonic development
of mice. J Reprod Fertil. 1997, 110(2): 263-266.
2. Tachibana M, Sparman M, Sritanaudomchai H, Ma H, Clepper L,
Woodward J, Li Y, Ramsey C, Kolotushkina O and Mitalipov S. Mitochondrial
gene replacement in primate offspring and embryonic stem
cells. Nature. 2009, 461(7262): 367-372.
3. Wang T, Sha H, Ji D, Zhang HL, Chen D, Cao Y, Zhu J. Polar body
genome transfer for preventing the transmission of inherited mitochondrial
diseases. Cell. 2014, 157(7):1591-1604.
4. Bartholomeusz R. Review of the longevity of the second polar body in
the mouse. Zygote. 2003, 11(1): 23-34.
5. Hino T, Kusakabe, H and Tateno H. Chromosomal stability of second
polar bodies in mouse embryos. J Assist Reprod Genet. 2013, 30(1):
91-98.
6. Fabian D, Cikos S, Rehak P, Koppel J. Do embryonic polar bodies
commit suicide? Zygote. 2012, 22(1): 10-17.
7. Montag M, K€oster M, Strowitzki T, Toth B. Polar body biopsy. Fertil
sertil; 2013, 100(3):603-607.
8. Hou Y, Fan W, Yan L, Li R, Lian Y, Huang J, Li J, Xu L, Tang F, Xie XS,
Qiao J. Genome analyses of single human oocytes. Cell. 2013, 155(7):
1492-1506.
Supported by: This study was Supported by grants from National Basic
Research Program of China (2014CB 81471512 to H.S.,2014CB 81370691
to C.Y.).
O-44 Monday, October 19, 2015 11:30 AM
ANEUPLOIDY AND RECOMBINATION IN IN VITRO FERTILIZED
EMBYROS (BLASTOCYSTS) UNDERGOING PREIMPLANTATION
GENETIC DIAGNOSIS (PGD). K. Ravichandran, a Z. K. Gunes, b
B. Bankowski, c A. M. Rosen, d S. H. Chen, e A. Hershlag, f B. Sandler, g
J. Grifo, h D. Wells, i M. Konstantinidis. a a Reprogenetics, Livingston, NJ;
b Reprogenetics UK, Oxford, United Kingdom; c Oregon Reproductive Medicine,
Portland, OR; d Mercy Hospital and Medical Center, Chicago, IL;
e IRMS at Saint Barnabas, Livingston, NJ; f North Shore LIJ Health System,
Cold Spring Harbor, NY; g Obstetrics, Gynecology and Reproductive Science,
Icahn School of Medicine at Mount Sinai, New York City, NY;
h NYU Langone Medical Center, NY, NY; i University of Oxford.
OBJECTIVE: To investigate aneuploidy and recombination in human preimplantation
embryos in an effort to advance knowledge regarding the occurrence
of these events during reproduction.
DESIGN: Single nucleotide polymorphism (SNP) arrays and array
comparative genomic hybridization (aCGH) were used in parallel to screen
blastocysts undergoing PGD.
MATERIALS AND METHODS: The Infinium Karyomapping assay (Illumina,
USA) was used to process 480 blastocysts derived from 133 couples.
Recombination data were obtained from 460 embryos. Aneuploidy data were
collected from 214 embryos determined to be either euploid or aneuploid
affected with trisomies of meiotic origin.
RESULTS: 49 embryos were found to carry meiotic trisomies. Almost
all of the meiotic trisomies were determined to be of maternal origin
(57/58). 66.1% of trisomies were determined to have occurred during
meiosis I and 33.9% during meiosis II. Chromosomes 5, 16, 21, and 22
were more likely to be affected by meiosis I errors while meiosis II errors
were more frequent in chromosomes 6 and 18. Recombination analysis
was carried out in 12,545 chromosomes and 19,769 recombination events
were recorded. For autosomes, the average recombination rate was
24.10.28 for male meiosis and 40.010.47 for female meiosis. Sites
of recombination tended to cluster towards the distal ends of the chromosome,
a pattern obvious in male meiosis. Female meiosis showed no patterns
in locations of crossover events but spikes in recombination activity
were noticed in the centromeric regions of chromosomes 16, 17, 18, 21,
and 22. Comparative analysis conducted between 165 euploid embryos
and 29 aneuploid embryos presenting meiotic trisomies showed no differences
(P>0.05) between the average recombination rates for male meiosis
(24.831.15 in aneuploid vs. 24.310.42 in euploid) or female meiosis
(37.171.64 in aneuploid vs. 40.890.69 in euploid). No differences in
the locations of crossover sites between euploid and aneuploid embryos
were observed.
CONCLUSIONS: This study, yielded unique data concerning recombination
and the origin of aneuploidy in human embryos. The dominance of female
meiotic errors, especially those occurring in meiosis I, was
confirmed. Certain chromosomes were most often affected during meiosis
I and others during meiosis II. This indicates that relative contribution of
mechanisms causing aneuploidy may differ for individual chromosomes.
Recombination rates as well as locations of crossover sites differed between
male and female meiosis but not between euploid and aneuploid embryos.
O-45 Monday, October 19, 2015 11:45 AM
MORPHOKINETICS AND NUCLEATION STATUS OF EMBRYOS
AS MARKERS OF IMPLANTATION POTENTIAL. J. A. Aguilar, a
E. Munoz, b E. Taboas, b M. Perez, b A. Delgado, c T. Viloria, c
M. Meseguer. d a IVF Laboratory, IVI Vigo, Vigo, Spain; b IVI Vigo, Vigo,
Spain; c IVI Valencia, Valencia, Spain; d Clinical Embryology, Valencia,
Spain.
OBJECTIVE: to study the nucleation status of in vitro human embryos in
the second cell cycle (2cells and 4 cells embryos), the effect of multinucleation
in 2cells in the implantation rate, the reversibility of multinucleation status
in the 4 cells embryos and the embryo morphokinetics their relationship
with embryo implantation.
DESIGN: Observational retrospective study conducted in IVI Vigo and
IVI Valencia, between 2011 and 2013. We analyzed 1679 embryos,
cultured in an Automated Time-lapse Incubator incubator in a 37 C,
6% CO2 and 20% de O2 atmosphere from 940 oocyte donation cycles.
The exact timing of the events cited below was evaluated in hours post
insemination by ICSI.
MATERIALS AND METHODS: Nucleation status was annotated according
to the number of nuclei present in each cell in the 2-cells and
4-cells embryos as follows: nMONO (mononucleated) where ‘n’ represents
the number of blastomeres in which a unique nucleus is seen. nBI
(binucleated) number of blastomeres in which two nuclei per cell are
visible. nMULTI (multinucleated) number of blastomeres in which
more than two nuclei are visible. This definition includes micronuclei.
The time for each embryo division was defined as t2(time to 2cells), t3
(3cells),t4 (4cells), t5(5cells), t8(8cells);cc2¼t3-t2; cc3¼t5-t3; s2¼t4-t3;
s3¼t8-t5 ANOVA test and c2-test were performed when applicable to
assess the influence of the length of S-phase in the implantation rate.
RESULTS: From 1602 embryos analyzed. 40.08% showed multinucleation
at 2cells, while it was present in 14,4% of 4 cells embryos. There
were no significant differences in implantation rate between mononucleated
embryos and multinucleated (one or both blastomeres) after analyzing multinucleation
at 2cells embryos(p¼0,07), on the contrary there were when
considering it at 4 cells embryos (p

potential of monosomy embryos is negligible compared to their trisomy
counterparts, yet the cause for this is unknown. Our previous investigation
of epigenetic modifiers uncovered a decreased expression of DNA methyltransferase
enzymes in monosomy blastocysts, as well as a hypomethylated
state limited to the monosomic chromosome. This study was thus extended to
determine the DNA methylation profiles of imprinting control regions (ICRs)
present on the affected chromosomes.
DESIGN: Research study.
MATERIALS AND METHODS: Surplus cryopreserved human blastocysts
were donated with IRB approval. Trophectoderm biopsies followed
by comprehensive chromosomal screening using qPCR (RMA-NJ) determined
chromosome constitution. Individual monosomy 11 (n¼10) or 15
(n¼10), trisomy 11 (n¼10) or 15 (n¼10), and euploid oocyte donor blastocysts
(controls; n¼20) underwent bisulfite conversion and PCR amplification
to determine the DNA methylation profiles of two maternally
methylated ICRs, KCNQ1OT1 (11p15.5) and SNRPN (15q11.2). Statistical
analysis was calculated using the Student’s t-Test, with significance at
p

with chromosome aneuploidy and embryo demise will assist in improving
clinical IVF protocols.
POLYCYSTIC OVARY SYNDROME
O-49 Monday, October 19, 2015 11:15 AM
ABNORMAL EXPRESSION OF GENES GOVERNING
ADIPOGENESIS AND EXTRACELLULAR MATRIX (ECM)
FORMATION IN SUBCUTANEOUS (SC) ABDOMINAL ADIPOSE
STEM CELLS (ASCS) OF LEAN POLYCYSTIC OVARY SYNDROME
(PCOS) WOMEN. G. D. Chazenbalk, a J. D. Phan, a V. Madrigal, a
X. Ding, b X. Li, b D. A. Dumesic. a a OB/GYN, UCLA, Los Angeles, CA;
b Pathology and Laboratory Medicine, UCLA, Los Angeles, CA.
OBJECTIVE: To identify differential expression of genes involved in adipogenesis
and ECM formation in subcutaneous (SC) abdominal adipose stem
cells (ASCs) of lean PCOS and age- and BMI-matched normoandrogenic
(NL) women.
DESIGN: Prospective Cohort Study.
MATERIALS AND METHODS: SC abdominal fat biopsy was obtained
from four lean (body mass index: 19-25 kg/m 2 )PCOSwomen,ages21-32
years, and four BMI- and age-matched NL women. ASCs were isolated
after digestion of SC abdominal adipose with collagenase and differential
centrifugation. RNA from ASCs was extracted using Qiagen miRNeasy
Mini Kit. Microarray and small non-coding RNA (miRNA) analysis
were performed using Affymetrix Human Genome U1.33 2.0 and Human
miRNA arrays, respectively. Data were analyzed using Ingenuity Pathway
Analysis.
RESULTS: Differential expression of >1.5 fold change between SC
abdominal ASCs of PCOS and NL women indicated 64 up-regulated
and 103 down-regulated genes (P

progesterone level>5 ng/ml. Blood samples were obtained during the
early follicular phase. TNF- a, IL-27, IL-35, IL-37, a-1 acid glycoprotein
were measured by a high sensitivity ELISA, while CRP, testosterone (T),
androstenedione (AE), SHBG, DHEA-S levels were measured by RIA.
Free androgen index (FAI) was calculated as percentage ratio of total
testosterone to SHBG values.
RESULTS: Levels of inflammatory mediators (CRP, TNF- a, a-1 acid
glycoprotein) were significantly increased in obese PCOS patients
compared to obese controls and in lean PCOS patients compared to
lean controls (all p

and tended to positively correlate with SC abdominal fat volume
(R 2 ¼0.29, P¼0.09) and waist size (R 2 ¼0.28, P¼0.08).
CONCLUSIONS: SC abdominal adipocyte size in lean PCOS and NL
women negatively correlates with Si as a predictor of abdominal fat content
rather than androgen excess per se, and is accompanied by a population of
small SC abdominal adipocytes in PCOS.
Reference:
1. Chazenbalk G, Singh P, Irge D, et al. Androgens inhibit adipogenesis
during human adipose stem cell commitment to predipocyte formation.
Steroids 2013;78:920-6.
References:
1. Chang WY, Clements D, Johnson SR. Effect of doxycycline on
proliferation, MMP production, and adhesion in LAM-related
cells. Am J Physiol Lung Cell Mol Physiol. 2010
Sep;299(3):L393-400.
2. Lewandowski, KC et al Increased circulating levels of matrix Metalloproteinase-2
and -9 in Women with the Polycystic Ovary Syndrome
JCEM 2006; 91:1173-77.
Supported by: NIH grant P50 HD071836.
O-54 Monday, October 19, 2015 12:30 PM
IMPACT ON OVULATORY PARAMETERS WITH THE USE OF
MMP INHIBITORS IN PCOS, MI-PCOS: A PILOT
STUDY. A. Light, a S. Alur, b S. Hammes, c K. Hoeger. b a Pathology and
Laboratory Medicine, University of Rochester Medical Center, Rochester,
NY; b Obstetrics and Gynecology, University of Rochester Medical Center,
Rochester, NY; c Endocrinology, University of Rochester Medical Center,
Rochester, NY.
OBJECTIVE: PolycysticOvary Syndrome (PCOS) is characterized in
part by anovulation and excess ovarian androgen mediated by excess
LH. Wepreviously reported LH induced steroid production in freshly
isolated murine ovarian follicles requires transactivation of the EGF
receptor following release of EGF-like ligands by matrix metalloproteinases
(MMPs). This steroidogenesis is suppressed by doxycycline
(DOXY), anagent known to inhibit MMP2/9(1). Furthermore, we
have shown that the addition of the MMP inhibitor Galardin to prenatally
androgenized mice improved cycling. Given the association of
LH induced steroidogenesis and evidence MMP2/9 activities are
elevated in PCOS(2), MMP-I may treat the abnormal endocrine state
in PCOS. In this pilot study, we examined the impact of MMP 2/9
inhibition with DOXY on ovulation and androgens in women with
PCOS.
DESIGN: Randomized placebo controlled pilot trial
MATERIALS AND METHODS: 13 women diagnosed with PCOS by
Rotterdam criteria were recruited and 10 met criteria and were randomized
to DOXY 100mg BID for 12 weeks or an identical placebo tablet
(PL) BID for 12 weeks and then no treatment for 12 weeks. At baseline,
total and free testosterone, and SHBG were obtained. Women collected
urine for urinary pregnanediol glucuronide (UDPG) assessment weekly
and were seen biweekly for serum progesterone (P4) and menstrual diary
collection for 24 weeks. Androgens were assessed at 12 and 24
weeks.
RESULTS: 5 subjects received DOXY and 5 PL. One DOXY subject
completed 4 weeks of UDPG and 9 P4. One PL subject completed 12
weeks. Missing data were assumed to be not ovulatory. Over 24 weeks,
DOXY had a mean of 2.4 ovulations versus 1.4 in PL (p¼0.12). 37%
of the possible cycles were ovulatory in DOXY versus 23% of the PL
(p¼0.13). 3/5 DOXY subjects had at least 3 ovulations versus 1/5 PL.
DOXY did not impact total or free testosterone measures or SHBG
compared to PL at 12 or 24 weeks.
CONCLUSIONS: DOXY did not reduce serum androgens in this pilot
study; however, there appeared to be a trend toward more ovulatory cycles
in the DOXY group, though this did not reach significance. If ovulation
is improved it may be through subtle changes in androgens that are
not detectable using our current techniques or may be via a different
mechanism than change in androgen production globally. A follow up
larger trial is needed.
Baseline Characteristics
PL
DOXY
Age (years, range) 27.4 (24-31) 28.2 (21-39)
BMI (kg/m2, range) 29.3 (24-37.6) 33.3 (24.6-47.7)
Total T (ng/dL) (SD) 49.6 (14.6) 50.6 (8.2)
Free T (ng/dL) (SD) 1.36 (0.39) 1.14 (0.57)
SHBG (nmol/L) (SD) 44.0 (13.5) 55 (13.2)
PROCEDURES AND TECHNIQUES
O-55 Monday, October 19, 2015 11:15 AM
EMBRYO SELECTION USING THE EEVA TEST, AN AUTOMATED
TIME-LAPSE QUANTITATIVE ANALYSIS, IN CONJUNCTION
WITH STANDARD MORPHOLOGY: A PROSPECTIVE
MULTI-CENTER PILOT STUDY. D. C. Kieslinger, a S. De Gheselle, b
P. De Sutter, b E. H. Kostelijk, a J. van Rijswijk, a C. B. Lambalk, a E. van
den Abbeel, b C. G. Vergouw. a a Department of Obstetrics and Gynecology,
VU University Medical Center, Amsterdam, Netherlands; b Centre for Reproductive
Medicine, University Hospital Gent, Gent, Belgium.
OBJECTIVE: To evaluate the impact of selecting embryos for transfer
using Eeva Test prediction scores combined with traditional morphology
on the pregnancy rate of IVF and ICSI patients undergoing Day 3 embryo
transfer.
DESIGN: Prospective, observational, multi-center pilot study. The analysis
involved 280 of 302 enrolled patients; 560 control patients were identified
from a cohort of 1777 patients (year 2011-2013) using propensity
matching based on age, cycle number, oocyte number and number of
fertilized oocytes. The majority of transfers (98%) were single embryo
transfers.
MATERIALS AND METHODS: Two academic hospitals (VUmc Amsterdam
and UZ Gent) enrolled patients

O-56 Monday, October 19, 2015 11:30 AM
COMPARISON OF A BENCHTOP INCUBATOR AND A TIME
LAPSE INCUBATOR FOR CULTURE OF HUMAN EMBRYOS:
IMPACT OF CULTURE DISH. R. L. Krisher, a C. Pospisil, b
A. F. Greene, a J. M. Stevens, b W. B. Schoolcraft, c J. E. Swain. d a National
Foundation for Fertility Research, Lone Tree, CO; b Fertility Labs of Colorado,
Lone Tree, CO; c Colorado Center for Reproductive Medicine, Lone
Tree, CO; d Fertility Lab Sciences, Lone Tree, CO.
OBJECTIVE: Much debate exists as to the optimal embryo culture incubator.
However, several variables exist that must be controlled to properly
examine the impact of the incubator itself, including the culture dish utilized
and the gas atmosphere. Our objective was to compare human embryo development
between two modern embryo culture incubators; the EmbryoScopeÔ
and the benchtop G185.
DESIGN: Prospective sibling embryo split
MATERIALS AND METHODS: Zygotes from 26 patients (average age
36.70.7) were placed individually into EmbryoSlideÔ dishes (25 mL under
oil). One EmbryoSlideÔ was placed into the EmbryoScopeÔ (Vitrolife,
n¼172 2PNs), while the other was placed into a G185 incubator (K-Systems,
n¼158 2PNs). Embryos were cultured in sequential media for 6 days. Both
incubators utilized 7% CO2, 5% O2. To examine the impact of the dish,
zygotes from 8 patients (average age 33.10.9) were randomly assigned to
culture in either an EmbryoSlideÔ (n¼58 2PNs) or to standard group microdrop
culture (up to 5 embryos per 30mL; n¼73 2PNs) and cultured in a G185
incubator.
RESULTS: Significantly more embryos developed to good quality day 5
blastocysts (>3BB) in the EmbryoSlideÔ when it was incubated in the
G185 benchtop incubator compared to the EmbryoScopeÔ (39.2% versus
28.5%, respectively; p¼0.039), although there was no difference in the number
of good quality blastocysts developing overall by day 6 (54.4% vs.
52.3%, respectively). When cultured in the same incubator (G185), the EmbryoSlideÔ
Supported good quality blastocyst development equal to that of
microdrop group culture (EmbryoSlideÔ- 25.9% day 5 and 58.6% overall;
group microdrop- 26.0% day 5 and 58.9% overall).
CONCLUSIONS: In patient embryo splits with a controlled gas atmosphere
and culture dish, the benchtop G185 incubator yielded more high
quality day 5 blastocysts compared to the EmbryoScopeÔ. The EmbryoSlideÔ
dish supports blastocyst development of individually cultured human
embryos equally well compared to group culture in standard
microdrops within the benchtop G185. Using sequential media and low oxygen,
modern benchtop incubators function as well as time-lapse incubators
and permit individual embryo culture, which is critical for the application of
biomarker analysis.
O-57 Monday, October 19, 2015 11:45 AM
PATIENT TREATMENT JOURNEYS, THEN AND NOW: 10 YEARS
OF CLINICAL PROGRESS?. A. Zgodic, a H. Karvir, a D. Parfitt, a
S. T. Globus, a A. B. Copperman, b P. Yurttas Beim. a a Celmatix Inc, New
York, NY; b Reproductive Medicine Associates of New York, New York, NY.
OBJECTIVE: As assisted reproductive technology (ART)-based therapies
are refined and clinical expertise is strengthened over time, improvements
are expected in live-birth outcomes for patients beginning
infertility treatment. To investigate this hypothesis, we analyzed trends in
clinical outcomes over a ten-year period at a large academic reproductive
medical center.
DESIGN: Retrospective analysis using de-identified ART cycles (10801
in-vitro fertilization (IVF) cycles, 1462 Egg Recipient (ER) cycles, and
33947 Non-IVF (NIVF) cycles) from 2003-2012.
MATERIALS AND METHODS: We performed trend analyses within:
1) All; 2) IVF; and 3) NIVF cycles. We used two-sided chi-square test to
investigate significant trends in live birth and multiples rates. The direction
and magnitude of these trends were assessed using a Binomial model with
treatment year as a predictor. For mean time-to-live birth (months), mean
cost of a singleton live birth, mean time between cycles (months), and
mean number of embryos transferred (IVF only), we used a linear model
to detect significant trends.
RESULTS: In all cycles, we observed no significant trends in odds of
achieving live birth or multiples (p

Augmented PR activity by AKAP13 in vivo was dependent upon the presence
of P4. These results suggest that in cells the interaction between
AKAP13 and PR might require the presence of P4. Further studies are needed
to characterize specific molecular mechanism(s) by which AKAP13 influences
PR function.
Supported by: HD 008737 to JHS.
O-59 Monday, October 19, 2015 12:15 PM
ART DOES NOT INCREASE THE MOLECULAR KARYOTYPE
ABNORMALITY RATE OF MISCARRIAGE. G. Li, W. Niu, Y. Su,
Y. Guo, Y. Sun. Reproductive Medical Center, The First Affiliated Hospital
of Zhengzhou University, Zhengzhou, China.
OBJECTIVE: To explore whether assisted reproductive technology (ART)
increase the molecular karyotype abnormality rate of miscarriage.
DESIGN: Retrospective study.
MATERIALS AND METHODS: IRB approval was obtained. Miscarriage
tissues underwent DNA extraction and 23-chromosome SNP microarray
analysis using humanCytoSNP-12 DNA beadchips and GenomeStudio software.
RESULTS: 618 coupes were enrolled. The mean age was 31.95.1
years. 144 experienced natural conception (NC) and 474 were pregnant
by ART. Of the 474 cases, 36 underwent artificial insemination (AI),
244 fresh in vitro fertilization-embryo transfer (IVF-ET), 66 fresh intracytoplasmic
sperm injection (ICSI)-embryo transfer and 128 thawed embryo
transfer. The total abnormal molecular karyotype rate was 70.9%
(438/618). There was no significant difference in the abnormal molecular
karyotype rate between ART (70.5%) and NC (72.%). The abnormal molecular
karyotype rate in the subgroup of ICSI was 65.6%,which was lower
than that of AI+NC( 71.7%) and IVF( 72.2%) ,However, there was no significant
difference between them. In the cases aged over 35 years, the
abnormal molecular karyotype rate was 83.3%,which was higher than
66.4% in the cases aged 35 years and under .
CONCLUSIONS: ART does not increase abnormal molecular
karyotype rate of abortion tissue. Moreover, in terms of fertilization
mode, compared with the IVF and AI+NC, ICSI does not increase
abnormal molecular karyotype rate of miscarriage. Abnormal molecular
karyotype rate is increased in the cases aged over 35 years in both NC
and ART. To our knowledge, this is the largest and most comprehesive
trial conducted evaluating SNP molecular karyotyping of spontaneous
miscarriage after ART.
O-60 Monday, October 19, 2015 12:30 PM
CYST ASPIRATION VERSUS GNRH ANTAGONIST
ADMINITRATION FOR OVARIAN CYSTS DETECTED AT
THE START OF FRESH IN VITRO FERTILIZATION
CYCLES. N. Pereira, a S. Amrane, b E. Hobeika, c J. Lekovich, a
P. Chung, d Z. Rosenwaks. d a The Ronald O. Perelman and Claudia Cohen
Center for Reproductive Medicine, New York, NY; b New York Presbyterian
Hospital, New York, NY; c North Shore LIJ at Staten Island, Staten
Island, NY; d Weill Cornell Medical College, New York, NY.
OBJECTIVE: To investigate effect of cyst aspiration versus GnRH antagonist
(GnRH-ant) administration for ovarian cysts detected at the start of
fresh in vitro fertilization (IVF)-embryo transfer (ET) on the outcomes of
the same IVF-ET cycles.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All patients undergoing fresh IVF-ET
between January 2008 and July 2013 who had transvaginal ultrasonogram-guided
cyst aspiration or treatment with GnRH-ant for ovarian
cysts detected at the start of IVF-ET prior to controlled ovarian simulation
(COS) were analyzed for potential inclusion. Exclusion criteria
included donor oocyte cycles, non-GnRH antagonist based cycles, or patients
having > 1 ovarian cysts at IVF start. Patients with known simple
ovarian cysts or endometriomas were excluded. Demographic and baseline
IVF characteristics were recorded from patient charts and included
age, gravidity, parity, body mass index (kg/m 2 ), estradiol (E 2 ) level at
cycle start (pg/mL), total days of ovarian stimulation, total dosage of
gonadotropins administered (IU), peak E 2 level (pg/mL), number of
mature oocytes retrieved, number of embryos transferred, and day
of ET. Pregnancy outcomes (clinical pregnancy rate, spontaneous miscarriage
rate, live birth rate) following ET were also noted. Student’s t-tests
and Chi-squared tests were used as indicated. Statistical analyses were
performed using STAT version 13 (College Station, TX: Stata Corp
LP). Statistical significance was set at P < 0.05.
RESULTS: 403 patients met inclusion criteria: 41 underwent cyst
aspiration and 362 were treated with GnRH-ant. Patients who underwent
cyst aspiration had a larger cyst size compared to GnRH-ant group i.e.,
31.1 5.71 mm versus 22.3 6.33 (P < 0.01). There was no difference
in the demographics or baseline IVF cycle characteristics of the two
groups. Patients treated with GnRH-ant had a longer duration of COS
(10.8 3.45 days vs. 9.05 4.06, P¼0.01) and required higher doses of
gonadotropins (3887.7 1097.8 IU vs. 3293.7 990.5; P¼0.003)
compared to the cyst aspiration group. The median number of mature
oocytes and embryos transferred were comparable between the groups.
There was no difference in the overall clinical pregnancy (43.9% vs.
41.4%), spontaneous miscarriage (9.76% vs. 8.01%) and live birth (34.1
vs. 33.4%) rates between the groups.
CONCLUSIONS: Our findings suggest that cyst aspiration or GnRH-ant
administration are equivalent modalities for management of solitary ovarian
cysts detected at the start of IVF-ET. However, in our study cohort, patients
treated with GnRH-ant had longer duration of COS and required higher
gonadotropin doses without adversely impacting the yield of mature oocytes
or pregnancy rates.
IN VITRO FERTILIZATION 1
O-61 Monday, October 19, 2015 11:15 AM
EFFECT OF ELEVATED PROGESTERONE ON THE DAY OF
TRIGGER AND PREGNANCY OUTCOMES IN GNRH AGONIST
TRIGGER CYCLES. M. T. Connell, a G. Patounakis, a M. W. Healy, a
A. DeCherney, a K. Devine, b E. A. Widra, c M. J. Hill. a a National Institutes
of Health, Bethesda, MD; b Shady Grove Fertility Center, Washington, DC;
c Shady Grove Fertility, Washington, DC.
OBJECTIVE: The current published data on progesterone elevation and
IVF outcomes have predominantly come from hCG trigger cycles. Trigger
medication has been shown to affect luteal phase hormone profiles. The
objective of this study was to determine whether elevated progesterone level
was similarly associated with clinical pregnancy outcome when GnRH
agonist trigger was used for IVF.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Fresh autologous IVF cycles from
2011-2013 were included. The primary outcome was clinical pregnancy.
GEE modelling with interaction testing was used to control for confounding
variables and multiple patient cycles.
RESULTS: 3226 cycles were evaluated of which 647 cycles utilized an
agonist trigger. The agonist trigger cohort were younger and had higher
oocyte yield, better quality embryos, and fewer embryos transferred
compared to the hCG trigger cohort (P

O-62 Monday, October 19, 2015 11:30 AM
SUCCESSFUL SURVIVAL IN CULTURE OF SPINAL CORD (SC)
DORSAL ROOT GANGLIA (DRG) NEURONS AND GLIA
CELLS AFTER AUTOMATIC VITRIFICATION. A. Arav, a
A. Shahar, b O. Ziv-Polat, b Y. Natan, c P. Patrizio. d a IVF Research lab, FertileSafe
Ltd., Nes Ziona, Israel; b NVR, Nes Ziona, Israel; c FertileSafe Ltd.,
Nes-Ziona, Israel; d Yale Fertility Center & Fertility Preservati, New Haven,
CT.
OBJECTIVE: To assess the survival and re-growth of neuronal cells after
cryopreservation with a new automatic vitrification system.
DESIGN: Experimental research with SC and DRG isolated from rat fetuses.
MATERIALS AND METHODS: Stationary organotypic DRG and SC
cultures were prepared from rat fetuses (gestational day 15, Lewis inbred,
Harlan, Israel). mmediately after dissection, the DRG and SC were cut
into small slices (400 micron) and seeded in 12 well culture plates containing
1 mL of NVR-Gel. Cultures were enriched once in gel with
GDNF conjugated iron oxide nanoparticles (10 ng/mL) and subsequently
with nutrient medium at each consecutive feeding. Monitoring of the
DRG-SC growth pattern was done by daily phase contrast microscopic
observations 24 hours after setting the cultures onward and by IF staining.
The various neuronal samples (SC, DRG, glia cells) were cryopreserved
with a new device for automatic vitrification (SarahÒ, Fertilesafe, IL),
consisting of a special capsule containing EM gold grids attached to
0.25ml straws (IMV,France). The straws loaded with neuronal tissue
were placed into the mixing chamber of the device attached to a syringe
pump dispensing timed and gradual increasing concentrations of equilibrium
solution for 12-25 min. and vitrification solutions for 3-5 min.,
before plunginginto liquid nitrogen slush (VitMaster, IMT IL). After 1
week the samples were rewarmed according to the Origio’s kit instructions.
To assess survival, the cells recovered were fixed (4% paraformaldehyde),
permeabilized with 0.1% Triton X-100 in PBS and then
immunoblocked with a 1% BSA in PBS for 1 h at RT prior to double
staining with mouse anti S-100 antibodies (1:80, Acris Antibodies, glial
cell marker) and rabbit anti neurofilament antibodies (NF, Novus Biologicals,
1:500, neuronal cell marker). The primary antibodies were diluted
in 0.1% BSA and 0.05% Tween-20 in PBS and incubated with the specimens
overnight at 4 C. After rinsing, the DRG specimens were incubated
for 1 h at RT with fluorescent secondary antibodies.
RESULTS: Organotypic cultures of SC, DRG neurons and glia cells
demonstrated a remarkable survival as well as nerve fiber regeneration after
cryopreservation/rewarming. The SC neurons maintained their multipolar
shape with re-growth of dendrites and axons. The round shaped DRG neurons
exhibited euchromatic nuclei with prominent nucleoli and an active regeneration
of nerve processes.
CONCLUSIONS: This is the first report on successful vitrification of SC
cells, DRG neurons and glia cells using an automatic freezing device. The
remarkable resumption of growth for neuronal cells will greatly expand
regenerative research opportunities.
O-63 Monday, October 19, 2015 11:45 AM
THE IN VITRO DEVELOPMENT OF HUMAN ZYGOTE
RECONSTRUCTED BY PRONUCLEAR TRANSFER. H. Liu, a
Z. Lu, a A. Chavez-Badiola, b P. Colls, c S. Munne, c J. Zhang. a a New
Hope Fertility Center, New York, NY; b New Hope Fertility Center Mexico,
Guadalajara, Jalisco, Mexico; c Reprogenetics, Livingston, NJ.
OBJECTIVE: Nuclear transfer techniques, including metaphase II
spindle transfer and pronuclear transfer (PNT), have been proposed as
potential therapeutic measures to prevent maternal mitochondrial (mt)
DNA disease transmission and to treat recurrent failure of implantation
due to ooplasmic dysfunction. Although PNT has been widely tested on
several animal models resulting in normal offsprings, the use of this
technique in humans is still limited due to safety and efficacy concerns.
This study aimed to examine the feasibility of PNT and its outcome
on human chromosome complements and pre-implantation blastocyst
development.
DESIGN: Two pronuclei (2PN) were exchanged between two zygotes
via PNT; the reconstructed zygotes were cultured to blastocysts followed
by aneuploidy testing using array comparative genomic hybridization
(aCGH).
MATERIALS AND METHODS: Oocytes and sperm from gamete donors
aged between 21 and 28 were collected and frozen for these experiments.
Donor oocytes and sperm were then thawed and cultured in vitro
for 3 hours before intracytoplasmic sperm injection (ICSI). Only zygotes
with 2PN at 15 hours after ICSI were used in this study and were subjected
to PNT via electric pulse to initiate membrane fusion between
the isolated karyoplasm and ooplasm. Reconstructed zygotes were
cultured in vitro until the blastocyst stage. The blastocysts were biopsied
by extracting 3-5 cells of the trophectoderm followed by aCGH testing
for aneuploidy. Blastocysts formed from fertilized donor oocytes and
sperm that were not subjected to PNT were used as controls (non-
PNT). Data were analyzed using chi-square test.
RESULTS: Thirty-two donor oocytes were thawed with 93.8% survival
rate, and 80% of the survived oocytes fertilized normally after ICSI with
thawed donor sperm forming a total of 24 zygotes (2PN). The zygotes
were then subjected to PNT, 37.5% of which developed to the blastocyst
stage compared to 53.7% blastocyst formation rate in the non-PNT group
(p>0.05). In addition, 7 out of 9 (77.8%) PNT blastocysts showed euploid
chromosomes following aCGH compared to 55% (n¼158) euploidy rate in
the non-PNT group (p¼0.6).
CONCLUSIONS: PNT is applicable in human zygotes without imposing
adverse impact onto the pre-implantation embryonic development and its
chromosomal content.
O-64 Monday, October 19, 2015 12:00 PM
IDENTIFICATION OF EMBRYO MARKERS PREDICTING
BLASTOCYST FORMATION BEFORE 1ST CLEAVAGE. Y. Kida,
N. Fukunaga, H. Kitasaka, T. Yoshimura, K. Nakayama, H. Ohno,
M. Takeuchi, M. Shimomura, S. Kounogi, Y. Asada. Asada Ladies Clinic
Medical Corporation, Nagoya, Aichi, Japan.
OBJECTIVE: To identify potential embryo markers predicting blastocyst
formation before 1st cleavage using EmbryoScopeÒ.
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: We examined 288 embryos resulting
from normal fertilization in 25 recipient patients undergoing ICSI between
March and July, 2013. The embryos were placed and cultured in the Embryo-
ScopeÒ in droplets of 30ml of Continuous Single Culture Medium (Irvine
Scientic:USA) till day 7. We analyzed the time of PB2 extrusion (tPB2),
appearance (tPNa) and fade (tPNf) of pronuclei. For tPNa, the time of
maternal pronuclei appearance was used. The following intervals were calculated:
PB2-PNa, PNa-PNf. We used Gardner’s classification to evaluate blastocysts.
Blastocysts evaluated more than 3BB were considered as goodquality
ones. T-test was used for comparison of mean timings and Chisquared
test for comparison of blastocyst rates. P-values

O-65 Monday, October 19, 2015 12:15 PM
A LONGITUDINAL STUDY OF FAMILIES CREATED
BY REPRODUCTIVE DONATION: FOLLOW-UP AT
ADOLESCENCE. E. C. Ilioi, V. Jadva, S. Golombok. Centre for Family
Research, University of Cambridge, Cambridge, United Kingdom.
OBJECTIVE: This study examines parental psychological well-being,
adolescent psychological adjustment and family functioning in families
that have used donor insemination (DI), egg donation (ED), surrogacy,
or natural conception (NC) to have a child. It is the first longitudinal
study to compare children conceived by different forms of reproductive
donation at adolescence, the time at which identity issues become
prominent and difficulties in parent-child relationships are likely to
increase.
DESIGN: The study represents the sixth phase of a longitudinal investigation.
Previous phases took place when the children were aged 1, 2, 3, 7, and
10 years. The study uses a multi-informant (mother and adolescent), and
multi-method (standardized interviews and questionnaires) approach to
data collection.
MATERIALS AND METHODS: Data were obtained from 31 DI families,
28 ED families, 29 surrogacy families, and 57 NC families when
the children were 13 to 14 years old. Questionnaires assessed parent psychological
well-being (the State-Trait Anxiety Inventory and the Edinburgh
Depression Scale), parenting and family functioning (the FAM
III Communication Scale, the Index of Family Relationships, the Rhoner
Short Parental Acceptance/Rejection Scale and the Rhoner Parental Control
Scale) and adolescent psychological adjustment (Strengths and Difficulties
Questionnaire).
RESULTS: Preliminary analyses of questionnaire data found few differences
between families who conceived through reproductive donation and
those who conceived naturally. In terms of disclosure rates since age 10, 3
DI families, 5 ED families, and 1 surrogacy family had told their child about
their conception resulting in 39% of DI families, 64% of ED families, and
86% of surrogacy families having disclosed this information to their child.
Overall, adolescents who were aware of their conception showed a clear understanding
of it, with the majority expressing a neutral or indifferent attitude.
CONCLUSIONS: Despite concerns about the psychological impact
of reproductive donation at adolescence, the findings indicate that the
families in this study were highly functioning in relation to parent psychological
well-being, and the quality of adolescents’ relationships with
their parents, irrespective of the specific method used in the child’s
conception. Initial findings indicate that parents who used reproductive
donation are not more likely to reject their child or have increased
strain in parent-child relationships at adolescence. Furthermore, adolescents
did not differ in their psychological adjustment, despite some of
them knowing they are not genetically related to one parent. Whilst
disclosure rates differed between groups, most of the adolescents who
were aware of their conception showed a clear understanding of it
regardless of whether they were told earlier in childhood or at adolescence.
Supported by: This research was Supported by the Wellcome Trust
[097857/Z/11/Z]. ECI is additionally funded by Pembroke College,
Cambridge.
O-66 Monday, October 19, 2015 12:30 PM
INTRALIPID FOR REPEATED IMPLANTATION FAILURE (RIF): A
RANDOMIZED CONTROLLED TRIAL. W. El-khayat a,b M. El
Sadek. a,b a Obstetrics and Gynecology, Cairo University, Cairo, Egypt;
b Middle East Fertility Centre, Giza, Egypt.
OBJECTIVE: To evaluate the effect of intravenous infusion of intralipid
20% on the outcome of intracytoplasmic sperm injection (ICSI) cycles in
women with history of repeated implantation failure (RIF).
DESIGN: An open label prospective randomized controlled trial
MATERIALS AND METHODS: This was a prospective randomized
controlled trial which included two hundred and three infertile couples
with a history of repeated implantation failure. Repeated implantation
failure defined as failure to achieve pregnancy after two to six ICSI cycles
with the transfer of more than ten high grade embryos. Sample size was
calculated as prior data (1) indicated that the pregnancy rate among controls
is 18%. If the pregnancy rate for experimental subjects will be
doubled to be 36%, we will need to study 94 subjects in each arm. We
will use an uncorrected chi-squared statistic to evaluate the null hypothesis.
We excluded patients with uterine fibroid, endometrial polyp, endometriosis
and hydrosalpinx, also disturbances of normal fat metabolism,
allergy to eggs, soybean oil, and finally liver disease, kidney disease,
lung disease and blood clotting disorder. The subjects were randomly
allocated to either the intervention group (group A) or the control group
(group B) with the use of computer-generated random number tables and
opaque sealed envelopes containing the participants’ group allocation.
Group A (n¼101) subjects were given Intralipid 20% ( Intralipid, Fresenius
Kabi AB, Uppsala, Sweden) by intravenous infusion between day
four and nine of ovarian stimulation during ICSI cycles & another dose
when got pregnant within the 1st week of positive pregnancy test, while
group B (n¼102) subjects were control group and not given Intralipid.
The primary outcome measure was the clinical pregnancy rate. Clinical
pregnancy was considered as the presence of a gestational sac with a fetal
heart activity. Comparison of quantitative variables between the study
groups was done with the use of Student t-test for independent samples.
For comparing categorical data, chi-square test was performed (Clinical
Trial Registration Number: NCT01540591).
RESULTS: There were statistically significant differences between both
groups regarding the clinical pregnancy rate, the implantation rate and
live birth rate. The clinical pregnancy rate was 35% in group A while it
was only 15% in group B (p ¼ 0.001) with RR 1.6 (1.25 - 2.09), the
implantation rate in group A was 13% while in group B it was 5%
(p ¼ 0.048) with RR 1.5 (1.09 - 2.08), and the live birth rate was 33%
in group A while it was 13% in group B (p ¼ 0.001) with RR 1.6
(1.28 - 2.13).
CONCLUSIONS: Intravenous Intralipid 20% infusion improved
significantly the clinical pregnancy rate, the implantation rate and the
live birth rate in ICSI cycles in women with history of repeated implantation
failure.
Reference:
1. Shohayeb A, El-Khayat W. Does a single endometrial biopsy regimen
(S-EBR)improve ICSI outcome in patients with repeated implantation
failure? A randomised controlled trial. Eur J Obstet Gynecol Reprod
Biol. 2012 Oct;164(2):176-9.
e26 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

REPRODUCTIVE SURGERY
O-67 Monday, October 19, 2015 11:15 AM
THE IMPACT OF UNILATERAL SALPINGECTOMY ON ANTRAL
FOLLICLE COUNT AND OVARIAN RESPONSE IN ICSI CYCLES:
COMPARISON OF CONTRALATERAL SIDE. B. Demir, a
G. Bozdag, b O. Sengul, c I. Kahyaoglu, d S. Mumusoglu, e D. Zengin. f
a Obstetrics and Gynaecology, Etlik Zubeyde Hanim Women’s Health Teaching
and Research Hospital, Ankara, Turkey; b Hacettepe University, School of Medicine,
Ankara, Turkey; c Yildirim Beyazid University, School of Medicine, Ankara,
Turkey; d Etlik Zubeyde Hanim Women’s Health Teaching and Research
Hospital, Ankara, Turkey; e Obstetrics and Gynaecology, Hacettepe University,
School of Medicine, Ankara, Turkey; f Hacettepe University, Ankara, Turkey.
OBJECTIVE: Salpingectomy is a treatment option in cases with hydrosalpinx
and ruptured ectopic pregnancy. Because of the close relationship
between mesosalpinx and ovarian blood supply, salpingectomy may compromise
ovarian reserve. However, salpingectomy related detrimental effect on
the ovarian reserve is still controversial. The purpose of this study is to determine
the impact of salpingectomy on the ovarian reserve. For this aim, antral
follicle count and ovarian response were compared with the contralateral side
in patients with unilateral salpingectomy undergoing ICSI cycles.
DESIGN: Retrospective multicenter study
TABLE 1. Antral follicle count and ovarian response of the patients
Operated
side
MATERIALS AND METHODS: Between January-2009 and April-2015,
all patients who had unilateral salpingectomy undergoing ICSI treatment in
ART Center of Etlik Zubeyde Hanim Women’s Health Teaching and
Research Hospital and Department of Obstetrics and Gynecology, Hacettepe
University were retrospectively reviewed from patient files and computer
based data. Inclusion criteria were history of unilateral salpingectomy, woman’s
age %40 years, no history of ovarian surgery. Women with bilateral salpingectomy
and a history of endometriosis were excluded from the study.
Ovarian reserve was evaluated with the antral follicle count (AFC). AFC
was measured at the beginning of the menstrual cycle. Controlled ovarian hyperstimulation
(COH) parameters and number of collected oocytes were used
for the evaluation of the ovarian response.
RESULTS: A total of 56 patients included in this study. The mean age of
the women is 31.64.7 years. The reason of the salpingectomy as follows:
hydrosalpinx 39.3% (n¼22), ruptured ectopic pregnancy 60.7% (n¼34).
The clinical pregnancy rate per embryo transfer is 30%. The AFC and
COH parameters were presented in Table.
CONCLUSIONS: The present study suggests that salpingectomy is not
associated with detrimental effects on the antral follicle count and ovarian
response.
O-68 Monday, October 19, 2015 11:30 AM
Non-operated
side
P value
Antral follicle count 5.914 6.113.7 NS
No. of follicle >17 mm in size 1.781.7 2.092.2 NS
on the hCG day
No. follicle >10-17 mm in size 6.234.4 5.923.8 NS
on the hCG day
No. of total collected oocytes 6.665.5 6.174.2 NS
THE COMPARISON OF ICSI-ET OUTCOMES IN SURGICALY
TREATED MINIMAL ENDOMETRIOSIS, ENDOMETRIOMA AND
TUBAL FACTOR WITHOUT UNDERLYING ENDOMETRIOSIS.
I. Guler, a A. Erdem, a M. Erdem, a N. Bozkurt, a M. Oktem, a M. F. Mutlu, b
Y. Oguz, a F. Cevher. a a Obstetrics and Gynecology, Gazi University Faculty
of Medicine, Ankara, Turkey; b Koru Hospital, Ankara, Turkey.
OBJECTIVE: To assess the effect of laparoscopic surgery in patients with
minimal endometriosis and endometrioma on the outcome of IVF-ET by
comparing with the results of patients with tubal factor without underlying
endometriosis that is confirmed by laparoscopy.
DESIGN: Retrospective case-control study
MATERIALS AND METHODS: From 2003 to 2013 retrospective data of
patients with minimal endometriosis or endometrioma who were diagnosed
and treated by laparoscopic surgery before ICSI-ET were reviewed for the
study. In all patients with endometrioma, laparoscopic unilateral or bilateral
cystectomy was performed. The control group was chosen from the patients
with the diagnosis of tubal factor who were underwent laparoscopy and not
diagnosed with underlying endometriosis. Patients with additional male factor,
thyroid disease, PCO in each group were also excluded. Statistical significance
was defined as p < 0.05.
RESULTS: A total number of 365 cycles of 206 patients, including 92
cycles of 47 patients with minimal endometriosis, 65 cycles of 30 patients
with endometrioma and 208 cycles of 129 patients with tubal factor were
analyzed. Mean ages of patients among three groups were comparable. Basal
antral follicle count, total dose of gonadotropins, duration of stimulation,
total number of retrieved oocytes, fertilization, embryo clivage and clinical
pregnancy rates were found to be significantly different between groups.
All results were summarized in the Table 1.
CONCLUSIONS: Even if minimal endometriosis or endometrioma was
treated by laparoscopy prior to IVF, clinical pregnancy rates tended to be
lower in endometriosis than tubal factor infertility that was not associated
with endometriosis. Laparoscopic removal of endometrioma was also related
with lower ovarian response and less number of oocyte retrieval.
TABLE 1. Comparison of ICSI outcomes among study and control groups
Variable
Minimal
Endometriosis
(n¼ 47, 92
cycles)
Supported by: Gazi University.
Endometrioma
(n¼30, 65
cycles)
Tubal
Factor
(n¼129,
208 cycles)
p
value
Age (years) 31.9 4.6 32.4 4.3 32.5 4.9 0.586
Total basal antral 8.9 5.4 6.1 3.9 6.8 5.4 0.003
follicle count
(n)
Day 3 Basal 6.7 2.8 8.8 4.4 8.4 8.2 0.125
FSH level
(IU/L)
Duration of 10.6 2.8 11.1 2.9 10.1 2.6 0.044
stimulation
(day)
Total dose of
gonadotropins
(IU)
3400.31
1903.98
4255.71
1601.21
3113.63
1424.09
0.000
Serum E2 on the
day of hCG
injection
(pg/mL)
Fertilization
rate (%)
Embryo cleavage
rate (%)
Total number of
retrieved
oocytes (n)
Total number of
retrieved M2
oocytes (n)
Clinical pregnancy
rate (%)
1635.52
1402.38
1287.13
1002.42
1741.19
1508.87
0.186
75.9 23.4 66.3 28.3 75.7 23.2 0.053
91.2 21.8 85.7 30.0 94.6 14.0 0.026
10.9 8.2 7.1 4.5 10.6 8.7 0.018
8.2 6.6 5.8 3.8 8.1 6.6 0.062
25.0 14.3 58.3 0.000
O-69 Monday, October 19, 2015 11:45 AM
MISDIAGNOSED SEPTATE UTERUS: DO WE REALLY NEED
THREE DIMENSIONAL TRANSVAGINAL ULTRASOUND (3D-US)
TO REEVALUATE ARCUATE UTERI DIAGNOSED BY
HYSTROSALPINGOGRAPHY (HSG)? A. M. Abdelmagied, a,b
M. A. Kamel, b A. M. Abuelhasan, b T. A. Farghaly, b A. A. Nassr, a,b
S. A. Shazly, a,b M. H. Makarem. b a Obstetrics and Gynecology, Mayo Clinic,
Rochester, MN; b Obstetrics and Gynecology, Women Health Hospital,
Assiut University, Assiut, Egypt.
OBJECTIVE: It is well known that septate uterus is a common cause of
reproductive failure and its correction has been linked to better prognosis.
Also, there is evidence that infertile women both with small and large septae
report decreased implantation and pregnancy rates. Our objective was to
assess the utility of 3D-US after HSG for the differentiation between arcuate
and septate uterus in infertile women.
FERTILITY & STERILITY Ò
e27

DESIGN: Prospective cohort study
MATERIALS AND METHODS: Infertile women with diagnosis of
arcuate and septate uterus based on HSG were recruited. All women
were examined by 3D-US on day 22 cycle to allow for better delineation
of the uterine contour. The outer and inner fundal contours and the length
of the fundal notch were examined by 3D-US in the mid-coronal view of
the multi-planar and multi-slice display of the uterus. The final diagnosis
of the anomalies was based on combined hysteroscopy/laparoscopy examination
(HL), the gold standard,. In cases of septate uterus, HL was accompanied
by hysteroscopic resection of the septum. ASRM description of the
congenital uterine anomalies was followed to categorize the findings. For
imaging and endoscopic analysis of the uterine morphology, standards in
the literature were utilized to appropriately define each anomaly. To calculate
the level of agreement between the diagnostic modalities, kappa statistics
were utilized.
RESULTS: Thirty eight women were included. Based on HL diagnosis,
3 groups of congenital uterine anolamlies were diagnosed: arcuate uterus
(n¼12,31.5%), septate uterus (n¼22,58%) and bicornuate uterus (n¼4,
10.5%). 3D-US showed perfect diagnostic accuracy (100%) and perfect
concordance (k ¼ 1.00, 95% CI ¼ 1.00-1.00) with HL in identifying all
anomalies in the 3 groups while HSG had 68.4% accuracy and fair concordance
(k ¼ 0.37, 95% CI ¼ 0.09-0.66) with HL. For septate uterus, 8 cases
were misdiagnosed by HSG as arcuate uterus (40%) while for bicornuate
uterus; HSG misdiagnosed all of them as septate. Septae missed by HSG
were less than 2 cm in length and shorter (Median, 17 vs. 29 mm) than those
correctly diagnosed.
CONCLUSIONS: In infertile women, diagnosis of arcuate uterus based on
HSG needs further evaluation by 3D-US in order to diagnose possible missed
small septae. Being in perfect concordance with HL, 3D-US averts the need
to endoscopy to differentiate between common uterine anomalies and offers
the preoperative confirmation of the anomaly that should be surgically corrected.
References:
1. Salim R, Woelfer B, Backos M, Regan L, Jurkovic D. Reproducibility
of threedimensional ultrasound diagnosis of congenital uterine anomalies.
Ultrasound Obstet Gynecol 2003; 21:578-82.
2. Ludwin A, Pity_nski K, Ludwin I, Banas T, Knafel A. Two- and three--
dimensional ultrasonography and sonohysterography versus hysteroscopy
with laparoscopy in the differential diagnosis of septate,
bicornuate, and arcuate uteri. J Minim Invasive Gynecol 2013; 20:90-9.
3. Bermejo C, Martınez Ten P, Cantarero R, Diaz D, P_erez Pedregosa J,
Barr_on E, et al. Three-dimensional ultrasound in the diagnosis of
mullerian duct anomalies and concordance with magnetic resonance
imaging. Ultrasound Obstet Gynecol 2010; 35:593-601.
OBJECTIVE: To determine if there is discrepancy between the measurement
of the mid fundal length of subtle incomplete uterine septum or arcuate
uterine anomaly on hysteroscopy and on trans-vaginal 3D ultrasound scan
(TV 3D US) with or without saline infusion sonohysterogram (SIH).
DESIGN: Retrospective study
MATERIALS AND METHODS: Two hundred sixty three patients who
had a subtle incomplete uterine septum (30.0%) or arcuate uterine anomaly
(70.0%) on hysteroscopy between 2010 and 2015 were studied. All patients
had TV 3D US with and without SIH as part of their work up for infertility
(65.0%), history of miscarriage (6.6%) or both (28.3%) and subsequently had
a hysteroscopy with or without laparoscopy. The length of the mid fundal
protrusion in the endometrial cavity was calculated on 3D TVUS with and
without SIH. If the 2 values were available, a mean value was obtained; otherwise
the one available value was used. At time of hysteroscopy the type of
uterine anomalies according to ASRM classification (Class IV b or
Class V) was documented and its mid fundal length was measured using
the tip of a straight resectoscope loop. If the incomplete uterine septum or
arcuate uterine anomaly was deemed clinically significant (length > 10
mm) it was divided at the same session. We compared the findings on hysteroscopy
and on 3D TVUS with and without SIH. Paired t-test and correlation
analysis were used.
RESULTS: Mean age was 32.6 + 5.2 years, mean body mass index was
27.6 + 6.6 kg/m 2, and 59.7% of patients had primary infertility. Overall
mean fundal length on TV 3D US (6.4 + 2.9 cm) was significantly lower
than mean actual length on hysteroscopy (13.5 + 3.3 cm), p

HEALTH DISPARITIES 1
O-73 Monday, October 19, 2015 11:15 AM
O-72 Monday, October 19, 2015 12:30 PM
COMPARISON OF OPERATIVE AND PERI-OPERATIVE
RESULTS FOR ROBOTIC AND LAPAROSCOPIC
MYOMECTOMIES. C. E. Miller, a K. Sasaki, b C. Steller, c S. Sulo, d
A. Cholkeri-Singh. e a The Advanced IVF Institute, Naperville, IL; b Advanced
Gynecologic Surgery Institute, Naperville, IL; c Advocate Lutheran General
Hospital, Park Ridge, IL; d Center for Advanced Care, Russell Institute of
Research and Innovation, Park Ridge, IL; e The Advanced Gynecologic Surgery
Institute, Naperville, IL.
OBJECTIVE: To compare operative and peri-operative outcomes between
laparoscopic and robotic myomectomies.
DESIGN: A review of prospectively maintained data in 2:1 matched cases
of 96 laparoscopic and 48 robotic myomectomies. The cases were matched
for number and size of fibroids removed, and timing of post-operative ultrasound.
Medical charts were abstracted for demographics, operative and perioperative
outcomes.
MATERIALS AND METHODS: Women over the age of 18 who underwent
a laparoscopic or robotic myomectomy from July 2009 through
December 2014.
RESULTS: The data was analyzed via Student t-test for continuous variables
and Chi-square or Fischer’s exact for categorical variables. A
Spearman correlation was performed for conversion rate with estimated
blood loss and operative time. A linear regression was performed for predictive
variables on number of post-operative fibroids found. No differences
were identified between the two cohorts with respect to age,
pregnancy history, prior pelvic surgery, trocar number, or number or
size of fibroids removed (p>.05). There were no differences in operative
time, estimated blood loss, complication or overnight admission rates. An
increased conversion rate in the robotic group to laparoscopy correlated
with an increased estimated blood loss (r¼0.237, p¼ .004) and operative
time (r¼.143, p¼ .038), due to case complexity. No differences in postoperative
ultrasound findings for fibroid number or size were found. Number
of fibroids removed intra-opeatively was a significant predictor of
increased post-operative fibroid number (b¼ .262, 95% CI .027-.113,
p¼ .002) for either cohort.
CONCLUSIONS: Laparoscopic and robotic myomectomies have similar
outcomes in skilled surgical hands. The only difference was an increased
conversion rate for complex cases in the robotic cohort. The number of
post-operative fibroids did not differ, thus demonstrating that although robotics
lacks haptic feedback, it does not limit a surgeon’s ability to identify
and remove fibroids.
Comparison of Operative and Peri-operative Outcomes
Laparoscopic
Myomectomy
N¼96
Robotic
Myomectomy
N¼48 p-value
Operative Time (min), 162.2 (80.2) 169.3 (87.2) .627
mean (SD)
Estimated Blood Loss 126.4 (189) 111.9 (171.9) .674
(EBL) (mL), mean (SD)
Conversion of Surgical 0 3 (6.3%) .035
Approach, N (%)
Complication Rate, N (%) 2 (2.1%) 3 (6.3%) .333
Admission Rate, N (%) 15 (15.6%) 11 (23%) .284
Number of fibroids removed 4.54 (4.0) 4.3 (4.2) .739
intra-operatively, mean (SD)
Number of fibroids
on post-operative
ultrasound, mean (SD)
0.4 (1.2) 0.3 (0.78) .591
EARLY LIFE EXPOSURE TO ESTROGEN-MIMICS INCREASE
THE OCCURRENCE OF UTERINE FIBROIDS VIA EXPANSION
OF MYOMETRIAL STEM CELL POPULATION. A. Mas, a
L. Elam, a C. Walker, b C. Simon, c M. P. Diamond, a W. Thompson, d
A. Al-Hendy. a a Department of Obstetrics and Gynecology, Georgia Regents
University, Augusta, GA; b Department of Obstetrics and Gynecology, Georgia
Regents University, Augusta, TX; c Fundacion Instituto Valenciano de Infertilidad
IVI-Universidad de Valencia-INCLIVA, Paterna (Valencia), Spain;
d Department of Obstetrics and Gynecology, Morehouse School of Medicine,
Atlanta, GA.
OBJECTIVE: Experimental animal studies have shown that exposure of
target tissues to xenoestrogens (XEs) during sensitive periods of development
can increase risk of disease in adult life, a process known as developmental
reprogramming. Normal uterine myometrium is sensitive to ovarian
hormones, making it a potential target for XEs. Developmental exposures
to XEs have been shown to cause alterations the epigenetic programming
of exposed myometrial cells, increasing risk for development of uterine
fibroids. Our goal was to determine if exposure to XEs during critical periods
of development altered the number and/or function of myometrial stem cells
in exposed tissues, contributing to fibroid tumorigenesis.
DESIGN: Laboratory research studies using a murine model of a human
disease.
MATERIALS AND METHODS: Eker rats (animal model for uterine fibroids)
were exposed to XEs diethylstilbestrol (DES) or vehicle control
(VEH) during postnatal days (PND) 10-12, and maintained until adulthood
at 5 months of age. Myometrial cells were isolated from the uterine horns
and cervix of adult Eker rats, and sorted by flow cytometry (FACS) using
Stro1/CD44 myometrial stem cell markers. Additionally, human myometrium
isolated from women without fibroids (MyoN) or women with fibroids
(MyoF) was examined by immunohistochemistry (IHC) for Stro1/CD44-
positive (stem) cells.
RESULTS: At 5 months of age, morphological and functional differences
were observed in the uterus of DES versus VEH treated rats. Quantification
of Stro1/CD44 myometrial stem cells by FACS revealed that the percentage
of myometrial stem cells in cervix (44.8%) and horns (23.4%) from DES
exposed animal was significantly higher than cervix (35.3%) and horns
(11.69%) from VEH rats (p%0.05). Quantification of Stro1/CD44 human
myometrial stem cells by IHC demonstrated that the percentage of Stro1/
CD44 positive cells was significantly higher in MyoF versus MyoN
(p

with sufficient certainty and accuracy. The imaging characteristics are shared
by the more common benign leiomyoma and potential serum biomarkers are
not reliable. Developing a diagnostic tool for LMS is a high priority and
would improve health care of women with suspicious myometrial mass.
we wanted to identify a LMS specific promoter that can potentially drive a
reporter gene to be expressed only in LMS cells (LMS-ON/Leiomyoma-
OFF)
DESIGN: Laboratory studies using human cells
MATERIALS AND METHODS: We screened several modified adenoviruses
encompassing various neoplasm-related promoters driving the
expression of luciferase reporter gene The Adenovirus construct that
showed highest expression potential in LMS, Ad-ANS-886, was selected
for further evaluation. In-vitro, we transfected 3 cell types, which are
SK-UT-1 (LMS), primary fibroid (1ry F) and myometrium (Myo F) cells
with Ad-ANS-886 at multiplicity of infection (MOI) 1, 5. Luciferase transactivation
was evaluated by both luciferase assay as well as xenogen camera
imaging of cells 24 and 48 hours after transduction. Further in vivo
evaluation of Ad-ANS-886 was performed by injecting adenovirus transfected
LMS cells (5^6 cells/mouse) both subcutaneous (SC) and intrauterine
(IU) in 6 weeks old female nude mice, followed by imaging the
animals with Xenogen camera. Control animals were implanted with
1ryF cells.
RESULTS: Ad-ANS-886 transfects LMS cells much more readily than
benign or normal myometrial cells as evidenced by luciferase assay that
showed 6-fold higher bioluminescence in LMS vs 1ryF, MyoF at MOI 5
with p < 0.0001. IVIS of LMS gave 100 fold higher total photon emission
per second (TPE/S) compared to 1ryF , MyoF (p< 0.0001). In-vivo studies
showed highly significant rise in TPE/S emission in LMS-based lesions vs
1ryF-related lesions at 48 hours post cell implantation both in SC as well
as IU locations (pA, p.G44D)] plasmid constructs containing a flag-tag
and a green fluorescence protein (GFP). Transfected cells were selected
with puromycin to generate stable cell populations which were characterized
to determine the effect of MED12 somatic mutations on expression levels of
Wnt4 and b-catenin.
RESULTS: In western blot analyses using lysates from the above stable
cells, we demonstrated that cells expressing mutant MED12 protein
exhibited higher levels of Wnt4 and b-catenin as compared with cells expressing
WT-MED12 protein. We also observed that the exogenous
mutant-MED12 protein can induce Wnt4 and b-catenin in both cytosolic
and nuclear compartments as compared with WT-MED12 protein. Cell-cycle
analyses by FACScan assay showed that the expression of mutant-
MED12 protein induced UtSMC cells into S-phase (22%, p

O-77 Monday, October 19, 2015 12:15 PM
THE IMPACT OF ALCOHOL CONSUMPTION ON OVARIAN
RESERVE IN REPRODUCTIVE-AGE AFRICAN AMERICAN
WOMEN. L. Hawkins Bressler, a L. A. Bernardi, b P. D. De Chavez, b
D. Baird, c M. Carnethon, d E. E. Marsh. e a Department of Obstetrics and
Gynecology, Northwestern University, Chicago, IL; b Northwestern University,
Chicago, IL; c Epidemiology, National Institute of Environmental Health
Sciences, Research Triangle Park, NC;
d Department of Epidemiology,
Northwestern University, Chicago, IL;
e Feinberg SOM - Northwestern
University, Chicago, IL.
OBJECTIVE: To evaluate the relationship between alcohol intake and
anti-m€ullerian hormone (AMH) levels as a marker of ovarian reserve in
reproductive-age African American women. A racial dichotomy exists in
the literature implying a differential impact of alcohol on AMH whereby consumption
does not affect AMH levels in white women (1-2) but is associated
with lower AMH levels in black women (3).
DESIGN: Cross-sectional analysis from an ongoing prospective study.
MATERIALS AND METHODS: A total of 1,654 African American
women (AAW) ages 23-35 completed questionnaires on the duration, frequency
and amount of their alcohol consumption and provided serum for
AMH measurement. Multivariable linear and logistic regression accounting
for age, body mass index (BMI), hormonal contraception (HC), irregular
menses and thyroid conditions were used to assess the relationship between
alcohol intake and AMH. Binge drinking was defined as four or more drinks
consumed on one occasion.
RESULTS: Most reproductive-age AAW (73.5%) drink alcohol. Of these,
33.9% report weekly or more frequent drinking in the last 12 months. The
majority (74.1%) report binge drinking, of which more than a fourth
(26.5%) report binge drinking multiple times per month in the last year.
Among AAW who drink alcohol, 83.7% report their heaviest consumption
occurred over the age of 19. Frequency of binge drinking appeared to demonstrate
an inverse association with AMH and approached significance
among AAW who binge drink twice weekly or more compared to never bingers
(b¼-0.27, P¼0.07). Ever drinking, years of regular drinking, frequency of
alcohol consumption, number of drinks per day, type of alcohol consumed
and age at heaviest consumption were not associated with AMH (all P>0.2).
CONCLUSIONS: Findings represent the most comprehensive assessment
to date of patterns of alcohol consumption on AMH levels. Among reproductive-age
AAW, alcohol intake does not appear to meaningfully impact AMH.
Additional investigation in the infertile population including functional
ovarian reserve testing would assess reproducibility and permit further characterization
of the impact of binge drinking on ovarian reserve.
References:
1. Dolleman, M., Verschuren, W. M., Eijkemans, M. J., Dolle, M. E., Jansen,
E. H., Broekmans, F. J., & van der Schouw, Y. T. Reproductive and
lifestyle determinants of anti-Mullerian hormone in a large populationbased
study. J Clin Endocrinol Metab, 2013;98(5):2106-2115.
2. Nardo, L. G., Christodoulou, D., Gould, D., Roberts, S. A., Fitzgerald,
C. T., & Laing, I. Anti-Mullerian hormone levels and antral follicle
count in women enrolled in in vitro fertilization cycles: relationship
to lifestyle factors, chronological age and reproductive history. Gynecol
Endocrinol, 2007;23(8):486-493.
3. Whitworth, K. W., Baird, D. D., Steiner, A. Z., Bornman, R. M., Travlos,
G. S., Wilson, R. E., & Longnecker, M. P. Anti-Mullerian Hormone
and Lifestyle, Reproductive, and Environmental Factors Among
Women in Rural South Africa. Epidemiology 2015;26(3):429-35.
Supported by: National Institutes of Health Grant R21HD077479-01; National
Institutes of Health Grant K12HD050121; Northwestern University
Women’s Reproductive Health (WRHR) Scholar Award; Harold Amos Medical
Faculty Development Award; Robert Wood Johnson Foundation.
O-78 Monday, October 19, 2015 12:30 PM
PREGNANCY RATES AMONG AFRICAN-AMERICAN WOMEN
UNDERGOING SINGLE EMBRYO TRANSFER (SET). T. Plowden, a
A. Christy, a A. DeCherney, a J. Csokmay. b a NIH, Bethesda, MD;
b WRNMMC, Bethesda, MD.
OBJECTIVE: One of the greatest risks of Assisted Reproductive Technologies
is iatrogenic multi-fetal gestation, which is associated with significant
fetal and maternal morbidity and mortality. In recent years, there is a national
impetus for IVF practices to move towards SET to decrease the rate of multiple
pregnancies. Previous IVF literature has shown that African-American
women undergoing IVF are less likely to become pregnant than their Caucasian
counterparts. To date, no studies have specifically explored the success
of SET in African-American women. Our objective was to explore clinical
pregnancy (IUP with cardiac activity) and spontaneous abortion (SAB) rates
in African-American women undergoing SET.
DESIGN: This study was a retrospective analysis of patients undergoing
treatment from Jan 2012-Dec 2014.
MATERIALS AND METHODS: Our institution routinely recommends
transfer of a single embryo for good prognosis women ( 0 was equal to or greater than .95 or equal to or less than
.05. Separate estimates for effects were calculated for men and women within
the same model and the correlation coefficient within a couple was calculated
for all sub-scale scores. In addition to the effect of time, age, infertility and
mental health treatment history (MHT) were added to the models. The impact
of miscarriage was measured using three sub-scales from the Revised Impact
of Miscarriage Scale (RIMS)(Huffman, Swanson, & Lynn, 2014): The RIMS
is comprised of three sub-scales: 1) Isolation and guilt (I/G), 2) Loss of baby
(LB), and 3) Devastating event (DE). Higher scores indicate greater impact.
RESULTS: 302 couples data were eligible for analysis.The mean age of
women was 32.62(SD ¼ 6.01) and 34.08( SD ¼ 6.77) for men, mean gestational
age at miscarriage was 9.9 weeks (SD ¼ 3.08). Seventy-four couples
had a history of infertility, 48% of women and 24% of men were positive
for MHT. When controlling for baseline scores, infertility had no effect on
I/G, DE, or LB scores over the course of a year. Women were no more
FERTILITY & STERILITY Ò
e31

likely to feel isolated or guilty than their fertile counter parts (pp of an effect
for infertility was .48 in women and .63 in men). Men with MHT were
more isolated/guilty during the first year than men without such history
(mean parameter estimate for MHT ¼ .62, pp ¼ .99). Older men and
women were less likely to view their miscarriage as the loss of a baby
than younger men and women, though this did not reach significance.
Time had a significant effect in reducing I/G, DE, and LB in women.
Men’s initial impact remained constant over the course of the year except
for devastation, which showed a small decrease. The couples scores were
significantly correlated, albeit, the correlation coefficients were low ( r ¼
.07 - .16, pp ¼ .96 -.99)
CONCLUSIONS: During the first year of loss, the impact of miscarriage is
not influenced by infertility. Men with a history of grief, depression, or anxiety
may experience more isolation and guilt. Women see a decrease in impact
over time, where as men only see a decrease in the devastation experienced.
References:
1. Swanson KM, Chen HT, Graham, JC, Wojnar DM, Petras A. Resolution
of depression and grief during the first year after miscarriage: a randomized
controlled clinical trial of couples-focused interventions. J
Womens Health (Larchmt), 2009; 18:1245-57.
2. Huffman CS, Swanson KM, Lynn M. Measuring the meaning of
miscarriage: revision of the Impact of Miscarriage Scale. J of Nurs
Measurement 2014; 22: 29-45.
Supported by: Funding for the original study was provided to K.M.S. by
the NIH, National Institute of Nursing Research, 5 R01 NR005343. Trial
registration number: NCT00194844.
O-80 Monday, October 19, 2015 11:30 AM
THE PREVALENCE OF ABNORMAL MAMMOGRAMS IN OVUM
RECIPIENTS DOES NOT CORRELATE WITH RECIPIENT
AGE. S. E. Yerkes, a J. Rodriguez-Purata, b J. A. Lee, a
M. C. Whitehouse, b M. Daneyko, a B. Sandler, b,c A. B. Copperman. a,c a Reproductive
Medicine Associates of New York, New York, NY; b Reproductive
Medicine Associates of New York, New York City, NY; c Obstetrics, Gynecology
and Reproductive Science, Mount Sinai School of Medicine, New
York, NY.
OBJECTIVE: Ovum recipients undergo endometrial preparation with estrogen
to mimic the natural menstrual cycle. Recipients R40 years are
required to complete a mammogram, but younger patients are not. The study
sought to compare if the prevalence of an abnormal mammogram in ovum
recipients increases with ovum recipient age.
DESIGN: Retrospective analysis.
MATERIALS AND METHODS: Patients who had a mammogram prior to
starting an ovum donation cycle from 2010 - 2014 were included. The following
Breast Imaging Reporting and Data System (BIRADS) categories were used for
reporting mammographic results: 0: incomplete (needs additional image evaluation
and/or prior mammograms for comparison); 1: negative; 2: benign; 3:
probably benign; 4: suspicious; 5: highly suggestive of malignancy; and 6:
known biopsy (proven malignancy). Patients were segregated into two groups
according to age (Group A:

for statistical significance. A logistical regression model was also utilized to
identify significance of multi-variables.
RESULTS: Responses (N¼117) were returned. The female rate was 66%
(N¼88). The majority,98%, were Christian (N¼112). The majority, 88%,
were college educated (N¼102) with 39% (N¼40) with some post-graduate
education. Sixty-eight percent (N¼79) had no knowledge of PGS prior to
their IVF cycle; however, after provider education, ninety-two percent
(N¼108) correctly identified that PGS was elective and 93% (N¼109) reported
their knowledge was sufficient to make an informed decision to accept
or decline PGS. The additional cost of screening (P¼0.004), the provider information
and influence (P¼0.0001), and social support or acceptance from
partner, family, friends (P¼0.03), were the three variables affecting the decision.
In a logistical regression model, additional cost (P¼0.003) and explanation
from the provider (P¼0.0003) were the only significant
determinants influencing the decision. Religious and ethical beliefs were
important (P¼0.20), but not statistically significant nor was information
regarding the disposition of abnormal embryos (P¼0.18) or the concerns
regarding cryopreservation and transfer at a later date (P¼0.79).
CONCLUSIONS: PGS use as an adjuvant therapy to aid in selection of the
embryo with the greatest potential for success, has become readily available
in many IVF clinics. Researchers have spent time, resources, and effort on the
clinical benefits and limitations of PGS, yet little research on the patient
perspective, knowledge, education, and awareness regarding the options
and ultimate decision making capabilities of the patient. This is the first study
to the authors knowledge to identify and assess the determinants of the patient
decision making process when presented with the choice of PGS in a
given IVF treatment cycle. Several factors contribute to the patient perceived
determinants when choosing to accept or decline PGS, including cost, religious/ethical
values, social influence, and the past experience of the patient.
O-83 Monday, October 19, 2015 12:15 PM
LESSENING THE BURDEN OF CARE: STEPPED VS. FIXED
ESTRADIOL PROTOCOL FOR FROZEN EMBRYO
TRANSFER. A. B. Tepper, a C. M. Bergh, a T. Molinaro, a,b
P. A. Bergh. a,b a Reproductive Medicine Associates of New Jersey, Basking
Ridge, NJ; b Obstetrics, Gynecology & Reproductive Sciences, Rutgers-
Robert Wood Johnson Medical School, New Brunswick, NJ.
OBJECTIVE: Simplification of cycle medication protocols without a
reduction in efficacy is a key priority in lessening the patient’s burden of
care. This study sought to compare frozen embryo transfer (FET) cycle outcomes
between estradiol therapies using a 1mg stepped protocol or 2mg fixed
protocol at cycle start.
DESIGN: Retrospective database study.
MATERIALS AND METHODS: Analysis was performed on 3,984 patients’
electronic medical records at Reproductive Medicine Associates of
New Jersey (RMANJ) from October 9, 1999 to April 30, 2015. FET cycles
using autologous oocytes and blastocyst transfers with a starting dose of
either 1 or 2mg were included in this study. Gestational Carrier cycles
were excluded. Chi-square and t-test were performed for our normally
distributed population. The study was powered to detect a 5% difference between
groups. Logistic regression was performed to control for confounders.
RESULTS: There were small statistical significant differences in endometrial
thickness, length of cycle and use of CCS. Implantation rates were
similar between both groups. When controlling for age, CCS use, number
of embryos transferred, and method of cryopreservation, there were no significant
differences in clinical pregnancy (OR 0.96; 95%CI 0.79 - 1.15)
nor clinical loss rate (OR 0.98; 95%CI 0.79 - 1.22).
Cycle Characteristics Stratified By Starting Estradiol Dose.
1mg. estradiol 2mg. estradiol
p value
Sample Size (n) 3102 882
Oocyte Age (years) 34.2 34.2 p ¼ 0.887
Endometrial Thickness (mm) 9.9 9.6 p ¼ 0.000
Length of Cycle (days) 12.5 11.8 p ¼ 0.000
Use of Comprehensive 53.5 57.4 p ¼ 0.041
Chromosome Screening
(CCS) (%)
Clinical Pregnancy Rate (%) 76.7 77.0 p ¼ 0.835
Implantation Rate (%) 68.7 69.5 p ¼ 0.618
CONCLUSIONS: Clinical pregnancy and loss rates are not impacted by
using a 2 mg fixed protocol for a FET cycle. Using a standardized fixed protocol
may reduce provider ordering and patient administration errors, number
of monitoring visits, and improve patient compliance. As described, a fixed
FET protocol may reduce the patient’s burden of care without compromising
outcomes.
O-84 Monday, October 19, 2015 12:30 PM
GAY SURROGACY-THE QUANDRY OF ACCESSING VERIFIABLE
FACTS. D. Smotrich, a A. Botes, a X. Wang, a M. Gaona, b D. Batzofin. a
a IVF, LaJolla IVF, La Jolla, CA; b Embryology/Andrology, LaJolla IVF, La
Jolla, CA.
OBJECTIVE: To bring attention to the lack of verifiable data pinpointing
clinics that are accommodating to LGBT patients as well as being able to
easily obtain LGBT friendly legitimate and objective statistical data
regarding the use and success of gestational surrogate (GS) egg donor
(ED) cycles to create their families.
DESIGN: Retrospective analysis.
MATERIALS AND METHODS: Retrospective review of IVF laboratory
database and patients’ charts was conducted on 529 consecutive fresh GS/ED
cycles performed for gay couples from January 2004 - December 2013 (a ten
year period) at a private clinic in California. After May 2005 all gamete providers
were subject to FDA regulations. Standard stimulation protocols,
monitoring, egg retrievals and embryo transfers were performed. Embryos
were created with ICSI (100%), PGD/S (75%) and blastocyst transfers
(80%). A review of a subset of 200 randomly selected gay intended parents’
charts from the 529 cycles was undertaken to analyze documented comments
by patients regarding their country of origin, from where they learned about
GS/ED as a treatment option and what additional information would have
been useful in their decision making.
RESULTS: From January 2004 - December 2013, 529 fresh GS/ED cycles
were performed for gay couples. A clinical pregnancy was confirmed in 454
GS for an 86% clinical pregnancy rate. 80% of the gay couples achieved a
live birth after one GS/ED cycle. Data obtained from the clinical chart review:
patients traveled from 54 countries and 47 US states; 80% learned
about GS/ED from the internet and media outlets, 7% from GS/ED agencies,
4% from peers and 9% from other patients. 96% stated some official statistics
in a report (along the lines of the CDC Clinic Success Rates Report) as being
the most authoritative guidance missing from their data search.
CONCLUSIONS: In experienced hands, gestational surrogacy/egg donation
is a highly effective treatment for gay couples in terms of family building.
These patients deserve and should be able to rely on official statistical
data in order to make a logical and informed decision in regards to their
choice of the most appropriate treatment facility for their needs, just as other
patients researching ART treatments are afforded.
EARLY PREGNANCY 1
O-85 Monday, October 19, 2015 11:15 AM
THE EFFECT OF A PUBLICLY FUNDEDED NORTH AMERICAN
IVF PROGRAM WITH MANDATED SINGLE EMBRYO TRANSFER
ON MATERNAL ANTENATAL ADMISSION RATES. M. Dahan, a
T. Shaulov, b S. Belisle. c a McGill University, Montreal, QC, Canada; b McGill
University Health Centre, Montreal, QC, Canada; c University of Montreal,
Montreal, QC, Canada.
OBJECTIVE: To determine the effect of a drastic drop in the multiple
pregnancy rate on antenatal admission of women who underwent IVF.
DESIGN: A retrospective study was performed by analyzing data concerning
IVF cycle outcomes and hospital admission in Quebec. Single payer government
health care with IVF coverage for all women enabled to track
outcome details which were gathered from the Ministry of Health and Social
Services ‘‘Pro-Assis’’ database and the hospital admission database (MED-
ECHO). The government program has mandated single embryo transfer in
most cases.
MATERIALS AND METHODS: Data is presented by financial year, and
involves more than 250 000 pregnancies. The government began covering
IVF in august 2010. Statistical analysis was performed using Chi squared
tests, odds ratios or correlation coefficients. Data is available from
FERTILITY & STERILITY Ò
e33

2009-2010, pre-program, to 2012-2013, the first year to reflect conceptions
achieved exclusively under government mandate.
RESULTS: The proportion of live births that are a result of IVF has
increased by 64% since 2009-2010, from 1.23% to 2.02% (p

(OR¼1.36 [1.09-1.69]), while it had no effect on EP in antagonist cycles
(OR¼0.99 [0.84-1.16], interaction test p¼0.01).
CONCLUSIONS: The GnRH antagonist protocol was associated with
elevated odds of EP as compared with the GnRH agonist protocol in fresh
autologous cycles. These findings highlight a possible role for extrapituitary
GnRH on the uterine environment and the need to account for multiple
maternal and treatment factors when evaluating the risk of EP after IVF.
O-88 Monday, October 19, 2015 12:00 PM
DON’T DISCRIMINATE: EVEN A DISCRIMINATORY ZONE AS
HIGH AS 4000 CAN RESULT IN INTERRUPTION OF AN
IUP. K. T. Barnhart, a M. D. Sammel, b A. Singer, a A. Reid, a
L. Taylor, b S. Senapati. a a Obstetrics and Gynecology, University of Pennsylvania
Perelman School of Medicine, Philadelphia, PA; b Biostatistics and
Epidemiology, University of Pennsylvania Perelman School of Medicine,
Philadelphia, PA.
OBJECTIVE: The discriminatory zone (DZ) is the level of hCG above
which one can be assured the pregnancy outcome will be ectopic pregnancy
(EP) or miscarriage (SAB) if a normal intrauterine pregnancy (IUP) is not
visualized by ultrasound (US). In this study, we assess how often the use
of a DZ may misclassify an IUP as a non-viable pregnancy.
DESIGN: Retrospective population-based cohort study.
MATERIALS AND METHODS: 4008 women with symptomatic first
trimester pregnancies of unknown location (PUL, defined as no intrauterine
or extrauterine gestational sac, yolk sac, or embryo) were evaluated between
January 1990 and August 2009. Subjects were stratified based on initial hCG
values at proposed DZ levels (R2000, R3000, R4000 mIU/mL) and final
outcomes of IUP, SAB, and EP. Subjects were further stratified based on
ultrasound findings that were suspicious for IUP (intrauterine echogenic
sac-like structure), suspicious for EP (adnexal mass or extrauterine sac-like
structure), or no ultrasound findings suspicious for IUP or EP. The proportion
of PUL that went on to be diagnosed as definitive IUP was computed for each
DZ threshold.
RESULTS: The median hCG level at presentation for care was 751, IQR:
198 - 2690 mIU/mL. The average age was 26.4 (sd¼6.9), 70.9% were
black and 15.6% were white. Final outcome was EP in 22%(N¼860),
IUP in 26%(N¼1054), and SAB in 52%(N¼2094) of the population.
30% had a previous pregnancy, 24% had a history of SAB, and 1% had
2 or more prior EP. 19-31% of the PUL population presented with an
hCG level above a DZ, depending on threshold (Table 1). Within this population,
most subjects were eventually diagnosed with a SAB. For those
subjects not diagnosed with a SAB, an ultrasound finding of suspected
IUP was more likely to result in a definitive IUP than an ultrasound finding
of suspected EP was to result in a definitive EP (74% vs. 50%, p or ¼ 3000 mIU/mL N¼923 (23%) IUP: 255 (28%)
EP: 187 (20%)SAB: 481 (52%)
hCG > or ¼ 4000 mIU/mL N¼763 (19%) IUP: 198 (26%)
EP: 154 (20%)
SAB: 411(54%)
IUP 253 (74%) 73 (21%) 15 (5%) 88 (7.2%)
EP 9 (3%) 125 (47%) 132 (50%)
IUP 198 (78%) 44 (17%) 13 (5%) 57 (6.2%)
EP 7 (4%) 82 (44%) 98 (52%)
IUP 155 (78%) 31 (16%) 12 (6%) 43 (5.6%)
EP 7 (5%) 66 (43%) 81 (52%)
FERTILITY & STERILITY Ò
e35

O-90 Monday, October 19, 2015 12:30 PM
FIRST TRIMESTER PREGNANCY LOSS FOLLOWING FRESH
AND FROZEN IN VITRO FERTILIZATION CYCLES. H. Hipp, a,b
S. Crawford, b J. F. Kawwass, a,b J. Chang, b D. M. Kissin, b
D. J. Jamieson. b a Gynecology and Obstetrics, Emory University School of
Medicine, Atlanta, GA;
b Centers for Disease Control and Prevention,
Atlanta, GA.
OBJECTIVE: To determine if there is an increased risk of first trimester
pregnancy loss in frozen embryo transfer cycles as compared to fresh cycles
following in vitro fertilization (IVF).
DESIGN: Retrospective cohort study using data from Centers for Disease
Control and Prevention National ART Surveillance System for 2007-2012.
MATERIALS AND METHODS: Multivariable log binomial regression,
stratified by maternal age (< 30, 30-34, 35-37, 38-40, >40 years), was performed
to compare age- specific risk of first trimester pregnancy loss between
fresh and frozen embryo transfers. The main analysis included fresh cycles
(n¼203,970) and frozen cycles for which maternal age at oocyte retrieval
could be ascertained (n¼45,660) that resulted in a pregnancy. A subgroup
analysis was performed to compare miscarriage risks of similar quality
embryos between fresh (n¼203,970) and frozen cycles (n¼7,885). Frozen
cycles for the subgroup analysis were restricted to those for which their originating
fresh cycle had not had an embryo transfer (i.e. the embryo(s) transferred
during the frozen cycle were the first embryos transferred from the
originating retrieval).
RESULTS: There was an increased risk of first trimester loss in pregnancies
achieved after frozen embryo transfers versus fresh cycles and the
adjusted relative risk remained significant among women less than 37 years
of age:

Supported by: NIH/NICHD R25 HD075737, U10 HD27049, U10
HD38992, U10 HD055925, U10 HD39005, U10 HD33172, U10
HD38998, U10 HD055936, U10 HD055942, and U10 HD055944, U54-
HD29834, U10HD03005-0851, U10HD055925-02W1.
O-93 Tuesday, October 20, 2015 11:45 AM
OVARIAN TUMOR RISK IN WOMEN AFTER ASSISTED REPRO-
DUCTIVE THERAPY (ART); 2.2 MILLION PERSON YEARS OF
OBSERVATION IN GREAT BRITAIN. A. G. Sutcliffe, a
C. L. Williams, a M. E. Jones, b A. J. Swerdlow, b M. C. Davies, c
I. Jacobs, d B. J. Botting. a a Institute of Child Health, University College London,
London, United Kingdom; b Institute of Cancer Research, Sutton, Surrey,
United Kingdom; c Reproductive Medicine Unit, University College
Hospital, London, United Kingdom; d University of New South Wales, Sydney,
Australia.
OBJECTIVE: To determine the risk of ovarian cancer, including malignant
and borderline ovarian tumors, in women who have been exposed to
ART.
DESIGN: Records from the Human Fertilisation & Embryology Authority
(HFEA) of all women who had ART in Britain between 1991-2010, were
linked to the National Health Service Central Registers (NHSCR) for England,
Wales and Scotland to obtain follow up for cancer outcomes, deaths
and emigrations. Reporting to the HFEA is mandatory.
MATERIALS AND METHODS: Cancer incidence in the cohort was stratified
by age and calendar period and compared with expectations derived
from annual age-specific national rates over the same period. Data were
also stratified for potential mediating/moderating factors such as repeated exposures,
age at first exposure, parity and subfertility diagnoses. Trends across
categories were evaluated using Poisson regression.
RESULTS: With 8.8 years average follow-up, 386 ovarian cancers
occurred in 255,786 women. An increased risk of developing an ovarian cancer
was observed in the cohort (standardized incidence ratio (SIR) 1.37; 95%
CI 1.24-1.51). No increased risk was found with increasing number of cycles
of ART (P trend ¼0.80). Increasing risk was found with decreasing parity
(P trend ¼0.002), with women who had no live births by the end of treatment
being at greatest risk (SIR 1.54; 95%CI 1.34-1.76). Risk was increased for
women with a ‘female factor’ cause of infertility (SIR 1.62; 95%CI 1.42-
1.84), especially endometriosis (SIR 2.35; 95%CI 1.80-3.07), but ‘male factor’
only infertility was not associated with risk (SIR 1.05; 95%CI 0.86-1.28).
Younger age at starting ART carried greater cancer risk (P trend

action. While additional studies to improve the efficacy of the approach to
100% are needed, this study supports further investigations of the approach
as a method of nonsurgical permanent female contraception.
Supported by: Bill and Melinda Gates Foundation OPP1025233,
P51OD011092, P51 OD011133, U54-055744-07.
O-96 Tuesday, October 20, 2015 12:30 PM
URINARY PHTHALATE METABOLITE CONCENTRATIONS
WERE ASSOCIATED WITH PREGNANCY LOSS AMONG WOMEN
CONCEIVING WITH MEDICALLY ASSISTED
REPRODUCTION. C. Messerlian, a B. Wylie, b P. Williams, c
J. B. Ford, a M. Keller, a A. M. Calafat, d R. Hauser. a,b a Department of Environmental
Health, Harvard T. H. Chan School of Public Health, Boston, MA;
b Department of Obstetrics and Gynecology, Massachusetts General Hospital,
Boston, MA; c Department of Biostatistics, Harvard T. H. Chan School of
Public Health, Boston, MA; d Centers for Disease Control and Prevention, Atlanta,
GA.
OBJECTIVE: Several phthalates are reproductive toxicants in animals and
human occupational exposure has been associated with decreased pregnancy
rates. We examined the association of pregnancy loss in relation to urinary
phthalate metabolites among women undergoing in-vitro fertilization and
ovarian stimulation with and without intrauterine insemination.
DESIGN: A prospective cohort of 249 women conceiving 294 pregnancies
from the Environment and Reproductive Health Study (EARTH) at the
Massachusetts General Hospital Fertility Center.
MATERIALS AND METHODS: Four di(2-ethylhexyl) phthalate (DEHP)
metabolites in urine were measured by the Centers for Disease Control and
Prevention at two time points per woman during each fertility treatment cycle
and the geometric mean urinary DEHP metabolite concentration was calculated.
We estimated adjusted risk ratios (RRs) and 95% confidence intervals
(CIs) for both biochemical and total pregnancy loss (

CONCLUSIONS: A nanocaged formulation of highly insoluble ORG
9935 improved bioavailability but did not maintain drug concentrations in
the range needed to prevent oocyte maturation. While the extended-release
preparation of NC35 prevented oocyte maturation, this improved PD performance
was associated with serum levels that varied widely and exceeded the
desired peak by 2-fold at the time of oocyte recovery. While promising,
further enhancements of the PK profile will be needed to improve feasibility
of this approach as a contraceptive in women.
Supported by: NIH U54 HD055744, P51OD011092.
O-99 Tuesday, October 20, 2015 11:45 AM
CANIDATE GENES AND PATHWAYS MEDIATING HORMONAL
REGULATION OF THE MACAQUE FALLOPIAN
TUBE. K. R. Bond a O. D. Slayden. b,c a Division of Reproductive & Developmental
Sciences, Oregon National Primate Research Center, Beaverton,
OR; b Oregon National Primate Research Center, Beaverton, OR; c Department
of Obstetrics and Gynecology, Oregon Health & Science University,
Portland, OR.
OBJECTIVE: The fallopian tube plays an important role in gamete transport
and fertilization, and may provide a target for reversible contraception.
The aim of this study was to identify differentially expressed, hormonally
regulated, genes and their related biological pathways in the rhesus macaque
fallopian tube during the menstrual cycle.
DESIGN: Affymetrix array analysis of macaque fallopian tube.
MATERIALS AND METHODS: Fifteen adult cycling rhesus macaques
were ovariectomized and treated sequentially with estradiol (E2) and then
E2 plus progesterone (P) in order to create artificial menstrual cycles. Fallopian
tubes were collected during the artificial proliferative phase (PP; E2
treatment alone), secretory phase (SP; E2 + P treatment), and the menstrual
phase (MP) cycle. RNA was isolated from the samples and analyzed using
Affymetrix GeneChip Ò Rhesus Macaque Genome Arrays. Transcript expression
for select targets were validated by real-time PCR with macaque specific
TaqMan Ò primer/probe sets and presented relative to expression of ribosomal
(S10) RNA expression.
RESULTS: Affymetrix analysis revealed 6297 genes that were significantly
regulated by steroid hormones (P

are four different PTGERs, our genomic data revealed that PTGER3 and
PTGER2 are the predominant PGE2 receptors expressed in the macaque
periovulatory follicle. Moreover, PTGER3 and PTGER2 mRNA levels
both increase significantly 12 hr after an ovulatory stimulus. Thus, the objective
of this study was to determine if PTGER3 is also directly involved in
C-OE or follicle rupture in rhesus monkeys.
DESIGN: Prospective non-human primate experimental study.
MATERIALS AND METHODS: Cumulus oocyte complexes (COCs)
aspirated from rhesus macaques undergoing an ovarian stimulation protocol
were cultured (n¼13-18/group; 3 experiments) in the presence or absence of
the PTGER3 agonist sulprostone (10 ng/mL) or PGE2 (500 ng/mL; positive
control). Bright field images were obtained at 0 hr and 30 hr and immunofluorescent
staining for hyaluronic acid (HA) expression (a marker for C-OE)
was performed following COC collection at 30 hr. In rhesus monkeys undergoing
a controlled ovulation protocol (n¼4/group), the dominant follicle was
injected with either the PTGER3 antagonist (L798106; 250 ng/follicle) or
vehicle (0.005% DMSO in saline) at the time the animals received an ovulatory
stimulus. Laparoscopy was performed 72 hr later to check for the presence
or absence of an ovulatory stigmata. Luteal phase length and circulating
progesterone levels were also assessed.
RESULTS: While PGE2 directly induced C-OE in vitro, sulprostone treatment
of macaque COCs did not result in cumulus expansion or HA synthesis.
Despite the absence of an effect on C-OE, intrafollicular injection of the
PTGER3-selective antagonist L798106 in the preovulatory follicle at the time
the animal received an ovulatory stimulus prevented (4 out of 4) the appearance
of a typical rupture site relative to controls. Additionally, antagonist treated animals
had an average luteal phase length of 18 days and an average peak progesterone
level of 3.47ng/mL, which was not significantly different from controls.
CONCLUSIONS: These findings demonstrate that PTGER3 plays an
important role in ovulation, but not C-OE, in primates.
Supported by: NIH T31HD007133 (JB), P51OD011092, and
U54HD55744 (JDH, RLS).
O-102 Tuesday, October 20, 2015 12:30 PM
RATES OF INTRAUTERINE DEVICES (IUD) UTILIZATION AND
STERILIZATION IN WOMEN OF REPRODUCTIVE
AGE. B. Tang, K. McCarthy, S. Eid, G. Strachan, M. Tjoa,
A. Pohlmeier, B. Howard. Teva Pharmaceutical, Frazer, PA.
OBJECTIVE: While there are a myriad of contraceptive choices available
to women, intrauterine devices (IUD) and female sterilization are two of the
most effective methods. The objective was to evaluate trends of IUD insertion
and sterilization rates in real world practice over the recent period of 2010 to
2013.
DESIGN: A retrospective study using US insurance administrative database
was conducted.
MATERIALS AND METHODS: Truven’s MarketScan Treatment Pathways
3.0 tool was used to analyze the rate of IUD insertion and sterilization
procedures among women from 2010 through 2013. Descriptive data on contraceptive
use and age group distribution was evaluated using R 2 to determine
the probability of change in rates predicted against time.
RESULTS: A total of 530,621 IUD insertions and 95,650 women who
chose sterilization were included in this study. While the overall trend
increased for IUD insertions over the analysis period, we observed the utilizations
of both IUD and sterilization decreased slightly in 2013 (R 2 ¼0.707).
The greatest proportions of IUD and sterilization users were between the ages
of 18-34 followed by the 35-44 age group. From 2010 to 2013, the proportion
of IUD insertions in women aged 45-54 increased from 0.9% to 5.0%.
CONCLUSIONS: Overall, IUD utilization rates of increased over time,
while sterilization rates remained stagnant.
Supported by: This study was Supported by Teva Pharmaceutical.
MENTAL HEALTH
O-103 Tuesday, October 20, 2015 11:15 AM
QUALITY OF PARENTING, MOTHER AND CHILD WELLBEING
AND ‘DADDY TALK’ IN SINGLE PARENT FAMILIES FORMED
THROUGH THE USE OF DONOR INSEMINATION. S. Zadeh, a
T. Freeman, b S. Golombok. c a Centre for Family Research, University of
Cambridge, Cambridge, United Kingdom; b University of Cambridge, Cambridge,
United Kingdom; c Centre for Family Research, Cambridge, United
Kingdom.
OBJECTIVE: The study examined parenting quality, mother and child
wellbeing and communication in single parent families formed through the
use of donor insemination (DI).
DESIGN: Semi-structured interviews and questionnaires were administered
to mothers at home.
MATERIALS AND METHODS: Data were obtained from 51 single
mothers and a comparison group of 52 partnered mothers with at least one
child aged 4-9 years. Mothers were interviewed using an adaptation of a
semi-structured interview designed to assess quality of parenting (Quinton
& Rutter, 1988) and to establish thoughts and feelings about motherhood.
Participants completed The Trait Anxiety Inventory (Spielberger, 1983), Edinburgh
Depression Scale (Thorpe, 1993) and the short form of the Parenting
Stress Index (Abidin, 1990). The Strengths and Difficulties Questionnaire
(SDQ) (Goodman, 1997) was administered to assess the presence of children’s
emotional and behavioural difficulties. Comparisons of parenting
quality and mothers’ wellbeing between the two family types were conducted
using multivariate analyses of covariance (MANCOVAs). Children’s psychological
adjustment was compared between family types using analyses of
variance (ANCOVAs). Hierarchical regression analysis was used to examine
factors associated with child adjustment problems within the solo mother
families. Qualitative analysis was used to supplement statistical findings.
RESULTS: All of the single mothers described their use of DI in ambivalent
terms. Most (n¼34, 67%) attributed their decision to use fertility treatment
to the lack of a suitable partner. Several (n¼28, 55%) expressed
concern about their child’s lack of a father. Different strategies were
described by mothers in discussing this issue with their child. There were
no significant differences in parenting quality or maternal wellbeing between
the solo mother and two-parent families. In terms of children’s adjustment,
there was no significant difference between family types for the total score
of the SDQ. Within the solo mother families, higher levels of financial difficulties
(F (2, 41) ¼ 5.00, p ¼ .01) and higher levels of parenting stress (F (7,
36) ¼ 2.55, p ¼ .03) were each associated with higher levels of children’s
emotional and behavioural problems.
CONCLUSIONS: Findings suggest that despite mothers’ reported preferences
and concerns, children in single parent DI families are no more likely
to experience emotional or behavioural difficulties than their two-parent
counterparts. Results indicate that single women seeking DI should not raise
specific concerns for clinicians with regards to psychological wellbeing,
quality of parenting, or child adjustment. As found in non-assisted single
parent families, the present study suggests that family process variables are
more influential in children’s adjustment than is family structure. ’Daddy
talk’ nevertheless remains central.
Frequency and Rates of Utilization of IUDs and Sterilization Over Time (Truven Marketscan Databases).
Year 2010 2011 2012 2013 Total
IUD (n, rate %) 109,504 (0.56) 129,457 (0.67) 151,699 (0.78) 139,961 (0.72) 530,621
Age %17 (%) 1.2 1.0 0.5 0
Age 18-34 (%) 66.7 66.2 66.3 63.6
Age 35-44 (%) 31.2 30.4 29.6 31.4
Age 45-54 (%) 0.9 2.4 3.5 5.0
Sterilization
24,305 (0.12) 25,827 (0.13) 25,708 (0.13) 19,810 (0.10) 95,650
(n, rate%)
Age %17 (%) 0 0 0 0
Age 18-34 (%) 58.4 58.4 58.8 57.8
Age 35-44 (%) 41.3 41.1 40.6 41.4
Age 45-54 (%) 0.3 0.4 0.06 0.8
e40 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

References:
1. Quinton, D., & Rutter, M. Parenting Breakdown: The making and
breaking of intergenerational links. Aldershot, UK: Avebury Gower
Publishing, 1988.
2. Spielberger, C. The Handbook of the State-Trait Anxiety Inventory.
Palo Alto, CA: Consulting Psychologists Press, 1983.
3. Thorpe, K. A study of the use of the Edinburgh Postnatal Depression
Scale with parent groups outside the postpartum period. Journal of
Reproductive and Infant Psychology, 1993;11:119-125.
4. Abidin, R. Parenting Stress Index Test Manual. Charlottesville, VA: Pediatric
Psychology Press, 1990.
5. Goodman, R. The Strengths and Difficulties Questionnaire: A research
note. Journal of Child Psychology and Psychiatry, 1997;38:581-586.
Supported by: This study was Supported by The Wellcome Trust.
O-104 Tuesday, October 20, 2015 11:30 AM
NOW THAT THEY ARE FROZEN-WHAT NEXT? STEPS TAKEN
AFTER OOCYTE CRYOPRESERVATION (OC) FOR DEFERRED
REPRODUCTION (DR). B. Hodes-Wertz, M. E. Fino,
K. N. Goldman, D. H. McCulloh, N. Noyes. NYU Langone Medical Center,
New York, NY.
OBJECTIVE: To assess post-treatment trends in patients completing OC
cycles for DR.
DESIGN: Retrospective cross-sectional study.
MATERIALS AND METHODS: Post-treatment electronic medical records
of all patients that underwent OC for DR from 2005-2014 were reviewed.
Patients completing OC for medical indications or due to lack of
sperm on day of egg retrieval (ER) and study patients were excluded. Any contact
with the patient regarding her stored oocytes or her fertility after the last
OC cycle was recorded (routine GYN matters were excluded). Post-ER treatments
included ovulation induction (OI), IVF and oocyte thaw (OT).
RESULTS: 1,394 patients underwent 1,759 cycles of OC for DR from
2005-2014 with 60% of cycles completed from 2012-2014. The average
age of patients that froze prior to 2012 was 383 y compared to 373 y
for those that froze from 2012-2014 (p

DESIGN: Randomized, controlled, prospective study.
MATERIALS AND METHODS: 166 women about to begin their first IVF
cycle were randomized to the self-administered CCRI or to a routine care
control group and then followed for 12 months
RESULTS: The 12-month pregnancy rate was similar for the RCC and
CCRI groups (OR¼1.02, 95% CI [.53 - 1.98]). Of the patients who were
not pregnant on the first cycle, 15.2% (n¼15/46) of patients assigned to
RC discontinued compared to 5.5% (n¼5/55) of patients assigned to the
CCRI (OR¼3.11, 95% CI [.756 - 12.80]. The CCRI group engaged in significantly
more positive reappraisal coping (OR¼.275, 95% CI [.16, .39] than
the Routine Care control group (OR¼.097, 95% CI [-.03, .23]). The CCRI
group had an improved Fertility Quality of Life (FertiQoL CORE,
OR¼4.07, 95% CI [2.07, 6.06]; FertiQoL Emotional, OR¼5.95 , 95% CI
[2.89, 9.00]) compared to the control group (Core OR¼ .67, 95% CI
[-1.55, 2.89], Emotional OR¼-.02, 95% CI [-3.36, 3.32]). The CCRI group
reported less global anxiety (OR¼ .275, 95% CI [.16, .39]) than the control
group (OR¼ .471, , 95% CI [-2.40, 3.34]). The CCRI reported positive evaluations
for the intervention (e.g., ease of use, helpfulness, perceived stress
reduction).
CONCLUSIONS: Use of the CCRI tool led to improved psychological status
but not significantly more treatment cycles or a higher pregnancy rate.
Supported by: This study as Supported by an unrestricted educational
grant from Merck & Co, Inc., Kenilworth NJ..
O-107 Tuesday, October 20, 2015 12:15 PM
STRESS MANAGEMENT AND RESILIENCE TRAINING (SMART)
THERAPY FOR COUPLES UNDERGOING IN VITRO FERTILIZA-
TION (IVF): A RANDOMIZED CLINICAL TRIAL (RCT). Z. Khan,
D. Fuehrer, C. Coddington, J. Bleess, G. Daftary, E. Stewart, J. Jensen,
A. Sood. Mayo Clinic, Rochester, MN.
OBJECTIVE: Infertility treatment is associated with high levels of stress
and anxiety. Some countries mandate routine counseling to couples during
infertility treatment. We conducted an RCT comparing the efficacy of Stress
Management And Resiliency Training (SMART) to stress reduction CD.
DESIGN: RCT
MATERIALS AND METHODS: Heterosexual couples undergoing a first
IVF cycle were eligible. Exclusion criteria were female age >40 years, recurrent
miscarriage and current or recent (6 months) psychotic episode. Couples
were randomized 1:1 to the SMART arm or CD. Those in the SMART arm
attended a single 90-minutes group session that taught skills in self-awareness,
and ways to strengthen attention, and incorporate greater gratitude,
compassion, acceptance and purpose. Breathing-based relaxation was also
incorporated. CD arm received a stress reduction CD. All participants
completed validated instruments: Perceived Stress Scale (PSS-10), Fertility
problem inventory (FPI), General Anxiety Disorder (GAD-7), Gratitude
Questionnaire (GQ-6), Satisfaction with Life Scale (SWL) and Subjective
Happiness Scale (SHS) at baseline, 6- and 12-weeks after enrollment. The
study had 80% power to detect a significant difference with 28 people per
group. Paired t-tests were used to compare the change in scores within a
group and 2-sample t-tests were used for between group comparisons.
RESULTS: Of 40 couples (n¼80) consented, 5 withdrew, 5 were lost to
follow-up and 30 (n¼60) had complete data. Baseline demographics were
comparable between the two groups. At 12 weeks, compared to baseline,
Comparison of Score Differences within and Between Groups.
Comparison within group
SMART Arm
Score
Difference
p-value
CD Arm
Score
Difference p-value
PSS-10 -6.57

Characteristics of Men Who Have Undergone Vasectomy without and with
Children.
No Children
National Estimate
(95% CI)
completed their family size and do not plan on having more children. While
uncommon, a childless man requesting a vasectomy can be ethically challenging
scenario for urologists. We hypothesized that men who had undergone
vasectomy prior to having children would have higher rates of
vasectomy reversal and family planning attitudes inconsistent with being
sterile.
DESIGN: Retrospective analysis of the National Survey for Family
Growth (NSFG).
MATERIALS AND METHODS: The NSFG is a survey periodically
administered in the United States by the National Center for Health Statistics.
The survey uses complex survey design with oversampling of to ensure a nationally
representative sample is queried on topics of sexuality and family
planning. The three cycles since 2002 have included 20,146 men aged 15-
44. We compared demographic information and family planning attitudes
between men who had undergone vasectomy with and without children.
We used the survey data analysis package in STATA to account for survey
design.
RESULTS: Of the 20,146 men surveyed, 696 (3.5%) reported undergoing
a vasectomy. Using the complex survey design to estimate the national
population, this is approximately 3,511,541 men aged 15-44 who have undergone
a vasectomy. Of the men reporting vasectomy, 3.5% (95% CI 2.4-5.1)
underwent the procedure without having children. Men without children
were older at the time of vasectomy (Table) and have a higher household income.
These men were less likely to have ever been married, and were more
likely to be non-religious. There were no differences in race, education,
employment status, or insurance status between the two groups (data not
shown). Whereas 1.3% (0.7-2.5%) of men with children underwent vasectomy
reversal during the follow-up, 0% of men without children underwent
reversal, p¼0.441. When asked how many children they intended to have,
men without children expected 0.00 children, whereas vasectomized men
with children expected 0.01 (0.0-0.2), p¼0.007. The two groups of vasectomized
men were of similar age at the time of the study, but the men without
children had shorter follow up compared to men with children [47.1 months
(30.8-63.4) vs. 80.8 (74.3-87.2), p

Number of veins ligated during varicocelectomy and improvement in pain post-surgery.
Varicocelectomy
(Number of Spermatic Veins Ligated)
Variable
All Subjects
1-4 veins 5-10 veins R11 veins
P-value
Mean Age (years, y) 35.34 9.22
N¼675 (100%)
Unilateral (Left) Varicocele 394 (73.2%)
n¼538
Bilateral Varicocele 144 (26.8%)
n¼538
Presenting Indication:
207 (30.8%)
With Pain
n¼673
Presenting Indication:
466 (69.2%)
Without Pain
n¼673
Outcome Indication:
94 (45.4%)
Pain Improvement
n¼207
Outcome Indication:
19 (9.2%)
No Pain
n¼207
Improvement
Outcome Indication:
94 (45.4%)
Unknown
n¼207
Outcome Indication (Only If Known): 94 (83.2%)
Pain Improvement
Outcome Indication (Only If Known):
No Pain Improvement
n¼113
19 (16.8%)
n¼113
36.47 10.91
n¼116 (20.0%)
74 (81.3%)
n¼91
17 (18.7%)
n¼91
29 (25.2%)
n¼115
86 (74.8%)
n¼115
18 (62.1%)
n¼29
2 (6.9%)
n¼29
9 (31.0%)
n¼29
18 (90.0%)
n¼20
2 (10.0%)
n¼20
32.98 8.56
n¼321 (55.4%)
219 (82.6%)
n¼265
46 (17.4%)
n¼265
118 (36.9%)
n¼320
202 (63.1%)
n¼320
53 (44.9%)
n¼118
12 (10.2%)
n¼118
53 (44.9%)
n¼118
53 (81.5%)
n¼65
12 (18.5%)
n¼65
38.32 8.08
n¼142 (24.5%)
47 (53.4%)
n¼88
41 (46.6%)
n¼88
40 (28.2%)
n¼142
102 (71.8%)
n¼142
18 (45.0%)
n¼40
3 (7.5%)
n¼40
19 (47.5%)
n¼40
18 (85.7%)
n¼21
3 (14.3%)
n¼21
P¼0.033 (C)
P¼0.033 (C)
P¼0.57 (C)
P¼0.57 (C)
P¼0.57 (C)
P¼0.81 (C)
P¼0.81 (C)
O-112 Tuesday, October 20, 2015 12:00 PM
PREGNANCY RATES OF FRESH VERSUS CRYOPRESERVED
SPERM OBTAINED BY PERCUTANEOUS TESTICULAR BIOPSY
IN MEN WITH OBSTRUCTIVE AZOOSPERMIA IS DEPENDANT
ON FEMALE AGE. S. Parsons, a J. A. Lee, a A. B. Copperman, a,b
N. Bar-Chama. a,c a Reproductive Medicine Associates of New York, New
York, NY; b Obstetrics, Gynecology and Reproductive Science, Icahn School
of Medicine at Mount Sinai, New York, NY; c Urology, Icahn School of Medicine
at Mount Sinai, New York, NY.
OBJECTIVE: A percutaneous testicular biopsy (Perc BX) is a noninvasive
technique of retrieving sperm for ART in men with obstructive azoospermia.
This minimally invasive technique allows for excess testicular tissue to be
cryopreserved for future IVF cycle(s). This study aims to determine factors
related to success in Perc BX cycles of IVF.
DESIGN: Retrospective cohort study
MATERIALS AND METHODS: Couples undergoing fresh IVF cycles
requiring a Perc BX for obstructive azoospermia by the same urologist
from May 2003 to March 2015 were reviewed. Testicular tissue was obtained
utilizing a 14 gauge core needle biopsy gun and typically under intravenous
sedation. Female cohorts were segregated by age (

(micro TESE) and intracytoplasmic sperm injection (ICSI) outcome of
AZFa, b, and c) in Japanese azoospermic patients.
DESIGN: Retrospective, multicenter AZF deletion study in infertile Japanese
men.
MATERIALS AND METHODS: A total of 1032 azoospermic patinets
men were examined genetic testing for AZF deletions by Promega Y Chromosome
AZF Analysis System (version 2.0) in 12 Japanese medical centers
from Octobar 2008 to March 2015. We analyzed sperm retrieval rate (SRR)
of the patiens with AZF microdeletion. In addition, we analyzed fertilization
and clinical pregnancy rates of those in whom the sperm retrieval was successful.
RESULTS: AZF microdeletions were found in 103 cases (9.8% of men
with azoospermia): 9 AZFa, 6 AZFb, 43 AZFc, 30 AZFb+c, and 15 AZ-
Fa+b+c. No spermatozoa were found with TESE or biopsy of the patients
with AZFa, AZFb, AZFb+c and AZFa+b+c deletions. Spermatozoa was
only obtained from patients with AZFc microdeletions. Of men with isolated
AZFc deletion, spermatozoa were found in 60.0% (24/40) by micro
TESE. Mean patient age was 34.8 4.7 years old. Testicular atrophy
was seen in almost all men (mean testicular volume: 11.9 4.5ml). Follicular
stimulating hormone (FSH), luteinizing hormone (LH), and testosterone
(T) levels before surgery was 19.5 10.5 mIU/ml, 6.9 4.0
mIU/ml, and 4.4 1.8 ng/ml, respectively. No correlation was found between
serum FSH, LH, and T level with the success of sperm retrieval. Patients’
age and testicular volume and also did not affect the SRR for micro
TESE. Fertilization rate was 60.3% and clinical pregnancy rate per embryo
transfer (ET) was 23.7%. Six children were safely delivered and 3 pregnancies
were ongoing.
CONCLUSIONS: The frequency of AZF deletions in Japan is similar to in
other countries. AZFc deletions are associated with severe dysregulations of
spermatogenesis, however, sperm retrieval is possible in more than half of the
patients with AZFc microdeletions by micro TESE. It is useful to obtain reliable
genetic information from azoospermic patients and to avoid unnecessary
micro TESE.
O-114 Tuesday, October 20, 2015 12:30 PM
MALE UNDERWEAR AND SEMEN QUALITY IN A POPULATION-
BASED PRECONCEPTION COHORT. K. J. Sapra, a S. Kim, a
M. Eisenberg, b Z. Chen, a G. M. Buck Louis. c a Division of Intramural Population
Health Research, Eunice Kennedy Shriver National Institute of Child,
Rockville, MD; b Stanford University, Stanford, CA; c NICHD, Rockville,
MD.
OBJECTIVE: To evaluate the relationship between type of male underwear
worn and semen quality in couples attempting pregnancy.
DESIGN: Population-based cohort with preconception enrollment.
MATERIALS AND METHODS: 501 couples were followed prospectively
from cessation of contraception until positive pregnancy test or 12
months of trying. At enrollment, male partners reported type of underwear
worn (briefs, boxer-briefs, boxers, none) during the day and to bed, separately.
Men also provided semen samples at baseline and one month later using
in-home collection protocols for 24-hour semen analysis of 35 semen
quality endpoints: 5 general, 8 motility, 6 sperm head, 14 morphology, and
2 SCSA endpoints. Linear mixed models were used to estimate beta (b) coefficients
and 95% CI for each Box-Cox transformed semen quality endpoint.
Models were adjusted for statistically significant confounders: race/ethnicity
and season of enrollment.
RESULTS: Semen analysis was available for 62 men wearing briefs
daytime/bed, 116 boxers-briefs daytime/bed, 129 boxers daytime/bed, 56
briefs daytime/boxers or none bed, 71 boxer-briefs daytime/boxers or
none bed, 29 boxers daytime/none bed. Compared with men wearing
briefs daytime/bed (i.e. presumably highest scrotal heat exposure), changing
to boxers or none for bed (i.e. lowering exposure) was associated with
decreased DNA fragmentation index (DFI) in men wearing boxer-briefs in
daytime (b -0.20, 95%CI -0.39, -0.01) though high DNA stainability
(HDS) was increased (b 0.24, 95%CI 0.05, 0.44). Men wearing boxers
in daytime and none to bed had decreased DFI (b -0.25, 95%CI -0.49,
-0.01).
CONCLUSIONS: Among men in the general population attempting pregnancy,
type of underwear worn during the day and to bed is associated with
semen quality. Reducing exposure for bed decreases DFI; better semen quality
parameters are observed in men wearing boxers during the day and none
to bed.
Supported by: Intramural Research Program Eunice Kennedy Shriver National
Institute of Child Health and Human Development.
OUTCOME PREDICTORS-CLINICAL: ART 2
O-115 Tuesday, October 20, 2015 11:15 AM
SIBLINGS CONCEIVED WITH ASSISTED REPRODUCTIVE
TECHNOLOGY: BIRTHWEIGHT AND GESTATION DIFFER-
ENCES IN FRESH VS FROZEN CYCLES. B. Luke, a M. B. Brown, b
E. Wantman, c J. E. Stern. d a Obstetrics, Gynecology, and Reproductive
Biology, Michigan State University, East Lansing, MI; b University of Michigan,
Ann Arbor, MI; c Redshift Technologies, New York, NY; d Geisel School
of Medicine at Dartmouth, Lebanon, NH.
OBJECTIVE: To evaluate birth outcomes of siblings conceived with ART
in fresh versus frozen cycles
DESIGN: Longitudinal cohort study
MATERIALS AND METHODS: Cycles reported to the Society for Assisted
Reproductive Technology Clinic Online Reporting System (SART
CORS) during 2004-12 were linked to individual women by the woman’s
birth date, first and last names, and social security number (when present);
linkages across clinics also included partner’s name and sequence of ART
outcomes. Included were cycles with at least one embryo transferred;
excluded were research cycles and those using gestational carriers. The
study population was limited to women with singleton live births conceived
with ART during the study period, both using the same oocyte source
(autologous or donor) but having live births from both fresh and frozen embryos
(the most recent live birth was chosen if there was more than one
eligible singleton birth). Mean differences in birthweight, birthweight z-
score (weight-for-gestation), and length of gestation between pairs of births
within each group were compared using Student’s t-test, after adjusting for
mother’s age, gravidity, infant gender and, when appropriate, length of
gestation.
RESULTS: The study population included 10,222 autologous pairs of siblings
and 2,240 donor pairs of siblings. In both groups, the frozen cycle was
approximately 2 years later than the fresh cycle. Siblings born from frozen
embryo transfers had shorter gestations (0.6 days for autologous and 1.1
days for donor, both p¼0.0003), heavier birthweights (160 grams for autologous
and 46 grams for donor, both p

ANOVA, Welch’s t-test and multivariate analysis. Significance was defined
as p

Embryo Transfer Practices and Outcomes in Mandated States.
CT HI IL MD MA NJ P Value
Day 5 transfer (%) 30.0 34.4 52.5 44.8 17.8 42.7

Science, Icahn School of Medicine at Mount Sinai, New York, NY; b Reproductive
Medicine Associates of New York, New York, NY; c RMANY-Mount
Sinai, New York, NY.
OBJECTIVE: The modern treatment of the infertile patient often includes
ovarian stimulation, oocyte retrieval, fertilization, biopsy and chromosomal
analysis (CCS), and vitrification. Embryos found to be normal can be transferred
in a subsequent FET cycle. We were able to control for embryo quality
by using only screened embryos and for synchronization by initiating progesterone
treatment five days prior to embryo transfer and were able to ask the
question of whether the number of days of estradiol preparation impacts the
likelihood of implantation.
DESIGN: Retrospective
MATERIALS AND METHODS: Patients who underwent an IVF cycle
with qPCR-based CCS that had R1 euploid embryo available for FET
were included. Fresh ETs were excluded from this analysis. Patients started
oral estradiol 2 mg twice daily for four days, then 2 mg three times daily.
Endometrial thickness was assessed weekly until a thickness of R7mm
was observed. Progesterone supplementation was then initiated, and five
days later, re-warming and ultrasound guided transfer was performed. A
Poisson regression model was used to predict if the total number of days
of estradiol impacted implantation rates. Statistical difference of p

TABLE 1. TEBX-PGS cycle outcomes using FR vs. frozen oocytes in women 39y at retrieval.
Age(y)
39y
groups
FR: Total No. Retrieved OC: Total No. 674 5798 292 4762
MI+MII Thawed
OC: Overall Survival 559 (83%) 238 (82%)
OC: Survived and Assessed as MII at Thaw 534 (79% ) 225 (77%)
No. Cycles Achieving BL Suitable for TEBX 39/47 (83%) 352 /407 (86%) 0.5 15/22 (68%) 339/407 (83%) 0.08
No. Biopsied BL/ 2PN Fertilization 145/373 (39%) 1911/3418 (56%) 0.0001 54/167 (32%) 1316/2628 (50%) 0.0001
No. Euploid BL 58/145 (40% ) 921/1911 (48%) 0.06 8/54 (15%) 254/1316 (19%) 0.5
No. Aneuploid BL 86/145 (60%) 955/1911 (50%) 0.03 44/54 (85%) 1031/1316 (78%) 0.7
No. TEBX Cycles with At Least One Euploid 28/39 (72%) 295/352 (84%) 0.07 7/15 (47%) 160/339 (47%) 1.0
BL
No. TEBX Cycles with All Aneuploid BL 11/39 (28%) 57/352 (16%) 0.07 8/15 (53%) 179/339 (53%) 1.0
Embryo Implantation Rate 14/22 (64%) 188/287 (66%) 0.8 2/3 (67%) 89/129 (69%) 1.0
Ongoing+Delivered Rate/Transfer 12/21 (57%) 154/276 (56%) 1.0 2/3 (67%) 72/123 (59%) 1.0
semen quality. All resultant zygotes were cultured for 5-7 days at which point
any BL reaching Stage R2BC was subjected to TEBX, then vitrified until
euploid ET. Thus, OC oocytes were subjected to freeze-thaw twice (oocyte+BL)
while in FR cycles, only biopsied BL were vitrified. In the OC
arm, average oocyte freezing duration was 3.42y. Data were compared
for OC vs. FR cycles in women ages < vs. R 40y. Fisher’s was used for statistical
comparisons.
RESULTS: See Table. In 54 (78%) OC and 691 (85%) FR cycles, at
least one 2PN-fertilized oocyte developed into a BL suitable for
TEBX (p¼ 0.2) with a mean of 3/cycle for OC & 4/cycle for FR. A total
of 199 BL (37% of 2PN fert) were suitable for biopsy in OC cycles vs.
3227 BL (53% of 2PN fert) in FR attempts (p¼ 0.0001). The latter difference
held across both age groups. Regardless of age, all but 12/423 (3%)
women had single- ET. EIR and pregnancy rates were not different between
the groups.
CONCLUSIONS: Adding OC to TEBX-PGS resulted in significantly
fewer 2PN-fertilized oocytes becoming BL suitable for TEBX, although
the total no. of cycles with at least 1 biopsiable BL was not different. Notably,
the euploidy rate for biopsied BL and implantation and ongoing/delivered
rates for transferred euploid BL were similar whether derived from OC or
FR oocytes, regardless of female age at retrieval. Thus, adding TEBX to
OC appears reasonable and potentially beneficial as long as patients are
counseled that OC results in a lower BFR, thus fewer biopsiable BL will
be available for chromosomal evaluation.
O-124 Tuesday, October 20, 2015 12:00 PM
PREGNANCY OUTCOMES AFTER FRESH VERSUS FROZEN-
THAWED BLASTOCYST TRANSFERS. N. Pereira, a A. C. Petrini, b
J. Lekovich, a G. L. Schattman, c Z. Rosenwaks. b a The Ronald O. Perelman
and Claudia Cohen Center for Reproductive Medicine, New York, NY;
b Weill Cornell Medical College, New York, NY; c Weill Medical College/
Cornell University, New York, NY.
OBJECTIVE: There has been a shift towards frozen-thawed embryo transfers
in the name of improved endometrial receptivity and pregnancy outcomes.
To investigate the effect of ovarian stimulation on endometrial
receptivity, we aim to compare pregnancy outcomes after fresh versus
frozen-thawed blastocyst transfer cycles at our center.
DESIGN: Retrospective single-center cohort study.
MATERIALS AND METHODS: All patients undergoing transfer of fresh
or frozen (vitrified)-thawed blastocysts after in vitro fertilization (IVF) between
January 2010 and September 2013 were analyzed for potential inclusion.
Cycles utilizing slow freezing protocols or pre-implantation genetic
screening were excluded from the analysis. Demographic characteristics
Comparision of Pregnancy Outcomes in Patients undergoing Fresh vs. Frozen-
Thawed ET.
Parameter
Fresh
Blastocyst
Transfer
(n¼918)
Frozen-Thaw
Blastocyst
Transfer
(n¼1273)
Age (years) 35.8 (5.09) 36.1 (5.28) 0.18
BMI (kg/m2) 23.2 (4.72) 23.4 (4.22) 0.60
Peak endometrial 10.7 (3.73) 10.5 (3.81) 0.19
stripe (mm)
Blastocele grading
1,2
3
151 (16.4%)
767 (83.6%)
220 (17.3%)
1053 (82.7%)
0.61
ICM grading
A
B
C
Trophectoderm
grading
A
B
C
61 (6.64%)
709 (77.2%)
148 (16.1%)
73 (7.95%)
701 (76.4%)
144 (15.7%)
60 (4.71%)
1098 (86.2%)
115 (9.03%)
62 (4.87%)
1111 (87.2%)
120 (9.43%)
compared were age and body mass index (kg/m 2 ), while baseline characteristics
analyzed included endometrial stripe thickness (mm), blastocele
grading (1-3), inner cell mass grading (A-C), and trophectoderm grading
(A-C). Implantation, clinical pregnancy, spontaneous miscarriage and live
birth rates were calculated. Student’s t-tests and Chi-square (c2) tests were
used as indicated. P

(5.28) years in the frozen-thawed group (P ¼0.18). There was no difference
in the mean BMI or peak endometrial stripe when comparing the groups.
There was also no difference in the grading of the blastocele expansion, inner
cell mass and trophectoderm grading of the blastocysts transferred in the
two groups. No differences in the rates of implantation (37.3% vs. 37.7%),
clinical pregnancy (50.2% vs. 49.4%), spontaneous miscarriage (7.30%
vs. 9.27%), or live birth (42.9% vs. 40.6%) were found between the two
groups.
CONCLUSIONS: Our study highlights that fresh blastocyst transfers
yield equivalent pregnancy outcomes as frozen-thawed blastocyst transfers.
While there continues to be a shift towards frozen-thawed ET cycles in the
name of improved pregnancy and perinatal outcomes, it is possible that
conservative stimulation protocols with fresh transfer offer equivalent benefits.
O-125 Tuesday, October 20, 2015 12:15 PM
SINGLE EUPLOID BLASTOCYST TRANSFER IS A VIABLE CLIN-
ICAL OPTION FOR AMA PATIENTS UP TO 42 YEARS: ANALYSIS
OF 1,000 CONSECUTIVE SINGLE FROZEN EMBRYO
TRANSFERS. M. Katz-Jaffe, a E. Surrey, a D. A. Minjarez, b
R. L. Gustofson, c L. A. Kondapalli, a J. M. Stevens, d W. B. Schoolcraft. a
a Colorado Center for Reproductive Medicine, Lone Tree, CO; b Colorado
Center for Reproductive Medicine, Denver, CO; c CCRM, Lone Tree, CO;
d Fertility Labs of Colorado, Lone Tree, CO.
OBJECTIVE: Women of advanced maternal age (AMA) presenting for
infertility treatment are considered to have poor prognosis. In order to increase
the likelihood of a clinical pregnancy, AMA women typically have
multiple embryos transferred. Often, this leads to undesirable, multiple gestations
that result in poorer obstetrical and neonatal outcomes. The objective
of this study was to evaluate the clinical efficacy of a single euploid frozen
blastocyst transfer for AMA women.
DESIGN: Research study
MATERIALS AND METHODS: A consecutive cohort of 1,000
infertility patients who received a single euploid, frozen blastocyst
transfer. All embryos were routinely biopsied at the blastocyst stage
for comprehensive chromosome screening prior to vitrification.
Standard protocols for a hormone replacement frozen embryo transfer
were employed. Analysis was performed to determine the probability
of achieving a live birth following the transfer of a single euploid blastocyst
in association with maternal age. Statistical analysis included
Fisher’s exact and Chi-square test for independence, with significance
at P

OVARIAN STIMULATION
O-127 Tuesday, October 20, 2015 11:15 AM
RANDOMIZED PROSPECTIVE TRIAL COMPARING THREE
DOSES OF GNRH ANTGONIST PROTOCOL IN PATIENTS WITH
POOR OVARIAN RESPONSE. E. Bastu, O. Dural, C. Yasa,
M. Ozsurmeli, I. Demiral, B. Baysal, F. Buyru. Department of Obstetrics
and Gynecology, Istanbul University School of Medicine, Istanbul, Turkey.
OBJECTIVE: To examine whether patients with poor ovarian response
(POR) according to the Bologna criteria, during conventional IVF/intracytoplasmic
sperm injection (ICSI) cycle, may benefit from different doses of gonadotropins
in ovulation induction protocols.
DESIGN: Randomized prospective trial carried out in an infertility clinic
of an university hospital.
MATERIALS AND METHODS: Only POR patients strictly defined according
to the Bologna criteria were included. 95 patients with POR were randomized
to receive three different doses of gonadotropins during ovulation
stimulation. Group 1 (31 patients) received 450 IU gonadotropins (225 IU
HP-hMG+225 IU rFSH); Group 2 (31 patients) received 300 IU gonadotropins
(150 IU HP-hMG+150 IU rFSH); Group 3 (33 patients) received 150 IU gonadotropins
(75 IU HP-hMG+75 IU HP-hMG+5mg letrozole). All transferred
embryos were Grade-1 according to the morphologic classification.
RESULTS: There were no statistically significant differences between the
age, body mass index (BMI), infertility period, day-3 E2, FSH, prolactin and
testosterone serum levels between the three groups (all p>0.05). Between the
three groups, there were no statistically significant difference in length of
ovulation induction, number of retrieved oocytes, number of MII oocytes,
number of fertilized oocytes, fertilization rate, number of transferred embryos,
cycle cancellation, chemical and clinical pregnancy rates (all
p>0.05). Although the difference was not statistically significant, rate of
chemical pregnancy was higher in the third group (75 IU HP-hMG+75 IU
rFSH+letrozole) compared to the first and second groups (0.160.37 vs.
0.160.38 vs. 0.210.42; p¼0.83).
CONCLUSIONS: Increasing the daily dosage of administered gonadotropins
is an intuitively appealing approach to the stimulation of the patients
with POR. However, increasing the dosage does not improve the number
of MII oocytes, fertilization rate, number of transferred embryos, chemical
and clinical pregnancy rates. Utilizing a mild stimulation with letrozole is
as effective as stimulation with higher doses.
Comparison of Outcome parameters
1st Group
(225 IU
HP-hMG
+ 225 IU
rFSH)
n¼31
References: The ClinicalTrials.gov identifier is NCT02293668.
O-128 Tuesday, October 20, 2015 11:30 AM
2nd Group
(150 IU
HP-hMG
+ 150 IU
rFSH)
n¼31
3rd Group
(75 IU
HP-hMG
+75IU
rFSH)
n¼33
Length of ovulation 8.771.65 9.652.74 9.881.82 0.097
induction, months
Gonadotropin dosage, IU 3929.03 2670.09 2309.09

presenting moderate/severe OHSS had significantly higher number of intermediate
but similar number of dominant follicles than women without OHSS
(respectively 124.9 and 7.25.3, p¼0.008; 4.44.1 and 4.33.7, p¼0.89).
A multivariable logistic regression revealed that the number of intermediate
follicles was the only statistically significant predictor of OHSS occurrence.
CONCLUSIONS: Shoview study shows that moderate/severe OHSS incidence
after COS with HP-hMG was 2.4% in this cohort where 41% of women
were considered at high risk of developing OHSS. Although some cases still
occurred, risk assessment and COS adaptation allowed satisfactory OHSS
risk management.
Supported by: Shoview cohort study was sponsored and conducted by Ferring
SAS, Gentilly, France.
O-130 Tuesday, October 20, 2015 12:00 PM
NEUTRALIZATION OF VASCULAR ENDOTHELIAL GROWTH
FACTOR (VEGF) BEFORE OR AFTER CG DURING COS CYCLES
RESTORES OVARIAN VASCULAR PARAMETERS SIMILAR TO
NATURAL MENSTRUAL CYCLES: A PILOT STUDY IN
NONHUMAN PRIMATES. C. V. Bishop, a X. Li, b D. M. Lee. c a Division
of Reproductive & Developmental Sciences, Oregon National Primate
Research Center, Beaverton, OR; b Advanced Imaging Research Center, Oregon
Health & Science University, Portland, OR; c Obstetrics & Gynecology,
Oregon Health & Science University, Portland, OR.
OBJECTIVE: To determine if VEGF neutralization either before or after
hCG administration would result in restoration of vascular parameters
similar to those observed in natural cycles in a nonhuman primate model
of ovarian hyperstimulation syndrome (OHSS).
DESIGN: Repeated measures in vivo pilot study.
MATERIALS AND METHODS: Rhesus macaque females (n¼8) underwent
baseline evaluation of ovarian vascular flow and blood volume (BV)
during normal menstrual cycles at mid luteal phase by contrast enhanced ultrasound
[1]. Ovarian vascular permeability to serum albumin was analyzed
on the same day by Dynamic Contrast Enhanced-MRI [2]. Females were then
treated with a Controlled Ovarian Stimulation [3] (COS) protocol with hCG
trigger, and then randomly assigned to treatment groups: Control (n¼3),
VEGF neutralizing agent Avastin 15-30hrs before hCG (single IV bolus;
10mg/kg; n¼3), and Avastin 3-4 days post-hCG (single IV bolus; n¼2).
Vascular flow, BV, and permeability of ovaries were analyzed 6-8 days
post-hCG (similar to mid-luteal phase in normal menstrual cycles).
RESULTS: OHSS symptoms developed in 2/3 Control, 0/3 Avastin prehCG,
and 1/2 Avastin post-hCG females: abdominal fluid was present at
time of oocyte retrieval and/or by MRI evaluation. Avastin administration
prior to hCG reduced serum progesterone levels compared to other groups
(P¼0.03); peak levels did not rise above 1.5 ng/ml. There was no significant
difference in ovarian vascular flow; however, increased BV occurred in COS
cycles (multiple CL) compared to natural cycles (single CL; P

DESIGN: Prospective randomized controlled trial at a university-affiliated
private IVF clinic between October 2009 and September 2013.
MATERIALS AND METHODS: Participants were IVF/ICSI candidates
considered at risk of POR. Since the trial had started prior to the publication
of the Bologna criteria, we had defined the risk of POR as the following: basal
FSH > 10 IU/L or AMH

for the evaluation of oocyte quality secondary to bidirectional communication
via gap junctions. The intent of this study was to investigate if differences
in the CC transcriptome, as determined by RNA-Seq, are associated
with embryo implantation.
DESIGN: Paired analysis.
MATERIALS AND METHODS: Patients with normal ovarian reserve
% 42 years of age were recruited for participation. COCs were dissected,
and CC RNA was isolated and then stored at -80 C. Embryo identity was
maintained, and blastocysts underwent CCS. Patients who had a double
embryo transfer resulting in a singleton live birth were potentially informative
and were selected for further analysis. DNA fingerprinting was utilized
to determine which embryo implanted. A cDNA library was
constructed from CC RNA and sequenced using a Life Technologies
(LT) Ion Torrent Sequencing platform. Filtered reads were aligned to the
hg19 AmpliSeq transcriptome reference (LT). Both negative-binomial
based and nonparametric methods were used to identify differentially expressed
genes with regards to live birth.
RESULTS: CC RNA sequencing of 38 samples (from 19 patients) generated
an average of 10.5 4.3 million reads/sample. 23 differentially expressed
genes between sibling embryos that resulted in a live birth and
those that did not were identified (P < 0.05). However, after correcting for
multiple testing none of the genes remained significantly differentially expressed
(false discovery rate < 0.05). 173 genes identified in previous studies
as predicting implantation, including RGS2, ANG, PTX3, and ZNF132, were
not differentially expressed in this analysis.
CONCLUSIONS: This study demonstrates that there were no candidate
biomarkers identified in the CC transcriptome that were differentially
expressed between euploid embryos resulting in a live birth and
those that failed. This is the first trial of this question to utilize paired
euploid sibling embryos which is the optimal study design for identifying
effective biomarkers within an individual patient’s cohort of
euploid embryos.
O-136 Tuesday, October 20, 2015 12:00 PM
MIRNAS ISOLATED FROM EXTRACELLULAR VESICLES IN
FOLLICULAR FLUID AND OOCYTE DEVELOPMENT
POTENTIAL. R. Machtinger, a R. Rodosthenous, b A. Mansour, c
M. Adir, c C. Racowsky, d R. Hauser, b A. A. Baccarelli. b a Obstetrics and Gynecology,
Sheba Medical Center and Tel-Aviv University, Tel Aviv, Israel;
b Harvard Chan School of Public Health, Boston, MA; c Obstetrics and Gynecology,
Sheba Medical Center and Tel-Aviv University, Ramat gan, Israel;
d Brigham and Women’s Hospital ART Center.
OBJECTIVE: Bidirectional communication between the oocyte and granulosa
compartment is critical for oocyte maturation, fertilization and embryonic
development. Recent studies have shown that extracellular vesicles
(EVs) within the ovarian follicle serve as vehicles for the transfer of mediators
such as miRNAs between the oocyte and the somatic cells. However, the
profile of these miRNAs remains largely unexplored. The aim of this study
was to characterize EV-encapsulated miRNAs from human follicular fluid
(FF) and to test their association with fertilization and day 3 embryo quality.
DESIGN: Prospective study.
MATERIALS AND METHODS: Follicular fluid (FF) samples were
collected from a single follicle of ICSI patients stimulated with GnRH antagonist
(n¼46). From this cohort, samples from non-severe male factor patients
and that contained a mature (MII) oocyte were analyzed (n¼40). Each
oocyte, its corresponding embryo and FF sample were tracked. Expression
profiles of EV-encapsulated miRNAs in FF were assessed using TaqMan
OpenArrayÒ. Only miRNAs that were detected in >50% of the samples
and exerted a Ct value %35 were included in the statistical analysis. Statistical
significance was adjusted for multiple comparisons using the Benjamini-Hochberg
method.
RESULTS: Of the 754 screened miRNAs, 215 were detected in all samples,
with several (e.g. miR-30d, miR-320b, miR-10b and miR-1291), being
detected in >85% of the samples. After excluding cases of abnormal fertilization
(n¼5), we compared 42 miRNAs from FF that contained normal
fertilized (n¼30) or failed to fertilize (n¼5) oocytes, and found 2.7-fold
and 1.5-fold higher expression of miR-202-5p (p¼0.001) and miR-7-1-3p
(p¼0.001) in EVs from follicles containing oocytes that fertilized normally
as compared those that failed to fertilize. Twenty nine embryos were evaluated
on day 3. No significant difference in expression of any EV-encapsulated
miRNA was observed in the miRNA profile (n¼35) for FF samples linked
with top quality embryos (n¼19) versus those linked to embryos of lesser
quality (n¼10).
CONCLUSIONS: Our preliminary results in this novel area of investigation
identified specific miRNAs carried within EVs in FF that may be associated
with fertilization potential. If our findings are Supported by further
studies, differential expression profiles of these miRNAs may prove to be
useful biomarkers for oocyte developmental potential.
Supported by: This study was funded by Grant Award no. RPGA1301 from
the Environment and Health Fund Israel and by grants P30ES00002 and
R21ES024236 from the National Institute of Environmental Health
Research.
O-137 Tuesday, October 20, 2015 12:15 PM
MICROBIOME AT THE TIME OF EMBRYO TRANSFER: NEXT
GENERATION SEQUENCING OF THE 16S RIBOSOMAL
SUBUNIT. J. M. Franasiak, M. D. Werner, C. R. Juneau, X. Tao,
J. N. Landis, Y. Zhan, N. R. Treff, R. T. Scott. RMA, NJ, NJ.
OBJECTIVE: Characterization of the human microbiome has benefited
from the application of powerful molecular tools utilizing the unique 16S ribosomal
subunit’s hypervariable regions to greatly increase sensitivity. The
microbiome of the lower genital tract can prognosticate obstetrical outcome
while the upper reproductive tract remains poorly characterized. Issues
outside of obstetrics are not well studied. Indeed, the microbiome at the
time of embryo transfer has only been evaluated utilizing culture based technology
which is known to capture only a fraction of dominant and major organisms.
This study analyzes the microbiome at the time of single embryo
transfer and characterizes the microbiome by reproductive outcome.
DESIGN: Prospective cohort.
MATERIALS AND METHODS: Consecutive patients undergoing
euploid, single embryo transfer were included in the analysis. Patients underwent
routine IVF care, oocyte retrieval, and embryo culture. The embryo
transfer was performed per routine. After transfer, the most distal 5mm
portion of the transfer catheter was sterilely placed in a DNA free PCR tubes.
Cell lysis and DNA purification were performed and DNA sequencing was
performed using Next Generation Sequencing of the bacteria specific 16S
ribosome gene. The sequences were compared to reference databases to procure
genus and species calls for microorganisms. This allowed for identification
and relative quantification of bacteria present in the sample.
RESULTS: Taxonomy assignments were made on 35 samples from 33 patients
and 2 E. coli controls. Of the 35 patients, 18 had ongoing pregnancies
and 15 did not. There were a total of 276 different genus calls present across
patient samples. The microbiome at time of transfer for those patients with
ongoing pregnancy vs. those without ongoing pregnancy is characterized
by top genera by sum fraction. Results are summarized in Table 1. Several
dominant species were present in both as would be expected; however, there
were varied major species by outcome.
CONCLUSIONS: The data presented here show the microbiome at the
time of embryo transfer may differ by pregnancy outcome. This novel
approach, both in specimen collection and analysis, is the first step towards
goal of determining physiologic from pathophysiologic microbiota. Further
studies will help delineate if differences in the microbiome at the time of embryo
transfer have a reliable impact on pregnancy outcome.
Microbiome at time of transfer which resulted in ongoing and non-ongoing
gestations after SET.
Ongoing
Pregnancy
(Genus)
Genus
Sum
fraction
Non-ongoing
Pregnancy
(Genus)
Genus
Sum
fraction
Flavobacterium 4.58 Flavobacterium 3.42
Lactobacillus 3.35 Lactobacillus 3.29
Limnohabitans 0.85 Limnohabitans 0.73
Polynucleobacter 0.78 Polynucleobacter 0.71
Bdellovibrio 0.68 Bdellovibrio 0.58
Chryseobacterium 0.64 Blvii28 0.53
Spirochaeta 0.63 Clostridium 0.52
Clostridium 0.55 Chryseobacterium 0.52
Blvii28 0.50 Spirochaeta 0.51
Paludibacter 0.39 Pseudomonas 0.31
e54 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

O-138 Tuesday, October 20, 2015 12:30 PM
DO THERMAL PROPERTIES OF A CULTURE DISH INFLUENCE
EMBRYO DEVELOPMENT?. L. B. Ramirez, a H. L. Shi, a E. Lvov, a
C. Racowsky, a,b C. L. Bormann. a,b a Department of Obstetrics, Gynecology
and Reproductive Biology, Brigham and Women’s Hospital, Boston, MA;
b Harvard Medical School, Boston, MA.
OBJECTIVE: Embryos display sensitivity to temperature fluctuations. As
rate of heat gain and loss may vary by type of culture dish, the question arises
whether dish-specific rate of temperature exchange (RTE) influences embryo
development during the short excursions of dishes from incubators to room
temperature (RT). If so, dish RTE consideration would be important when
choosing the type of dish used in clinical IVF. This study tested the hypothesis
that high dish RTE negates any adverse effect of temperature fluctuation
on embryo development.
DESIGN: Experimental study.
MATERIALS AND METHODS: Thirteen commercial dishes containing
medium and pre-equilibrated to 37 C were assessed for their RTE’s by moving
the dishes from 37 C to RT and then back to 37 C. The RTE’s were determined
from times required for a 5 C temperature change after these
excursions. The two dishes selected for embryo experiments were at the extremes
for RTE values (Falcon 1007, low RTE¼1.360.04 C/100s; Vitro-
Life micro-droplet dish, high RTE¼1.710.16 C/100s). A total of 464
fresh 2-cell mouse embryos were randomly allocated to 4 groups (Falcon
control [F-C], Falcon treatment [F-T], Vitrolife control [V-C], and Vitrolife
treatment [V-T]) in triplicate experiments. Embryos were cultured in 25 ml
drops of Global TotalÒ, overlaid with 9 ml OvoilÔ at 37 C in 5% O2,
6.2% CO2 and balanced N2. All dishes were removed twice a day from
the incubator and placed on a RT surface (treatments) or 37 C surface (controls)
for 5 min, and then returned to the incubator. Development was assessed
on day 4.5, blastocyst number and quality were recorded, and
embryos were stained with Hoescht for total cell counts. Groups were
compared using Fisher’s Exact and t-test where appropriate, with p

O-141 Tuesday, October 20, 2015 11:45 AM
DISTRIBUTION OF THE FMR1 GENE IN FEMALES BY RACE-
ETHNICITY: WOMEN WITH DIMINISHED OVARIAN RESERVE
VERSUS WOMEN WITH NORMAL FERTILITY (SWAN
STUDY). L. Pastore, a S. L. Young, b A. Manichaikul, c V. Baker. d a Stony
Brook Medicine, Stony Brook, NY; b UNC School of Medicine, Chapel Hill,
NC; c University of Virginia, Charlottesville, VA; d Stanford University, Stanford,
CA.
OBJECTIVE: Premutation level trinucleotide repeat lengths in the
FMR1 gene (CGG 55-199) are associated with primary ovarian insufficiency
before age 40. However, an association between the repeat length
with other ovarian aging phenotypes is not established. We examined
whether reported associations between the FMR1 CGG repeat lengths
in the intermediate, high normal, or low normal range differentiate
women diagnosed with diminished ovarian reserve (DOR) from those
with normal reproductive histories, and whether associations vary by
race-ethnic group.
DESIGN: DOR cases (n¼129) enrolled from 5 US fertility clinics vs.
normal fertility female controls (n¼803) from the US-based Study of
Women’s Health Across the Nation (SWAN).
MATERIALS AND METHODS: Cases’ (95 Caucasians, 22 Asian, 12
other) and controls’ (386 Caucasians, 219 African-Americans, 102 Japanese,
96 Chinese) banked DNA was analyzed for FMR1 CGG repeat
lengths. Cases were clinically diagnosed with DOR, with regular menses
and no fragile X syndrome family history. Controls had >¼1 menstrual
period in the 3 months pre-enrollment, >¼1 pregnancy, no history of infertility
or hormonal therapy, and menopause R46 years. In a previous analysis
the SWAN Chinese and Japanese groups had similar FMR1 CGG
repeat lengths, so those two groups were combined. We used Fisher’s exact
tests to analyze data.
RESULTS: We found fewer CGG trinucleotide repeats in the lower of the
two FMR1 alleles (i.e., allele with fewer repeats) in DOR cases relative to
controls among Caucasians (p&lt0.0001), but not Asians (p¼0.24). Caucasian
DOR cases were more likely to have fewer CGG repeats in the lower
allele compared with Asian DOR cases (p¼0.027). No significant differences
were found in the CGG repeat length distribution of the higher FMR1 allele
among DOR cases compared with controls (p&gt0.20), or by race/ethnic
group (p¼0.41).
CONCLUSIONS: This study did not find an association between DOR and
high normal/intermediate repeats, but there was an association between DOR
and low normal repeats (

membrane permeability and compromises oocyte quality. These findings
contribute to understanding of the premature ovarian insufficiency and infertility
seen commonly in women with classic galactosemia.
O-144 Tuesday, October 20, 2015 12:30 PM
SELECTING EMBRYOS WITH OPTIMAL CLEAVAGE KINETICS
IMPROVES ONGOING PREGNANCY RATE FOLLOWING BLAS-
TOCYST TRANSFER IN A MOUSE MODEL. R. S. Weinerman, a
T. S. Ord, a R. Feng, b C. Coutifaris, a M. A. Mainigi. a a Division of Reproductive
Endocrinology and Infertility, University of Pennsylvania, Philadelphia,
PA; b Department of Biostatistics and Epidemiology, University of Pennsylvania,
Philadelphia, PA.
OBJECTIVE: To develop a model predicting blastocyst (blast) formation
based on early cleavage parameters in the mouse and to determine if the
model predicts pregnancy outcome following blast transfer. Microarray analysis
was performed to identify genes associated with early embryo development.
DESIGN: Laboratory research.
MATERIALS AND METHODS: 313 2-pronuclear (PN) embryos were
collected from superovulated (SO) female mice and were cultured to blast
in 5% (n¼180) and 20% (n¼133) oxygen (O2). Time-lapse videos were
collected in the EevaÔ System (Progyny). Classification and regression
tree analysis (CART) was used to predict blast formation based on 4 cleavage
parameters: 1st cytokinesis duration, and time from 2-3 cell, 3-4 cell, and 4-5
cell stages. The model was built in the 20% O2 cohort and validated in the 5%
O2 cohort. For the transfer experiments, 2PN embryos were collected
following mating of SO females to transgenic male mice heterozygous for
green-fluorescent protein (GFP) and cultured in the EevaÔ system. Blasts
were rated as having ‘‘optimal’’ (Opt) or ‘‘suboptimal’’ (Subopt) timing based
on the CART model. 10 blasts were transferred into each of 10 pseudo-pregnant
recipients, using GFP status to tag the embryos as Opt or Subopt. Pregnancy
outcomes were assessed at mid-gestation (E10.5). Differences in
outcomes were assessed using Fisher’s exact test. Single-embryo microarray
analysis was performed using the Affymetrix GeneChip Mouse Gene ST 1.0
Array on Opt and Subopt blasts.
RESULTS: The final model utilized 2 parameters, cleavage times from 2-3
cell and 3-4 cell. The model predicted blast formation with a 97.5% sensitivity
and 62.5% specificity in 5% O2. Following transfers, 8 mice became
pregnant and were included for analysis. The overall implantation rate per
embryo transferred did not differ significantly between groups (86% Opt
vs. 77% Subopt, p¼0.3). However, the ongoing pregnancy rate was higher
for the Opt compared to Subopt embryos [60% (35/58) vs. 32% (7/22)
(p¼0.03)]. The resorption rate (implantation sites with placental tissue
only) among implanted embryos was significantly higher in the Subopt
compared to Opt group [59% (10/17) vs. 30% (15/50)(p¼0.04)]. Gene
expression differences were seen in 74 genes between the Opt and Subopt
groups (FD

Comparison according to blastocelic expansion
Gardner 4-5
(n¼112)
Gardner 6
(n¼122)
Chi
Square
Oocyte Age 36.64.2 35.95.0 NS
Day 3 FSH 5.72.9 6.13.2 NS
End. thickness 9.71.9 9.91.9 NS
(mm) at surge
Peak estradiol 2292.71072.2 2453.91243.3 NS
Oocytes retrieved 17.29.0 17.89.6 NS
MII 13.18.0 13.97.8 NS
Number of 2PN 10.66.8 11.97.7 NS
Implantation rate 56.3% (63/112) 51.2% (66/129) NS
Biochemical PR 65.2% (73/112) 64.8% (79/122) NS
Miscarriage rate 54.5% (61/112) 53.3% (65/122) NS
Miscarriage rate 30.1% (22/73) 27.8% (22/79) NS
O-147 Tuesday, October 20, 2015 11:45 AM
BLASTOCYST TRANSFER IN WOMEN AGED 40 YEARS OR
OLDER: HOW MANY IS TOO MANY?. K. S. Richter, J. E. O’Brien,
M. J. Tucker, R. J. Stillman. Shady Grove Fertility, Rockville, MD.
OBJECTIVE: To evaluate blastocyst transfer outcomes for women aged
40 to 44 years according to the number and quality of embryos transferred.
DESIGN: Retrospective review.
MATERIALS AND METHODS: From 2008-2013 all autologous embryo
transfers performed on day 5 or 6 after oocyte retrieval among women 40 to
44 years were reviewed. Embryos were categorized as ‘‘A’’ blastocysts (grade
AA or BA), ‘‘B’’ blastocysts (grade AB or BB), ‘‘C’’ blastocysts (ICM and/or
trophectoderm grade C or insufficiently expanded to grade), or ‘‘D’’ embryos
(pre-cavitation). Generalized estimating equations (GEE) analysis was used
to account for multiple cycles per patient and adjust for age and transfer day.
RESULTS: 1,145 cycles by 932 patients were available for analysis.
Adjusted GEE analysis of the independent contributions of embryos in
each category estimated that live born children resulted from 21.2% of
‘‘A’’ blastocysts (p20 weeks of
pregnancy).
DESIGN: Prospective randomized controlled trial (RCT) started in June
2012 with recruitment closed in December 2014. Patients were allocated
through computer-generated randomization into two groups: conventional
blastocyst transfer or preimplantation genetic screening (PGS). Sample
size calculated for the endpoint of ongoing pregnancy rates per cycle and delivery
rates was 120 patients per arm with ovum pick-up.
MATERIALS AND METHODS: Women between 38-41 years, %3 implantation
failures and %3 previous miscarriages, and R5 MII oocytes
from one or two stimulation cycles. Embryo culture and fresh blastocyst
transfer was performed in the non-intervention arm. Cleavage stage embryo
biopsy and array-CGH were performed in the intervention arm with fresh
blastocyst transfer of euploid embryos. Individual blastomeres underwent
whole genome amplification using the Sureplex Kit (Illumina). Amplified
DNA and reference DNAwere labeled and co-hybridized in 24sure arrays (Illumina).
After washing, slides were scanned and analyzed by BlueFuse Multi
software. Statistical comparisons were performed using two-sided Fishersexact
test and t-Student test.
RESULTS: A total of 215 cycles have been completed, 113 cycles in the
conventional blastocyst transfer group and 102 in the PGS group. Mean female
age (39.42.8 and 39.53); and mean number of MII oocytes
(9.63.9 and 9.83.5) were similar in both groups. Mean number of transferred
embryos was significantly higher in the blastocyst group compared to
the PGS group (1.80.6 vs. 1.30.7; p

effective for good prognosis patients. This study aim to answer the question:
Does multiple pregnancy rate decrease in good prognosis patients who failed
in fresh elective SET (eSET) and received an elective double frozen embryo
transfer (eDFET) compared with patients receiving fresh elective double embryo
transfer (eDET)?
DESIGN: Retrospective observational study.
MATERIALS AND METHODS: This study evaluated 723 ICSI cycles using
standard conventional protocol, at a private Assisted Reproduction Center
during last 10 years. eDETwas carried out in 639 patients, and 84 patients
received eSET and failed in first fresh cycle and had an subsequent eDFET
(eSET+eDFET). It was defined as eSET and eDET patients who transferred
one or two top quality embryos, respectively, and had at least two spared top
quality embryos cryopreserved.
RESULTS: Patients results for eDET and eSET+eDFET groups were: age
(35.24.1 x 34.03.1; p¼0.016), basal FSH (6.24.7 x 7.110.6;
p¼0.207), FSH dose administered (1881.1516.4 x 1708.2238.9;
p¼0.003) and oocytes collected (13.05.8 x 10.75.3; p¼0.001). Groups
presented similar pregnancy (36.2% x 31.2%; p¼0.385) and multiple pregnancy
rates (31.1% x 25.0%; p¼0.541).
CONCLUSIONS: Limiting the number of embryos transferred can benefit
the bottom line by decreasing the incidence of multiple pregnancy. We hypothesized
that patients who failed in fresh eSET could present worst prognosis
and hence had lower multiple pregnancy rates in a second attempt with
two embryos transferred. However, in this sample, the second transfer with
two elective embryos, after a failed elective transfer of one embryo, was
not effective to protect this population of multiple gestation, suggesting
that for this purpose the appointment of new elective transfer of an embryo
would be the most appropriate for good prognosis patients, even after a failure
Supported by: Not applicable.
O-150 Tuesday, October 20, 2015 12:30 PM
THE IMPACT OF TRANSFER TIME ON CLINICAL PREGNANCY IN
FROZEN EMBRYO TRANSFER CYCLES. M. S. Lee, E. R. Cardozo,
A. E. Karmon, D. L. Wright, T. Toth. Vincent Obstetrics and Gynecology,
Massachusetts General Hospital, Boston, MA.
OBJECTIVE: To identify the critical aspects of embryo transfer that
impact clinical pregnancy rates, specifically the impact of transfer time.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All consecutive frozen embryo transfer
(FET) cycles using cryopreserved blastocysts from 2007 - 2014 were reviewed.
All blastocysts were of high quality, all cycles used a uniform
cryopreservation technique and controlled hormone replacement protocol,
and all transfers were done under ultrasound guidance. Each transfer was
scored on a set of standardized metrics including cohort score (morphologic
appearance at transfer), ease of mock transfer, bends placed in catheter,
presence of blood or mucus, and overall difficulty of transfer.
Transfer time (total seconds from when the catheter was loaded until
the embryo(s) was expelled into the uterine cavity) was recorded.Logistic
regression models using generalized estimating equations were fit to
investigate the relationship between transfer time and clinical pregnancy
rate (CPR) while controlling for confounders and within-person correlations
in cycle outcome. Multivariate results were adjusted for age, diagnosis,
cohort score, number of embryos transferred, use of an outer
sheath, type of catheter used, blood inside the catheter, and physician performing
the transfer.
RESULTS: 467 women undergoing 571 FET cycles were included. Mean
age was 35.9 years (22-51 4.4). Overall CPR was 47.1% . The cohort was
divided into tertiles by transfer time (seconds) (T1: 33-55; T2: 57-81; T3: 82-
582) with mean transfer times (SD) of 47.4 (5.7), 67.1 (7.3) and 121.9 (55.1)
seconds. Crude CPR was 43.9% , 48.7% and 48.7% among respective tertiles.In
univariate analysis, worse cohort score (p

MATERIALS AND METHODS: One-cell embryos from F1 mice were
cultured in 20% oxygen either individually, or in groups of ten, in media
G-1/ G-2 with 5mg/ml HSA, in the presence or absence of 10mM carnitine/10mM
cysteine/5mM lipoate. Embryo development was analysed
through time-lapse and embryo transfers performed. Intracellular levels of
reduced glutathione (GSH) were assessed using fluorometric analysis.
RESULTS: There was no effect of treatment on blastocyst formation.
However, compared to controls, the three antioxidants significantly
increased embryo cell numbers when used individually (P

O-155 Tuesday, October 20, 2015 12:15 PM
CRYOPRESERVATION METHODS AND COMPARISON OF OUT-
COMES IN THE HUMAN OOCYTE PRESERVATION EXPERIENCE
(HOPE) REGISTRY. Z. P. Nagy, a T. Elliott, a C. Chang, a B. Hayward, b
M. C. Mahony. b a Reproductive Biology Associates, Atlanta, GA; b EMD
Serono, Inc., Rockland, MA.
OBJECTIVE: To compare IVF outcomes by cryoprotectant use and cryopreservation
methodologies for donor oocytes cryopreserved by either slowfreezing
(SF) or vitrification (VIT) in the HOPE Registry.
DESIGN: Post hoc analysis of data from the Phase IV, prospective, multicenter
(16 US centers), observational HOPE Registry.
MATERIALS AND METHODS: Clinical and laboratory data were
collected on ART cycles that used cryopreserved oocytes (autologous and
donor) from women aged 18-50 years enrolled between June 2008 and
September 2010. Six centers (involving 85/145 [59%] patients) were audited;
regular monitoring across all centers ensured clean patient data. We
compared implantation rates (IR) using analysis of variance and clinical
pregnancy rates (CPR) with Fisher’s exact tests in SF and VIT cycles using
donor oocytes for various cryopreservation parameters.
RESULTS: Data from 136 patients receiving donor oocytes cryopreserved
by either SF (n¼41, 302 oocytes) or VIT (n¼95, 704 oocytes) were analyzed.
For VIT cycles, container type had a significant effect on IR (p¼0.0194) and
CPR (p¼0.0374); CryotopÒs were used most often (n¼71) and associated
with the highest CPR (70.4%). Culture media used in the cryosolution also
had a significant effect on IR for VIT cycles (p¼0.0200); homemade
(n¼30) and protein (20%)/homemade/HEPES-based/amino-acid based
(n¼38) were used most often and were associated with IRs of 63.9% and
58.3%, respectively. For SF cycles, 1-2 oocytes/container vs R3 (both
n¼20) was associated with respectively: significantly greater IR (25.1% vs
2.9%, p¼0.0049) and CPR (55.0% vs 5.0%, p¼0.0012); a cooling rate
( C/minute) of 2vs 3or 0.3 was associated with IRs 22.8% vs 6.3%
vs 0% (p¼0.0511) and CPR 50% vs 10% vs 0% (p¼0.0088); a plunging temperature
( C) of < 40 vs 30 to 35 vs 36 to 40 was associated with
greater IR (23.9% vs 3.3% vs 0%, p¼0.0352) and CPR (52.4% vs 5.9% vs
0%, p¼0.0041).
CONCLUSIONS: Container type (CryotopÒ best) and culture media type
in the cryosolution had significant effects on VIT outcomes. SF outcomes
were affected by number of oocytes per container (1-2 best), cooling rate
( 2 C/minute best) and plunging temperature (< 40 C).
References: An abstract reporting data on VIT vs SF use and outcomes
(oocyte survival rate and implantation rates) will be reported at the ESHRE
2015 Congress (Nagy ZP, Hayward B, Mahony MC. Evaluation of the effect
of various cryoprotectants and protocols on donor oocyte survival and embryo
viability based on HOPE Registry data. Accepted for presentation at
ESHRE 2015). This ASRM abstract includes important different data on
cryoprotectant methodology and ART outcomes.
Supported by: The study and abstract development were Supported by
EMD Serono, Inc., Rockland, MA, USA, a subsidiary of Merck KGaA,
Darmstadt, Germany.
O-156 Tuesday, October 20, 2015 12:30 PM
THE EFFICACY OF SYSTEMIC ADMINISTRATION OF GRANU-
LOCYTE COLONY STIMULATING FACTOR (GCSF) ON THE IN
VITRO FERTILIZATION (IVF) SUCCESS IN WOMEN WITH
REPEATED IMPLANTATION FAILURE. Z. Abedi Asl. Shariati IVF
Center, Tehran, Iran, Islamic Republic of.
OBJECTIVE: To evaluate the efficacy of systemic subcutaneous Granulocyte
Colony Stimulating Factor (GCSF) administration on In Vitro Fertilization
(IVF) successin infertile women with repeated IVF failure.
DESIGN: Multi-centric, prospective, randomized, open label, controlled,
trial.
MATERIALS AND METHODS: 100 infertile women with normal endometrial
thicknesswho had R 2 implantation failure after IVF cycles who
were referred for IVFto Infertility Departments of Shariati Hospital, Bahman
Hospital (two tertiary referral centers) and Omid infertility clinic, Tehran,
Iran. Sealed, numbered, opaque envelopes assigned 50 patients to receive
subcutaneous 300mg GCSF before implantation and 50 in the control group.
The outcomes were the implantation (number of gestational sacs on the total
number of transferred embryos) chemical pregnancy (positive serum b-
HCG), and clinical pregnancy (gestational sac and fetal heart)rates.
RESULTS: The implantation rate in the intervention group was significantly
higher than the control group (15.3% vs. 7.2%, p ¼ 0.04),the chemical
pregnancy rate was significantly higher in the intervention group than the
control group (40% vs. 20%, p ¼ 0.02),and the clinical pregnancy rate was
higher in the intervention group than the control group, however this was
not statistically significant (28% vs. 16%, p ¼ 0.14). There were no adverse
events in any of the study groups.
CONCLUSIONS: Administration of single-dose systemic subcutaneous
GCSF before implantation significantly increases the IVF success, implantation
and pregnancy rates in infertile women with repeated IVF failure.
ASSISTED REPRODUCTIVE TECHNOLOGY - GENERAL 1
O-157 Tuesday, October 20, 2015 11:15 AM
GNRH AGONIST INDUCES TH1 AND TH17 IMMUNITY, WHILE
GNRH ANTAGONIST SUPPRESSES TH1 IMMUNITY. N. Sung, a
M. D. Salazar Garcia, a S. Dambaeva, b J. Kwak-Kim. a a Rosalind Franklin
University of Medicine and Science, Vernon Hills, IL; b Microbiology and
Immunology, Rosalind Franklin University of Medicine and Science, North
Chicago, IL.
OBJECTIVE: Inflammatory immune responses have been reported in
women with multiple implantation failures after IVF cycles. In this study,
we aim to investigate the effect of GnRH analogues to peripheral blood lymphocytes,
in-vitro.
DESIGN: Experimental study.
MATERIALS AND METHODS: PBMCs were treated with various concentration
of GnRH agonist (Buserelin acetate) and antagonist (Cetrorelix
acetate) of 0.1, 1, 5 and 10mM for 4 hours to 48 hours. Th1, Th2 and Th17
cells were analyzed by flow cytometry. Anti-CD45, anti-CD3 and anti-
CD8 were used to isolate T cell subpopulation and intracellular cytokine
staining was made to analyze Th1, Th2 and Th17 cell population using
anti-TNF-a, anti- INF-g, anti-IL-10 and IL-17.
RESULTS: The proportion of TNF-a and IFN-g producing Th cells were
increased in dose dependent manner with GnRH agonist culture for 24 hours.
The proportions of TNF-a producing Th cells with 5 and 10 mM GnRH
agonist (P¼0.046 each) were significantly higher than that with 1 mM
GnRH agonist. There were no significant differences in IL-10 producing
Th cells after 24 hour culture with GnRH agonist. The proportion of Th17
cells treated with 1 mM GnRH agonist were significantly higher than those
of controls (without GnRH agonist) (P¼0.008), with 5 mM (P¼0.028) and
10 mM (P¼0.028) GnRH agonist. TNF-a/IL-10 and IFN- g/IL-10 ratios
treated with GnRH agonist for 24 hours (0.1, 1 and 5 mM) were not different
from those of controls. Th17 cells treated with 1 mM GnRH agonist was
significantly higher than those without GnRH agonist (p¼0.008) after 48
hour culture. TNF-a/IL-10 and IFN-g/IL-10 secreting Th cell ratios in
PBMCs treated with GnRH agonist for 4 hours (0.1, 1, 5 and 10 mM) were
increased as compared to controls, and those treated with GnRH antagonist
(0.1, 1 and 5 mM) were decreased as comparison to controls. Th17 cells
cultured with 1 mM GnRH agonist was significantly decreased in comparison
to controls (P¼0.019). Th17 cells treated with 10 mM GnRH antagonist was
significantly increased as compared to controls (P¼0.023).
CONCLUSIONS: GnRH analogues affect T helper cell subpopulations
including Th1 and Th17 cells. GnRH agonist seems to induce Th1 and
Th17 immunity. Further investigation of GnRH analogues will be necessary
to understand inflammatory immune responses during ovulation induction
cycles.
O-158 Tuesday, October 20, 2015 11:30 AM
INHIBITION OF AUTOPHAGY BY SERA FROM WOMEN WHOSE
IVF CYCLE RESULTED IN A SPONTANEOUS ABORTION OR
BIOCHEMICAL PREGNANCY. G. Sisti, T. Kanninen, I. Ramer,
S. Witkin, S. D. Spandorfer. Cornell University Medical Center, New
York City, NY.
OBJECTIVE: Autophagy is an intracellular process whereby degraded or
foreign proteins and defective organelles become surrounded by a structure
called an autophagosome. By subsequent fusion with a lysosome, the enclosed
macromolecules are degraded and its component parts returned to
the cytoplasm for reutilization. Induction of autophagy at various stages of
FERTILITY & STERILITY Ò
e61

pre- and post-implantation fetal growth in animal models has been shown to
enhance embryo development and survival. We previously reported that sera
from pregnant women induced autophagy in peripheral blood mononuclear
cells (PBMCs). In the present study we evaluated whether the extent of autophagy
induction by sera of pregnant women would differ according to the
outcome of their IVF cycle.
DESIGN: We evaluated the extent of autophagy induction by available
sera from a retrospective cohort of 94 women who completed an IVF cycle
at our institution. The breakdown was 28 women with a term pregnancy,
16 who delivered preterm, 11 with a SAB, 12 with a BC pregnancy, 16
who did not become pregnant and 11 who had an ectopic pregnancy (EP).
MATERIALS AND METHODS: Donor PBMCs were isolated from
reproductive age healthy female volunteers by Ficoll-Hypaque gradient
centrifugation and added to wells of a sterile microtiter plate (5x105 cells
per well) that contained 35 ml RPMI culture medium and 15 ml sera from
the study subjects. All sera were from day 28 of the IVF cycle, obtained
at the time of the initial pregnancy test. As a positive control parallel cultures
also contained 4800 mM rapamycin, an autophagy inducer. Following
incubation for 48 hours (37 C, 5% CO2) the cells were centrifuged, lysed in
the presence of protease inhibitors and stored at -80oC until assayed. The
extent of autophagy was determined by measuring the concentration of
p62 by ELISA. p62 is a protein whose intracellular concentration is
inversely proportional to the extent of autophagy induction. The non-parametric
Kruskal Wallis test with Dunn’s correction was used to determine
associations.
RESULTS:
Autophagy induction in donor PBMCs by sera from women with different IVF
outcomes
OUTCOME
Median ng/mL p62
No rapamycin
(range)
+ rapamycin
Term birth 7.5 (

O-161 Tuesday, October 20, 2015 12:15 PM
AN FSH-LOWERING ACTIVIN DISRUPTING THERAPY PRE-
VENTS EGG CHROMOSOME AND SPINDLE MISALIGNMENTS
THAT PREDISPOSE TO ANEUPLOIDY, AND INCREASES
FERTILITY, IN A MOUSE MODEL OF MIDLIFE REPRODUCTIVE
AGING. L. R. Bernstein, a,b,c,d A. C. Mackenzie, b C. L. Chaffin, e
I. Merchenthaler. b a Pregmama, LLC, Montgomery Village, MD; b Epidemiology
and Public Health, University of Maryland School of Medicine, Baltimore,
MD;
c Gynecology and Obstetrics, Johns Hopkins School of
Medicine, Baltimore, MD; d Veterinary Integrative BioSciences, Texas A &
M College of Veterinary Medicine, College Station, TX; e Obstetrics, Gynecology,
and Reproductive Sciences, University of Maryland School of Medicine,
Baltimore, MD.
OBJECTIVE: Women of advanced maternal age women (AMA, >age 35)
have increased risk of oocyte & embryo aneuploidy, infertility, miscarriages,
and trisomic pregnancies (collectively ‘‘egg infertility.’’). Egg infertility increases
markedly with age due to elevated rates of egg aneuploidy. It is a significant
public health problem, with 1 in 5 US women now attempting her
first pregnancy after 35. Elevated FSH is one of the first signs of ovarian aging.
We hypothesize that high FSH is a cause of egg infertility, that elevating
FSH activity for the period of oocyte growth will increase egg infertility, and
that lowering FSH for the period of oocyte growth will prevent egg infertility.
DESIGN: We developed SAMP8 mice as model with human-like midlife
female reproductive aging characteristics, including elevated FSH, increased
rates of oocyte spindle misalignments, and diminished fertility by midlife
(age 7 months). A regimen to raise FSH activity with chronic PMSG treatment
was given to one test group of midlife SAMP8 for 3 weeks, the period
of oocyte growth. An FSH lowering regimen was developed using ActRIIB:Fc,
an activin decoy receptor that sequesters activin and suppresses activin
signaling. This was given for 3 weeks to a second test group. A third test
group was comprised of untreated control mice.
MATERIALS AND METHODS: Chromosome and spindle misalignments
of ovulated oocytes are highly predictive of impending aneuploidy,
were scored in fluorescence microscopy. Fertility was compared between ActRIIB:Fc-treated
and untreated midlife SAMP8 groups by quantitation of
litter sizes after mating with young proven SAMP8 males.
RESULTS: PMSG increased rates of chromosome misalignments from
32/193 oocytes (16.5%) to 38/121 oocytes (31.4%; P¼0.0013), and
increased rates of spindle misalignments from 13/192 (6.77%) to 14/98
(14.3%; P¼0.0331). ActRIIB:Fc lowered FSH in midlife SAMP8 to the
level of young SAMP8. ActRIIB:Fc decreased chromosome misalignments
from 32/193 (16.5%) to 11/159 (6.9%; P¼0.0030), and decreased spindle
misalignments from 13/192 (6.77%) to 4/155 (2.58%; P¼0.0182). Rates
of chromosome and spindle misalignments were lowered to those of young
mice. ActRIIB:Fc restored nearly 40% of the fertility lost with age,
increasing litter sizes from 5.06 to 6.29 pups/litter (P¼0.0305, vs. 8.22 in
young SAMP8).
CONCLUSIONS: These data provide supportive evidence that FSH may
play a role in egg aneuploidy and infertility. Hormone normalization therapy
(‘‘HNT’’) to lower FSH and disrupt activin signaling shows promise as a
novel therapeutic intervention to prevent oocyte aneuploidy and infertility,
miscarriages, and trisomies.
Supported by: 1. Maryland Industrial Partnerships Grant (MIPS). 2. Technology
Development Corporation of Maryland (TEDCO). 3. Max and Victoria
Dreyfus Foundation Grant. 4. Bernstein and Pine Families. 5. Pregmama.
6. University of Maryland School of Medicine Funds. 7. Indiegogo
campaign.
O-162 Tuesday, October 20, 2015 12:30 PM
GNRH AGONIST LEUPROLIDE ACETATE NEITHER ACTIVATES
ANTI-APOPTOTIC GENES NOR PROTECTS HUMAN OVARY
AND GRANULOSA CELLS FROM DNA DAMAGE AND APOPTOSIS
INDUCED BY CYCLOPHOSPHAMIDE. G. Bildik, a N. Akin, a
F. Senbabaoglu, a Y. Guzel, b U. Ince, c B. Balaban, b B. Urman, d,b
O. Oktem. d,b a Reproductive Biology, Koc University Graduate School of
Health Sciences, Istanbul, Turkey; b Women’s Health Center Asssited Reproduction
Unit Fertility Preservation Program, American Hospital, Istanbul,
Turkey; c Pathology, Acibadem University, Istanbul, Turkey; d Obstetrics
and Gynecology, Koc University School of Medicine, Istanbul, Turkey.
OBJECTIVE: Inconsistent results of randomized controlled trials (RCTs)
and lack of a proven molecular mechanism of action with ovarian protection
with co-administration GnRH agonists (GnRHa) with chemotherapy places
GnRHa under scrutiny as a fertility preservation strategy. We aimed in this
study to provide molecular evidence for-or-against the role of GnRHa in
the prevention of cyclophosphamide induced damage in human ovarian tissue
samples and granulosa cells.
DESIGN: A translational research study.
MATERIALS AND METHODS: Ovarian cortical pieces (n¼15, age
14-37) and human mitotic non-luteinized (COV434, HGrC1) and nonmitotic
luteinized (HLGC) granulosa cells were treated with 4-hydroperoxy
cyclophosphamide (in vitro active metabolite of cyclophosphamide
used at 50 and 100 mM) with and without GnRHa leuprolide acetate
(50 ng/mL: peak intraovarian concentration of the drug) for 24 hrs.
Cell proliferation (real-time quantitative assessment by xcelligence system),DNA
damage (p-histone H2AX), apoptosis (cleaved caspase-3,
YO-PRO-1), follicle counts, hormonal markers of ovarian function and
reserve (estradiol, progesterone and AMH), and the expression of antiapoptotic
genes (bcl-2, bcl-xL, bcl-2L2, Mcl-1, BIRC-2 and XIAP)
were compared among control, chemotherapy and chemotherapy+GnRHa
groups.
RESULTS: GnRH receptor expression and its activation by GnRHa were
validated with qRT-PCR and measuring intracellular cAMP level, respectively.
Exposure to cyclophosphamide resulted in massive follicle loss, arrested
cell growth, increased DNA damage/apoptosis and decreased
hormone productions in the tissue samples and granulosa cells. The coadministration
of GnRHa with cyclophosphamide did not prevent or attenuate
any of these cytotoxic effects. Furthermore, GnRHa did not up-regulate
the anti-apoptotic genes compared to control and cyclophosphamide treated
samples.Mcl-1 and BIRC2 expressions were further decreased after cyclophosphamide+GnRHa
(Table).
CONCLUSIONS: GnRH agonist leuprolide acetate does not offer any protection
against cyclophosphamide induced damage in human ovary and granulosa
cells via its cognate receptors.
The Impact of Cyclophosphamide (Cyc)GnRHa on Ovarian Tissue Samples
and Granulosa Cells
Control Cyc Cyc+GnRHa P value
Ovarian Tissue: Follicle reserve, anti-apoptotic gene expression and
hormone productions
Primordial/mm2
Preantral/mm2
2.540.5
0.60.2
0.330.2
0.120.05
0.330.1
0.140.1
p¼0.006 control vs.
cyc+GnRHa
p¼0.3 cyc vs.
Bcl-2
Bcl-xL
Bcl-2L2
Mcl-1
BIRC2
XIAP
AMH(ng/mL)
E2(pg/mL)
P(ng/ml)
10.06
10.03
10.04
10.02
10.04
10.01
1.20.09
78898
1.760.4
0.600.07
0.970.03
0.600.03
0.930.03
0.750.01
0.860.02
0.460.01
0.920.01
0.510.01
0.810.01
0.590.01
0.720.05
0.30.02
14843
0.310.06
p
cyc+GnRHa
0.30.03
p¼0.004
18516
control vs.
0.30.02
cyc+GnRHa
p¼0.2
cyc vs. cyc+GnRHa
Luteinized Granulosa Cells (HLGCs): Hormone productions and apoptosis rate
E2(pg/mL)
P(ng/mL)
Apoptosis (%)
1560112
59695
3%
27128
15018
89%
20731
10516
91%
Mitotic granulosa cells: Proliferative index and apoptosis rate
Proliferative
index
Apoptosis(%)
1.20.2
3%
0.140.01
88%
p¼0.009 control vs.
cyc+GnRHa
p¼0.3 cyc vs.
cyc+GnRHa
p
p¼0.9 cyc vs.
cycGnRHa
(for proliferative
index)
p
p¼0.8 cyc vs.
cyc+GnRHa
(for apoptosis
rate)
FERTILITY & STERILITY Ò
e63

GENETIC COUNSELING 1
O-163 Tuesday, October 20, 2015 11:15 AM
ADVANTAGES OF TRIPLET REPEAT EXPANSION DETECTION IN
BLASTOCYST BIOPSY FOR PREIMPLANTATION GENETIC
DIAGNOSIS OF FRAGILE X SYNDROME. R. Prates, a S. Jaroudi, b
A. Jordan, a N. Goodall, a B. Chu, a V. Tecson, a A. Hershlag, c
M. Garrisi, d F. Licciardi, e B. Witt, f M. Konstantinidis. a a Reprogenetics, Livingston,
NJ; b Reprogenetics Middle East, Al Ain, United Arab Emirates;
c North Shore LIJ Health System, Cold Spring Harbor, NY; d IRMS-NJ, Livingston,
NJ; e New York University Langone Medical Center, New York, NY;
f Greenwich Fertility and IVF Center, Greenwich, CT.
OBJECTIVE: To evaluate the utilization of a comprehensive diagnosis
strategy for preimplantation genetics diagnosis (PGD) of fragile X syndrome
(FXS) in order to enhance selection of embryos suitable for transfer and increase
therefore, the chances of patients achieving a pregnancy.
DESIGN: Trophectoderm biopsies obtained from blastocysts preimplantation
embryos were processed for direct FXS repeat size determination and
linkage analysis via Karyomapping (Illumina, USA). When requested,
comprehensive chromosome screening (CCS) via array comparative
genomic hybridization testing was also performed in parallel to PGD for
FXS.
MATERIALS AND METHODS: Trophectoderm biopsies were sent from
27 different IVF clinics between December 2013 and March 2015 to a single
PGD laboratory for analysis. A total 168 blastocysts from 34 PGD cycles (4.9
embryos per cycle) were assessed for FXS direct mutation test (average
maternal age: 34.8).
RESULTS: Diagnostic results were obtained for 96.4% of the samples
(162/168). The proportion of embryos found to be available for transfer based
on the inheritance of the normal allele via linkage analysis was 53.7% (87/
162). Among the 46.3% (75/162) categorized as having inherited the affected
allele, direct triplet repeat sizing allowed the identification of 24.1% (39/162)
additional blastocysts suitable for transfer (carriers of premutation or intermediate
expansions) (p43). The aneuploidy rate for the each male-female age group interaction
was computed, with 95% confidence intervals calculated by Clopper-Pearson
method. The aneuploidy rate was modeled by logistic regression. The youngest
age groups were considered reference factors. The model was assessed
by chi-square of ANOVA with significance at p

Table 1: Incidence and types of chromosome errors in aneuploid blastocysts
(*p

for 8511 patients. Individuals were screened for up to 213 recessive genetic
diseases using Illumina’s Infinium HD Genotyping Platform. Carrier counts
were tallied for each disease screened, and overall number of carriers was
analyzed for 3 disease panels of increasing scope: (1) diseases currently recommended
for screening by professional society guidelines; (2) diseases
considered to be high-impact for reproductive decision making; and (3) a
broad panel of 213 diseases that range in severity.
RESULTS: On the most limiting panel, which only screened for diseases
currently included in professional society guidelines, 10.7% of patients were
identified as carriers. When compared to a larger panel inclusive of 188 high
impact diseases, our results indicated that limited screening failed to identify
26.5% of carriers in our patient population. An additional 5% of patients
were identified as carriers when the panel was expanded further to include
a total of 213 diseases of varying impact.
CONCLUSIONS: New technologies have provided the ability to simultaneously
screen for a multitude of mutations at a reduced cost. Concurrently,
genetic diversity has increased due to the admixture of different ethnic groups.
These cultural and technological changes have altered the clinical approach to
genetic carrier screening, and expanded carrier panels with greater than 100
diseases are increasingly being offered to patients. Data from our ethnically
diverse patient population demonstrate that screening for only traditionally
recommended diseases failed to identify over a quarter of carriers. Such oversight
might contribute to decreased detection of carrier couples, and thus the
inability for patients to make fully informed reproductive decisions. Furthermore,
an additional 5% of patients were identified as carriers when including
moderate impact conditions, which may not affect reproductive decisions but
may provide helpful information to couples about their future child’s health.
Overall, our data support that expanded carrier panels are more effective at
identifying patients at risk of having children with genetic conditions who
thus may benefit from increased reproductive options. The continued support
of genetic counselors will be critical to guiding all patients through the
screening process, regardless of carrier status or disease severity.
References:
1. Edwards JG, Feldman G, Goldberg J, et al. Expanded Carrier Screening
in Reproductive Medicine_Points to Consider: A Joint Statement of the
American College of Medical Genetics and Genomics, American College
of Obstetricians and Gynecologists, National Society of Genetic
Counselors, Perinatal Quality Foundation, and Society for Maternal-
Fetal Medicine Obstet Gynecol. 2015;125(3):653-662.
MALE REPRODUCTION
O-169 Tuesday, October 20, 2015 11:15 AM
INTRACYTOPLASMIC SPERM INJECTION REDUCES TOTAL
FAILED FERTILIZATION RATE BUT DOES NOT IMPROVE PREG-
NANCY OR LIVE BIRTH RATES IN UNEXPLAINED INFERTILITY:
ANALYSIS OF OVER 20,000 CYCLES FROM THE SART
DATABASE. L. Johnson, a,b M. D. Sammel, c A. Dokras. b a Reproductive
Endocrinology Associates of Charlotte, Charlotte, NC; b University of Pennsylvania,
Philadelphia, PA; c Univ. of Pennsylvania, Perelman School of
Medicine, Philadelphia, PA.
OBJECTIVE: In vitro fertilization (IVF) with conventional insemination
is an effective therapy for unexplained infertility, but total fertilization failure
(TFF) remains high. A recent meta-analysis suggests that intracytoplasmic
sperm injection (ICSI) improves fertilization rates and decreases TFF in couples
with unexplained infertility. (1) However, pregnancy data are lacking.
We aim to determine if ICSI improves clinical pregnancy rate and live birth
rate in couples with unexplained infertility using the SART Database.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: All IVF cycles reported to the Society
for Assisted Reproductive Technology (SART) between 2004 to 2011 were
examined. IVF cycles were included if unexplained infertility was the only
diagnosis. The first fresh cycle for each patient was analyzed. Exclusion
criteria included: first fresh cycle before 2004, both ICSI and conventional
insemination in the same cycle, second-day ICSI, and missing data for
age, BMI, and smoking. Linear and logistic regression models were utilized
to estimate fertilization rate, TFF rate, transfer rate, number of embryos
transferred and cryopreserved, clinical pregnancy rate, miscarriage rate,
and live birth rate. Adjusted models controlled for age, BMI, smoking,
FSH dose, and number of oocytes retrieved. This study has 95% power to
detect a 10% increase in clinical pregnancy and live birth rates. A p value
of

O-171 Tuesday, October 20, 2015 11:45 AM
DELETERIOUS EFFECTS OF MALE OBESITY ON REPRODUC-
TIVE PARAMETERS AMONG A LARGE MULTI-INSTITUTIONAL
COHORT. J. M. Bieniek, a J. A. Kashanian, b C. M. Deibert, c
E. D. Grober, a R. Brannigan, b J. I. Sandlow, c K. A. Jarvi. a a University of
Toronto, Toronto, ON, Canada; b Northwestern University, Chicago, IL;
c Medical College of Wisconsin, Milwaukee, WI.
OBJECTIVE: Obesity has been suggested as a contributing factor in male
subfertility but correlations with semen parameters have been mixed, therefore,
this study was designed to analyze the relationships between body mass
index (BMI) and measures of male fertility among a large multi-institutional
cohort.
DESIGN: Retrospective review of prospectively-collected demographic
data to calculate measures of obesity from three North American male
infertility clinics with correlation to semen and reproductive hormonal
parameters.
MATERIALS AND METHODS: Self-reported or measured height and
weight were captured in review board-approved databases at male infertility
clinics since 2002. Men with semen analysis or reproductive hormone parameters
at initial evaluation were included to reduce effects of infertility-related
treatments. BMI was calculated for all patients with comparisons to reproductive
parameters performed utilizing ANOVA, chi squared, and Spearman’s
rank correlations as appropriate with p< .05 reported as significant.
RESULTS: Complete height and weight data to calculate BMI was available
for 4440 patients with a mean age of 36.1 (7.6) years. Based on WHO
definitions, 30.9% of the cohort was normal weight (BMI 18.5-24.9), 45.1%
overweight (25-29.9), and 23.3% obese (>30). Semen analysis and reproductive
hormone results were collected for 4236 (95.4%) and 2973 (67.0%) men,
respectively. The gonadotropins FSH and LH demonstrated weak positive
correlations with BMI with only LH reaching significance (r¼0.04,
p¼0.06; r¼0.06, p¼0.01). Testosterone (r¼ -0.27, p

discarded (egg donation) and male factor is diagnosed with abnormal protamine
ratio compared to previous ejaculate egg donation cycles.
DESIGN: Unicentric, retrospective study (Jan.2014-Apr.2015) including
couples who underwent at least one previous egg donation cycle with sperm
ejaculate. Protamine ratio was studied when poor or null blastocyst rate was
observed and no pregnancy was achieved. Cases with altered protamine ratio
were indicated for TESA in the following cycle.
MATERIALS AND METHODS: 38 couples from egg donation treatment
were included. Gardner’s classification was used for embryo
grading. Protamine ratio was measured through FertiCertTM. TESA
was performed in subsequent cycle if altered protamine ratios were diagnosed.
To compare cycle outcome, fertilization, blastocyst, AA blastocyst
and pregnancy rates were studied in both. Accepted by our Institutional
Review Board.
RESULTS: Fertilization rate was significantly higher in TESA compared
to ejaculate (85.0% vs 69.3%; p¼0.001). Blastocyst formation rate was also
significantly higher in TESA group than in ejaculate (47.6% vs 21.3%;
p¼0.000). Regarding AA quality blastocyst formation rate, this was significantly
different as well (30.9% vs 9.3%; p¼0.001). When implantation rate is
analyzed, it increases from ejaculate (12.5%) to TESA group (75%) being
this difference significantly different (p¼0.0018). T-Student and c2 tests
with SPSS were used to analyze data.
CONCLUSIONS: Improved cycle outcomes were observed in terms of
fertilization, blastocyst formation and implantation rates when TESA was
used for fertilization, compared to previous ejaculate egg donation cycles
in patients with altered protamine ratios. More cases are needed. A randomized
controlled trial would be difficult due to TESA is an invasive technique
and should not be performed without firm indication. The improvement
observed is difficult to determine whether it comes from unprotected DNA
or epigenetic factors. This study provides a new proceeding strategy for couples
who are not prepared for donor sperm. These data corroborate the relationship
between altered protamine ratios and infertility, but also suggest that
a different sperm sources can result in an improvement of fertilization and
blastocyst rates and thus in the cycle outcome.
O-174 Tuesday, October 20, 2015 12:30 PM
PREGNANCY LOSS IS SIGNIFCANTLY INCREASED IN MALE
FACTOR OLIGOASTHENOZOOSPERMIA PATIENTS DESPITE
THE TRANSFER OF A EUPLOID BLASTOCYST. J. M. Stevens, a
S. McCormick, a A. Schneiderman, a W. B. Schoolcraft, b M. Katz-Jaffe. b
a Fertility Laboratories of Colorado, Lone Tree, CO; b Colorado Center for
Reproductive Medicine, Lone Tree, CO.
OBJECTIVE: Male factor (MF) infertility contributes to half of all infertility
cases worldwide. Compromised quality of the male gamete is known
to impact fertilization potential, embryo quality and recurrent miscarriage
(RM). Studies have shown that men with MF or RM have increased sperm
aneuploidy compared to controls. This study explored the influence of oligoasthenozoospermia
(sperm motility

OBJECTIVE: The role of the endocannabinoid system (ECS) in the pathogenesis
of endometriosis is still under investigation. In this study, the
changes on ECS related to apoptosis and autophagy processes have been
analized in human granulosa cells (GCs) from women affected by endometrioma
DESIGN: This prospective non-randomized study has been carried out
from January to September 2014 on women undergoing a controlled ovarian
hyperstimulation for an IVF treatment. In particular, GCs were collected
from both ovaries of 10 women with a diagnosis of unilateral ovarian endometrioma
at the time of oocytes retrieval. 9 women with male, idiopathic or
tubal-factor infertility diagnosis were selected for the control group. The
three experimental groups matched for female age (36.24.1 vs 35.42.6)
MATERIALS AND METHODS: GCs obtained from follicles aspirates
were isolated from red blood cells and follicular fluid by density gradient
centrifugation. Q-PCRs were performed with the SYBR green method in
an iQ5 iCycler thermal cycler using bactin and GAPDH as reference genes.
Data are presented as mean S.D. Two-Way ANOVA followed by Tukey
test as Multiple comparisons test, was used for comparison among experimental
groups. All statistical analyses were performed using the statistical
software package Prism5 (Graphpad Software, Inc. USA) with significance
accepted at P

O-179 Tuesday, October 20, 2015 12:15 PM
STEROIDOGENESIS IN ENDOMETRIOSIS: THE IMPORTANCE OF
GATA6. L. A. Bernardi, S. E. Bulun, M. T. Dyson. Obstetrics and Gynecology,
Feinberg School of Medicine, Northwestern University, Chicago, IL.
OBJECTIVE: The production of steroids from cholesterol is initiated by
the concerted effort of multiple factors coordinated in a tissue specific
pattern. The same steroidogenic factors are also essential for the development
of endometriosis. The PKA pathway and signaling from steroidogenic factor
1 (NR5A1) critically regulate steroidogenic gene expression, and these along
with GATA6 have been implicated in the pathogenesis of endometriosis. This
study evaluated the combinatorial impact of GATA6 and NR5A1 on steroidogenesis
in diseased cells, and expanded these findings to demonstrate how
these pathways permit healthy cells to acquire the steroidogenic potential
that drives endometriosis.
DESIGN: Basic science, in-vitro.
MATERIALS AND METHODS: Ectopic endometriotic fibroblasts
(OSIS), obtained from the cyst walls of endometriomas, and normal eutopic
endometrial fibroblasts (NoEM), obtained from the endometrium of subjects
without endometriosis, were isolated in accordance with IRB-approved protocols.
Human foreskin fibroblasts (BJ) were purchased commercially.
NR5A1 and GATA6 were depleted in OSIS by the reverse transfection of
gene-specific silencer select siRNAs in comparison to a scrambled siRNA.
NoEM and BJ cells, which express little endogenous NR5A1 or GATA6,
were transduced with adenoviruses bearing CMV-driven NR5A1, GATA6,
or an empty CMV-null control. Expression of steroidogenic genes including
steroidogenic acute regulatory protein (StAR), cholesterol side chain cleavage
enzyme (CYP11A1), 3-beta-hydroxysteroid dehydrogenase type 2
(HSD3B2), 17a-hydroxylase (CYP17A1), and aromatase (CYP19A1),
were assessed using RT-qPCR and immunoblotting.
RESULTS: Depleting NR5A1 in OSIS significantly reduced the expression
of StAR and CYP11A1, and attenuated HSD3B2, CYP17A1 and
CYP19A1 as compared to controls. Depleting GATA6 in OSIS attenuated
the PKA-dependent induction of CYP17A1 and CYP19A1 compared to untreated
controls. In both NoEM and BJ cells, the expression of StAR and
CYP11A1 was induced by NR5A1 alone; however, expression was significantly
higher in NoEM than in BJ cells. In contrast, HSD3B2, CYP17A1,
and CYP19A1 were synergistically increased by NR5A1 and GATA6
compared to either gene alone in NoEM, while in BJ cells expression of
HSD3B2, CYP17A1 and CYP19A1 was minimal.
CONCLUSIONS: Silencing of NR5A1 and GATA6 in endometriotic tissue
leads to decreased expression of key steroidogenic enzymes. In non-endometriotic
cells the ability of NR5A1 and GATA6 to work synergistically to
increase steroid production depends on a tissue’s inherent potential for steroidogenesis.
In conclusion, GATA6 appears to play an important role in
the regulation of steroid production and ultimately in the pathogenesis of
endometriosis.
Supported by: Research Supported by NIH R37HD038691-12S1 and a
grant from the Friends of Prentice at Northwestern Memorial Hospital.
O-180 Tuesday, October 20, 2015 12:30 PM
PATIENTS WITH ENDOMETRIOSIS DO NOT HAVE A HIGHER
RATE OF ANEUPLOIDY. C. R. Juneau, J. M. Franasiak,
M. D. Werner, E. J. Forman, T. Molinaro, R. T. Scott. RMA, NJ, NJ.
OBJECTIVE: The diminution in outcomes which impact women with
endometriosis undergoing IVF have variably been attributed to alterations
in endometrial receptivity, decreased oocyte quality, and premature depletion
of follicular reserve. Little data are available to determine if the changes in
ovarian reserve experienced by women with endometriosis adversely impacts
oocyte maturation ultimately resulting in an increased risk of embryonic
aneuploidy. This study seeks to address that question.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: Patients participating in their first IVF
cycle at a single infertility clinic from 2009-2014 with comprehensive chromosome
screening (CCS) were included. Patients with endometriosis were
identified through surgical diagnosis or by ultrasound findings consistent
with persistent space-occupying disease whose sonographic appearance is
consistent with endometriosis. Patients were divided into the SART age
groups, and aneuploidy rates in each group were compared to those in
3301 age-matched patients in the general IVF population undergoing CCS.
While the control group included all patients in the general IVF population,
including patients with endometriosis, the proportion of patients with endometriosis
only accounts for 1% of this population and is therefore not likely
to confound results. Statistical analysis was performed using c2 test of proportions.
An alpha error of 0.05 was accepted.
RESULTS: There were 253 patients with endometriosis who produced
1283 blastocysts that met criteria for inclusion. Mean age of patients with
endometriosis was 35.7 4.0 years (range 24-46 years). The median antimullerian
hormone level was 1.70 ng/mL (range 0.16-15.36 ng/mL). When
aneuploidy rates in patients with endometriosis and aneuploidy rates in the
general IVF population were stratified by SART age groups and compared,
there was no statistical difference (table 1).
CONCLUSIONS: Patients with endometriosis undergoing IVF have aneuploidy
rates equivalent to their age-matched peers in the general IVF population.
The rate of aneuploidy is not increased in patients with endometriosis
undergoing IVF.
SART age
group
Rate of aneuploidy
in patients with
endometriosis
Rate of aneuploidy
in general IVF
population
p-value
26.7% (160/600) 28.9% (1974/6819) 0.2556
35-38 years 33.3% (128/384) 37.4% (1369/3659) 0.1285
38-40 years 54.0% (129/239) 52.7% (1629/3084) 0.6581
41-42 years 72.9% (35/48) 71.2% (857/1203) 0.9288
>42 years 75.0% (9/12) 84.6% (342/404) 0.6141
FIBROIDS
O-181 Tuesday, October 20, 2015 11:15 AM
ELAGOLIX FOR THE MANAGEMENT OF HEAVY MENSTRUAL
BLEEDING (HMB) ASSOCIATED WITH UTERINE FIBROIDS
(UF): RESULTS FROM A PHASE 2A PROOF-OF-CONCEPT
STUDY. D. F. Archer, a K. Chwalisz, b R. Feldman, c E. A. Stewart, d
A. S. Lukes, e J. North, b J. Gao, b L. A. Williams, b R. Jain. b a Department
of Obstetrics & Gynecology, Eastern Virginia Medical School, Norfolk,
VA; b AbbVie, Inc., North Chicago, IL; c Miami Research Associates, Pinecrest,
FL; d Mayo Clinic, Rochester, MN; e Consulting Teaching Professor,
Duke University Med, Chapel Hill, NC.
OBJECTIVE: Evaluate safety and efficacy of elagolix (an oral GnRH antagonist)
alone and combined with low-dose estrogen + progestin add-back therapy
regimens in premenopausal women with HMB associated with UF.
DESIGN: Phase 2a study with 3 randomized, double-blind, placebo
(PBO)-controlled (RPC) and 3 open-label (OL) cohorts. Three elagolix
dosing regimens were used, alone and in combination with 2 low-dose
add-back therapy regimens for 3 mo (Table). Primary efficacy endpoint
was mean change in menstrual blood loss (MBL) from baseline to the final
month (last 28 d on treatment); percentage change from baseline in MBL
was a secondary HMB efficacy endpoint.
MATERIALS AND METHODS: Premenopausal women aged 20-49 y
with UF (R2 cm), regular menses every 24-35 d, and HMB (MBL >80
mL during 2 screening cycles) were eligible. MBL was assessed at screening
and monthly during treatment as measured with the alkaline hematin method.
Between-group comparisons were tested using analysis of covariance, with
treatment as a factor and baseline as a covariate.
RESULTS: 271 women were randomized; 228 completed. Approximately
75% of women were black, with a mean age of 41.8 y. A substantial reduction
in MBL (ie, improvement in HMB) at the final month was observed in all elagolix-containing
treatment groups (Table). Changes from baseline in MBL
with all elagolix doses were statistically significant (P%0.001) vs PBO in
RPC cohorts. Overall, adverse event (AE) rates were similar across treatment
groups, except hypoestrogenic events such as hot flush. Hot flush was the
most frequent AE in elagolix-only treatment groups (45%-63%) vs PBO
(6%-19%); rates were 26% (E2:NETA) and 19% (E2:P) with add-back therapy.
AEs resulted in study drug discontinuation in 24 women (elagolix alone,
18/160 [11%]; elagolix + add-back therapy, 2/61 [3%]; PBO, 4/50 [8%]).
CONCLUSIONS: All elagolix-containing treatment regimens substantially
improved HMB associated with UF; 300 mg BID was the most
efficacious dose. Both low-dose add-back regimens reduced hot flush by
40%-50% vs corresponding elagolix-only dose groups, with minimal
impact on HMB efficacy endpoints.
e70 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

Table. Key Efficacy Parameters by Total Daily Dose of Elagolix
Elagolix
Daily Dose
0mg
(PBO)
Study
Type/Elagolix
Dosing/
Add-Back
Therapy/n
Mean
(SD) BL
MBL, mL
n¼50 251.7
(160.3) to
200 mg RPC/100
mg BID/
none/n¼33
400 mg RPC/200 mg
BID/none/
n¼35
400 mg RPC/400
mg QD/none/
n¼32
400 mg OL/200
mg BID/
E2:NETA/
n¼34
600 mg RPC/300
mg BID/
none/
n¼30
600 mg OL/600 mg
QD/none/
n¼30
600 mg OL/300 mg
BID/E2:P/
n¼27
Mean
(SE) MBL, Change From
Baseline to Final Month
(Last 28 d on Treatment)
mL %
349.2 (424.1)
-114.8
(32.6) to
8.9 (36.7)
-7.7
(11.1) to
-41.2 (6.6)
269.4 (163.2) -181.0 (25.2) -72.0 (7.7)
335.1 (322.7) -252.8 (24.4) -80.2 (13.6)
213.7 (108.1) -197.0 (25.5) -83.3 (7.7)
247.7 (177.7) -192.3 (191.5) -79.6 (43.6)
206.3 (125.1) -237.0 (21.3) -98.0 (4.6)
215.6 (122.8) -189.1 (151.2) -88.6 (39.6)
258.9 (207.5) -217.1 (157.5) -85.4 (28.1)
exosome treatment to measure level of proliferation of the treated cells vs
vehicle control.
RESULTS: Hypoxia induced dramatic increase in exosomes production
by hFSC (up to 6 fold increase) compared to normoxic-grown hFSC (P ¼
0.005). The hypoxic and normoxic exosomes were similar in size about
200 nm. The hypoxic exosomes induced significantly higher rate of proliferation
in primary fibroid cell proliferation compared to normoxic exosomes
(P ¼ 0.02) after 48 hours of treatment. Further characterization of effect of
hypoxia on exosome contents (such as miRNA profile) is ongoing in our laboratory.
CONCLUSIONS: Our data presents, for the first time, a mechanism by
which hypoxic-exposed tumor forming fibroid stem cells would enhance tumor
formation. Hypoxic hFSCs exert pro-proliferative paracrine stimulatory
effect on primary fibroid cells by producing abundant amount of exosomes
with robust oncogenic potential. Further understanding of the cross-talk between
human fibroid tumor-forming stem cells and surrounding differentiated
tumor cells can provide novel therapeutic targets for treatment of
symptomatic uterine fibroids.
O-183 Tuesday, October 20, 2015 11:15 AM
WITHDRAWN
BID¼twice daily; BL¼baseline; E2:NETA¼continuous/combined estrogen
(0.5 mg) + norethindrone acetate (0.1 mg); E2:P¼1 mg continuous estrogen
+ 200 mg oral progesterone on days
17-28; MBL¼menstrual blood loss; OL¼open label; PBO¼placebo;
QD¼once daily; RPC¼randomized PBO-controlled; SD¼standard deviation;
SE¼standard error.
Supported by: AbbVie, Inc.
O-182 Tuesday, October 20, 2015 11:30 AM
EXOSOMES FROM HYPOXIA-DRIVEN HUMAN FIBROID STEM
CELLS ACCELERATE TUMOR GROWTH. S. Brakta,
S. M. Shalaby, M. P. Diamond, A. Zimmerman, I. Helwa, Y. Liu,
S. A. Mohamed, L. Gavrilova-Jordan, A. Al-Hendy. GRU, Augusta, GA.
OBJECTIVE: Exosomes are nanovesicles, the smallest extracellular vesicles
identified so far. They are secreted from a variety of cell types and play
an important role in intercellular communication, often inducing physiological
changes in recipient cells by transferring bioactive lipids, nucleic acids,
and proteins. They involve many physiologic processes such as cell growth,
neuronal communication, immune response activation, and cell migration. In
the case of cancer, exosomes may transfer angiogenic proteins, oncogenes or
oncogenic miRNA from tumor forming stem cells to surrounding cells and
promote tumor growth. We have recently characterized human tumor forming
stem cells from uterine fibroid lesions using specific surface markers. In
this work, we wanted to investigate if human fibroid stem cells do indeed produce
exosomes, factors that regulate this process (such as hypoxia) and
possible effect on surrounding differentiated fibroid cells.
DESIGN: Laboratory research studies using human cells.
MATERIALS AND METHODS: For exosome isolation, human fibroid
stem cells were cultured in media with exosome-free FBS for 72 hours at normoxic
or hypoxic (2% O2) conditions. Ten ml of culture media was collected
from each flask and Exoquick TC reagent was added to isolate exosomes according
to manufacturer’s protocol. Electron microscopy and ZetaView were
used to characterized human fibroid stem cell (hFSC) derived hypoxic and
normoxic exosomes. .For proliferation studies, primary human (differentiated)
fibroid cells were cultured in 96 well plates. Media was then changed
to exosome free-FBS and the cells were treated with hypoxic versus
normoxic exosomes. MTT assay was carried out at 24 and 48 hours post
O-184 Tuesday, October 20, 2015 12:00 PM
EXTRACELLULAR MATRIX PRODUCTION DECREASES IN ULI-
PRISTAL ACETATE (UPA) TREATED HUMAN LEIOMYOMAS IN
VITRO AND IN VIVO RESULTING IN DECREASED FIBROID
SIZE. J. Cox, a M. Malik, b J. L. Britten, b A. Patel, b L. Nieman, a
W. H. Catherino. b a National Institutes of Health, Bethesda, MD; b Uniformed
Services University of the Health Scienc, Bethesda, MD.
OBJECTIVE: Multiple prospective, randomized trials have demonstrated
that UPA significantly reduced fibroid volume. In our trial, study patients underwent
hysterectomy and surgical specimens were collected. Using these
tissues, we examined the impact of UPA treatment on extracellular matrix
FERTILITY & STERILITY Ò
e71

(ECM) expression in leiomyomas compared to placebo treated patients. To
further characterize the impact of UPA, we used 3D leiomyoma and myometrial
cultures to assess ECM.
DESIGN: Sub-analysis of randomized placebo controlled trial and laboratory
analysis of clinically-relevant pharmacologic concentrations of UPA on
3D cell cultures.
MATERIALS AND METHODS: A total of 10 patients (5 placebo, 5
treated with 10mg UPA) that underwent hysterectomy and tissue preservation
were identified. Pre and post fibroid volume was evaluated by MRI measurements.
Proteins related to ECM production were evaluated: fibronectin
(FN), versican (VER), and collagen 1A1 (COL). For in vitro analysis, leiomyoma
cells were grown in collagen gel and treated with different concentrations
of UPA for 48hrs. RNA was extracted for qRTPCR analysis.
Protein concentration and distribution was analyzed using immunohistochemistry
(IHC) and Western Blot.
RESULTS: Of treated patients, 80% had decrease or no change in fibroid
volume over 3 months compared to placebo, with a mean decrease of 16%.
Though increased expression of ECM genes was observed in treated surgical
specimens, 80% of the treated specimens demonstrated a significant decrease
in VER protein as compared to placebo. FN protein was decreased in 60% of
treated samples but no consistent alteration in COL was found compared to
placebo surgical specimens. 3D leiomyoma cultures exhibited a UPA
concentration-dependent decrease in VER (1.62+/- 0.13-fold) and FN
(2.58+/-0.22-fold) proteins as compared to untreated cells. This effect was
highlighted by IHC, where both surgical specimens and 3D cell cultures
demonstrated decreased amount of FN and VER in UPA treated leiomyoma
samples.
CONCLUSIONS: UPA treatment decreased fibroid volume in placebocontrolled,
randomized trials. Gene expression and protein production are
altered in leiomyoma tissue in vivo and in vitro compared to placebo controls.
Proteoglycans like VER are known to contribute to the increased water content
of tissue and the large decrease of VER may thus contribute to decreased
fibroid volume on UPA treatment.
Supported by: Intramural grant from Uniformed Services University of
Health Sciences, R21 grant R085193713, and intramural research of the Program
in Reproductive and Adult Endocrinology, NICHD, NIH.
O-185 Tuesday, October 20, 2015 12:15 PM
ALTERED DNA REPAIR GENES IN HUMAN UTERINE FIBROIDS
ARE EPIGENETICALLY REGULATED VIA EZH2 HISTONE
METHYLTRANSFERASE. Q. Yang, L. Elam, A. Laknaur,
L. Gavrilova-Jordan, J. Lue, M. P. Diamond, A. Al-Hendy. Georgia Regents
University, Augusta, GA.
OBJECTIVE: Uterine fibroids (UFs) are benign smooth muscle neoplasms
affecting up to 80% of women. Treatment of symptomatic UFs is a significant
economic burden. Etiology and pathogenesis of UFs is complex. Several genetic
abnormalities related to pathogenesis of UFs have been investigated
including deletions in 7q, trisomy of chromosome 12, rearrangements in
the HMGA2 gene, and mutations in exon 1 and 2 of the MED12 gene. Previous
findings suggest that a low DNA damage repair function may be
responsible for UF formation. The objective of this study is to investigate
whether the DNA damage repair system is altered in UFs and thereby
contribute to an increased risk of UFs development.
DESIGN: Laboratory research studies using human tissues and human
cell lines.
MATERIALS AND METHODS: Surgically removed fresh human
fibroid and adjacent myometrial tissues were collected and primary cells
isolated from fibroids and adjacent myometrial tissues were used for in
vitro studies. The primary cells were grown in media containing inhibitors
of DNMT, HDAC, and EZH2. RNA was isolated using Trizol extraction
method and subjected to cDNA synthesis using Superscript III cDNA
synthesis kit. cDNA was loaded onto the DNA damage signaling pathway
PrimerPCR array plate (H96). RNA levels of over 80 genes were detected
on Bio-Rad CFX96 real-time PCR system. Chromatin Immunoprecipitation
assay was performed to determine the enrichment of H3K27me3
and H3K4me3 in the promoter region of MSH2. Quantitative PCR was
performed to determine the mRNA expression levels of DNA damage
repair genes.
RESULTS: PrimePCR array assay indicated that the expression levels of
over 20 DNA repair genes are altered between fibroid and adjacent myometrial
tissue. Among them, the expression levels of MSH2, a DNA mismatch
repair gene, were significantly downregulated in human fibroid tissue as
compared to the adjacent myometrial tissue. A similar finding was
observed in primary fibroid cells as compared with adjacent myometrial
cells. The fibroid primary cells treated with DZNep (inhibitor of EZH2) exhibited
a marked increase in MSH2 expression in a dose dependent manner.
ChIP assay demonstrated that the enrichment of H3K27me3 in the promoter
region of MSH2 were significantly decreased in DZNep-treated
fibroid cells as compared to vehicle control. In contrast, the recruitment
of H3K4me3 in MSH2 promoter region was not altered in response to
DZNep treatment. These data suggest that EZH2 decreases expression
levels of DNA repair gene MSH2 through epigenetic mark H3K27me3 in
uterine fibroids.
CONCLUSIONS: Identification of the novel EZH2 targeted gene and
MSH2 expression downregulation in fibroid tissue may explain the impaired
DNA repair and high frequency of genetic abnormalities found in fibroid tissue
comparing to normal myometrium. We found that EZH2 decreases the
expression of MSH2 through epigenetic mark H3K27me3. Understanding
of a complex pathogenesis of UF may lead to effective targeted treatment
of symptomatic in the future.
References:
Yang Q, Mas A, Diamond MP, et al. The Mechanism and Function of Epigenetics
in Uterine Leiomyoma Development. Reprod Sci. 2015 Apr
28. pii: 1933719115584449.
Supported by: This work was Supported in part by the Georgia Regents
University Startup package and the National Institutes of Health grant
HD04622811.
O-186 Tuesday, October 20, 2015 12:30 PM
SYNERGISTIC EFFECT OF ESTROGEN AND PROGESTERONE
ON MYOMETRIAL STEM CELL EXPANSION IN VIVO. A. Mas, a
L. Elam, a M. P. Diamond, a C. Simon, b A. Al-Hendy. a a Department of Obstetrics
and Gynecology, Georgia Regents University, Augusta, GA; b Fundacion
Instituto Valenciano de Infertilidad/University of Valencia, INCLIVA,
Paterna (Valencia), Spain.
OBJECTIVE: Uterine fibroids are common gynecologic tumors stimulated
by estrogen (E2) and progesterone (P4), and likely originate from
abnormal myometrial stem cells. We have recently characterized human
myometrial stem cells using specific surface markers, and we demonstrated
lower levels of E2 receptor a and P4 receptors A/B in these cells
compared to differentiated myometrial cells. However, it has also been reported
that these myometrial stem cells are still influenced by E2 and P4
effects in vitro, via a paracrine pathway by the surrounding differentiated
myometrial cells. Our goal in this work is to evaluate the effect of both
E2 and P4 on the number and distribution of myometrial stem cells in
Long Evans rats.
DESIGN: Laboratory research studies using a murine model.
MATERIALS AND METHODS: Steroid hormone deprivation was
achieved by the ovariectomy of 16 female Long Evans rats at 4 months of
age, followed by a 4 weeks rest period, when part of the myometrial tissues
was collected for further analysis. For the hormone supplement experiments,
ovariectomized rats were implanted with hormone pellets subcutaneously for
4 weeks (60-d time- release, 1.5mg b-estradiol pellet; and/or 60-d timerelease,
200mg progesterone pellet). In addition, 4 hormonally intact rats
were used as controls. Stem cell isolation was performed by digesting endometrium-free
myometrial tissues collected from different locations (cervix
and horns) followed by flow cytometry (FACS) analysis. At that time, the percentage
(%) of Stro1/CD44 stem cells was measured in rats supplemented
with E2 or P4 alone or combined, versus control rats.
RESULTS: Hormonal deprivation (by ovarectomy) caused an 8-fold drop
in the number of myometrial stem cells in the cervical region and a 6-fold
drop in the horns, compared to hormonally intact control (p

REPRODUCTIVE ENDOCRINOLOGY: CLINICAL
O-187 Wednesday, October 21, 2015 11:15 AM
DOES A FROZEN EMBRYO TRANSFER AMELIORATE THE EF-
FECT OF ELEVATED PROGESTERONE ON THE ENDOMETRIUM
SEEN IN FRESH TRANSFER CYCLES: A PAIRED AND UNPAIRED
ANALYSIS. M. W. Healy, a G. Patounakis, a M. T. Connell, a K. Devine, b
A. DeCherney, a M. Levy, b M. J. Hill. a a NIH, Bethesda, MD; b Shady Grove
Fertility Reproductive Science Center, Rockville, MD.
OBJECTIVE: Many ART programs are cryopreserving embryos when
progesterone (P) is elevated in a fresh IVF cycle, but there is minimal evidence
of the effect of P on oocyte quality and thus embryo performance in
subsequent frozen embryo transfer (FET) cycles. Our objective was to
demonstrate the effect of P on the day of trigger in fresh IVF transfer cycles
compared to subsequent FET cycles.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Autologous IVF cycles with fresh transfers
and FET cycles from 2011-2013 were included if P was measured on the
day of trigger in the initial fresh cycle. The primary outcome was clinical
pregnancy. Analysis 1 was a paired analysis of patients who underwent
both a fresh transfer and subsequent FET. Analysis 2 included unpaired comparisons
of data from all fresh transfers and all FET transfers. GEE modeling
was performed to account for patients with multiple cycles and to control for
age, embryo quality, embryo stage, and number of transferred embryos.
Interaction testing was performed to determine if the effect of P was similar
in fresh transfer and FET cycles. P was treated as a continuous variable
except in additional analyses comparing P threshold of 2ng/mL. Subgroup
analysis was performed in good quality blastocyst transfers.
RESULTS: P was negatively associated with pregnancy in fresh transfers
but not FET in both analysis 1 and 2. Interaction testing of P and cycle type
indicated P had a different association with pregnancy outcomes in fresh
transfers versus frozen transfers. When P was R 2ng/mL, pregnancy was
more likely in FET versus fresh transfers in analysis 1 (56% vs 5%,
P

draw and 52% completed a post-test survey. The mean AMH was 4.3 3.5
ng/mL, range

DESIGN: A prospective, randomized controlled trial.
MATERIALS AND METHODS: A prospective, randomized controlled
trial was conducted to evaluate the outcomes in three different hormone
replacement protocols for thawed blastocyst transfer. A total of 330
women (median age 38.2 years) who were undergoing IVF at our clinic
were enrolled. The trial registration number was UMIN000016919. Participants
were separated into groups via computer-generated randomization
as follows. Premarin (Ò) group (n¼110; 2.49 mg/day; from the
second day of menstruation to the fifth day and 4.98 mg/day after the
sixth day); Estrogel (Ò) group (n¼110; 2.16 mg/day in Estrogen conversion
after the second day of menstruation); and Estrana tape (Ò) group
(n¼110; 0.72 mg, 1 piece per 2 days). Statistical analyses were investigated
using univariate regression. Statistical significance was defined
as p

O-195 Wednesday, October 21, 2015 11:45 AM
HIGH FREQUENCY OF INTERMEDIATE ALLELES OF FRAGILE
X MENTAL RETARDATION 1 GENE (FMR1) IN CANDIDATES
FOR OOCYTE DONATION. J. Guillen, R. Vassena, V. Vernaeve,
A. Rodriguez. Clinica EUGIN, Barcelona, Spain.
OBJECTIVE: The availability of preconceptional screening for X-
linked and recessive diseases offers the opportunity to lower the risk to
conceive an affected child. Genetic matching of donors and patients
further improves ART service; however, the donor selection process might
alter expected frequencies of certain diseases, potentially leading to variations
of pretest risk. Fragile X syndrome (FXS) is the most common
cause of inherited intellectual disability affecting approximately 1:4000-
1:6000 births. FXS is caused by the expansion of CGG repeats in the
FMR1 gene, and depending on CGG repeats number the population has
been classified into: normal (N) 5-44 repeats, intermediate allele (IA)
45-54, premutation (PM) 55-200, and full mutation (FM) >201. People
with PM or FM could develop clinical symptoms and transmit the disease
to the next generation. IA alleles do not confer an increased FXS risk for
the very next generation, however, the 50-54 range may show instability,
with potential expansions in subsequent generations. The aim of the study
is to assess carrier status and ethnic variation of the FMR1 gene in oocyte
donation (OD) candidates.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: 336 consecutive OD candidates aged 18
to 35 years were tested for FXS (Asuragen Amplidex FMR1) in January-
April 2015. Women with a family history of FXS, mental retardation, chromosomal
abnormalities, genetics disorders or neurological conditions were
excluded from further screening. All women diagnosed with 45 o more
CGG repeats were referred to genetic counseling. Relative frequency of IA
status by ethnicity was assessed by Chi2 test, while the association between
IA status and ovarian reserve was assessed using a logistic regression model,
adjusted by age.
RESULTS: Women ethnicities were: Mediterranean 127 (37.8%), European
103 (30.6%), Latino American 61 (18.2%), Caribbean 27 (8.0%), other
ethnicities 18 (5.4%). There were 17 carriers (1:20 overall), all of them IA; 13
carriers had 45-49 CGG, while 4 had 50-54 CGG. The frequency of IA was
7.4%, 7.8%, and 5.5% in women of Caribbean, European, and Mediterranean
ethnicity, respectively (p>.05). IA status was not associated with the woman
ovarian reserve (p>.05).
CONCLUSIONS: Screening of potential donors for family history of
FXS-like phenotypes lowers the frequency of PM and FM alleles in accepted
donors. The high frequency of IA alleles in Caribbean, Europeans, and Mediterranean
ethnicity is likely due to ethnic and geographical variability, a phenomenon
described in other subgroup population.
O-196 Wednesday, October 21, 2015 12:00 PM
DO YOU REALLY KNOW YOUR ETHNICITY? COMPARING
SELF-REPORTED AND GENETICALLY-PREDICTED ETHNICITY:
CLINICAL IMPLICATIONS. R. Shraga, a S. L. Bristow, a
A. Manoharan, a S. Yarnall, a A. Bisignano, a N. Kumar, a J. L. Frattarelli, b
S. Ghadir, c J. Grifo. d a Recombine, New York, NY; b Fertility Institute of Hawaii,
Honolulu, HI; c Southern California Reproductive Center, Beverly Hills,
CA; d NYU Langone Medical Center, New York, NY.
OBJECTIVE: Current guidelines published by professional societies
recommend genetic carrier screening be offered on the basis of ethnicity.
However, as the genetic pool homogenizes people are less aware of, or less
likely to identify with, a specific ethnicity. Our goal was to investigate the accuracy
of self-reported ethnicity as a basis for making clinical decisions.
DESIGN: Retrospective.
MATERIALS AND METHODS: Self-reported ethnicity was evaluated in
1442 patients who received expanded carrier screening. Reports were gathered
from patient requisition forms and during genetic counseling appointments.
Comparisons were made to ancestral origin as predicted by a
statistical model based on 672 SNPs validated using samples from the
1000 Genomes Project. Documented informed consent was obtained from
all patients.
RESULTS: We found several discrepancies when comparing self-reported
ethnicities on requisition forms, self-reported ethnicity during genetic counseling
consults, and genetic ancestry. For example, only 33.3% of individuals
who would be considered to be of Mediterranean ancestry based on genetic
counseling consults self-reported this background. Further, in 27.2% of
cases, patients predicted to be of South Asian descent by the statistical model
and confirmed during consults self-reported a different ancestry. Finally, individuals
who reported Latin American ancestry demonstrated a high degree
of genetic admixture; the 3 highest contributing ancestral origin groups were
European (0.5090.126), Native American (0.2340.185), and African
(0.1010.183).
CONCLUSIONS: Our comparison has demonstrated inconsistencies between
self-reported ethnicities on requisition forms, self-reported ethnicity
during genetic counseling consults, and genetic ancestry, highlighting the unreliability
of patient self-reports. Basing carrier screening on patient-reported
ethnicity may result in failure to screen for all appropriate genetic conditions
and thus failure to identify individuals at high reproductive risk. This emphasizes
the importance of offering pan-ethnic expanded carrier screening to all
patients. Moreover, as identification of patient ethnicity is critical to the accurate
calculation of residual reproductive risk, it may be worthwhile to factor
both self-reported and genetic ancestry in these calculations. Finally, in
the clinical setting, considerations should be made in how ethnic groups
are defined for patient reporting purposes.
O-197 Wednesday, October 21, 2015 12:15 PM
REPRODUCTIVE DECISION-MAKING IN PATIENTS DIAGNOSED
WITH BRCA MUTATIONS. J. Chan, a L. N. Johnson, a L. DiGiovanni, b
C. Voong, a M. D. Sammel, c S. M. Domchek, b C. Gracia. d a Reproductive
Endocrinology and Infertility, Hospital of the University of Pennsylvania,
Philadelphia, PA; b Hematology Oncology, Hospital of the University of
Pennsylvania, Philadelphia, PA; c Univ. of Pennsylvania, Perelman School
of Medicine, Rose valley, PA; d University of Pennsylvania.
OBJECTIVE: Despite our understanding of the medical implications of a
BRCA mutation, there is little data on how knowledge of carrier status influences
decisions about reproduction and parenthood. We aim to investigate
how knowledge of BRCA carrier status impacts women’s decisions about relationships,
childbearing and the use of preimplantation genetic diagnosis
(PGD) and prenatal diagnosis.
DESIGN: Cross-sectional survey.
MATERIALS AND METHODS: BRCA mutation carriers were identified
through the University of Pennsylvania’s Cancer Risk Program and through a
national advocacy group for hereditary ovarian and breast cancers. Demographics,
medical and social history, pregnancy and fertility history, and
age at BRCA testing were obtained from a survey. Participants answered
questions regarding how BRCA status influenced decisions about marriage
and relationships, childbearing and fertility treatment, as well as attitude towards
technologies such as PGD and prenatal diagnosis. Demographic characteristics
were summarized and questionnaire responses compared using
ranksum and Chi-square for continuous and categorical variables as appropriate.
RESULTS: In the 1081 women who completed the survey, the mean age at
the time of BRCA diagnosis was 44.1. 387 (36%) had a history of cancer. The
majority was partnered at the time of learning of their BRCA mutation
(84%). Amongst those who were not partnered, 37% felt pressure to find a
partner after learning of their carrier status, 24% had more desire to get married
and 46% reported that carrier status influenced what characteristics they
were looking for in a partner. 66% had biological children and 64% reported
that their families were complete at the time of diagnosis. Amongst women
whose families were not complete, 38% reported that knowledge of their
BRCA status impacted their decision to have biological children. Younger
women (

O-198 Wednesday, October 21, 2015 12:30 PM
PREIMPLANTATION GENETIC DIAGNOSIS (PGD) FOR SINGLE
GENE DEFECTS (SGD): THE SIGNIFICANT VALUE OF CONCUR-
RENT ANEUPLOIDY SCREENING. T. G. Nazem, a K. N. Goldman, a
A. S. Berkeley, a J. Grifo. b a NYU School of Medicine, New York, NY;
b NYU Langone Medical Center, New York, NY.
OBJECTIVE: To evaluate outcome differences between patients undergoing
trophectoderm (TE) biopsy and PGD for SGD with and without 24-chromosome
aneuploidy screening.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients undergoing their first cycle of
blastocyst culture, TE biopsy, and PGD with and without aneuploidy screening
(array comparative genomic hybridization, aCGH) from July 2010 to August
2014 were included. Cycles were excluded if performed for gender selection,
HLA matching, or translocation. Primary outcomes included number (no.) and
percentage (%) of blastocysts affected/unaffected/carrier of SGD, aneuploid/
euploid blastocysts, and blastocysts eligible for transfer. FET cycle outcomes
were analyzed: implantation rate (IR), spontaneous abortion rate (SABR),
and live birth rate (LBR). Data were analyzed using student’s t-test and Fisher’s
exact t-test and presented as mean S.D. (p

O-201 Wednesday, October 21, 2015 11:45 AM
MALE OBESITY IS AN INDEPENDENT RISK FACTOR FOR
TREATMENT FAILURE AMONG COUPLES WITH UNEX-
PLAINED INFERTILITY IN THE ASSESSMENT OF MULTIPLE IN-
TRAUTERINE GESTATIONS FROM OVARIAN STIMULATION
(AMIGOS) TRIAL. A. J. Polotsky, a A. A. Allshouse, a S. Krawetz, b
N. Santoro, a E. Eisenberg, c H. Zhang, d M. P. Diamond. e a University of Colorado,
Aurora, CO; b Wayne, Detroit, MI; c NICHD, Bethesda, MD; d Yale,
New Haven, CT; e Georgia Regents University, Augusta, GA.
OBJECTIVE: Couples with unexplained infertility often seek advice on
lifestyle changes but evidence-based medicine is limited. We sought to determine
the impact of male body mass on the chance of pregnancy and live birth
after fertility treatment.
DESIGN: Secondary analysis of a randomized clinical trial (1).
MATERIALS AND METHODS: 900 couples with unexplained infertility
were randomized to ovulation induction with gonadotropins, or letrozole, or
clomiphene citrate, followed by intrauterine insemination (IUI) for up to 4
cycles. Females were 18-40 years old, had regular manses, at least 1 patent
fallopian tube, normal uterine cavity and a male partner with at least 5 million
total motile sperm (TMS) count. Analysis was limited to subjects for whom
complete male partner information was available (n¼836). Logistic regression
was conducted with clinical pregnancy and live birth as the outcomes.
Each model was adjusted for male and female body mass index (BMI), treatment
arm, and TMS.
RESULTS: Two hundred and forty-five couples achieved clinical pregnancy
(29.3%) and 214 women had a live birth (25.6%) in our analytic sample.
Primary trial results have been presented (2). 272 male partners were
obese with BMI greater than 30 k/m2 (32.5%) and 134 couples were
concordant for male and female obesity (16.2%). Overall, there was a significant
but very weak inverse correlation between male BMI and TMS
(rho¼-0.08, p¼0.02) and a significant and moderate positive correlation between
male and female BMI (rho¼0.41, p 30), n¼272 0.58 (0.37-0.91) 0.60 (0.38-0.96)
CONCLUSIONS: In the AMIGOS trial, male obesity decreased the likelihood
of live birth after fertility treatment by 40% as compared to couples
with normal male weight. This effect was seen independent of female obesity,
treatment arm and sperm parameters. Patients with unexplained infertility
should be informed that obesity in the male partner reduces success of
fertility treatment.
References:
1. Diamond et al. Fertil Steril 2015;103:962-973.
2. Diamond et al Fertil Steril 2014;102, p. e39.
Supported by: This work was Supported by National Institutes of Health
NIH)/Eunice Kennedy Shriver National Institute of Child Health and Human
Development (NICHD) Grants U10 HD39005 (to M.P.D.), U10 HD38992 (to
R.S.L.), U10 HD27049 (to C.C.), U10 HD38998 (to R.A.), HD055944 (to
P.R.C.), U10 HD055936 (to G.M.C.),U10HD055925 (to H.Z.); and U10
U54-HD29834 (to the University of Virginia Center for Research in Reproduction
Ligand Assay and Analysis Core of the Specialized Cooperative
Centers Program in Reproduction and Infertility Research). Most importantly,
this research was made possible by the funding by American Recovery
and Reinvestment Act. The content is solely the responsibility of the authors
and does not necessarily represent the official views of the NICHD or NIH.
O-202 Wednesday, October 21, 2015 12:00 PM
PREVALENCE OF TESTICULAR MICROLITHIASIS IN 382 NON-
VASECTOMIZED, AZOOSPERMIC MEN. J. Fedder. Centre of Andrology
& Fertility Clinic, Odense University Hospital, Odense, Denmark.
OBJECTIVE: To determine the prevalence of testicular microlithiasis
(TM) in unselected azoospermic men and to relate the findings to specific
etiological subgroups and to risk of malignancy.
DESIGN: A retrospective study including 382 nonvasectomized, azoospermic
men consecutively referred to our fertility clinic.
MATERIALS AND METHODS: All ultrasonographic examinations were
performed by JF and the prevalence of TM summarized in total and for specific
etiological subgroups of azoospermia. TM was classified as extensive
TM, universally distributed (uTM) or collected in plaques (pTM), and finally
borderline TM (bTM) with about 5 (3 to 7) TM elements in one or both testicles.
Representative testicular biopsies were taken in 300 men using a
TruCut needle, Ch.14 (Angiotech, USA). Frequencies of Carcinoma In
Situ (CIS) testis/malignancy in men with different categories of TM were
compared to men without TM using Fisher’s exact test.
RESULTS: UTM was found in 11 men (2.9%). In four of these (36%), the
pattern was found bilaterally. In 3 cases (0.8%) including 2 with Klinefelter’s
syndrome (KS), bTM was detected in the contralateral testis. PTM was detected
in 8 cases (2.1%), and except for one KS man, the condition was in
all cases unilaterally. BTM was detected in 32 cases (8.4%), bilaterally in
17 (53%). Unilateral bTM was found in further 3 cases with uTM in the other
testis, and these cases were categorized as having uTM. A frequent overlapping
between the most relevant etiological groups was found, e.g. 10 (23%)
of the 44 men with KS and 4 (20%) of the 20 men with Y microdeletions had
a history of cryptorchidism. Of the men with KS, 5 had extensive TM, 3 (7%)
uTM and 2 (5%) pTM, while 14 (32%) showed bTM. Of 101 men with a history
of cryptorchidism, 3 (3%) had uTM, 4 (4%) pTM, and 13 (13%) bTM.
Compared to a frequency of CIS testis/testicular malignancy of 1 of 266
found in men without TM, the frequencies of CIS testis/testicular malignancy
in men with uTM was 1 of 6 (p¼ 0.04), in men with pTM 1 of 7 (p¼0.05), and
in men with bTM 1 of 21 (p¼0.14).
CONCLUSIONS: A total prevalence of TM, extensive and borderline, was
found in 13.4% of 382 unselected, nonvasectomized, azoospermic men. The
relatively high frequency of bilateral uTM and bTM supports a hypothesis of
a general defective spermatogenesis as background of TM. The risk of testicular
malignancy was increased in men with extensive TM.
O-203 Wednesday, October 21, 2015 12:15 PM
A CASE FOR BANKING SPERM DUE TO INCREASED AGE:
DECLINE IN SPERM COUNT AND MOTILITY IN YOUNG ADULT
MEN. G. M. Centola, a A. Blanchard, b J. L. Demick, c S. Li, d
M. L. Eisenberg. d a New England Cryogenic Center, Newton, MA; b New England
Cryogenic Center, Brookline, MA; c Laboratory, New England Cryogenic
Center, Brookline, MA; d Urology, Stanford University School of
Medicine, Stanford, CA.
OBJECTIVE: Controversy exists as to the stability of semen quality over
the past half century with numerous papers reporting a decrease, increase or
stable parameters in heterogeneous populations. Varied methodology and geography
of studies make definitive conclusions challenging. The purpose of
the current study was to examine semen parameters of young adult men who
provided multiple semen specimens over a 10 year period at the same facility.
DESIGN: Retrospective analysis of semen data for the years 2003 - 2013.
MATERIALS AND METHODS: Semen parameters, including volume,
count, and motility prior to and after cryopreservation were analyzed for a
total of 9425 semen specimens from 489 individuals. Demographic information
was obtained from an extensive social and medical history questionnaire
for all participants. Following 2-3 days of abstinence, the specimens were
collected by masturbation at the lab facility, and were assessed by the
same two laboratory technicians, using a standard Makler semen analysis.
Specimens were frozen with a glycerol-based cryopreservative in liquid nitrogen.
A vial was thawed after 48 hours and count and motility determined
post-thaw. The data was analyzed using generalized linear regression after
adjustment for age, days of abstinence and for repeated semen samples, as
well as by the Cochran-Armitage trend test. All p values were two sided
with p < 0.05 considered statistically significant.
e78 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

RESULTS: There was a significant decline in initial sperm count (p <
0.001), sperm motility (p < 0.001), total count (p < 0.001) and total motile
count (p < 0.001) during the period 2003-2013. There was no significant
change in semen volume (p ¼ 0.2). Following cryopreservation, the mean
post-thaw motility significantly (p < 0.001) decreased with time. There
was a significant decline in age (p trend ¼ 0.003) and alcohol use (p trend
¼ 0.005), as well as an increase in college GPA (p trend ¼ 0.02). However,
BMI (p trend ¼ 0.73) educational attainment (p trend ¼ 0.2), race/ethnicity
(p trend ¼ 0.53), and lifestyle habits (weekly exercise, p trend ¼ 0.21;
smokers p trend ¼ 0.99; marital status, p trend ¼ 0.85) remained constant
over the period of study.
CONCLUSIONS: There has been a decline in semen quality among young
men presenting for sperm donation during the past 11 years. Uniform technicians
and techniques over the study period make measurement bias unlikely.
Subject characteristics also remained constant during the study
period. The decrease in sperm count and motility following cryopreservation
suggests that the sperm were not as vital since cryosurvival appears to be
compromised over the ten-year poor of time. This report demonstrates a
decline in sperm count in a young adult male population. Although absolute
values did not decline into the abnormal or sub fertile range, it may be prudent
for men to consider sperm cryopreservation if fertility is being delayed
until later years/advanced age.
O-204 Wednesday, October 21, 2015 12:30 PM
MALE PARTNER SEXUAL ABSTINENCE LESS THAN 2 OR
GREATER THAN 7 DAYS IS NOTASSOCIATED WITH INCREASED
ANEUPLOIDY RATE. J. Gingold, a,b M. C. Whitehouse, c M. Gerson, d
S. Parsons, d J. A. Lee, d A. B. Copperman, d,a N. Bar-Chama. d,e a Obstetrics,
Gynecology and Reproductive Science, Icahn School of Medicine at Mount
Sinai, New York, NY; b Obstetrics/Gynecology and Women’s Health Institute,
Cleveland Clinic Foundation, Cleveland, OH; c Reproductive Medicine
Associates of New York, New York City, NY; d Reproductive Medicine Associates
of New York, New York, NY; e Urology, Icahn School of Medicine at
Mount Sinai, New York, NY.
OBJECTIVE: Males are routinely counseled to maintain 2-7 days sexual
abstinence prior to processing and use during an in vitro fertilization (IVF)
cycle(s) in order to enhance semen quality. Although obtaining an adequate
concentration of motile sperm is an important objective, an adverse deviation
rarely limits a couple from moving forward in their cycle. The ultimate goal
is to maximize the number of euploid embryos prior to transfer selection.
This study evaluates if duration of male sexual abstinence is associated
with an increase in embryonic aneuploidy.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Couples who underwent an IVF cycle
and utilized pre-implantation genetic screening (PGS) from June 2010 -
March 2015 were included. Oocyte age was recorded (A: %35; B: (35-
38]; C: (38-41]; D: (41-43]; and E: >43). Duration of male sexual abstinence
was recorded. Aneuploidy rate for each group was computed, with 95% confidence
intervals calculated by Clopper-Pearson method. Aneuploidy rate
was modeled by logistic regression using oocyte age and days of abstinence.
The youngest oocyte age group (

Supported by: European Union Seventh Framework Programme DEER
grant 212844, and NIH grants P30 DK046200 and T32 DK007703-16.
O-206 Wednesday, October 21, 2015 11:30 AM
WITHDRAWN
OBJECTIVE: To develop an anatomically accurate fluid dynamics model
in order to assess the anatomic causes of elevated testicular vein pressures.
DESIGN: Finite element analysis was used to model the fluid dynamics of
human testicular venous drainage.
MATERIALS AND METHODS: A two dimensional model was created to
represent the inferior vena cava, gonadal and renal veins. The geometry was
constructed to correspond to a representative cross-section of a male patient’s
CT scan that was obtained for non-urologic causes. The geometry was subsequently
meshed into contiguous triangular elements. Finite element analysis
was employed to solve for the stationary Navier-Stokes equations
using Comsol Multiphysics 3.3 (Comsol, Inc., Burlington, MA). Boundary
conditions, such as renal vein and inferior vena cava flow rates and pressures,
were set to known physiologic values reported in the literature. The dynamic
viscosity of blood was set to 0.01 Pa-sec. Gravity was simulated to correspond
to an upright posture of the patient. The final solution was subsampled
to provide for flow rates and pressures along both testicular veins.
RESULTS: The model accurately predicted physiologic flow rates within
both the renal and testicular veins. Without left renal vein impingement by
the superior mesenteric artery (SMA), the left and right venous pressures
at the inguinal canals were predicted to be 41 and 40 cmH2O respectively
- approximately half the pressure that has previously been reported. Turbulence
due to the perpendicular insertion of the left gonadal vein into the
left renal vein contributed minimally to elevated left sided pressures. Simulated
SMA impingement was modeled with up to 75% occlusion of the left
renal vein. Left gonadal vein pressures rose to only 47 cmH2O despite the
significant change in renal vein diameter.
CONCLUSIONS: Gonadal vein pressures can be accurately modeled using
finite element analysis. The perpendicular insertion of the left gonadal vein is a
negligible source of flow resistance to testicular drainage. Marked compression
(beyond a 75% decrease in diameter) of the left renal vein may be required to
manifest a clinically meaningful pressure elevation (e.g., Nutcracker syndrome)
in the distal left testicular vein. The principle contributor to elevated
left testicular vein pressures, and possibly to the greater prevalence of left sided
varicoceles, is the added flow resistance of the longer left gonadal vessel.
O-208 Wednesday, October 21, 2015 12:00 PM
MELANOMA ANTIGEN PROTEIN MAGEC1 MUTATION
IDENTIFIED IN FAMILIAL NON-OBSTRUCTIVE
AZOOSPERMIA. A. W. Pastuszak, a c. cengiz, b M. Bekheirnia, c
D. J. Lamb. d a Scott Department of Urology, Baylor College of Medicine,
Houston, TX; b Technician, Houston, TX; c Department of Genetics, Baylor
College of Medicine, Houston, TX; d Baylor College of Medicine, Houston,
TX.
O-207 Wednesday, October 21, 2015 11:45 AM
HYDRODYNAMICS OF THE HUMAN GONADAL VEIN: A FINITE
ELEMENT MODELTO EXPLORE ELEVATED LEFT TESTICULAR
VENOUS PRESSURES RELATED TO VARICOCELE. R. P. Hayden,
C. Tanrikut. Massachusetts General Hospital, Boston, MA.
OBJECTIVE: Many currently unrecognized genetic etiologies of male
infertility likely exist, and we sought to identify genes influencing male infertility
using next-generation sequencing approaches. The melanoma antigen
(MAGE) family of genes may function in germ cell maintenance and fertility,
and herein we present genomic data associating the melanoma antigen gene
MAGEC1 with male fertility.
DESIGN: Genomic study comparing infertile and fertile men.
MATERIALS AND METHODS: Whole-exome sequencing of genomic
DNA from 2 Saudi Arabian brothers with non-obstructive azoospermia
(NOA) and Sertoli cell only (SCO) histology was performed. Sequence data
were filtered against several genomic databases and deleterious SNPs common
to both brothers identified. Sanger sequencing confirmed SNPs of interest and
assessed their presence in other NOA men. Array comparative genomic hybridization
(aCGH) analysis of 22 NOA men with normal karyotypes and Y-
chromosome microdeletion assays assessed for gene dosage changes due to
copy number variants (CNVs), and the presence of additional CNVs in genes
of interest was evaluated using qPCR. Immunohistochemical staining of testis
sections used commercially available antibodies and standard protocols.
RESULTS: An A>T SNP at position 3530 in exon 4 of the MAGEC1 gene
resulting in a potentially deleterious R1082W mutation was identified in a set
of 6 SNPs mapping to 6 unique genes selected from over 300,000 SNPs shared
between the two brothers’ genomes. Sanger sequencing confirmed the SNP in
the two brothers but did not identify it in 171 additional NOA and 34 fertile
control men. aCGH of 22 NOA men did not identify CNVs in MAGEC1,
although qPCR based analysis of 57 additional NOA men identified microduplications
in 2/57 (3.5%). In contrast, the frequency of MAGEC1 CNVs in the
general population (including fertile and infertile individuals) is approximately
0.2%. Immunohistochemical localization of MAGEC1 in human testis
from a male with normal spermatogenesis revealed cytoplasmic staining of
spermatocytes, which was less prominent in testis from a male with hypospermatogenesis,
and was absent in the testis from an SCO male.
CONCLUSIONS: A potentially damaging SNP and microduplications in
MAGEC1 were identified in a cohort of infertile NOA men. Future work
elucidating the molecular role of MAGEC1 in male fertility using cell-based
assays and animal models will further our understanding of the MAGE genes
in male germ cell maintenance and fertility.
Supported by: AWP is a National Institutes of Health (NIH) K12 Scholar
Supported by a Male Reproductive Health Research Career (MRHR) Development
Physician-Scientist Award (HD073917-01) from the Eunice Kennedy
Shriver National Institute of Child Health and Human Development
(NICHD) Program (to DJL). MRB is a National Institutes of Health (NIH)
K12 Scholar Supported by a Multidisciplinary K12 Urologic Research
(KURe) Career Development Physician-Scientist Award (K12
DK0083014) (to DJL).
e80 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

O-209 Wednesday, October 21, 2015 12:15 PM
DIETARY COENZYME Q10 INTAKE AND SEMEN PARAMETERS
IN A SUBFERTILE POPULATION. B. C. Tiseo, a A. J. Gaskins, b
J. E. Chavarro, b R. Hauser, b C. Tanrikut. c a Division of Urology, Hospital
das Clinicas, University of Sao Paulo Medical School, S~ao Paulo, Brazil;
b Harvard T.H. Chan School of Public Health, Boston, MA; c Massachusetts
General Hospital, Boston, MA.
OBJECTIVE: To investigate whether dietary coenzyme Q10 (CoQ10)
intake is associated with semen parameters in a cohort of men from a fertility
clinic.
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: Dietary intake data were obtained via
validated food frequency questionnaires from 155 male partners in subfertile
couples. CoQ10 intake was derived by summing specific contributions across
all food items. A total of 338 semen samples were evaluated. Multivariable
linear mixed models were used to examine the relation between quartiles
of CoQ10 intake and semen quality parameters while adjusting for age,
race, smoking, body mass index, abstinence time and male factor diagnosis
as well as for caloric intake, caffeine, alcohol and total fat intakes and accounting
for within-person correlations.
RESULTS: Mean dietary CoQ10 intake was 7.1mg/day (range¼1.5 to
20.2mg/day). No subjects were taking supplemental CoQ10. The adjusted
mean sperm motility percentage was calculated as 43.1% for the lowest quartile
of CoQ10 intake and 39.3% for the highest quartile(p-trend: 0.489). Evaluating
specifically progressive sperm motility the adjusted means were
24.2%, 25.7%, 25.5% and 21.3% for increasing quartiles of CoQ10 intake
(p-trend: 0.365). There were no associations between dietary CoQ10 intake
and sperm concentration, total motility or morphology (See Table 1).
CONCLUSIONS: Our findings indicate that dietary CoQ10 intake is not
related to semen quality parameters. The mean dietary intake of CoQ10
was 25-fold lower than the supplemental 200-300 mg daily dose used in
controlled trials which have demonstrated improved sperm motility. The
relative low dietary intake of CoQ10 in our cohort may explain the discrepancy
between our results and the findings of the supplemental trials. Dietary
CoQ10 intake alone may not be sufficient to optimize semen parameters.
Seminal parameters marginal means by CoQ10 intake groups.
Quartile 1
(Lowest)
N ¼ 38
Quartile 2
N ¼ 39
Quartile 3
N ¼ 39
Quartile 4
(Highest)
N ¼ 39
CoQ10 intake (mg/day) 1.45-4.63 4.80-6.36 6.56-8.83 8.99-20.15
Semen Samples (N) 83 85 86 84
Mean Mean Mean Mean p-trend
Abstinence time (hrs) 149.6 133.4 133.8 126.9 0.989
Sperm concentration (x10⁶/mL) 28.2 36.8 34.4 31.2 0.518
Sperm motility (%) 43.1 46.0 45.8 39.3 0.365
Progressive sperm Motility (%) 24.2 25.7 25.5 21.3 0.489
Normal sperm morphology (%) 6.4 6.7 7.0 5.5 0.129
Note: N: number of subjects
Results shown as marginal means of the population adjusted for age, race, smoking,
body mass index, abstinence time, male factor diagnosis, calorie intake,
caffeine intake, alcohol intake and total fat intake using linear mixed models.
Supported by: National Institutes of Health grants ES009718, ES022955,
ES000002, P30 DK046200 and T32 DK007703-16.
O-210 Wednesday, October 21, 2015 12:30 PM
SPERM QUALITY AND FERTILITY ARE COMPROMISED IN
BRCA1 MUTANT MALE MICE. R. Stobezki, a S. Titus, a B. Musul, a
F. Moy, a K. H. Oktay. a,b a Obstetrics & Gynecology, New York Medical College,
Valhalla, NY; b Innovation Institute for Fertility Preservation and IVF,
New York, NY.
OBJECTIVE: BRCA1 and 2 are DNA double strand break (DSB) repair
genes and their mutations result in cancer predisposition. Presence of
BRCA mutations have been reported to affect ovarian reserve. We have previously
shown that BRCA1 gene mutations may also affect sperm DNA
integrity. In this study, we sought to determine if BRCA1 mutations affected
fertility and embryo quality in a transgenic mouse model.
DESIGN: Experimental study.
MATERIALS AND METHODS: We inbred a transgenic BRCA1-mutant
mouse colony, which carries a deletion of 330 base pairs (bp) in intron 10 plus
407 bp in exon 11 of the BRCA1 gene. This deletion causes the gene product
to be retained in the cytoplasm, which severely compromises its nuclear functions.
BRCA1 mutant mice were studied at three months of age. Sperm were isolated
from the vas deferens to observe sperm concentration. Litter size data were
determined from mating BRCA1-mutant and wild type (WT) males with WT female.
The same crosses were performed to determine embryos reaching the blastocyst
stage. Prior to the setup of these breeding crosses, WT females received
PMSG (pregnant mare’s serum gonadotropin, 5 IU) followed by HCG (human
chorionic gonadotropin, 5 IU) 72 hrs later. The morning after the setup, vaginal
plugs were checked and those females who have plugs were separated. On day 5,
embryos were flushed from the uterus and observed for morphology.
RESULTS: BRCA1-mutant male mice had lower sperm counts than the
WT (1.8x10 6 0.1x10 6 vs. 2.1 x10 6 0.07, n¼8; p¼0.045). Upon mating
with WT females, BRCA1-mutant males resulted in a smaller litter size
than when the WT males which were mated with the same (3.22.3 vs.
6.91.3, n¼9; p¼0.0008). Embryos produced by crossing BRCA1 mutant
male mice with WT females had a higher percentage of arrested embryos
(32.14%0.128 vs. 8.33%0.066, n¼7; p¼0.026) as compared to the WT.
CONCLUSIONS: These results indicate that BRCA1 mutations may be associated
with decline in sperm quantity and quality in mice, which results in compromised
embryo development and implantation. Our findings also underscore the
importance of DNA DSB repair in maintenance of sperm quality. Further molecular
studies are needed to understand the mechanism of arrest in embryos resulting
from BRCA1-mutant male mice. Translational studies are warranted in men with
BRCA mutations to determine the clinical significance of these findings.
Supported by: NIH R01HD053112.
HEALTH DISPARITIES 2
O-211 Wednesday, October 21, 2015 11:15 AM
THE ONTOGENY OF MYOMETRIAL STEM CELLS IN OCT4-GFP
TRANSGENIC MICE. S. Brakta, A. Mas, M. P. Diamond,
S. M. Shalaby, A. Al-Hendy. GRU, Augusta, GA.
OBJECTIVE: Somatic stem cells (SSC) are master stem cells that through
asymmetric division retain their ability to self-renew while producing
daughter cells. Similarly, tumor initiating cells are a subset of cells within
a tumor cell population, which retain the ability to sustain tumors through
asymmetric division. It is thus believed that tumors such as uterine fibroids
originate from a single transformed SSC of the myometrium followed by
expansion and propagation in a steroid-dependent manner. Our group has
recently identified two specific cell surface markers (CD44 & Stro1) as human/rat
myometrial stem cell markers. In this work, we aim to identify myometrial
stem cells from mice at different age points by using stem cell
markers Oct4 and CD44.
DESIGN: Laboratory studies using transgenic mice.
MATERIALS AND METHODS: Twenty four female mice, B6, CBA-Tg
(Pou-5fl-EGFP) 2Mnn/j mouse strains, 1 week to 24 weeks old were purchased
from Jackson laboratory. These mice were homozygous for the
Pou5fl/OCT4 transgenic insert and expressed enhanced green fluorescent
protein (EGFP) in the uterus under the control of POU domain, class 5, transcription
factor 1 (Oct4) promoter and distal enhancer. Uterine blocks were
shipped at ages 1, 3, 4, 8, 12 and 24 weeks. Immunohistochemistry (IHC) and
immunofluorescence studies were performed in order to co-localize the specific
cell surface marker CD44 with the well-known stem cell marker Oct4.
Image J was used to quantify the stem cells as a percentage of total myometrial
cells. Three random high power fields were selected from each uterine
age and the average of stem cell percentage was determined. To correlate
these numbers with possible response to the effects of ovarian sex steroid hormones,
we also evaluated the uterine expression of estrogen receptor a and
progesterone receptors A&B at ages 1, 3 and 4 using inverted microscopy.
RESULTS: The frequency of myometrial stem cells was significantly
lower (2.0%1.2) at 1 week of age compared to sexually more mature
mice (P < 0.05). The frequency of myometrial stem cells at ages 3, 4, 8,
12, and 24 weeks was 13.0%5.4, 11.3%1.5, 10.4%4.5, 6.6%0.6,
and 10.4%3.7 respectively. Interestingly, abundant positive staining for estrogen
receptor a and progesterone receptors A&B were detected in the myometrium
ages 1, 3, and 4 weeks.
FERTILITY & STERILITY Ò
e81

CONCLUSIONS: Myometrial stem cells are present in the myometrium at a
significantly lower percentage at the pre-sexual age of 1 week than at the sexually
mature ages of 3 to 24 weeks. This is likely due to lack of the estrogen hormone
ligand at that early age rather than the availability of the steroid receptors
which are similarly expressed in neonatal and adult myometrium. These results
suggest that stem cells are steroid dependent and increase in number with reproductive
maturity at 3-4 weeks of age in mice. Our findings emphasize the
vulnerability of neonatal myometrium to environmental xeno-estrogen exposure
which can potentially lead to permanent reprogramming of myometrial
stem cells leading to adult onset of diseases such as uterine fibroids.
O-212 Wednesday, October 21, 2015 11:30 AM
FREE, BIOAVAILABLE, AND TOTAL 25 HYDROXY VITAMIN D IN
VIABLE PREGNANCIES: COMPARISONS ACROSS RACE AND
OVER TIME. S. Senapati, a T. Adedji-Fajobi, b M. D. Sammel, c
C. Coutifaris, a M. S. Bartolomei, a S. Butts. c a University of Pennsylvania,
Philadelphia, PA; b Obstetrics and Gynecology, Perelman School of Medicine,
Philadelphia, PA; c Perelman School of Medicine, University of Pennsylvania,
Philadelphia, PA.
OBJECTIVE: Mounting evidence supports an association between
vitamin D deficiency and reproductive outcomes, however, the optimal
method for characterizing maternal vitamin D status, especially in African
Americans, is controversial. Our objective was to characterize differences
in total, free, and bioavailable vitamin D at peri-conception (PC) and midgestation
(MG) in women with viable pregnancies by race.
DESIGN: Prospective cohort of women attempting to conceive naturally
or following IVF treatment.
MATERIALS AND METHODS: Total 25 (OH)D was measured by liquid
chromatography-tandem mass spectrometry at two timeponts [1] within one
month of conception (PC) and [2] at 18-22 weeks gestational age (MG). Free
(non-protein bound) and bioavailable (free plus albumin-bound fraction)
25(OH)D was calculated with total 25(OH)D, vitamin D binding protein
(VDBP), albumin and relevant binding constants by established methods.
Subjects were classified by Vitamin D status (VitD) as sufficient (¼> 75
nmol/L), insufficient (¼50nmol/L) or deficient (< 50 nmol/L). Kruskal
wallis, Wilcoxon Rank Sum, c2, and Fisher exact testing was used to evaluate
differences in demographic characteristics by VitD status, and the association
between race and change in markers of vitamin D during pregnancy.
RESULTS: Of the 110 women studied, 74.5% were White, 15.5% were African
American, 8.2% were Asian, and 1.8% were other/mixed race. 33.6%
were VitD sufficient, 45.5% were VitD insufficient and 21% were VitD deficient
peri-conception. PC VitD deficiency was associated with non-White
race (p¼0.01) and median PC total 25(OH)D was lower in African American
(55.4 nmol/L) than White subjects (70.6 nmol/L) (p

95% CI 1.05-15.2, p

(evaluated by HSCORE) and real-time RT-PCR (evaluated for relative
expression using the delta-delta Ct method). An optimal HSCORE threshold
of 1.4 was previously established by ROC analysis. Differences in proportions
of positive staining were analyzed by Chi-square, while RNA data
were evaluated by ANOVA and T-test.
RESULTS: Age and BMI were similar between the three groups.
HSCORE was significantly different (p < 0.0001) (Table 1). Endometrial
BCL6 immunostaining was positive (above the 1.4 HSCORE threshold) in
2/28 (7%) fertile control subjects as compared to 103/116 (88.7%) UI subjects
and 32/43 (74.4%) uRPL subjects. Mean and median expression of
BCL6 RNA was lowest in the fertile controls, intermediate in the uRPL
and highest in UI, with ANOVA analysis approaching significance
(p¼0.07). In simple pairwise comparison, mRNA levels in UI were 2-fold
of those in controls (p¼0.04). Subsequent surgical exploration of a subset
of subjects demonstrated endometriosis or hydrosalpinges in 98% and 89%
in those with positive BCL6 immmunostaining and UI or uRPL, respectively.
CONCLUSIONS: Endometrial immunostaining for BCL6 closely correlated
with the fertility status of those tested, with fertile women demonstrating
low levels of nuclear BCL6 and the majority of women with UI
and uRPL expressing high levels. Endometrial BCL6 mRNA expression
showed similar trends, but differences between groups were smaller. Additionally,
high BCL6 immunostaining strongly predicted the presence of
endometriosis or, occasionally, hydrosalpinges. Early pregnancy loss and
Infertility appear to share a common pathophysiological feature, but may
differ by severity of the defect.
Demographics
Characteristic
Normal Fertile
Controls
(n¼ 28)
Unexplained
Infertility
(n ¼ 116)
Unexplained
Recurrent
Pregnancy
Loss (n ¼ 43)
AGE
32.78 2.6 33.04 4.2 34.65 4.7
(mean STD)
BMI 25.6 4.7 25.23 5.5 25.23 6.6
HSCORE 0.43 0.69 2.8 1.1 2.4 1.4
Supported by: NIH R01 HD067721 (S.L.Y. and B.A.L.).
O-218 Wednesday, October 21, 2015 11:30 AM
SIZE MATTERS: SINGLE NUCLEOTIDE POLYMORPHISM (SNP)
BASED CHROMOSOME ANALYSIS OF PRODUCTS OF CONCEP-
TION (POC) SAMPLES IDENTIFIES CLINICALLY SIGNIFICANT
DELETIONS/DUPLICATIONS BELOW KARYOTYPE
RESOLUTION. M. K. Maisenbacher, a K. Merrion, b S. Sigurjonsson, a
K. G. Paik, b M. J. Young, a B. Pettersen. c a Natera, San Carlos, CA; b Natera,
Inc., San Carlos, CA; c Genetic Counseling, Bend, OR.
OBJECTIVE: Report on products of conception (POC) samples with deletions
and duplications below the resolution of traditional karyotyping (<
10 Mb). These segmental abnormalities are clinically relevant based on
size, location and/or correlation with a known genetic syndrome.
DESIGN: Retrospective analysis.
MATERIALS AND METHODS: Review of 17,424 consecutive fresh
POC samples sent to a reference lab along with maternal blood samples. Genotyping
was performed using Illumina CytoSNP-12b microarray and bioinformatics.
RESULTS: 2600 cases (14.9%) had maternal cell contamination, 17 cases
(0.01%) had incomplete results, and 14,807 cases (85%) had fetal results. Of
these, 100 cases (0.7%) had deletions/duplications less than 10 Mb without
aneuploidy or uniparental disomy (UPD) of other chromosomes identified:
22 cases (22%) had a single isolated deletion or duplication (11 deletions
and 11 duplications), 43 cases (43%) had an additional deleted or duplicated
segment that was either >10 Mb or

O-219 Wednesday, October 21, 2015 11:45 AM
LIFESTYLE AND PREGNANCY LOSS AMONG WOMEN USING
HOME PREGNANCY TEST KITS, LIFE STUDY. G. M. Buck Louis, a
K. J. Sapra, a E. Schisterman, b C. D. Lynch, c J. Maisog, a K. L. Grantz, a
R. Sundaram. b a Division of Intramural Population Health Research, NICHD,
Rockville, MD; b Eunice Kennedy Shriver National Institute of Child, Rockville,
MD; c Department of Reproductive Medicine, Ohio State University
Medical Center, Columbus, OH.
OBJECTIVE: Recent cohorts of reproductive aged women now detect
their pregnancies early using sensitive home pregnancy tests, replacing clinical
diagnosis as the gold standard. This prompted us to assess whether the
incidence or risk factors for pregnancy loss have changed relative to guidance
based upon clinically recognized pregnancies.
DESIGN: Prospective cohort with preconception enrollment.
MATERIALS AND METHODS: 501 couples were recruited upon discontinuing
contraception and followed for a year of trying and through pregnancy.
Couples were interviewed upon enrollment and completed daily journals about
usage of vitamins, cigarettes, and caffeinated and alcoholic beverages until 7
post-conception weeks, then monthly until delivery. Women used ClearblueÒ
digital pregnancy tests on the day of expected menses or with bleeding. Pregnancy
loss was identified by conversion to a negative test, onset of menses or
clinical confirmation depending upon gestation. Using proportional hazards
regression and accounting for truncation and right censoring, we estimated hazard
ratios and 95% confidence intervals (HR; 95% CI) for partners’ ages and
BMI, females’ history of pregnancy loss and couples’ lifestyle (alcohol,
caffeine, smoking, vitamins) for two time intervals: 1) enrollment through early
pregnancy and 2) only during early pregnancy.
RESULTS: Among 344 women with a singleton pregnancy, 98 (28%)
experienced a pregnancy loss. In adjusted models for enrollment through
early pregnancy, we observed significant associations: female age R35 years
(2.19; 1.28,3.76), female smoking or alcohol usage (4.63; 2.23, 9.60), female
>2 daily cups of caffeinated beverages (1.86; 1.02, 3.42), and female vitamin
usage (0.42; 0.22, 0.80). When restricting time to only during pregnancy, female
age (2.04; 1.21,3.44) and smoking or alcohol usage (5.00; 2.57, 9.73)
remained significant.
CONCLUSIONS: Lifestyle during sensitive windows was associated with
pregnancy loss corroborating earlier results, although with higher risks. Of
note was the reduced risk associated with vitamin use, a finding not previously
reported. These findings support encouraging preconception guidance
for reproductive aged couples.
Supported by: Intramural Research Program of the Eunice Kennedy
Shriver National Institute of Child Health and Human Development.
O-220 Wednesday, October 21, 2015 12:00 PM
IS THYROID AUTOIMMUNITYASSOCIATED WITH PREGNANCY
LOSS?. T. Plowden, a E. Schisterman, a S. Zarek, a R. M. Silver, b
N. Galai, c A. DeCherney, a S. L. Mumford. a a NICHD, NIH, Bethesda,
MD; b Unversiy of Utah, Salt Lake City, UT; c Haifa University, Haifa, Israel.
OBJECTIVE: Overt thyroid dysfunction has been associated with infertility,
early pregnancy loss and other adverse obstetrical outcomes. However,
results of studies assessing the relationship between thyroid antibodies and
pregnancy loss have varied. Thus, our objective was to examine the association
between pre-pregnancy anti-thyroid antibodies and pregnancy loss.
DESIGN: Prospective cohort study from a multi-center randomized, placebo-controlled
trial of preconception low-dose aspirin to prevent pregnancy
loss among healthy fertile women (n¼1228). Women with abnormal fT4 at
baseline were excluded.
MATERIALS AND METHODS: At baseline, TSH, fT4, anti-thyroglubulin
antibody (anti-TG) and anti-thyroid peroxidase antibody (anti-TPO)
levels were measured. Relative risk (RR) and 95% confidence intervals
(CIs) for pregnancy loss were estimated using generalized linear models adjusting
for age and body mass index.
RESULTS: 155 women in our study had thyroid autoimmunity (+anti-TG
antibodies and/or +anti-TPO antibodies) compared to 902 without thyroid
immunity. During the study period, 114 women with thyroid autoimmunity
became pregnant while 639 women without thyroid immunity conceived. After
adjusting for age and BMI, women with thyroid autoimmunity who
became pregnant during the study had similar rates of clinical pregnancy
loss (RR 0.80; 95% CI 0.50, 1.27) and live birth (RR 1.05; 95% CI 0.95,
1.16) compared to those without thyroid autoantibodies.
CONCLUSIONS: Among healthy fertile women with a history of 1 or 2
losses, presence of anti-TG or anti-TPO antibodies was not associated with
pregnancy loss.
Clinical Pregnancy Loss and Live Birth Rate in Women With and Without
Thyroid Autoimmunity.
Absence of
Thyroid Antibodies
Presence of
Thyroid Antibodies
N(%) 649 (85.1) 114 (14.9)
Clinical Pregnancy Loss 1.00 (ref) 0.80 (0.50,1.27)
Live Birth Rate 1.00 (ref) 1.05 (0.95,1.16)
Supported by: Intramural Research Program, DIPHR, PRAE, NICHD,
NIH
O-221 Wednesday, October 21, 2015 12:15 PM
GALECTIN 14 PROTECTS HUMAN TROPHOBLAST CELLS
FROM OXIDATIVE STRESS AND PROMOTES EXTRAVILLOUS
DIFFERENTIATION. R. Fritz, a B. Kilburn, a H. Kohan-Ghadr, a
S. Drewlo, a D. Armant. a,b a Wayne State University, Detroit, MI; b National
Institute of Child Health & Human Development, Bethesda, MD.
OBJECTIVE: In the first trimester, invasion of extravillous trophoblast
(EVT) cells into maternal spiral arteries with subsequent endovascular
plug formation is necessary for healthy placentation by maintaining low oxygen
concentrations at the implantation site (1). Failure to form an EVT plug
within the spiral arteries is strongly linked to early pregnancy loss (EPL) (2).
Hypoxia/re-oxygenation injury (H/R) in the first trimester placenta could
contribute to adverse pregnancy outcomes, including EPL. The aim of this
study was to evaluate the effects of galectin 14, a rarely studied placental specific
galectin, on EVT cell survival, invasion, and proliferation using a human
first trimester trophoblast cell line.
DESIGN: Laboratory prospective study
MATERIALS AND METHODS: The HTR-8/SVneo first trimester human
trophoblast cell line was cultured at 20% oxygen (O2), at 2% O2,
or exposed to H/R injury, induced by culture for 2 hours at 2% O2 followed
by 6 hours at 20% O2. Expression of galectin 14 was quantified by immunocytochemistry
(ICC) and image analysis. To evaluate rescue from
apoptosis, 10 ng/ml recombinant galectin 14 was added during re-oxygenation.
Cell death was quantified using terminal deoxynucleotidyl transferase
dUTP nick end labeling (TUNEL). Effects of galectin 14 on
invasion were measured using a Matrigel-based assay, counting trophoblast
cells in the lower chamber after culture for 72 hours. Integrin switching
from a6b4 toa1b1 is associated with extravillous differentiation to an
invasive phenotype, and was assessed by ICC, evaluating the percentage
of cells positive for a6b4 and a1b1 . Proliferation was analyzed using
ICC for Ki-67. All experiments were conducted in triplicates and alongside
negative controls. Statistical analysis was performed with the non-parametric
Kruskal-Wallis test, using open-source R software (http://www.rproject.org/).
P < 0.05 was considered significant.
RESULTS: Expression of galectin 14 significantly increased after treatment
with 2% O2, and significantly decreased after H/R injury. H/R injury
significantly increased cell death; however, galectin 14 rescued trophoblasts
from cell death. Supplementation with galectin 14 significantly increased
trophoblast invasion through Matrigel and caused a significant switch from
integrin a6b4toa1b1. Galectin 14 had no effect on trophoblast proliferation.
CONCLUSIONS: Galectin 14 contributes to trophoblast survival under
hypoxic conditions, and rescues cell death due to H/R injury. Furthermore,
galectin 14 promotes trophoblast invasion and differentiation to the extravillous
phenotype. Galectin 14 could serve as a therapeutic agent for early pregnancy-related
placental disorders, such as recurrent EPL.
References:
1. Burton, G.J. and E. Jauniaux, Placental oxidative stress: from miscarriage
to preeclampsia. J Soc Gynecol Investig, 2004. 11(6): p. 342-52.
2. Hempstock, J., et al., The contribution of placental oxidative stress to
early pregnancy failure. Hum Pathol, 2003. 34(12): p. 1265-75.
Supported by: Grants from the NIH (HD071408), the March of Dimes, the
W.K. Kellogg Foundation, PerkinElmer Health Sciences, and, in part, by the
Intramural Research Program of the NICHD, NIH.
FERTILITY & STERILITY Ò
e85

O-222 Wednesday, October 21, 2015 12:30 PM
DISCRIMINANT ANALYSIS FORECASTING MODEL OF FIRST
TRIMESTER PREGNANCY OUTCOME FOLLOWING 5054 INFER-
TILE PATIENTS AFTER IVF-ET. Y. Yi, a X. Li, b Y. Ouyang, c G. Lu. c
a Institute of Reproduction & Stem Cell Engineering, Central South University,
Changsha, China; b Reproductive and Genetic hospital of CITIC-Xiangya,
Chang-sha, China; c Central South University, Changsha, China.
OBJECTIVE: To investigate whether ultrasound indicators of embryo on
27th -29th day after IVF-ET are different between 1st trimester miscarriage
group and ongoing pregnancy group and to build discriminant analysis forecasting
model to predict 1st trimester pregnancy outcome.
DESIGN: Prospective study.
MATERIALS AND METHODS: 5054 single pregnancy female were recruited
after IVF-ETover a period of 18 months. Maternal age, infertility duration
and number of IVF cycles of these patients were collected. Transvaginal
ultrasound was performed for these patients to observe MSD (Mean Sac Diameter
), CRL (Crown-Rump Length), YSD (Yolk Sac Diameter), embryonic cardiac
activity and fluid collection around GS (Gestational Sac) on 27th -29th day
after IVF-ET. Then 1st trimester pregnancy outcome of these patients were followed
up. Characteristics and Ultrasound parameters of these patients were
compared by Two Sample t Test or Chi-Squared Test between 1st trimester
miscarriage group (n¼679) and ongoing pregnancy group (n¼4375). Forecasting
model was built by discriminant analysis to predict 1st trimester pregnancy
outcome. The difference was statistically significant when P < 0.05.
RESULTS: Maternal age (32.575.06 VS 30.364.29 years ), infertility
duration (6.014.13 VS 5.273.50 years) and number of cycle
(1.330.81 VS 1.190.55) were all significant higher in 1st trimester
miscarriage group than those in ongoing pregnancy group (P

DESIGN: Prospective cohort study.
MATERIALS AND METHODS: The Longitudinal Investigation of
Fertility and Environment Study (2006-2010) enrolled 501 couples attempting
pregnancy. Couples were followed until they had a positive human chorionic
gonadotropin pregnancy or for up to 12 months of trying. Residences were geocoded
and the distance to the nearest major roadway was determined in meters
(

DESIGN: Cross sectional observational study.
MATERIALS AND METHODS: Women aged between 20-38 years (300
infertile couples) presented to a university affiliated fertility center were approached
to participate in the study. Only 150 couples that underwent ICSI
for male factor infertility accepted to participate and 94 of them had retrieved
enough FF samples suitable for laboratory testing. The FF sample was obtained,
centrifuged, and stored in liquid nitrogen. The concentrations of
four PCBs 28, 52, 138, 180 were estimated in the obtained FF samples using
Gas Chromatography/ Mass Spectrometry. SPSS statistical analysis program
(version 17) was used. Multiple regression analysis was used to correlate the
PCBS to ICSI outcomes.
RESULTS: The mean age of study participants was 31.5 6.2 years. Their
body mass index (BMI) was 26.19 6.84, baseline FSH was 7.6 2.65, and
Estradiol 59.3 10.5. There were significant negative correlations between
the concentrations of the 4 PCBS and the endometrial thickness (Adjusted r
0.2, P ¼ .0001). The higher PCB 28 concentration in the FF was associated
with lower number of retrieved oocytes (adjusted r 0.07, P ¼ 0.0001). Moreover,
fertilization and embryo cleavage rates were negatively correlated with
PCB 180 FF concentration (adjusted r 0.07, P ¼ 0.001 and adjusted r 0.1,
P¼0.002, respectively). The number of the implanted embryos was negatively
correlated with PCB 52 FF concentration (adjusted r 0.2, P¼ 0.02).
However the clinical pregnancy rate did not reach the level of significance.
CONCLUSIONS: Higher concentrations of any type PCBs are associated
with thinner endometrium. The higher the level of PCB 28 and 180 the lower
the retrieval, fertilization and embryo cleavage rates. High PCBs concentration
in the FF adversely affect embryological ICSI outcome. However, more
data is needed to evaluate the PCBS effect on the clinical outcome.
FERTILITY PRESERVATION
O-229 Wednesday, October 21, 2015 11:15 AM
CO-TRANSPLANTATION OF ENDOTHELIAL CELLS WITH XEN-
OTRANSPLANTED HUMAN OVARIAN TISSUE IMPROVES PRI-
MIRDIAL FOLLICLES SURVIVAL. L. Man, a Z. Rosenwaks, b
G. L. Schattman, c D. James. a a CRMI, Weill Cornell Medical College,
NYC, NY; b Weill Cornell Medical College, New York, NY; c Weill Medical
College/Cornell University, New York, NY.
OBJECTIVE: Improved treatments have raised cancer survival rates and
increased the number of patients needing fertility preservation. A major
obstacle to efficient ovarian tissue autotransplantation is graft ischemia.
Endothelial cells (ECs) exhibit the unique capacity to auto-assemble neovascular
networks following in vivo transplantation of single cell suspensions.
Here, we show that co-transplantation of exogenous ECs improve survival
and follicular reserve of xenografted ovarian tissue by accelerating anastomosis
of blood vessels at the interface of host and graft.
DESIGN: Xenograft of human ovarian tissue into NSG mice with cotransplantation
of endothelial cells.
MATERIALS AND METHODS: Tissue-specific mouse ECs, were
labeled with green fluorescent protein and modified by the adenoviral gene
fragment E4-ORF1. Human ovarian tissue was co-transplanted with GFP-
ECs into the gluteus maximus of NSG oopherectomized mice aged 10-12
weeks. Each mouse served as its own control with a contralateral site of
engraftment without ECs. Engrafted tissue was harvested 1-2 weeks after
transplantation. The contribution of exogenous ECs to functional vessels,
amount of surviving follicles as well as the extent of necrosis were analyzed
in histologic sections using light and confocal microscopy.
RESULTS: At 1 and 2 weeks we demonstrated the same proportion of
follicular survival between the ovarian tissue co-transplanted with ECs, in
comparison to ovarian tissue without ECs. At 2 weeks after co-transplantation
we found a three-fold increase in the survival of primordial follicles at
the ECs group in comparison to the control; 73% vs 27%, P

give them the greatest chance of a live-born baby. We set out to answer this
question by calculating the theoretical number of MII oocytes necessary to
result in one baby born (BB).
DESIGN: Retrospective cohort study and theoretical model.
MATERIALS AND METHODS: Data from 160 autologous OC thaw
(AOCT) cycles were reviewed. Cycles did not include medically-indicated
or those with Day-3 transfer. To calculate the no. of oocytes needed to result
in one BB, the following data were assessed: total no. of thawed and survived
MII oocytes, 2-pronuclear (2PN) fertilization, and development of morulae
and blastocysts (BL). BL formation yield (BFY) was calculated as total
no. BL/no. MII thawed, and total embryo rate (TER) was calculated as total
no. morula+BL./no. MII thawed. Rate of babies born (RBB) was calculated
as no. BB from morula or BL ET/total no. embryos transferred. The theoretical
number of oocytes needed to result in one BB was calculated as 1/
(TERxRBB). To calculate the RBB, data from PGS cycles was excluded.
RESULTS: See Table. Of 1,438 AOCT oocytes that survived, the 2PN
fertilization rate was 68% and of those, 41% formed BL. The estimated
no. of MII needed to achieve a BB was 8, 10, 14, and 50, and the likelihood
of BB per MII oocyte was 13%, 10%, 7%, and 2% for the age groups 25-34y,
35-37y, 38-40y, and 41-42y respectively.
CONCLUSIONS: As OC is increasingly relied upon for fertility preservation,
our findings provide realistic and important counseling data for women
electing to freeze gametes for usage at a later time. Women in the 25-34, 35-
37, and 38-40 age groups may only require one OC cycle, whereas women in
the 41-42 age group should be counseled extensively and provided realistic
expectations regarding the number of cycles needed to achieve success.
Overall, the calculations likely underestimate the efficiency of each oocyte,
as cycles involving PGS were excluded from final RBB calculations and PGS
cycles resulted in an implantation rate of 64% and live birth rate of 58%.
TABLE. MII oocyte development following AOCT by age.
Age (y)
25-34
(mean 31.5y;
n¼22)
35-37
(mean 36.3y;
n¼39)
38-40
(mean 39.1y;
n¼74)
41-42
(mean 41.6y;
n¼25)
Oocytes retrieved 500 751 1185 367
MII thawed 343 431 735 234
MII survived 286 348 610 194
No. 2PN fertilized 228 259 358 132
No. Morula + 40 + 94 26 + 99 72 + 153 13 + 56
No. BL
BFY 94/343
(27%)
99/431
(23%)
153/735
(21%)
56/234
(24%)
TER 134/343
(39%)
125/431
(29%)
225/735
(31%)
69/234
(29%)
OP+LB from
morula ET
1/7 (14%) 1/8
(13%)
5/27
(19%)
0/7
(0%)
OP+LB from
BL ET
11/29
(38%)
5/9
(56%)
3/10
(30%)
1/10
(10%)
RBB 12/36 6/17 8/37 1/17
No. MII oocyte to
achieve BB
(33%) (35%) (22%) (6%)
8 10 14 50
O-232 Wednesday, October 21, 2015 12:00 PM
CURRENT TRENDS IN PUBLIC SUPPORT FOR ELECTIVE
OOCYTE CRYOPRESERVATION. E. I. Lewis, a,b S. A. Missmer, a,b,c
L. V. Farland, a,b,c E. Ginsburg. a,b a Center for Infertility and Reproductive
Surgery, Dept of OBGYN, Brigham and Women’s Hospital, Boston, MA;
b Harvard School of Medicine, Boston, MA; c Harvard School of Public
Health, Boston, MA.
OBJECTIVE: As oocyte cryopreservation (OC) is now an established
method to preserve fertility for women due to health, aging or other factors,
our aim was to determine whether there is public support for this treatment
and if support varies by demographic factors.
DESIGN: Cross-sectional electronic survey.
MATERIALS AND METHODS: A nationally representative sample of
1064 people aged 18-75 years completed an online questionnaire. Supporters
of OC for various indications were compared with those participants who
were neutral or in opposition using log binomial regression. Statistical
models were adjusted for confounding factors including gender, ethnicity,
age, income, sexual orientation, education, marital status, state political party
affiliation and history of being a parent, to yield relative risk ratios (95% confidence
intervals; Wald 2-sided p-values) of support.
RESULTS: Of the 1,383 recruited, 17% declined to take the survey and 5%
were against any in-vitro fertilization and were disqualified. OC for cancer patients
was the indication most Supported (89%), followed by delayed childbearing
for career advancement (72%), current lack of a partner (63%), and
insufficient funds for child-rearing (58%). Despite considerable support for
OC, only 37% of participants agreed that employers should fund egg freezing
for female employees, with those never having a child more likely to be in favor
(p¼0.03). Younger participants (18-44 years old) were more likely to support
OC for women currently with insufficient funds for child-rearing (p¼0.03).
Younger age (p

Effectiveness of this method was tested by histology and whole ovary transplantation.
MATERIALS AND METHODS: 1) Ovary preparation. Umodl1-Tg
ovaries aged 3 weeks were collected and encapsulated in 0.5% alginate in
aMEM/ITS+0.25mM ascorbic acid. Cross-linking was done in 50mM
CaCl 2 /130mM NaCl solution. The aggregates were then grafted subcutaneously
under abdominal wall of SCID mice.2) Histological and marker gene
analyses. Preserved ovaries (Pr.) were dissected out at 2, 4 and 6 months after
grafting for H&E or IHC-staining. Antibodies against GCNA, FSHR, Laminin
and CD34 were used to examine ovarian ultrastructures.3) Fertility test.
Ovaries collected at 4 months after grafting were first treated with alginate
lysase. Ovaries from one side of the CD1 females were removed from bursa
sacs and replaced with the Pr. ovaries. Undisturbed ovaries on the other side
were used as internal fertility control. After one-month recovery, CD1 hosts
were primed with PMSG/hCG. Ovulated eggs were collected for lacZ-staining
to distinguish Umodl1-Tg from WT oocytes.Equal numbers of agematched
WT ovaries (nR 3) were included as controls in each assay. Statistical
significance was analyzed by unpaired t-test.
RESULTS: Extensive vascular network had already been established between
transplants and host subcutaneous tissue to support transplant survival
among 2-month old transplants. All of the preserved ovaries examined exhibited
excellent morphologies with distinct structures including ovarian follicles
and corpus luteum. Moreover, these cells showed correct expression of
their lineage-specific markers. Follicle counts on 4-month old tissue sections
(n¼8) showed no significant difference between WT and the Pr. groups
(WT,13745 vs. Pr, 11936/section; p¼0.40). Unlike the WT controls in
which follicles of various stages were observed, the preserved follicles in
the cortex seems to be synchronized at primordial/early preantral stages.
When re-transplanted back to the host bursa sacs, these quiescent ovaries
could be reactivated and produce comparable numbers of oocytes as the
WT controls (Pr., 7.251.28 vs. WT, 8.882.85/mouse; p¼0.16).
CONCLUSIONS: Ectopic transplantation of alginate-enclosed ovaries
can minimize the impact from the disturbed Umodl1-Tg endocrine system,
and thus maintain the ovaries at a quiescent state. Once implemented into
an appropriate milieu, their ovarian functions can be fully restored.
O-234 Wednesday, October 21, 2015 12:30 PM
EFFICACY OF STANDARDIZED NURSING FERTILITY COUN-
SELING ON SPERM BANKING RATES IN CANCER
PATIENTS. K. L. Rotker, a H. T. Vigneswaran, b D. Omil-Lima, b
G. Baird, c M. Sigman, d K. Hwang. b a Urology, Brown University, Cranston,
RI; b Urology, Brown University, Providence, RI; c Lifespan Biostatistics
Core, Brown University, Providence, RI; d Brown University and Lifespan,
Providence, RI.
OBJECTIVE: Chemotherapy often damages spermatogenesis, leading to
transient or permanent infertility. However, many clinicians fail to consistently
incorporate fertility preservation prior to treatment. The current study
examines sperm banking amongst patients undergoing formalized fertility
counseling prior to initiating chemotherapy.
DESIGN: A retrospective chart review was performed for two institutions
in Providence, RI. Men aged 18-50 with newly diagnosed cancer, from 1998
to 2003, prior to initiation of chemotherapy were evaluated. A standardized
nursing education session including directed fertility counseling was implemented
at one of the institutions in 2007.
MATERIALS AND METHODS: Patient characteristics including age,
cancer type, chemotherapy and/or radiation regimen, marital status, offspring
status, desire for information on sperm banking were recorded. Records of
the area’s only sperm bank were reviewed. Using bivariate statistical analysis,
rates of sperm banking amongst patients who received counseling
were compared to those without counseling at institution A. Multiple logistic
regression was used to model the relationship between time, offspring status
and age at both institutions.
RESULTS: 775 male patients were identified with a mean age of 40.98
years old. The most common cancer type treated was lymph node (19.3%).
At the time of diagnosis, 59.7% of patients already had children, 55.4%
were married, 30.6% were single and 7.5% were divorced. Of the 775 patients
studied, 411 (53%) were treated at institution A and of those 90
(23.4%) received fertility counseling. Of those who received counseling 15
(16.67%) underwent sperm banking compared to 20 (6.2%) of the 321
who did not receive counseling. This represented a significant increase in
sperm banking rates (p

Toronto, ON, Canada; f Department of Gynecology, Women’s College Hospital,
Toronto, ON, Canada.
OBJECTIVE: The ability to expand testicular biopsy-derived spermatogonial
stem cells (SSC) ex-vivo may be required for the development male fertility
preservation strategies. Our aim was to characterize and assess the potential use
of first trimester human umbilical cord perivascular cells (HUCPVC) as a source
of human feeder cells for the xeno-free ex-vivo expansion of SSCs.
DESIGN: Paracrine testicular niche-related growth factors were measured
in HUCPVC cultures. The ability of HUCPVC feeders to support ex-vivo
murine germ cell expansion was compared to mouse embryonic fibroblast
(MEF) feeders. In addition, the molecular properties of HUCPVCs were
compared to human testicular somatic cells (TSC), to evaluate the capability
of HUCPVCs to support human SSC propagation.
MATERIALS AND METHODS: We established mitotically inactivated
HUCPVC feeder cultures from previously established male lines in a manner
analogous to MEF feeder cultures. Paracrine factors were measured in
HUCPVC-conditioned media (CM) using ELISA (n¼3 independent lines).
CD-1 male murine neonatal germ cells were seeded on HUCPVCs or
MEFs (7500 cells/cm2) and expanded in complete SSC media. Colony number
and size were compared between the 2 feeder conditions. A BrdU incorporation
assay and immunocytochemistry was used to confirm the presence
of proliferating GPR125+ve and DAZL+ve germ cells. Targeted RNA
AmpliseqÔ was used to compare levels of 89 testicular cell lineage-associated,
growth factor and ECM transcripts in CD90+ve HUCPVCs and human
testicular somatic cells (TSCs) derived from orchiectomies.
RESULTS: FGF2, GDNF, LIF and BMP4 levels were detected at high
levels in HUCPVC-derived CM. A 3-fold increase in number SSC-like colonies
was observed in conditions where germ cells were seeded on HUCPVCs
when compared to MEFs after 2 passages (representing > 28 days in culture,
P

O-239 Wednesday, October 21, 2015 12:15 PM
CUMULUS-DERIVED FEEDER CELLS MAINTAIN THE PLURIPO-
TENCY POTENTIAL AND IMPROVE SELF-RENEWAL PERFOR-
MANCE OF HUMAN PLURIPOTENT STEM CELLS. O. Ait-
Ahmed, a S. Assou, a E. Pourret, a A. Ferrieres-Hoa, b S. Hamamah. b a Human
Early Embryonic Development and Pluripotency, INSERM U1203, Montpellier
University (UM), Montpellier University Hospital (CHRU), Montpellier,
France; b Human Early Embryonic Development and Pluripotency,
INSERM U1203, Montpellier University (UM), Montpellier University Hospital
(CHRU), ART/PGD Department, Montpellier, France.
OBJECTIVE: Based on the assumption that the microenvironment is
essential in pluripotency induction and maintenance, our objective was to
examine the capacity of cells cultured from the human metaphase II oocyte
CCs niche (hCC), to effectively support the propagation of human pluripotent
stem cells (PSC) in comparison to human foreskin fibroblasts (hFF).
DESIGN: Human cumulus cells collected from patients referred to our
center for intra-cytoplasmic sperm injection (ICSI) for male infertility
were cultivated in xeno-free medium and used as feeders for the growth of
hiPS cells.
MATERIALS AND METHODS: Human cumulus cells were dissociated
mechanically from the MII oocyte on collection day and adapted to growth
on a human collagen substrate for a long period in a xeno-free defined medium.
Affymetrix Human Genome U133 Plus 2.0 DNA chips were performed
to analyze the transcriptome of hCC and hiPS cells. Pluripotency/
differentiation markers were examined by RT-qPCR, immunofluorescence
and flow cytometry. Chromosomal abnormalities were analyzed using array
comparative genomic hybridization (aCGH).
RESULTS: We have optimized isolation and long-term culture of hCC in
xeno-free conditions. This new feeder is able to support growth of hiPS cells
during long term passaging. The Flow cytometric analysis showed that hiPShCC
retained expression of surface markers (CD24, SSEA4, TRA-1-81 and
TRA-1-60), demonstrating successful preservation of stemness. Moreover,
hiPS-hCC also improved the competence to differentiate into the three
germ layers in vitro (embryoid bodies) as well as in vivo (teratoma formation).
Interestingly the self-renewal rate was higher for hiPS-hCC than for
hiPS-hFF, suggesting that hCC has a growth-promoting effect. Additionally,
hiPS-hCC did not exhibit detectable sub-chromosomal aberrations, confirming
their genetic stability. A comparative gene expression study of hCC and
hFF revealed significant differences in expression of cellular matrix components
and an up regulation in hCC of genes known to be important players in
cell proliferation such as interleukin 6 gene (IL6).
CONCLUSIONS: This study shows that hCC have intrinsic properties that
make them better feeders to support pluripotent stem cells growth. The upregulation
of secreted factors encoding genes in hCC opens new perspectives
for investigating the impact of the microenvironment in the pluripotency
competence of the oocyte.
Supported by: This work was partially Supported by Ferring Pharmaceuticals.
O-240 Wednesday, October 21, 2015 12:30 PM
ESTABLISHMENT OF GYNOGENETIC HAPLOID EMBRYONIC
STEM CELLS AND GENERATION OF CLONED OOCYTES IN
MICE. M. Kobayashi, A. Yoshida. Kiba Park Clinic Research Center, Tokyo,
Japan.
OBJECTIVE: The establishment of embryonic stem cells (ESCs) with one
set of maternal genome will be able to provide the valuable cell source for the
genetic analysis of female genome, the production of genetically modified
animals, and the regenerative medicine. However, available information
about these ESCs is very limited. The aim of this study was to establish gynogenetic
haploid ES cells (GHESCs) with maternal haploid genome in
mice. In addition, we performed the generation of cloned oocytes by using
nuclear transfer with GHESCs.
DESIGN: ES-like cell lines from mouse GH embryos were established and
characterized. The developmental ability of cloned oocytes reconstituted
with GHESCs was studied.
MATERIALS AND METHODS: GH embryos were produced by removal
of male pronucleus from fertilized BDF1 oocytes. Proliferated cells from inner
cell mass (ICM) were picked, dispersed and cultured on fresh feeder cells.
Once ESC-like colonies were formed, they were passaged by Trypsin-EDTA
treatment. After several passages, marker expressions for ESCs were examined
with immunohistochemical staining. The DNA content of the cells was
measured by flow cytometry, and karyotype analysis was also performed.
Next, for reconstitution of cloned oocytes, GHESCs were synchronized the
cell cycle at G2/M phase with Nocodazoleand transferred into the perivitelline
space of enucleated mouse oocyte with Sendai virus. After the membrane
fusion between GHESCs and enucleated oocytes, the cloned oocytes were
fertilized by ICSI and cultured in KSOM.
RESULTS: Blasturation rate for GH embryos was 18.9%, and the ICM
outgrowth was observed in approximately 60% of these. In these cells, the
spherical colony formation and proliferation were maintained up to 4
months. By immunohistchemical staining, the ESC-like cells were positive
for ES cell markers, such as Oct4, Nanog, and Rex1. Further, flow cytometric
analysis and karyotyping revealed that these cells had a haploid set of 20
chromosomes (19 autosomes and the X chromosome). Next, in generation
of cloned oocytes, high rate (95.7%) of membrane fusion between GHESC
and enuclated oocyte was observed. After ICSI to the cloned oocytes reconstituted
by using nuclear transfer with GHESCs, 88.5% of these cleaved to 2-
cells and 17.3% formed blastocyst.
CONCLUSIONS: ESC-like cells with a maternal haploid chromosome set
derived from GH embryos were able to establish and maintain them for a
long-term period. Further, we showed clearly that the cloned oocytes reconstituted
by nuclear transfer with GHESCs were able to fertilize and developed
to blastocyst stage.
OUTCOME PREDICTORS - CLINICAL: ART 3
O-241 Wednesday, October 21, 2015 11:15 AM
PERINATAL OUTCOMES WITH AND WITHOUT ART: A POPULA-
TION-BASED STUDY OF LINKED SIBLINGS IN 12
STATES. B. Luke, a M. B. Brown, b L. G. Spector. c a Obstetrics, Gynecology,
and Reproductive Biology, Michigan State University, East Lansing,
MI; b Biostatistics, University of Michigan, Ann Arbor, MI; c Pediatrics, University
of Minnesota, Minneapolis, MN.
OBJECTIVE: To evaluate perinatal outcomes of siblings, conceived with
and without ART.
DESIGN: Longitudinal cohort study.
MATERIALS AND METHODS: Cycles during 2004-09 from the Society
for Assisted Reproductive Technology Clinic Online Reporting System
(SART CORS) resulting in live births were linked to their respective birth
certificates in 12 States (CA, CO, CT, FL, MI, NC, NJ, NY, OH, PA, TX,
VA) (ART children). All other live births to the same woman were identified
(ART siblings), as well as a 10:1 sample of non-ART deliveries (Control children).
All children were also linked to their respective State Cancer Registries,
and death certificates through one year of age. Pairs of singleton
siblings were identified in which one child was in SART CORS and the other
was not. Differences in length of gestation and birthweight were compared
between siblings using Student’s t-test, after adjustment for the effects of
mother’s age, weight, weight gain, parity and infant gender observed in the
controls. Percent distribution of ART factors were compared using the chisquare
test.
RESULTS: The study population included 3,213 pairs in which the ART
child was born after the sibling (ART later) and 4,597 pairs in which the sibling
was born later (SIB later). The difference between siblings was 3.01.4
years (meanSD) for ART later and 1.91.0 years for SIB later. The ART
birth was 33512 grams heavier in ART later pairs and 83488 grams heavier
in SIB later pairs. The ART birth was 2.117.2 days earlier in ART later
pairs but there was no difference in SIB later pairs. Analysis of a subset of
pairs in which the sibling had no indicators of infertility treatment on their
birth certificate (1,085 pairs with the ART child was born later and 2,881
pairs with the sibling was born later) showed similar results. Factors used
in ART that were associated with significantly less likelihood of having a sibling
after an ART birth included the diagnoses of diminished ovarian reserve
and tubal ligation, the use of donor oocytes, thawed embryos, and the use of
ICSI or assisted hatching. The occurrence of childhood cancer or infant mortality
in either the ART child or the sibling was associated with a significantly
greater likelihood of having a subsequent child.
CONCLUSIONS: Differences in birthweight and length of gestation between
siblings were very small, regardless of method of conception. The
ART factors were associated with a less optimal environment for a spontaneous
birth. This suggests that factors inherent in the woman, rather than
those attributable to infertility treatments, may explain the lower birthweight
and shorter gestation associated with ART in prior studies which compared to
the general population.
e92 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

O-242 Wednesday, October 21, 2015 11:30 AM
DO SERUM ANTI-MULLERIAN HORMONE LEVELS CORRE-
LATE WITH PREGNANCY OUTCOMES IN PATIENTS WITH
DIMINISHED OVARIAN RESERVE UNDERGOING IN VITRO
FERTILIZATION?. N. Pereira, a J. Lekovich, a Z. Rosenwaks, b
S. D. Spandorfer. c a The Ronald O. Perelman and Claudia Cohen Center for
Reproductive Medicine, New York, NY; b Weill Cornell Medical College,
New York, NY; c Cornell University Medical Center, New York, NY.
OBJECTIVE: Previous studies have suggested that patients with serum
markers of diminished ovarian reserve (DOR) have higher rates of embryonic
aneuploidy and lower rates of clinical pregnancy. To test this hypothesis, we
investigate the correlation between pregnancy outcomes in patients < 35
years of age with good quality embryos and serum anti-mullerian hormone
(AMH) levels as a marker for DOR.
DESIGN: Retrospective cohort study. Patients were sub-grouped, a priori,
based on serum AMH levels: < 1or> 1 ng/mL and < 0.5 or > 0.5 ng/mL.
MATERIALS AND METHODS: All patients < 35 years of age undergoing
fresh in vitro fertilization (IVF) - embryo transfer (ET) cycles between
January 2006 and June 2013 were analyzed for potential inclusion. Patients
who had 2, 8-cell, day-3 embryos transferred with grades 1, 1.5 or 2 were
only included in the analysis. Cycles cancelled prior to oocyte retrieval, or
those utilizing donor oocytes were also excluded. Within each AMH subgroup,
demographic, baseline IVF characteristics and controlled ovarian
stimulation (COS) response parameters were compared. Demographic characteristics
included age, gravidity, parity, and body mass index (kg/m 2 ). COS
parameters included protocol type (GnRH antagonist vs. GnRH agonist), total
days of ovarian stimulation, total dosage of gonadotropins administered
(IU), peak estradiol (E 2 ) level (pg/mL), peak endometrial stripe (mm), total
number of oocytes retrieved, and total number of mature oocytes. Clinical
pregnancy (CP), spontaneous abortion (SAB) and live birth (LB) rates
were compared between AMH sub-groups. Student’s t-tests and Chi-square
(c2) tests were used as indicated. Statistical significance was set at P < 0.05.
RESULTS: A total of 1005 patients met inclusion criteria. Overall, there
was no difference in baseline demographic or IVF characteristics. Patients
in the > 1 ng/mL group required lesser gonadotropins (1952.1 968.4
IU) compared to the < 1 ng/mL group (2517.1 1056.8 IU) and the < 0.5
ng/mL group (3328.2 1765.9 IU). More oocytes were retrieved from the
same group 13.8 (5.51) compared to the latter two (P < 0.001). The CP,
SAB and LB rates in the < 1 ng/mL group was 49.2%, 5.97% and 43.2%,
respectively. Similarily, the corresponding rates were 50.5%, 6.93%,
43.6% in the < 0.5 ng/mL group. Overall, these rates were comparable to
the CP (50.2%), SAB (6.86%), and LB (43.3%) rates in the > 1 ng/mL group.
CONCLUSIONS: Serum AMH levels correlate well with number of oocytes
retrieved in patients < 35 years of age with DOR. However, in patients
with DOR who have good quality embryos, the correlation between serum
AMH levels and clinical pregnancy, spontaneous miscarriage, and live birth
rates is very limited.
O-243 Wednesday, October 21, 2015 11:45 AM
MONOZYGOTIC TWINNING IN IVF: WHY DO THEY
CLUSTER?. D. A. Vaughan, a R. Ruthazer, b A. Penzias, c E. Norwitz, a
D. Sakkas. d a OBGYN, Tufts Medical Center, Boston, MA; b Clinical and
Translational Research Department, Tufts Medical Center, Boston, MA;
c Beth Israel Deconness Hospital/Boston IVF, Waltham, MA; d Boston IVF,
Waltham, MA.
OBJECTIVE: The incidence of monozygotic twinning (MZT) is greatly
increased among IVF patients compared to the general population (0.7-
13% vs. 0.45%, respectively), but the reason for this remains unclear1.To
further investigate this association, we sought to ascertain whether MZT
events occur in clusters and, if so, to explore possible explanations for this
clustering.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Using an established and validated IVF
database from a single large urban IVF center, consecutive fresh IVF cycles
(autologous and oocyte donation) resulting in a viable clinical pregnancy
(confirmation of a gestational sac(s) and presence of a fetal pole with a heartbeat(s)
on ultrasound) from Jan 2002 to Dec 2013 were retrospectively reviewed.
The incidence of MZT overall and separately for each 6-month
interval was calculated as the total number of MZT events divided by the
number of viable clinical pregnancies. 6-month intervals with a MZT incidence
rate >2 standard deviations (SD) higher than the overall rate were regarded
as a high risk interval. Logistic regression modeling was then used to
adjust for both time and non-time related risk factors for MZT events within
our cohort (Table 1).
RESULTS: Over the 12 year study period, 25,502 fresh IVF cycles were
performed, resulting in 8,598 clinical pregnancies. Of these, 95 cycles
(1.1%) resulted in MZ twins. Median patient age (SD) in the MZT cohort
was 35.4 years (4.4). The % of MZTwas >2SD higher than the overall % of
MZT in 4 of the 24 6-month intervals. PGD, extended embryo culture (R4
days), and more recent cycle (2005 or later) were independent risk factors
for MZT. Conversely, increasing numbers of embryos transferred appeared
to decrease the risk of MZT. Use of multivariable logistic regression
modeling to control for risk factors for MZT did not correct for this clustering
effect, with both high-risk interval (clustering) and extended embryo culture
remaining significant after adjustment (Table 1.)
CONCLUSIONS: This study supports our hypothesis that MZ occurs in
clusters, and that this clustering effect could not be explained by demographics
and cycle characteristics alone. Our study would suggest that
external factors both clinical (such as type of stimulation) and laboratory
(type or lots of cultured media, days in culture) may be involved.
Table 1. Risk Factors for MZT events among 8598 fresh IVF cycles from Jan
2002 through Dec 2013.
MZT
(N¼95)
Non-MZT
(n¼8503)
Un-adjusted
p-value
Adjusted
OR
(95% CI)
and p-value
High-risk
6-month interval
35.8% (34) 15.5% (1321) < .0001 2.37 (1.53, 3.66)
p¼ .0001
Embryo Biopsy 12.6% (12) 3.5% (298) < .0001 1.63 (0.82, 3.23)
p¼ .16
Assisted Hatching 7.4% (7) 11.1% (940) 0.2537 –
Donor cycle 8.4% (8) 8.7% (743) 0.9133 –
ICSI 31.6% (30) 38.5% (3277) 0.1655 –
Year 2005 or later 86.3% (82) 72.9% (6197) 0.0033 1.56 (0.84, 2.91)
p¼0.16
Extended Culture 45.3% (43) 18.1% (1541) < .0001 2.68 (1.66, 4.33)
p< .0001
Oocyte provider
>35 years
46.3% (44) 48.6% (4136) 0.6519 –
Age at cycle start 35.4 4.4 35.9 4.6 0.3753 –
>2 embryos
transferred
20.0% (19) 31.7% (2692) 0.015 0.75 (0.45, 1.40)
p¼0.42
References:
1. Vitthala S et al. The risk of monozygotic twins after assisted reproductive
technology: a systematic review and meta-analysis. Hum Reprod
2009;15:45-55.
O-244 Wednesday, October 21, 2015 12:00 PM
KNOWLEDGE OF EMBRYONIC PLOIDY STATUS IS ASSOCIATED
WITH DECREASED TRANSFER ORDER AND INCREASED ELEC-
TIVE SINGLE EMBRYO TRANSFER (ESET). J. M. Franasiak,
M. D. Werner, C. R. Juneau, R. Barnett, K. H. Hong, E. J. Forman,
R. T. Scott. RMA, NJ, NJ.
OBJECTIVE: To evaluate how the knowledge of embryonic ploidy status
affects embryonic transfer order and the rate of elective single embryo transfer.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: All first time IVF cycles from 1/2012 to
12/2014 resulting in either a fresh or frozen embryo transfer were selected for
review. Patients with recurrent pregnancy loss or undergoing PGD for single
gene disorders or translocations were excluded. During this period of time
Comprehensive Chromosomal Screening (CCS) was routinely offered to
all patients as a way to increase pregnancy rates and decrease miscarriage
rates. Data collected included use of CCS or not, number of aneuploid/
euploid embryos if CCS was performed, number of embryos transferred,
and number of embryos cryopreserved. eSET was designated when there
were embryos cryopreserved in excess of those transferred. As per practice
standards, maximum transfer order was two embryos. Thus functionally, patients
selected between a one or two embryo transfer.
RESULTS: During the study timeframe there were 3683 unique patients
identified. Of these 1778 (48%) underwent CCS and 1905 (52%) did not.
Of those undergoing CCS, the average number of embryos biopsied was
FERTILITY & STERILITY Ò
e93

5.6 (1-46), number euploid was 3.7 (0-27), number aneuploid was 1.8 (0-21),
and 114 (6.4%) had no euploid for embryo transfer. Of the 3569 who underwent
transfer, the mean number of embryos transferred was 1.3 with CCS and
1.6 without CCS (p

OUTCOME PREDICTORS - LAB: ART
O-247 Wednesday, October 21, 2015 11:15 AM
CIRCULATING MICRORNAS, AS POWERFUL TOOLS TO PRE-
DICT IVF/ICSI OUTCOMES. E. Scalici, a,b,c T. Mullet, a,c A. Gala, a
C. Vincens, a T. Anahory, a S. Belloc, d S. Hamamah. a,b,c a ART-PGD
Department, Montpellier, France; b INSERM U1203, Montpellier, France;
c Montpellier 1 University, UFR of Medicine, Montpellier, France; d Eylau-
Unilabs Laboratory, Paris, France.
OBJECTIVE: Our aim was to investigate if circulating microRNAs (miR-
NAs) in human follicular fluid (FF) could be used as helpful biomarkers for
predicting IVF/ICSI outcomes.
DESIGN: In this prospective study, a pool of FF was retrieved for each patient
during IVF/ICSI procedure. A total of 91 FF samples from women with
normal ovarian reserve (n¼91) were collected at oocyte retrieval day.
MATERIALS AND METHODS: For each patient, all follicles were aspirated
and all FF samples were pooled. MicroRNAs were extracted from each
FF pool and quantified by RT-qPCR, using TaqMan technology. The expressions
of miR-320a, let-7b and miR-29a were analyzed in FF and related to
IVF/ICSI outcomes.
RESULTS: FF pools related to low number of mature oocytes (%2) contained
significant lower miR-320a expression levels than those related to high
number of mature oocytes (>2), respectively (p¼0.03). Moreover, significant
high let-7b levels were found in FF pools related to embryo cohorts with a
high total blastomere number/total embryo number ratio at day 3 (>8, ie
accelerated development) than in those with normal developmental kinetics
(ratio between 6 and 8) (p¼0.02). Interestingly, we found a significant and
negative correlation between FF let-7b expression levels and blastulation
rate (r¼-0.33, p¼0.003). The Receiving Operator Curve (ROC) analysis indicated
that the performance of FF let-7b in predicting the expanded blastocyst
development was 0.67 (0.54-0.79), with 70% sensitivity and 64.3% specificity
(p¼0.02). In addition, the area under the ROC curve (AUC), evaluating
the potential of FF miR-29a in predicting clinical pregnancy outcome
reached 0.68 [0.55-0.79] with a high sensitivity of 83.3% and a specificity
of 53.5% (p¼0.01).
CONCLUSIONS: Our results suggest that circulating miRNAs constitute
non-invasive powerful tools in IVF process to predict embryo development
and clinical pregnancy outcomes, in order to promote personalized IVF strategy.
Supported by: This work was Supported by the University-Hospital of
Montpellier, INSERM and by a grant from the Ferring Pharmaceutical Company.
The authors of the study have no competing interests to report.
O-248 Wednesday, October 21, 2015 11:30 AM
CELL-FREE DNA AND PREGNANCY OUTCOME: NEW BIO-
MARKER OF FOLLICULAR MICRO-ENVIRONMENT. T. Mullet, a
E. Scalici, a A. Gala, b A. F. Ferrieres-Hoa, c V. loup, a C. Brunet, d
S. Hamamah. a a ART PGD Department, Montpellier, France; b CHU Montpellier,
Montpellier, France; c CHRU Montpellier, Montpellier, France; d Endocrinologist,
Montpellier, France.
OBJECTIVE: The aims of this study were to investigate if cell-free DNA
(cfDNA) in follicular fluid (FF) could be related to women’s ovarian reserve
status and to their response to controlled ovarian stimulation (COS) protocols,
and thus constitute a potential biomarker of oocyte micro-environment
to predict clinical pregnancy outcome.
DESIGN: This prospective study included 94 women with normal ovarian
reserve, 6 with ovarian insufficiency (OI) and 27 with polycystic ovary syndrome
(PCOS). For each patient, FF samples were collected during IVF/
ICSI procedure.
MATERIALS AND METHODS: At oocyte retrieval day, the FF samples
of the same patient were collected and pooled. CfDNA concentration in
pooled FF samples was evaluated by ALU quantitative PCR.
RESULTS: CfDNA level was significantly higher in follicles from women
with impaired ovarian reserve (OI and PCOS), compared to women with
normal ovarian reserve (2.7 2.7 ng/mL versus 1.7 2.3 ng/mL, p¼0.03,
respectively). Likewise, FF cfDNA levels were significant more elevated in
women who received long ovarian stimulation protocols (> 10 days) or
high total dose of gonadotropins (R 3000 IU/L) than in women who
received short stimulation protocols (7-10 days) or total dose of
gonadotropins < 3000 IU/l (2.4 2.8 ng/mL versus 1.5 1.9 ng/mL,
p¼0.008; 2.2 2.3 ng/mL versus 1.5 2.1 ng/mL, p¼0.01, respectively).
FF cfDNA level was an independent and significant predictive factor for
pregnancy outcome (adjusted odds ratio¼0.69 [0.5; 0.96], p¼0.03). In
multivariate analysis, the Receiving Operator Curve (ROC) analysis showed
that the performance of FF cfDNA in predicting clinical pregnancy reached
0.73 [0.66-0.87] with 88% specificity and 60% sensitivity.
CONCLUSIONS: Our results suggest that cfDNA quantification in FF
pools could provide a new non-invasive and easy method to assess the quality
of the oocyte microenvironment and to predict the clinical pregnancy
outcome. Moreover, in the case of IVF failure, FF cfDNA quantification
could be used to improve the personalized patient’s care and thus to increase
the chance of success of the next IVF cycle by selecting embryo for replacement
issue from follicle which contains low cfDNA level.
O-249 Wednesday, October 21, 2015 11:45 AM
IMPACT OF TIME-LAPSE AND REDUCED OXYGEN CULTURE
ON LIVE BIRTH RATE AND ITS CORRELATION WITH INFER-
TILITY DIAGNOSIS. N. Zaninovic, Q. Zhan, R. Clarke, Z. Ye,
N. Pereira, Z. Rosenwaks. CRM, Weill Cornell Medical College, New
York, NY.
OBJECTIVE: To evaluate the impact of implementing time-lapse with
reduced oxygen for all IVF patients on live birth (LB) rate. To determine
the correlation between culture conditions with specific patient clinical diagnosis.
DESIGN: A retrospective analysis of IVF patient outcomes from 2012-
2013 cultured in standard incubator (S; n¼1199) vs. time-lapse incubator
(E; n¼1318) was performed. LB rate was used to calculate ongoing pregnancy
(OP) and implantation (IMPL) rates for D3 and D5 ET patients; pregnancy
data was analyzed by maternal age groups (SART). Logistic regression
analysis was used to correlate patient diagnosis with clinical outcome.
MATERIALS AND METHODS: Patients were cultured either in standard
incubator (S, 20% oxygen, Thermo Forma, USA) or EmbryoScopeÒ (E, 5%
oxygen, Vitrolife) using in-house sequential medium.
RESULTS: A significant increase in OP, IMPL and live birth cycles (LBC)
was observed In EmbryoScopeÒ vs. standard incubator (38.7% vs. 29.6%;
19.5% vs. 13.4%; 38.1% vs. 29.3%, respectively; p

MATERIALS AND METHODS: ICSI videos were quantified for the
following parameters: time of needle exposure to the ooplasm, intracellular
needle movement, cytoplasmic leakage, needle touching the membrane adjacent
to the holder, sperm position, volume of cytoplasm removed during injection,
as well as oocyte morphological characteristics. Various endpoints
were evaluated [degeneration, fertilisation, two-pronucleate(2PN), selection
for embryo transfer, blastocyst formation, embryo quality from day 2 to 6,
known implantation data]. Parameters found to significantly affect outcome
were compared between four ICSI practitioners, five needle types (Humagen
SI,SLM,35 or Wallace 45V,50V) and two ICSI rigs (Integra-Ti and Integra-
3). Data were analysed using ANOVA, Kruskal Wallis or Chi-square as
appropriate.
RESULTS: Fertilisation rates were not significantly affected by
practitioner(range71-78%), equipment(74-74%) or needle type(75-78%).
Of all the endpoints assessed, only degeneration, 2PN and
fertilisation rates were affected by ICSI events. ICSIs associated with
intracellular needle movement(66%, n¼ 153) were significantly less
likely to fertilise than controls(77%, n¼373, p

CLINICAL FEMALE INFERTILITY AND GYNECOLOGY
O-253 Wednesday, October 21, 2015 11:15 AM
AN INTERDISCIPLINARY LIFESTYLE INTERVENTION IMPROVES
CLINICALLY RELEVANT FERTILITY OUTCOMES IN OBESE
INFERTILE WOMEN - PRELIMINARY RESULTS. K. Duval, a,b
M. Belan, a,b F. Jean-Denis, b J. Baillargeon. a,b a Medicine, Division of Endocrinology,
Universite de Sherbrooke, Sherbrooke, QC, Canada; b Centre de
Recherche du Centre Hospitalier Universitaire de Sherbrooke, Sherbrooke,
QC, Canada.
OBJECTIVE: To assess the impact of a lifestyle intervention targeting
obese infertile women on fertility outcomes.
DESIGN: Randomized controlled trial.
MATERIALS AND METHODS: Participants were followed for 18
months or until the end of pregnancy. They were randomized into the intervention
(interdisciplinary lifestyle intervention without fertility treatments
for the first six months) or control group (standard fertility treatments
only). The lifestyle intervention consisted in individual’s encounters with a
dietician and a kinesiologist every 6 weeks, and weekly group sessions (at
least 12 different ones). Pregnancy was defined by a positive serum pregnancy
test. A total of 105 women were randomized, 21 were excluded/withdrawn
(13 intervention; 8 control), 29 are active in the study and 55
completed (24 intervention; 31 control). Only the results of participants
who completed the study are presented.
RESULTS: Mean age of participants was 30.2 4.7 yrs, BMI is 40.1 7.5
kg/m 2 , waist circumference is 118 16 cm, and 67% had polycystic ovary
syndrome. As compared with the control group, women in the intervention
group had significantly higher pregnancy rate (79.2% vs. 41.9%, p¼0.003)
and spontaneous pregnancy rate (50.0% vs. 12.9% of all women,
p¼0.003). There was no significant difference between groups for pregnancy
following assisted reproductive technology (29% in both groups). Importantly,
a tendency to a higher rate of life birth was observed in the intervention
group (62.5 % vs. 38.7%, p¼0.08). At any time during the study, 42% of
women in the intervention group and 36% in the control group lost R5%
of their initial body weight (p¼NS). Furthermore, 50% of women in the intervention
group and 36% in the control group lost R5 cm of their initial waist
circumference (p¼NS). The median weight loss at time of pregnancy in
women who delivered (n¼28) was -2.5% [-6.9 - -1.4%] (p¼0.0001) in the
intervention group and -1.6% [-2.4 - +1.0] (p¼0.22) in controls (p¼0.05 between
groups).
CONCLUSIONS: These results demonstrate that even a modest weight
loss following an interdisciplinary lifestyle intervention in obese infertile
women could improve significantly their pregnancy rate, mainly the occurrence
of a spontaneous pregnancy. They also suggest that the rate of living
babies may also be improved. Such interdisciplinary lifestyle intervention
may improve fertility by other factors than weight or waist loss alone.
Supported by: Quebec Ministry of Health and Social Services, and Canadian
Institutes of Health Research.
O-254 Wednesday, October 21, 2015 11:30 AM
CLOMID STAIR-STEP PROTOCOL MAY SHORTEN THE TIME TO
OVULATION BUT NOT TO PREGNANCY: A RANDOMIZED CLIN-
ICAL TRIAL. L. B. Craig, a J. D. Peck, b D. Zhao, b K. R. Hansen. a a Dept
Ob/Gyn, Section Reproductive Endocrinology & Infertility, University of
Oklahoma Health Sciences Center, Oklahoma City, OK; b Dept of Biostatistics
& Epidemiology, University of Oklahoma Health Sciences Center, Oklahoma
City, OK.
OBJECTIVE: A novel dosing regimen for clomiphene citrate that
involved increasing the dose (stair-step protocol) without inducing a
period with progestin has been retrospectively described. The objective of
this study was to prospectively compare length of time to ovulation and
pregnancy with escalating doses of clomiphene administration in the
traditional protocol versus the stair-step approach in women with ovulatory
dysfunction.
DESIGN: Unblinded, randomized clinical trial
MATERIALS AND METHODS: Women ages 18-45 with anovulatory
infertility were randomized in a 1:1 ratio at baseline ultrasound to two
protocols for ovulation induction. The traditional protocol included
clomiphene followed by progestin withdrawal if anovulatory before
increasing the clomiphene dose. Under the stair-step protocol, clomiphene
dose was increased cycle day 11-14 without a progestin withdrawal if no
follicle >12mm. Clomiphene dose started at 50mg and was increased up
to 150mg per protocol. Primary outcome was time to ovulation. Secondary
outcome was time to pregnancy. Outcomes were compared between the
two treatment groups using the log-rank test and the proportions of ovulation/pregnancy
versus time were plotted using the Kaplan-Meier method.
The proportion of ovulation/pregnancy between the two treatment groups
was analyzed with the chi-square test.
RESULTS: A total of 120 patients (60 stair-step, 60 traditional) were
randomized. The groups were similar in age, BMI, ethnicity, years of
infertility, and smoking. There was no significant difference in proportion
that ovulated between the stair-step and traditional groups (p¼0.77, 90.0%
vs 88.3%). In addition, there was no significant difference in time to first
ovulation between the two groups (p¼0.65). However, when comparisons
were limited to the 49 patients (34 stair-step, 15 traditional) who did not
ovulate at clomiphene 50mg, time to first ovulation was significantly
decreased (p $50,000 USD
(OR¼2.07, CI¼1.27-3.38), duration of infertility of >24 months
(OR¼0.65, CI¼0.47-0.89), and history of a prior pregnancy loss
(OR¼1.59, CI¼1.12-2.27) were significantly associated with the probability
of live-birth. Other baseline demographic, biochemical, and lifestyle characteristics
were not associated with outcomes.
CONCLUSIONS: While age and duration of infertility were significant
predictors of all pregnancy outcomes, most other baseline characteristics
FERTILITY & STERILITY Ò
e97

were not. The identification of income as a predictor of outcomes independent
of race and education may be reflective of differences in the underlying
etiologies of infertility between the groups as well as disparities in access to
fertility and/or obstetrical care.
References: The content is solely the responsibility of the
authors and does not necessarily represent the official views of the NICHD
or NIH.
Supported by: The Eunice Kennedy Shriver National Institute of
Child Health and Human Development (NICHD) Grants for AMIGOS:
U10 HD39005, U10 HD38992, U10 HD27049, U10 HD38998, U10
HD055942, HD055944, U10 HD055936, U10HD055925 PPCOSII: U10
HD27049, U10 HD38992, U10HD055925, U10 HD39005, U10 HD38998,
U10 HD055936, U10 HD055942, U10 HD055944. This research
was made possible by the funding by American Recovery and Reinvestment
Act.
O-256 Wednesday, October 21, 2015 12:00 PM
AUTOIMMUNE PROGESTERONE DERMATITIS: CLINICAL PRE-
SENTATIONS AND MANAGEMENT. THE BRIGHAM AND
WOMEN’S HOSPITAL EXPERIENCE. D. Foer, a K. Buchheit, b
D. Lynch, b A. R. Gargiulo, c M. Castells, b P. Wickner. b a Internal Medicine,
Brigham and Women’s Hospital, Boston, MA; b Rheumatology, Immunology
and Allergy, Brigham and Women’s Hospital, Boston, MA; c Brigham and
Women’s Hospital, Boston, MA.
OBJECTIVE: Autoimmune progesterone dermatitis (APD) is a poorly
recognized complex syndrome that can lead to difficulty tolerating fertility
treatment. It is commonly associated with hypersensitivity reaction to exogenous
or endogenous progesterone, often with cyclical symptoms correlating
with the menstrual cycle. Symptoms can range from dermatitis to anaphylaxis,
and an increasing number of cases have been reported after the advent
of in vitro fertilization (IVF), during which women are exposed to supraphysiological
levels of progesterone. We present the largest case series of
APD with successful outcomes following progesterone desensitization
including pregnancy and symptom resolution.
DESIGN: Case series from a large academic hospital Immunology
and Allergy practice, managed in conjunction with a reproductive
endocrinologist.
MATERIALS AND METHODS: Twenty-two cases of APD seen at BWH
were evaluated. Symptom presentation, diagnostic modalities, desensitization
protocols, and outcomes were analyzed. Patient follow-up was conducted
to assess for long-term effects of desensitization including fertility
treatment and quality of life.
RESULTS: Symptoms were heterogeneous and included cyclical dermatitis,
infertility, urticaria, angioedema, and bronchospasm. In nine patients
(41%) the symptoms followed menarche or pregnancy, while in 13 patients
(59%), these seem to have been triggered by exogenous progesterone/progestin
used for contraception or fertility treatment. Nine patients were desensitized
to progesterone. All nine patients had successful desensitizations, 3
patients were desensitized prior to embryo transfer with an IM progesterone
protocol. Desensitization resulted in significant symptom relief in 6 cases and
at least 2 pregnancies through IVF.
CONCLUSIONS: APD presents heterogeneously and may be misdiagnosed.
Women with cyclical allergic symptoms and infertility should be evaluated
jointly by an allergist and a reproductive endocrinologist, for possible
diagnosis of APD and desensitization/endocrine manipulation. This is the
largest case series of APD reported to date, and demonstrates that progesterone
desensitization is successful, reproducible, and can result in pregnancy
via IVF and symptom resolution with improved quality of life.
References:
1. Jenkins, J, Geng, A, Robinson-Bostom. Autoimmune progesterone
dermatitis associated with infertility treatments, Journal of American
Academy of Dermatology, (2008) 58: 353-354.
2. Farah, FS, Shbakli, Z. Autoimmune progesterone urticaria, Journal of
Allergy and Clinical Immunology (1971) 48:257-261
3. Snyder, JL, Krishnaswamy, G. Autoimmune progesterone dermatitis
and its manifestations as anaphylaxis: a case report and literature review,
Annals Allergy, Asthma, and Immunology (2003) 90:469-477.
4. Prieto-Garcia, A, Sloane, DE, Gargiulo, AR, Feldweg, AM, Castells,
M. Autoimmune progesterone dermatitits:clinical presentation and
management with desensitization for successful in vitro fertilization,
Fertility and Sterility (2011).
O-257 Wednesday, October 21, 2015 12:15 PM
PASSIVE FERTILITY PREDICTION USING A NOVEL VAGINAL
RING AND SMARTPHONE APPLICATION. W. Webster, a
E. M. Godfrey, b L. Costantini, c J. Katilius. c a Emergency Department, EvergreenHealth
Medical Center, Kirkland, WA; b Obstetrics & Gynecology, University
of Washington, Seattle, WA; c Prima-Temp, Inc, Boulder, CO.
OBJECTIVE: Continuous core temperature (CCT) monitoring provides a
reliable marker of circadian rhythm and a statistical model for identifying the
circamensal nadir (CN) in a woman’s menstrual cycle. We have developed a
self-inserted flexible vaginal ring that continuously monitors true core body
temperature and passively communicates CCT to a smartphone app. Diurnal
changes in CCT associated with the transition of the follicular phase to luteal
phase include a transient drop in the mesor (mean temperature) with an
altered period and acrophase (time of peak temperature) for each cycle
that characterizes the CN. The CN occurs within 1 to 2 days before the LH
surge. In this study, we evaluate the safety and reliability of this sensor,
and collect data to further define the predictive algorithm for identifying a
woman’s fertile window.
DESIGN: The study compares the ovulation predicted by this
temperature sensor compared with: 1) daily oral basal body temperature
(BBT) measures, 2) urine LH ovulation prediction kit, 3) serum progesterone
levels, and 4) transvaginal ultrasound. The study also identifies user
acceptance and ease of use. Throughout the menstrual cycle, the sensor
measures CCT every 6 minutes, and data are transferred wirelessly to an
iPhone app every 2 hours. This observational, multi-site, randomized study
is designed to evaluate the sensor in healthy volunteers or women who are
currently trying to conceive.
MATERIALS AND METHODS: 20 female subjects use the sensor for 3
menstrual cycles. Subjects keep an electronic diary of daily oral BBT and
urinary LH tests. Statistical analysis is performed by paired t-test and
multilevel model approach. Ultrasound is used to ascertain ovulation. The
resulting algorithm both predicts and identifies ovulation in comparison
to once a day oral temperature, urinary LH detection and progesterone
methods.
RESULTS: Comparison between all methods is analyzed by measuring the
temporal relationship of a positive test given by each method from the true
day of ovulation as recorded by ultrasound. The resulting novel algorithm
correctly predicts ovulation.
CONCLUSIONS: This innovative vaginal ring and its associated smartphone
app are a novel method to address the needs for passively predicting
ovulation. CCT will provide an improved algorithm for ovulation prediction
from a complete model of circadian rhythm resulting in a safe, convenient
and more accurate method of ovulation prediction.
Supported by: Prima-Temp, Inc.
O-258 Wednesday, October 21, 2015 12:30 PM
UTILITY OF SONOHYSTEROGRAPHY FOR TUBAL PATENCY
ASSESSMENT IN THE PREGNANCY IN POLYCYSTIC OVARY
SYNDROME II TRIAL. M. S. Christianson, a R. Legro, b S. Jin, c
E. Eisenberg, d M. P. Diamond, e K. R. Hansen, f W. Vitek, g A. K. Styer, h
P. R. Casson, i C. Coutifaris, j G. M. Christman, k R. Alvero, l
E. E. Puscheck, m A. Christy, n H. Zhang, c A. J. Polotsky, l N. Santoro. l
a Johns Hopkins University School of Medicine, Baltimore, MD; b Penn State
University College of Medicine; c Yale School of Public Health, New Haven,
CT; d NICHD, Bethesda, MD; e Georgia Regents University, Augusta, GA;
f University of Oklahoma Health Sciences Center, Oklahoma City, OK; g University
of Rochester Medical Center, Rochester, NY; h Massachusetts General
Hospital/Harvard Medical Sch, Boston, MA; i Northeastern Reproductive
Medicine, Colchester, VT; j University of Pennsylvania, Philadelphia, PA;
k University of Florida School of Medicine, Gainesville, FL; l University of
Colorado, Aurora, CO; m Wayne State University School of Medicine, Detroit,
MI; n NIH, Rockville, MD.
OBJECTIVE: To compare saline infusion sonohysterography (SIS) versus
hysterosalpingogram (HSG) for confirmation of tubal patency.
DESIGN: Secondary analysis of the randomized controlled trial, Pregnancy
in Polycystic Ovary Syndrome II (PPCOS II).
MATERIALS AND METHODS: 750 infertile women (18-39 years old)
with polycystic ovary syndrome (PCOS) were randomized to up to 5 cycles
of letrozole or clomiphene citrate. Tubal patency was determined by HSG,
e98 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

the presence of pelvic free fluid during SIS, laparoscopy, or evidence of an
intrauterine pregnancy within the previous 3 years. Logistic regression was
conducted with clinical pregnancy as the outcome and HSG or SIS as the
key independent variable. Results are reported as adjusted odds ratios
(OR) and 95% confidence intervals.
RESULTS: Tubal patency was confirmed in 511 subjects (68.1%) with SIS
and 217 subjects (28.9%) with HSG (minimum of 1 patent tube). Unilateral
occlusion was observed in 32 subjects (2.9%) during HSG. After adjustment
for treatment arm, body mass index, duration of infertility, smoking and education,
the likelihood of clinical pregnancy was similar with HSG or SIS
(OR 1.16, 95% CI 0.80,1.70, p¼0.440). Clinical pregnancy rates remained
similar between groups following sensitivity analysis for ovulatory cycles
(n¼619). Ectopic pregnancy occurred more often in subjects with tubal
patency confirmed by HSG compared to SIS (2.8% versus 0.6%, p¼0.02).
Two out of 32 women (6.3%) with unilateralocclusion on HSG were diagnosed
with ectopic pregnancy.
CONCLUSIONS: In this large cohort of women with PCOS, there was no
significant difference in clinical pregnancy rate between subjects who had
tubal patency confirmed by SIS versus HSG. SIS is an acceptable imaging
modality for assessment of tubal patency in this population. The increased
incidence of ectopic pregnancy with unilateral occlusion on HSG merits
further evaluation.
Supported by: NICHD R25HD075737
IN VITRO FERTILIZATION 3
O-259 Wednesday, October 21, 2015 11:15 AM
COST ANALYSIS OF GLOBAL VERSUS SELECTIVE SCREENING
STRATEGIES FOR CAVITARY LESIONS IN WOMEN SCHEDULED
FOR IN-VITRO FERTILIZATION (IVF). A. M. Abdelmagied, a,b
M. A. Kamel, b A. M. Abuelhasan, b T. A. Farghaly, b A. A. Nassr, a,b
S. A. Shazly, a,b M. H. Makarem. b a Obstetrics and Gynecology, Mayo Clinic,
Rochester, MN; b Obstetrics and Gynecology, Women Health Hospital, Assiut
University, Assiut, Egypt.
OBJECTIVE: Although it is a common practice to screen all IVF women
by office hysteroscopy (OH), it is not always a cost effective approach especially
in low resource settings. The objective of this study was to provide a
cost analysis to support global versus selective use of OH to detect cavitary
lesions (CL) in women scheduled for IVF.
DESIGN: Prospective cohort study and subsequent cost modeling
MATERIALS AND METHODS: Infertile women scheduled for IVF were
examined by trans-vaginal ultrasound (TVUS) then by OH, the gold standard,
to evaluate the endometrial cavity. To assess the utility of OH after
TVUS, two suggested OH screening strategies were compared in terms of
diagnostic accuracy and cost benefits: global for all women and selective.
The selective strategy included women with abnormal TVUS and/or significant
clinical predictors for CL. Statistical methods used for comparisons
included t-test, Chi-square test, Wilcoxon rank sum test and logistic regression.
The diagnostic accuracy of the selective strategy (index method) was
compared to the global strategy (reference method) using McNemar’s test.
A Cost-by-Detected Case Cost Model was utilized to compare the cost benefits
of both strategies.
RESULTS: Out 120 women included, 34 (28.3%) had CL by OH. TVUS
detected only 21 CL with sensitivity, specificity and accuracy: 62%, 93% and
77% respectively. Women with CL had older maternal (Median(IQR),
31.5[5.25] vs. 29[7], P¼0.009) and paternal (MeanSD, 38.8 6.5 vs
35.5 5.6, P¼ 0.007) age and were more likely to have metrorrhagia
(17.6% vs.1.2%, P¼0.002) and a previous significant endometrial
procedure (23.6 vs 3.5%, P¼0.002) than women with no CL. Using
multivariate regression analysis, only metrorrhagia (aOR ¼ 20.15, 95%
CI¼2.15- 189.08, P¼0.009) and a previous procedure (aOR ¼ 9.16, 95%
CI¼2.13 - 39.3, P¼0.003) were significant. Based on that, a predictive model
including abnormal TVUS, metrorrhagia and/or a previous endometrial
procedure was used for the selective screening strategy. Selective screening
using this model would have had a Negative Predictive Value (NPV) of 93%
and would have missed only 6 cases (17.6 %). These 6 cases would be 4
arcuate uteri and 2 small endometrial polyps. Both selective and global
strategies were comparable in terms of diagnostic accuracy (P¼0.33).
Considering OH cost of $2153 (according to Medicare 2015 National Fee
Estimate), the cost of global screening was $7598.8/detected case, and the
cost of selective screening would be $2998.8/detected case.
CONCLUSIONS: Selective screening based on abnormal TVUS, metrorrhagia
and/or past significant endometrial procedure was comparable to
global screening in terms of diagnostic accuracy with cost benefits utilizing
the selective strategy. The suggested selective strategy is particularly valuable
in low resource setting for CL screening before IVF.
Supported by: Research support in university hospitals
O-260 Wednesday, October 21, 2015 11:30 AM
ABNORMAL IMPLANTATION IN DAY 3 VERSUS DAY 5 EMBRYO
TRANSFERS IN FRESH IVF CYCLES: AN ANALYSIS FROM THE
SOCIETY FOR ASSISTED REPRODUCTIVE TECHNOLOGY
DATABASE. A. Kathiresan, a E. T. Wang, b N. Greene, b
C. J. Alexander, c M. D. Pisarska. b a UCLA, Los Angeles, CA; b Cedars Sinai
Medical Center, Los Angeles, CA; c Southern California Reproductive Center,
Beverly Hills, CA.
OBJECTIVE: Abnormal implantation may be an indicator of a non-receptive
uterine environment. This could be on the spectrum of placentation abnormalities
that lead to adverse pregnancy outcomes associated with IVF. In
order to better understand if timing of embryo transfer plays a role, our objective
was to determine if there was a difference in abnormal implantation in
day 3 (D3) versus day 5 (D5) fresh embryo transfers (ET) as reported to
the Society of Assisted Reproductive Technology (SART).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Data was obtained from the SART national
database. First autologous non-PGD IVF cycles involving D3 or D5
fresh embryo transfers in the United States between 2004 and 2009 were
included in the analyses. A total of 339,837 cycles fulfilled the inclusion
criteria. Parametric and nonparametric statistical analyses were used to evaluate
differences between D3 and D5 ET in all cycles. In a subset analysis of
193,218 cycles in which conception occurred, logistic regression was used to
determine if day of transfer was associated with a higher risk of abnormal implantation.
Abnormal implantation was defined as biochemical pregnancy,
ectopic pregnancy, or pregnancy loss.
RESULTS: When divided into D3 and D5 ET groups, the D3 ET group
consisted of women who were older (35.6 4.7 vs. 33.7 4.3, p

clinic, or if embryo transfer occurred on days other than 3 or 5. Among the 40
states with more than one clinic, 6 have legislation requiring insurance
coverage for at least one IVF cycle and were designated ‘‘mandated:’’ CT,
HI, IL, MA, MD, and NJ. The remaining 34 states were designated ‘‘nonmandated.’’
Regression models with a mandate*year of cycle interaction
term were used to examine the effect of mandate status on changes in multiple
birth rate per live birth, the proportion of transfers with elective single
embryo transfer (eSET), and the mean number of embryos transferred over
time.
RESULTS: A total of 173,968 cycles were included in the analysis. The
multiple birth rate was lower in mandated than non-mandated states (P <
0.001) and decreased over time in both groups (P < 0.001). Similarly, the
proportion of transfers with eSET was higher in mandated than nonmandated
states (P < 0.001), and increased over time in both groups (P <
0.001). The mean number of embryos transferred was lower in mandated
than non-mandated states (P < 0.001), and decreased along a similar trajectory
over time when embryo transfer was performed on day 5 (P < 0.001).
Conversely, there was a significant interaction between mandate status and
time among cycles with day-3 embryo transfer (P < 0.001). The mean number
of embryos transferred decreased over time in both groups (P < 0.001),
but the trajectory of decline was steeper for non-mandated states between
2007 and 2009 (P < 0.001). Between 2010 and 2011, the trajectory of decline
was similar between the groups (P ¼ NS).
CONCLUSIONS: Multiple birth rates have decreased over time in both
mandated and non-mandated states. Although the gap between these two
groups has narrowed with respect to the mean number of embryos transferred
on day 3, providers in mandated states still transfer fewer day-5 embryos and
are more likely to perform eSET.
O-262 Wednesday, October 21, 2015 12:00 PM
DIFFERENCE IN BIRTH WEIGHT OF CONSECUTIVE SIBLING
SINGLETONS IS NOT FOUND IN OOCYTE DONATION
COMPARING FRESH VERSUS FROZEN EMBRYO
REPLACEMENTS. D. Galliano, a N. Garrido, b V. Serra, c A. Pellicer. c
a IVI Foundation. IVI Barcelona, Barcelona, Spain; b Istituto Universitario
IVI, Valencia, Spain; c IVI Valencia, Valencia, Spain.
OBJECTIVE: First, to assess if there are any differences in birth weight
or gestational length in newborns from egg donation pregnancies delivering
singletons, originating from either fresh or frozen/thawed embryos, when
they were developed and delivered within the same mothers. Second, to
determine if there are any clinical, phenotypical or laboratory factors influencing
this relationship, including the origin of the oocyte (same or
different donor), the order of the children (first fresh or first frozen/thawed
embryo transfer), embryo freezing technique (vitrification or slow
freezing), the in-vitro embryo culture length, and the duration that embryos
remained frozen.
DESIGN: Retrospective cohorts study.
MATERIALS AND METHODS: 360 women undergoing oocyte donation
(OD), delivering (>28 weeks) at least two babies, each one from a single
pregnancy, originating from at least one fresh and one frozen-thawed embryo
transfer, controlling maternal and laboratory characteristics, to test the effect
of embryo freezing on children size (n¼731).
RESULTS: From fresh vs. thawed embryos, respectively, mean birth
weight of children was 3183.7g (95%CI: 3115.0-3252.4) vs. 3226.4g (95%
CI: 3166.3-3243.2), gestational age (days) was 272.1 days (95%CI: 270.1-
274.0) vs 268.8 days (95%CI: 263.1-274.5), and mean percentiles were
47.6th (95%CI: 44.5-50.8) vs 50.1th (95%CI: 46.8-53.3). The proportion
and corresponding odds ratio (OR) from fresh vs. thawed embryos,
respectively, were for children LGA: 13.6% vs. 11.3%
(OR¼0.81(0.52-1.27)); SGA: 9.4% vs 12.5% (OR¼1.37(0.85-2.2)); ONR
23.1% vs. 23.8% (OR¼1.04(0.74-1.47)); and macrosomy 0.3% vs 0.8%
(OR 3.1(0.3-29.7)). Mean Z-scores were 0.28(-0.13 to 0.09) vs 0.04(-0.06
to 0.14), respectively. After adjusting for clinically relevant variables, the
Adj(OR)LGA¼0.96(0.50-1.87); Adj(OR)SGA¼1.40(0.72-2.71); Adj(OR)
ONR¼1.20(0.73-1.97), and Adj(OR)MS¼not computable. None of the
stated measures were significantly different. Also, independent analyses
run on the origin of the oocytes, cryopreservation technique, cleavage stage
of the embryos, and time range of embryos remaining frozen did not reveal
any significant trends.
CONCLUSIONS: This study comparing siblings from OD cycles, and
eliminating the independent variables which affect early events in pregnancy,
revealed no difference in duration of gestation and live birth weights between
fetuses obtained after the replacement of fresh or frozen embryos. Moreover,
no clinical, phenotypical or laboratory factors appeared relevant, once statistically
controlled.
O-263 Wednesday, October 21, 2015 12:15 PM
TRENDS AND OUTCOMES OF ASSISTED REPRODUCTIVE
TECHNOLOGY (ART) CYCLES USING GESTATIONAL CARRIERS
IN THE UNITED STATES, 1998-2012. K. Perkins, S. Boulet,
D. M. Kissin, D. J. Jamieson. Centers for Disease Control and Prevention,
Atlanta, GA.
OBJECTIVE: To determine trends in gestational carrier (GC) cycles from
1998 to 2012 and compare reproductive outcomes of GC and non-GC cycles
DESIGN: Retrospective cohort study
MATERIALS AND METHODS: We used data from the National ART
Surveillance System and included all ART cycles with transcervical embryo
transfer performed during 1998-2012. We calculated the absolute number
and percent of all ART cycles using GCs over the study period. Use of
GCs among non-US residents was assessed due to high utilization in this population.
Temporal trends were assessed using the Cochran-Armitage trend
test. For the most recent years of data (2007-2012), we used log binomial
regression with generalized estimating equations for correlated outcomes
within clinics to calculate adjusted relative risks (aRRs) and 95% confidence
intervals (CIs) for the association between reproductive outcomes and use of
a GC among fresh cycles, stratified by oocyte status (donor versus nondonor).
RESULTS: Of 1,595,089 cycles performed from 1998 to 2012, 28,228
(1.8%) used a GC, and increased from 733 (1.1%) in 1998 to 3,200 (2.4%)
in 2012 (p

consistent when analyzed per cycle start and per embryo transfer. Using
AMH as a sole predictor of pregnancy outcome, a sensitivity analysis demonstrated
lower clinical pregnancy rates in those with AMH

the ultimate outcome being a child born alive and surviving the neonatal
period. Survival rates and pregnancy lengths were calculated for white
and non-white infants. Differences were determined using the Wilcoxon
signed rank test.
RESULTS: Compared with other months, infant survival was highest in
non-whites (p

matrix scaffolds are reflective of the native tissue. These scaffolds have the
potential to serve as a novel 3-dimensional platform for leiomyoma and myometrium
research in the future.
Supported by: National Institutes of Health Grant R21HD077479-01; National
Institutes of Health Grant K12HD050121; Northwestern University
Women’s Reproductive Health (WRHR) Scholar Award; Harold Amos Medical
Faculty Development Award; Robert Wood Johnson Foundation
O-270 Wednesday, October 21, 2015 12:30 PM
LEUKEMIA INHIBITORY FACTOR (LIF) GENE POLYMOR-
PHISM T>G (RS929271) PREDICTS IMPLANTATION AND PREG-
NANCY AFTER IVF/ICSI INDEPENDENT OF TP53 GENE
POLYMORPHISM. J. A. Oliveira, a,b L. D. Vagnini, b A. Renzi, b
G. R. Oliveira-Pelegrin, b C. G. Petersen, a,b A. L. Mauri, a,b
F. C. Massaro, a,b F. Dieamant, a,b M. Cavagna, a,b,c R. Baruffi, a,b
J. G. Franco, Jr., a,b a Center for Human Reproduction Prof. Franco Jr, Ribeirao
Preto, Brazil; b Paulista Center for Diagnosis Research and Training, Ribeirao
Preto, Brazil; c Women’s Health Reference Center Perola Byington Hospital,
Sao Paulo, Brazil.
OBJECTIVE: Leukemia inhibitory factor (LIF) plays a critical role in
embryo development and implantation. The literature provides evidence
that a LIF T/G gene polymorphism (rs929271) is associated with female
fertility. However, studies on LIF polymorphisms are still scarce. On the
other hand The TP53 gene plays a critical role in regulating blastocyst implantation
function mediate by genes involved in the TP53 pathway,
including the LIF gene. Studies have demonstrated that the genotype of
the TP53 codon 72 polymorphism (rs1042522; encoding arginine [Arg]
or proline [Pro]) is associated with the expression of LIF. This study
analyzed whether the LIF T/G gene can predict pregnancy outcomes in
ART and, if so, whether this is independent of the TP53 codon 72 polymorphism.
DESIGN: Prospective
MATERIALS AND METHODS: A total of 411 women who had undergone
cycles of IVF/ICSI were recruited. DNA was extracted from peripheral
blood samples that were taken from each participant, and LIF T/G TP53
codon 72 Arg/Pro single nucleotide polymorphisms (SNP) were genotyped
using real-time PCR. All procedures were performed under the same clinical/laboratory
conditions. The cumulative results, including fresh and frozen
cycles, were analyzed.
RESULTS: Hardy-Weinberg genotype distributions of the entire sample
indicated concordance between the observed and expected frequencies.
Characteristics such as age, infertility, etiology, number of
transfers and number of transferred embryos were not significantly
different (P>0.05) between the groups. The distribution of SNP genotypes
at codon 72 of the TP53 gene was similar among the three genotypes
of LIF SNP T/G (P>0.05).The G/G LIF genotype was associated
with increased implantation and ongoing pregnancy rates after IVF/
ICSI. However, no correlation was observed between TP53 codon 72
polymorphism genotypes and clinical outcomes after IVF/ICSI.The table
below summarizes the data.
CONCLUSIONS: LIF SNP T/G (rs929271) appears to be a susceptibility
biomarker that is capable of predicting implantation efficiency and pregnancy
outcomes after IVF/ICSI. This influence appears to be independent
of TP53 codon 72 polymorphism (rs1042522) genotypes.
Supported by: Merck Serono grant (GFI-2014-16). The funders had no
role in study design, data collection and analysis, or preparation of the abstract
NUTRITION
O-271 Wednesday, October 21, 2015 11:15 AM
MALE DIETARY TRANS FAT INTAKE IS INVERSELY ASSOCI-
ATED TO FERTILIZATION RATES. M. Arvizu, a C. Tanrikut, b
R. Hauser, c M. Keller, c J. E. Chavarro. c a Nutrition, Harvard T.H. Chan
School of Public Health, Boston, MA; b Massachusetts General Hospital,
Boston, MA; c Harvard T.H. Chan School of Public Health, Boston, MA.
OBJECTIVE: Saturated and trans fat intake has been related to lower
sperm counts, but it is not known whether this leads to lower reproductive
success among couples undergoing infertility treatment. To address this
issue, we evaluated the association between dietary fat intake and fertilization
rate among couples undergoing assisted reproductive technologies
(ART).
DESIGN: Prospective cohort study
MATERIALS AND METHODS: We followed 141 men from couples presenting
to an academinc fertility center whose partners underwent ART
(n¼246 cycles). Diet was estimated before treatment with a validated food
frequency questionnaire and outcome data were extracted from medical records.
Generalized linear mixed models with random intercepts to account
for multiple cycles per woman were used to evaluate the association of fat
intake with fertilization rate while adjusting for male and female age, total
Fresh Cycles
Cumulative(Fresh+frozen/thawed) Cycles
GENOTYPE
Implantation rate
Ongoing
pregnancy
rate/transfer
Ongoing pregnancy
rate/patient
Implantation rate
Ongoing
pregnancy
rate/transfer
Ongoing
pregnancy
rate/patient
WOMEN’S GENOTYPES
LIF T/G (RS929271) GROUPS
T/T
n:168
16.8% a
(87/519)
19.6%a
(44/225)
26.2%a
(44/168)
15.9%a
(100/630)
18.5%a
(53/287)
T/G
17.8%b 22.4%b 31.7%
16.2%b
20.2%b
n:202 (107/601) (64/286) (64/202)
(121/747) (73/362)
G/G
30.1% a,b 38.8%a,b 46.3%a
27.0%a,b
36.7%a,b
n:41
(31/103) (19/49) (19/41)
(34/126)
(22/60)
P
0.002 a
0.007a 0.02a 0.004a
0.003a
0.006 b 0.01b
0.005b
0.007b
GENOTYPE Fresh Cycles Cumulative
(Fresh+frozen/thawed)
Cycles
Implantation rate
WOMEN’S GENOTYPES
TP53 CODON 72 ARG/
PRO (RS1042522) GROUPS
Ongoing
pregnancy
rate/transfer
Ongoing
pregnancy
rate/patient
Implantation
rate
Ongoing pregnancy
rate/transfer
Ongoing pregnancy
rate/patient
31.5%a
(53/168)
36.1%
(73/202)
53.7%a
(22/41)
0.01a
Arg/Arg
n:198
18.8%
(112/595)
22.5%
(63/280)
31.8%
(63/198)
17.9%
(130/728)
21.1%
(74/350)
37.4%
(74/198)
Arg/Pro
n:182
18.5%
(100/540)
23.8%
(56/235)
30.8%
(56/182)
16.8%
(111/661)
21.7%
(65/300)
35.7%
(65/182)
Pro/Pro
n:31
14.8%
(13/88)
17.8%
(8/45)
25.8%
(8/31)
12.3%
(14/114)
15.3%
(9/59)
29.0%
(9/31)
P ns ns ns ns ns ns
Values within columns containing the same letter were significantly different. ns: not significant.
FERTILITY & STERILITY Ò
e103

calories, male body mass index, race, male and female smoking status, infertility
diagnosis, protocol type and dietary patterns.
RESULTS: Men’s median age was 36.9 years and median total fat intake
was 32% of calories/day. Fertilization rates were lowest in couples with the
highest male partner intake of trans fats (median 1.20 % calories/day, IQR
1.21,1.46%). The multivariate-adjusted fertilization rates (95% Confidence
Interval) in couples in increasing tertiles of men’s trans fat intake were
77% (70-84%), 78% (72-84%) and 64% (55-72%) (p, linear trend ¼ 0.05).
This association was stronger in cycles with conventional insemination
where the corresponding adjusted fertilization rates were 83% (69-91%),
70% (54-83%), and 47% (30-64%) (p, linear trend ¼ 0.005), and less pronounced
in ICSI cycles (p, linear -trend¼0.11).
CONCLUSIONS: Higher male-partner intake of trans fat was associated
with lower fertilization rates in couples undergoing ART, particularly in conventional
insemination cycles.
Supported by: NIH grants R01ES009718, R01ES022955, P30ES000002,
P30DK46200.
Ln mice (Table 1). There were no differences in the levels of IL-6, IL-10, and
interferon-gamma between the groups.
CONCLUSIONS: 10-week exposure to HFD causes significant reduction
in primordial follicles, compromised fertility, higher pro-inflammatory cytokine
levels, and increased ovarian macrophage infiltration, independent of
obesity. In addition, obesity worsens the effects of HFD alone. The negative
effects of HFD on primoridal follicles may be mediated by increased ovarian
inflammation. To the best of our knowledge, this is the first time that HFD
was found to be detrimental to fertility and ovarian function independent
of obesity in an interventional study. Further studies are needed to elucidate
the mechanisms behind these findings.
References:
1. Norman JE. The adverse effects of obesity on reproduction. Reproduction.
2010;140:343-345.
Supported by: 5T32HD040135-13A National Training Program in Reproductive
Medicine (MSW)
O-272 Wednesday, October 21, 2015 11:30 AM
HIGH-FAT DIET CAUSES COMPROMISED FERTILITY AND
INCREASED PRO-INFLAMMATORY CYTOKINES INDEPEN-
DENT OF OBESITY. M. E. Skaznik-Wikiel, A. J. Polotsky,
J. L. McManaman. OB/GYN, University of Colorado Denver, Aurora, CO.
OBJECTIVE: Female obesity is associated with ovarian dysfunction and
subfertility (1). However, there are no studies to date to assess whether
high-fat diet (HFD) alone (without obesity) causes reproductive dysfunction.
The goal of this study was to determine if HFD impacts ovarian function,
fertility, and markers of inflammation independent of obesity.
DESIGN: Prospective animal study.
MATERIALS AND METHODS: 5-week old mice were fed either low fat
diet containing 10% fat (control group-LF-Ln) or HFD containing 60% fat.
After 10 week feeding trial HFD-fed mice were divided into three groups
based on body weight (BW): group 1: BW >25 g - high fat obese (HF-
Ob), group 2: BW

increased with aging from 20w to 32w for both the NC (2.5 0.48 vs. 45.9
4.7, p¼0.01 and 1.4 0.18 vs 10.2 1.4, p¼ 0.02, respectively) and the HF
mice converted to NC (5.0 0.75 vs. 27.9 4.7, p¼ 0.01 and 3.6 0.67 vs
11.6 2.6, p¼ 0.02, respectively).
CONCLUSIONS: Mouse ovarian macrophage marker gene expression
increases with high fat diet, obesity and aging. The pro-inflammatory M1
macrophage phenotype expression significantly increased with HF diet
while there was no change in the anti-inflammatory M2 macrophage phenotype.
As the mice aged, there was an increase in both overall macrophage
marker expression and M1 macrophage marker expression. This increase
in overall and proinflammatory macrophage infiltration in the ovary may
contribute to the reproductive dysfunction seen with obesity, high fat diet
and aging.
O-274 Wednesday, October 21, 2015 12:00 PM
OMEGA 3 FATTY ACID SUPPLEMENTATION DOES NOT
IMPROVE RELATIVE HYPOGONADOTROPIC HYPOGONADISM
IN OBESE WOMEN. Z. A. Al-Safi, a H. Liu, b J. Chosich, a
M. Harris, c A. P. Bradford, a C. Robledo, a R. E. Eckel, d N. Carlson, b
A. J. Polotsky. a a Obstetrics and Gynecology, University of Colorado, Aurora,
CO; b Biostatistics and Informatics, Colorado School of Public Health,
Aurora, CO; c Food Science and Human Nutrition, Colorado State University,
Fort Collins, CO; d Medicine, University of Colorado, Aurora, CO.
OBJECTIVE: Female obesity is associated with relative hypogonadotropic
hypogonadism and chronic pro-inflammatory state. Dietary omega-3
fatty acids (FA) exhibit strong anti-inflammatory properties. We hypothesized
that dietary supplementation with omega-3 FA would result in normalization
of obesity-linked hypogonadotropism via improvement in chronic
inflammation.
DESIGN: Prospective interventional study.
MATERIALS AND METHODS: Regularly menstruating obese women
(34.8 +/-1.2 year-old) underwent frequent blood sampling q 10 min for 8
hours. A 75 ng/kg intravenous bolus of gonadotropin releasing hormone
(physiologic dose) was given at 6 hours [1]. Testing was done in the early
follicular phase BEFORE and AFTER 1 month of omega-3 FA supplementation.
At the completion of baseline studies, a 4 g/day omega-3 FA supplementation
was started on day 1 of the subsequent menses. Luteinizing (LH)
and follicle-stimulating hormones (FSH) were assayed by immunofluorometry
(DELFIA). Plasma FA composition were analyzed by gas liquid chromatography.
A custom 10-cytokine array kit (Raybiotech) was used. Paired t and
Wilcoxon tests were used for normally distributed and skewed data, respectively.
RESULTS: 15 women (BMI 38 +/- 2 kg/m2) completed the study. While
the omega-6/omega-3 ratio has been significantly reduced after the intervention,
it still remained higher than 1:1 ratio that was previously shown to be
beneficial for reproductive function in mice [2]. While pro-inflammatory cytokines
were significantly lower after the intervention, no change was
observed for LH or FSH parameters.
Omega-3 FA Supplementation Improves Inflammation but Not Reproductive
Hormones in Obese Women.
Baseline
Following 1 month
of omega-3 FA
supplementation
LH mean serum level, IU/L 3.3 (0.3) 3.3 (0.3) 0.84
FSH mean serum level, IU/L 4.4 (0.4) 4.2 (0.2) 0.51
LH response to GnRH, 6.6 (1.0) 6.7 (1.2) 0.82
mean serum level, IU/L
FSH response to GnRH, 5.1 (0.4) 4.9 (0.3) 0.71
mean serum level, IU/L
Plasma
8.6 (0.4) 3.9 (0.4)

M. S. Christianson, b W. Shen. a a Obstetrics and Gynecology, Johns Hopkins
O-276 Wednesday, October 21, 2015 12:30 PM
TRANSITION. T. S. Robinson, a T. Gaines, a J. Wu, a MCM8[a-b]. The primary endpoint was time to menopause, defined as R
University School of Medicine, Baltimore, MD; b Johns Hopkins University
VITAMIN D ATTENUATES THE ADVERSE EFFECT OF
ADVANCED GLYCATION END PRODUCTS ON HUMAN GRANU-
School of Medicine, Lutherville, MD.
LOSA CELLS: IMPLICATIONS FOR WOMEN WITH OBJECTIVE: To evaluate the quality of life and experiences of female patients
PCOS. Z. Merhi, a A. Fadiel, a E. Buyuk, b F. Naftolin, a M. Cipolla. c a Obstetrics
with HIV (human immunodeficiency virus) during the menopause tran-
and Gynecology, NYU School of Medicine, New York, NY; b Obstetrics
& Gynecology and Women’s Health, Albert Einstein College of
Medicine / Montefiore M, Bronx, NY; c Obstetrics, Gynecology and Reproductive
Sciences, University of Vermont, Burlington, VT.
sition.
DESIGN: Interview survey of patients at a HIV-clinic.
MATERIALS AND METHODS: We conducted an in-person survey of female
patients at a HIV clinic over 18 months. Inclusion criteria included
women 40 to 50 years old who had at least one menstrual period within
OBJECTIVE: Women with PCOS commonly have elevated levels of the the last 6 months. Interviews utilized the Greene Climacteric scale, a validated
menopausal questionnaire that surveys 21 items on a 4 point Likert
pro-inflammatory advanced glycation end products (AGEs) and low levels
of vitamin D (VD). AGEs are highly reactive molecules that form when lipids scale (0-3) covering 3 domains, including psychological, somatic, and vasomotor
symptoms. Additionally, we queried patients on: (1) if they were in-
or proteins become glycated after exposure to glucose. AGEs accumulate in
the serum and ovaries of women with PCOS potentially altering ovarian forming their primary care physician of menopausal symptoms and (2) if
function. We have shown that: 1) 1,25-dihydroxyvitamin D3 (VD3) supplementation
increases serum levels of the ‘‘anti-inflammatory’’ soluble receptor demographic data was obtained from patient chart review including age,
these symptoms were being addressed and treated by their providers. Basic
for AGEs (sRAGE) in women with PCOS, and 2) VD3 induces a state of race, time of diagnosis and marital status.
luteinization in human cumulus granulosa cells (CCs). We thus hypothesized RESULTS: A total of 23 women ages 40-50 years old agreed to participate
that AGEs adversely affect ovarian function and induce abnormal steroidogenesis
and follicular development. We further hypothesized that VD3 will viewed were African American. The average length of HIV diagnosis was
in an interview. The mean age was 47.1 (SD 2.48). All of the patients inter-
attenuate the AGE-induced ovarian dysfunction.
13.4 years (SD 7.28). The majority of patients, 87% (n¼20), reported experiencing
at least one menopausal symptom with intense frequency, having
DESIGN: Basic science.
MATERIALS AND METHODS: Experiment #1: 72 women who underwent
IVF were enrolled. Follicular fluid (FF) was collected from the first viewed, 100% percent (n¼23), reported experiencing hot flashes to some de-
extreme detrimental effects on their quality of life. Of the women inter-
large follicle and was tested by ELISA for sRAGE, AGEs (pentosidine and gree, ranging from infrequent to persistent episodes. Difficulty in sleeping
N ε-carboxymethyllysine [CML]), 25 hydroxyvitamin D (25 OHD), insulin, was reported by 78% (n¼18) of interviewed women, and on average was
glucose, and SHBG. Experiment #2: CCs (n¼6) were cultured in media (control)
human glycated albumin (HGA; 0.2-0.4 mg/mL) as a source of AGEs of women, 78% (n¼18), reported feeling tired or lacking in energy with mod-
experienced with moderate frequency, 2-3 occurrences a week. A majority
VD3 (50-100 nM) for 48 h after which mRNA was compared using RT- erate frequency. When asked if they told their primary care physician about
PCR for LH receptor (LHR), anti-Mullerian hormone (AMH) and its receptor their menopausal symptoms, 87% (n¼20), reported doing so. Of these patients,
only 20% (n¼4) received treatment for their symptoms, whereas
(AMHR-II), RAGE (pro-inflammatory membranous receptor for AGEs), and
VD receptor (VDR). In addition, RAGE protein was assessed by immunofluorescence.
Experiment #3: KGN granulosa cell line was treated with recom-
CONCLUSIONS: These findings suggest that HIV-infected women under-
80% (n¼16) of these patients went without treatment.
binant AMH (rAMH) with or without HGA VD3 after which going the menopause transition experience intense symptoms that severely
immunofluorescence for SMAD 1/5/8 phosphorylation (AMH signaling impact their quality of life. Although the majority of women reported experiencing
menopausal symptoms to their providers, many of these patients
pathway) was assessed. Data were expressed as % change.
RESULTS: In FF, sRAGE correlated positively with pentosidine (r¼0.24), remain untreated. An opportunity exists to educate providers for this population
on menopause medicine to effectively treat this group and enhance their
CML (r¼0.32), 25 OHD (r¼0.27), SHBG (r¼0.37), and negatively with insulin
(r¼-0.28) and glucose (r¼-0.39) (p

12 months of amenorrhea after the end of chemotherapy. For survivors who
did not undergo chemotherapy, time to menopause was defined as R 12
months of amenorrhea after diagnosis. The association between SNPs and
time to menopause was assessed using Cox proportional hazards models.
An omnibus variable summing the total number of risk alleles that affected
each participant was created for analysis.
RESULTS: Our cohort had a median enrollment age of 40.5 years [range
20.6 - 46] with 76.2% receiving chemotherapy. Median follow up time was
1.77 years [range 0.06 - 5.8 years], and 39 (23%) of participants met criteria
for amenorrhea. Time to menopause was associated with age (HR 1.2 [95%
CI 1.05, 1.26]), BMI (HR 0.92 [0.85, 0.99]) and cyclophosphamide-based
chemotherapy (HR 3.31 [1.17 - 9.40]). No significant association with
time to menopause was found for any SNP (p-value range: 0.22 - 0.88) (Table
1). Restricting to white participants or testing the omnibus SNP variable did
not change findings.
CONCLUSIONS: Thirteen previously identified SNPs associated with
natural time to menopause in genome wide association studies were not
related to timing of menopause in this limited cohort of breast cancer patients.
Supported by: MRSG-08-110-01-CCE, HD058799; T32 HD007203
FEMALE REPRODUCTIVE ENDOCRINOLOGY
P-3 Tuesday, October 20, 2015
GDF-8 DECREASES STAR EXPRESSION THROUGH ALK5-MEDI-
ATED SMAD3 AND ERK1/2 SIGNALING PATHWAYS IN HUMAN
GRANULOSA CELLS. L. Fang, a H. Chang, b J. Cheng, b Y. Yu, a
P. C. Leung, b Y. Sun. a a Reproductive Center, The First Affiliated Hospital
of Zhengzhou University, Zhengzhou, China; b Department of Obstetrics
and Gynaecology, University of British Columbia, Vancouver, BC, Canada.
OBJECTIVE: The aim of this study was to investigate the effects of GDF-
8 on steroidogenic enzyme expression and investigate the potential mechanisms
of action in human granulosa cells.
DESIGN: An established immortalized human granulosa cell (SVOG) was
used as the study model to examine the steroidogenic enzyme expression,
phosphorylation of Smad2/3 or ERK1/2 and progesterone accumulation
following exposure to GDF-8. Signaling pathway involvement was investigated
with inhibitors (SB431542 and U0126) and small interfering RNAs targeting
activin receptor-like kinase (ALK)4, ALK5, Smad2, Smad3 and
Smad4. The concentrations of GDF-8 and progesterone were examined in
the follicular fluid or conditioned medium.
MATERIALS AND METHODS: RT-qPCR and Western blot were used to
measure mRNA and protein levels, respectively. GDF-8 concentrations and
progesterone accumulation were measured by enzyme immunoassay
(ELISA).
RESULTS: Treatment of GDF-8 significantly down-regulated steroidogenic
acute regulatory protein (StAR) expression and decreased progesteron
production. The suppressive effect of GDF-8 on StAR expression was abolished
by the inhibition of TGF-b type I receptor, SB431542. In addition, the
treatment of GDF-8 activated both Smad2/3 and ERK1/2signaling pathways.
Furthermore, knockdown of activin receptor-like kinase (ALK)5 reversed the
effects of GDF-8 on Smad2/3 phosphorylation and StAR down-regulation.
The inhibition of Smad3 or ERK1/2 signalings attenuated the GDF-8-
induced down-regulation of StAR and production of progesterone. Interestingly,
the concentrations of GDF-8 were negatively correlated with those of
progesterone in follicular fluid.
CONCLUSIONS: GDF-8 down-regulates StAR expression and decreases
progesterone production in human granulosa cells, most likely through
ALK5-mediated Smad3 and ERK1/2 signaling pathways.
Supported by: This work was Supported by an operating grant from the
National Natural Science Foundation of China (31471404) to Y.P.S. and
the Canadian Institutes of Health Research to P.C.K.L.
P-4 Tuesday, October 20, 2015
ABORTIFACIENT EFFECT OF THE AQUEOUS EXTRACT OF
STRIGA HERMONTHECA IN FEMALE WISTAR
RATS. I. G. Bako, a H. N. Madugu, b A. Adamu, a I. M. Maje. c a Human
Physiology, Ahmadu Bello University, Zaria, Nigeria; b Obstetrics & Gynaecology,
Ahmadu Bello University, Zaria, Nigeria; c Pharmacology, Faculty of
Pharmaceutical Sciences, Ahmadu Bello University, Zaria, Nigeria.
OBJECTIVE: Large percentage of the world population still rely on herbs
for their primary reproductive health care needs. Striga Hermontheca which
is readily available, is used locally among the rural dwellers for abortion or
induction of labour.
DESIGN: The abortifacient effect of Striga Hermontheca was investigated
by evaluating the serum progesterone level in female pregnant wistar rats after
treatment with the aqueous extract of Striga Hermontheca in varying
doses for three days. The toxicity of the herbs was accessed to ascertain its
safety for consumption.
MATERIALS AND METHODS: The study was compiled with ethical
committee guidelines of Ahmadu Bello University Teaching Hospital, Zaria
with registration number ABUTH/PGO/COMM/0138. The sample of Striga
Hermontheca was collected in Katsina in August 2014 and it was identified
in the Herbarium section of the Department of Biological sciences, with
voucher number 1845. The herbs was extracted using maceration method in
the Department of Pharmacognosy and drug development. Twenty female wistar
rats were randomly grouped into four groups of five rats each (n¼5). Female
wistar rats in their pro-estrous cycle were mated for twenty four (24) hours and
presence of sperm cells in vaginal smear confirmed copulation and day one of
pregnancy. Group I received (control) 10 mg/kg normal saline, Group II
received 200 mg/kg Misoprostol, Groups III and IV received 250 mg/kg and
500 mg/kg of the aqueous extract of Striga Hermontheca respectively. Normal
saline, Misoprostol and aqueous extract of Striga Hermontheca were administered
orally from day five of conception. All data are expressed as Mean SEM
and analyzed using t - test Student’s test, SPSS package 20.0 and post hoc test
for multiple comparison. The (P14 mm in diameter. Categorical variables were assessed by chi-square or
FERTILITY & STERILITY Ò
e107

Fisher’s exact test for small frequencies, with significance at a p-value of
14mm
(p5mg>2.5mg). Because the small number of cases in Group A, statistical
comparison was only carried out between Group B and Group C. Both the
biochemical (15.5% vs. 11.6%) and the clinical (12.8% vs. 9.9%) PRs
were statistically significant increase in Group B when compared to Group
C. The multiple PR and the miscarriage rate were similar between groups.
CONCLUSIONS: Letrozole has been shown to be an efficient and effective
agent in inducing both ovulation and superovulation. There has been
debate as to the optimal starting dose of letrozole. The 5mg-daily yields
higher clinical PRs (p

for treatment management. We embarked on a multi-center initiative to identify
possible subclinical, molecular drivers of idiopathic infertility and treatment
failure.
DESIGN: Multi-center retrospective cohort study.
MATERIALS AND METHODS: Genomic DNA was extracted from
whole blood collected at geographically distinct infertility centers from
women with idiopathic infertility, who had attempted but not achieved a
pregnancy that had progressed beyond week 12 with either timed intercourse
or non-IVF fertility treatments. Medical history and treatment-level clinical
metrics were linked to whole-genome sequence data (30x mean coverage)
generated using Illumina HiSeq sequencers. Novel and rare genetic variants
predicted to be deleterious were identified and filtered using a specially designed
bioinformatics pipeline, incorporating a novel fertility-centric knowledgebase
of >3,000 genetic loci categorized by their role in particular
reproductive processes. Principal component analysis and sequence kernel
association tests were used to analyze the variant signatures of patients in
relation to their infertility treatment histories.
RESULTS: We detected associations between the genetic signatures of patients
and different features of their infertility treatment journeys. For
example, mutations in genes regulating egg quality and early embryo development
were among those most strongly associated with increases in the
number of non-IVF and IVF treatment cycles undertaken before achieving
live birth (LB). We also defined a subset of patients who, after several unsuccessful
non-IVF cycles, achieved live birth after just one IVF treatment.
Analysis revealed that these patients were genetically distinguishable from
those requiring multiple rounds of IVF to achieve LB, as well those experiencing
multiple pregnancy loss.
CONCLUSIONS: Genetic signatures provide novel, subclinical insights
into fertility potential and treatment response that could be a powerful counseling
tool for helping patients progress more quickly to treatment strategies
that will result in a LB outcome.
Supported by: Celmatix Inc
P-9 Tuesday, October 20, 2015
RELATIONSHIP BETWEEN EXPRESSION OF SIRT1 AND SIRT6
GENES AND THE RESPONSE TO OVARIAN
STIMULATION. A. Palumbo, a,b D. Rotoli, c,d R. Gonzalez-Fernandez, d
J. Hernandez, a J. Avila. d a Reproductive Endocrinology, Centro de Asistencia
a la Reproduccion Humana de Canarias, La Laguna, Spain; b Department of
Obstetrics and Gynecology, New York University, New York, NY; c Istituto di
Endocrinologia ed Oncologia Sperimentale (IEOS), National Research
Council (CNR), Naples, Italy; d UDI Bioquimica y Biologia Molecular, Universidad
de La Laguna, La Laguna, Spain.
OBJECTIVE: Sirtuins (SIRTs) are a family of proteins with NAD+-dependent
deacetylase activity with an important function in the response to
different types of metabolic stress or genome injury. SIRT1 and SIRT6
play an important role in basic cellular functions such as genomic stability,
metabolic homeostasis, survival in stress conditions and cellular differentiation.
The objective of this work was to study the expression of Sirt1 and Sirt6
genes in granulosa-lutein cells (GL cells) from IVF patients and its relationship
with the infertility diagnosis and ovulation induction.
DESIGN: Analysis of Sirt1 and Sirt6 gene expression levels by qRT-PCR
in GL cells of young egg donors and IVF patients.
MATERIALS AND METHODS: 82 women were studied, including: 17
egg donors (ED); 15 women >40 years old (yo) with tubal or male factor
and no ovarian pathology (>40); 16 poor responders (PR); 18 endometriosis
cases (EM) and 16 polycystic ovarian syndrome patients (PCOS). After ultrasound
guided egg retrieval, GL cells were isolated from pooled follicular
fluids from each woman using a percoll gradient and anti-CD45 immunobeads
to eliminate WBCs; viability was assessed by trypan blue. After total
RNA isolation and cDNA synthesis, Sirt1 and Sirt6 gene expression levels
were measured by RT-PCR. Expression data were analyzed using the DCt
method and statistical analysis was performed with SPSS using Pearson’s
Correlation coefficient and Student t-test.
RESULTS: Expression levels of Sirt1 in GL cells of patients >40 yo were
higher than in ED and EM, PR and PCOS patients (P40 yo (P40
yo compared to young ED and other diagnostic groups may support a role of
Sirt1 gene in aging. 2) The increased expression level of Sirt6 in EM and PR
may be an indicator of a much higher level of DNA damage and oxidative
stress in both groups. 3) The positive correlation of Sirt1 and Sirt6 with gonadotropins
doses observed only in EM suggests a role of DNA damage and
oxidative stress in the lower response to FSH stimulation observed in EM patients.
Supported by: FIS PI12/00729, Instituto Salud Carlos III, Spain
P-10 Tuesday, October 20, 2015
THE EFFECT OF ORAL TESTOSTERONE UNDECANOATE THER-
APY ON CONTROLLED OVARIAN STIMULATION AND IVF
OUTCOME IN POOR RESPONDERS. G. Mskhalaya, a E. Eltsova, a
V. Zaletova, b E. Lubimkina, b S. Kalinchenko. c a Andrology and Endocrinology,
Center for Reproductive Medicine MAMA, Moscow, Russian Federation;
b Gynecology, Center for Reproductive Medicine MAMA, Moscow,
Russian Federation;
c Endocrinology, People’s Friendship University of
Russia, Moscow, Russian Federation.
OBJECTIVE: To evaluate the efficacy of treatment with oral form of
testosterone undecanoate (TU) before controlled ovarian stimulation
(COS) in poor ovarian responders undergoing in vitro fertilization (IVF)/intracytoplasmic
sperm injection (ICSI).
DESIGN: Prospective controlled trial
MATERIALS AND METHODS: 89 women with poor ovarian response,
defined based on ESHRE consensus/the Bologna criteria of low ovarian
response were included. Patients were randomly divided into 2 groups: TU
treatment group (44 women) or control group (45 women). For TU group
Testosterone undecanoate oral form 40 mg was given daily for at least 40
days (mean - 48 days) preceding COS for IVF. Primary outcome measures
were clinical pregnancy and live birth rates. Statistical research was made using
a software package statistics (StatSoft Inc. U.S., version 12). Quantitative
data is presented as median and quartile range. When comparing the quantitative
data of two independent groups Mann-Whitney U test and Fisher exact
two-tailed test were used. Values were considered statistically significant if p

symptoms were assessed up to 4 times per cycle via questionnaires. Linear
mixed models evaluated the association between serum vitamin A, C, and
E concentrations and individual and grouped PMS symptoms scores and
severity, and generalized linear models were used to evaluate associations
with classification of PMS, adjusted for age, body mass index, and total energy
intake.
RESULTS: Increased levels of serum antioxidant vitamins were not significantly
associated with decreased prevalence or severity of PMS symptoms
either individually or by category (e.g. vitamin C (mg/dL): psychological
symptoms, beta: 0.13, 95% CI: -0.95,1.21; hydration and craving, beta:
-0.11, 95% CI: -0.88,0.67; physical pain, beta: 0.05, 95% CI: -0.73,0.83).
Similarly, increased levels of antioxidant vitamin concentration did not
decrease the chances of moderate or severe PMS classification (e.g. vitamin
C: 5+ moderate/severe symptoms, OR: 1.07, 95% CI: 0.68, 1.69). We also
observed no significant associations between vitamins A and E and PMS
symptoms.
CONCLUSIONS: These data do not support the hypothesis that antioxidant
vitamin concentrations are associated with presence or severity of
PMS symptoms. Therefore, information indicating dietary changes to control
PMS symptoms may be unjustified.
Supported by: Intramural Research Program, DIPHR, NICHD, NIH.
number of metaphase II oocytes per oocyte pick-up. Cancellation rate was
similar in all groups (p¼0.787); however, the probability of cancelling a cycle
was a little bit higher with the corifollitropin alfa (OR¼1.463; 95%IC
0.484-4.424).
CONCLUSIONS: Stimulation with a long-acting gonadotropin yields
similar results in a stimulation protocol in terms of metaphase II oocytes.
The observed differences in the endocrine profile of follicular fluid could
be explained by the structure of the corifollitropin alfa, which induces high
levels of FSH, resulting in a higher concentration of estradiol. In serum,
the higher estradiol concentrations observed in the HP-hMG group might
be explained by the presence pf LH. The absence of significant differences
in the rate of apoptosis confirms the suitability of this treatment in assisted
reproduction cycle
P-13 Tuesday, October 20, 2015
WITHDRAWN
P-12 Tuesday, October 20, 2015
IS THERE ANY DIFFERENCES IN SERUM AND FOLLICULAR
FLUID ENDOCRINE PROFILE AS WELL AS APOPTOSIS RATE
BETWEEN A LONG-ACTING GONADROTOPIN AND HP-
HMG? M. Cruz, a A. Pacheco, a D. Collado, a M. Munoz, b P. Alama, c
A. Requena. a a IVI Madrid, Madrid, Spain; b IVI Alicante, Alicante, Spain;
c IVI Valencia, Valencia, Spain.
OBJECTIVE: To determine the endocrine profile in follicular fluid and the
effectiveness in terms of metaphase II oocytes of a long-term gonadotropin
versus a conventional dose daily in women participating in an oocyte donation
program.
DESIGN: Multicentre prospective randomized study
MATERIALS AND METHODS: 120 oocyte donors have been included.
Subjects were assigned to receive 100 mg of corifollitropin alfa which was
potentially followed by daily administration of recombinant FSH (rFSH)
from day 8 if instructed by the researcher (n¼40) or daily doses of 150 UI
rFSH (n¼40), or 225 UI HP-hMG (n¼40). Daily doses of GnRH antagonist
(0.25 mg/day) were started on day 5 of stimulation in all groups and a single
dose of GnRH agonist (0.2 mg) was administered for triggering oocyte final
maturation. Additionally, it was determined the apoptosis rate by flux cytometry
in cumulus cells as well as the hormonal profile (estradiol and progesterone)
in follicular fluid.An ANOVA test were applied to detect statistical
differences; p

P-14 Tuesday, October 20, 2015
GENETIC INFLUENCES ON ENDOCRINE SIGNALING IN
WOMEN. S. L. Bristow, a R. Shraga, a N. Kumar, a S. Yarnall, a
S. Rodriguez, a A. Bisignano, a S. Munne, b R. Allen, c S. H. Chen. d a Recombine,
New York, NY; b Reprogenetics, Livingston, NJ; c Santa Barbara Fertility
Center, Santa Barbara, CA; d IRMS at Saint Barnabas, Livingston, NJ.
OBJECTIVE: The hypothalamic-pituitary-ovarian (HPO) axis is a critical
signaling pathway in female reproduction, including reproductive hormones
such as gonadotropin releasing hormone (GnRH), follicle stimulating hormone
(FSH), luteinizing hormone (LH), estradiol, and progesterone. Previous
studies have shown that endocrine levels in women are affected by
single nucleotide polymorphisms (SNPs). This study aimed to identify
SNPs impacting the HPO by exploring links between SNPs and baseline
serum hormone levels in women referred from fertility centers.
DESIGN: Retrospective
MATERIALS AND METHODS: Clinical and genetic data was collected
for 143 women less than 40 years of age. Day 2/3 serum FSH, LH, estradiol,
and progesterone levels and AMH levels were collected by chart review. Genetic
polymorphisms within FSH receptor (FSHR), FSH beta polypeptide
(FSHB), anti-Mullerian hormone (AMH), and AMH receptor type II
(AMHR2) were measured using Illumina’s Infinium HD Genotyping assay.
Welch’s t-test was used to test 150 associations between hormone levels
and SNPs. A p-value of pT
(rs10835638; p¼0.034). Finally, LH levels were associated with two AMH variants:
c.-649C>T (rs4807216; p¼0.019) and p.S49I (rs10407022; p¼0.03).
CONCLUSIONS: This study has demonstrated the impact of SNPs on the
HPO signaling pathway, particularly FSHR, FSHB, and AMH. The link between
FSH levels and FSHR SNPs and between progesterone levels and
FSHB SNPs have been shown before, demonstrating our ability to validate previous
findings. We also identified a novel interaction between LH levels and
AMH SNPs, indicating a potential interaction between AMH and LH that has
yet to be characterized. While individual interactions are significant, the combination
of these and other SNPs have not been studied. This endocrine signaling
pathway is highly interconnected. Thus, multiple SNPs inherited together may
influence signaling efficiency in more complex ways than previously thought.
Further and larger studies designed to investigate the possibility of multifactorial
influences on endocrine signaling in female reproduction will lead to a deeper
understanding of the clinical implications of hormone levels on fertility.
P-15 Tuesday, October 20, 2015
WHOLE EXOME SEQUENCING IN MONOZYGOTIC TWINS
DISCORDANT FOR PREMATURE OVARIAN
INSUFFICIENCY. N. Banks, a A. Martinez, b L. Brown, c J. Hughes, c
A. DeCherney, a D. Page, c S. J. Silber, d M. Muenke. b a National Institute
of Child Health and Human Development, National Institutes of Health, Bethesda,
MD; b National Human Genome Research Institute, National Institutes
of Health, Bethesda, MD; c Whitehead Institute, Howard Hughes
Medical Institute, Cambridge, MA; d St. Luke’s Hospital, St. Louis, MO.
OBJECTIVE: Premature Ovarian Insufficiency (POI) affects 1% of
women under the age of 40. Evidence from animal models, the high prevalence
of familial POI (15% of total POI), and the identification of several genetic
causes, all support the hypothesis that POI is largely a genetic disease.
Monozygotic twins (MZ) discordant for the POI phenotype allow for study of
somatic mutations in an otherwise identical genetic background. Novel causative
de novo somatic mutations may be detected by analyzing exome
sequencing in discordant MZ twin pairs.
DESIGN: Whole Exome Sequencing (WES) analysis
MATERIALS AND METHODS: Seven monozygotic twin pairs with one
sibling with POI were consented for WES to IRB approved protocol 11-HG-
0093. Exome sequencing was carried out by the National Intramural
Sequencing Center (NISC, NIH/NHGRI). Capture utilized the SureSelect
Human All-Exon System. Flow cell preparation and 76-bp paired end read
sequencing were performed as per the GAIIx Sequencer protocol. Exome
analysis was conducted using the DNA-Seq module of Golden HelixÒ
SVS 8.3.1 software. Data were filtered for exonic or splicing variants, read
depth greater than 10 and quality score greater than 20.
RESULTS: Each twin pair contained two to 23 variants per pair, with a total
of 81 variants identified. Six variants were in pseudogenes. Twenty-nine
variants were found in known highly polymorphic genes. Forty-six remaining
variants require further analysis and confirmation via Sanger sequencing.
CONCLUSIONS: Based upon our study design and inclusive filtering
strategy, we anticipate a high false positive rate for gene variants detected
with WES. Further analysis and variant confirmation are required. WES of
seven MZ twin pairs discordant for POI offers a unique opportunity to
analyze the role of somatic mosaicism in the pathogenicity of POI and the
potential to identify novel genetic causes.
Supported by: NICHD, NHGRI Intramural Research Programs
P-16 Tuesday, October 20, 2015
RESISTIN DECREASES THE EXPRESSION OF ENDOMETRIAL
RECEPTIVITY RELATED FACTORS THROUGH BINDING TO
TOLL-LIKE RECEPTOR 4 IN ENDOMETRIAL EPITHELIAL
CELLS. J. Yang, a Y. Lu, b Q. Lyu, a Y. Kuang. a a The Ninth Hospital,
Shanghai Jiao Tong University School of Medicine, Shanghai, China; b Department
of Obstetrics and Gynecology, Women’s Hospital, Zhejiang University
School of Medicine, Shanghai, China.
OBJECTIVE: To study the impact of resistin, an adipose-derived peptide
hormone, on the expression of endometrial receptivity related factors in human
endometrial epithelial cells (HEECs) in vitro.
DESIGN: Cross-sectional study and in vitro experiment.
MATERIALS AND METHODS: We detected serum resistin, an adiposederived
peptide hormone, in a group of 93 clinically well-defined infertile patients
with pathology of the fallopian tubes: 42 patients with BMI > 24 and
51 patients with 18.5%BMI %24. The HEECs were isolated from hysterectomy
specimens of 15 premenopausal women undergoing hyerterectomy for
benign reasons, treated with both estrogen and progesterone in vitro and incubated
with or without resistin. Resistin and leukocyte inhibitor factor(LIF) in
serum and supernatant were measured with enzyme-linked immune sorbent
assay (ELISA), and the expression of integrin b3, claudin-4 and dickkopf-
1(DKK-1) were detected with quantitative real-time PCR and western-blotting.
The expression of toll-like receptor 2 (TLR2), toll-like receptor 4
(TLR4) and adenylate cyclase associated protein 1(CAP1) were suppressed
by using specific siRNA transfection.
RESULTS: The clinical data demonstrated that serum elevated resistin
was associated with high BMI in patients who always have lower fertility.
The estrogen combined with progesterone induced expression of endometrial
receptivity related factors including LIF, integrin b3, claudin-4 and DKK-1
could be inhibited by resistin. And TLR4 knockdown also inhibited the
expression of endometrial receptivity related factors, whereas suppression
of TLR2 or CAP1 had no impact on the expression of endometrial receptivity
related factors in HEECs.
CONCLUSIONS: Resistin might bind to TLR4 to perform its inhibition
effect on the expression of endometrial receptivity related factors including
LIF, integrin b3, claudin-4 and DKK-1 in HEECs.
P-17 Tuesday, October 20, 2015
PREDICTING PREGNANCY AMONG WOMEN WITH NO KNOWN
FERTILITY PROBLEMS. K. A. Hahn, a L. A. Wise, b E. E. Hatch, b
K. J. Rothman. c a Epidemiology, Boston University School of Public Health,
Boston, MA; b Boston University School of Public Health, Boston, MA; c RTI
International, Newton, MA.
OBJECTIVE: To create a statistical model for women with no known
fertility problems that provides individual predictions of the probability of
conception based on the women’s characteristics and habits.
DESIGN: Prospective cohort study of women trying to conceive from the
U.S., Canada, and Denmark.
MATERIALS AND METHODS: We pooled participants from two prospective
Internet-based cohort studies of pregnancy planners, including
835 women from the Pregnancy Study Online (PRESTO) and 3785 women
from Snart Gravid and Snart Foraeldre (SG/SF). Eligible women had been
trying to conceive for less than 3 months and had no history of infertility.
We included only those terms in a logistic regression model that made a substantial
difference to goodness of fit, assessed using the likelihood ratio test
p

women aged 25 years who had intercourse 4 or more times a week, were
timing their pregnancy attempts, were non-smokers without a history of vaginitis,
had a cycle length of 28 days, and had normal BMI and were not using
hormonal birth control as their last method of conception to about 8% for nulligravid
women aged 35 years who have intercourse

with accompanying histone modification, we examined C/EBPb binding to
the promoter region of Cyp11a1. The binding of C/EBPb which was analyzed
by ChIP assay was significantly higher at 12 h than 0 h.
CONCLUSIONS: Changes of histone modification status and chromatin
structure in the Cyp11a1 promoter region in addition to DNA hypomethylation
status of the promoter are closely associated with the rapid increase of Cyp11a1
mRNA expression in GCs undergoing luteinization during ovulation. In addition,
it’s suggested that accompanied by binding of C/EBPb to chromatin structural
change, the change of Cyp11a1 mRNA expression is controlled.
P-22 Tuesday, October 20, 2015
A RANDOMIZED TRIAL OF WEB-BASED FERTILITY-TRACKING
SOFTWARE AND FECUNDABILITY. L. A. Wise, a E. E. Hatch, b
J. Stanford, c C. J. McKinnon, a A. Wesselink, b K. J. Rothman. d a Department
of Epidemiology, Boston University School of Public Health, Boston, MA;
b Boston University School of Public Health, Boston, MA; c University of
Utah, Salt Lake City, UT; d RTI International, Newton, MA.
P-21 Tuesday, October 20, 2015
CHANGES IN HISTONE MODIFICATION, DNA METHYLATION
AND C/EBPB BINDING OF THE CYP11A1 PROMOTER REGION
IN RAT GRANULOSA CELLS UNDERGOING LUTEINIZATION
DURING OVULATION. M. Okada, a H. Asada, b H. Tamura, c
N. Sugino. a a Yanaguchi University Graduate School of Medicine, Yamaguchi,
Japan; b Department of Obstetrics and Gynecology, Yamaguchi, Ube,
Japan; c Yamaguchi University Graduate School of Mrdicine, Ube, Japan.
OBJECTIVE: The ovulatory LH surge rapidly alters the expression of steroidogenesis-related
genes such as StAR, Cyp19a1 and Cyp11a1 in granulosa
cells (GCs). Cyp11a1 codes P450scc and plays an important role in
production of progesterone. The ovulatory LH surge induces rapid up-regulation
of Cyp11a1 in granulosa cells (GCs) undergoing luteinization during
ovulation. Recent evidence has shown that epigenetic mechanisms such as
histone modification and DNA methylation are essentially involved in the
regulation of gene expression.
DESIGN: In this study, we investigated whether epigenetic mechanisms
including histone modifications and DNA methylation are involved in the
rapid change of Cyp11a1 expression after LH surge and also investigated
whether transcription factor is associated with the change.
MATERIALS AND METHODS: 21-day-old immature rats were injected
with eCG followed by hCG injection 48 h later. The ovaries were removed
and luteinized GCs were collected before hCG (0 h), and 4 h, 8 h, and 12
h after hCG injection.
RESULTS: 1) In RT-PCR analysis, Cyp11a1 mRNA levels rapidly increased
after hCG injection, reached the peak at 4 h, and remained high level until 12 h.
2) Histone modification status in the Cyp11a1 promoter regionwas analyzed by
chromatin immunoprecipitation (ChIP) assay. The level of trimethylation of
histone-H3 lysine-4, which is an active chromatin marker, was increased and
significantly higher at 12 h than 0 h. The level of trimethylation of histone-
H3 lysine-9 and trimethylation of histone-H3 lysine-27, which is a suppressive
chromatin marker, was decreased and significantly lower at 4 h and 12 h than
0 h. 3) DNA methylation status was analyzed from -1427 bp to -73 bp around
Cyp11a1 promoter region by sodium bisulfite sequencing. 5 CpGs were demethylated
while the other 3 CpGs, which locate at relatively distal region, were
methylated. This DNA methylation profile did not change during luteinization
induced by hCG injection. 4) Chromatin accessibility assay showed that the
chromatin condensation of the Cyp11a1 promoter region decreased after
hCG injection. 5) Since we previously found C/EBPb, which is transcription
factor, is involved in the expression of other steroidgenesis-associated genes
OBJECTIVE: To evaluate whether randomization to use of web-based
fertility-tracking software is associated with improved fecundability.
DESIGN: Randomized Trial.
MATERIALS AND METHODS: The Boston University Pregnancy Online
Study (PRESTO) is a prospective cohort study of female pregnancy planners
aged 21-45 years in the U.S. and Canada. Women were recruited using
internet-based advertisements promoting an incentive to win a premium
membership to FertilityFriend.com (FF), a web-based software application
that records data on menstrual cycles and fertility signs. At baseline, participants
completed a health history questionnaire and were randomized with
50% probability to FF. Women were followed every 8 weeks for up to 12
months or until reported pregnancy. The analysis was restricted to women
who were eligible for FF randomization (i.e., non-users of FF) and who
had been attempting to conceive for %6 cycles at study entry (N¼1,238). Using
an intent-to-treat analysis, we estimated the proportion of women who
became pregnant over follow-up and used proportional probabilities regression
to estimate fecundability ratios (FR) and 95% confidence intervals (CI).
RESULTS: Baseline characteristics were evenly distributed between the
two randomization groups. Median follow-up among women randomized
to FF was 6.6 months (interquartile range: 3.5-9.4 months) and 6.6 months
among non-randomized women (interquartile range: 3.4-10.3 months).
Among the 629 women who were randomized to FF, 392 pregnancies
(62.3%) were reported during follow-up. Among the 609 women who were
not randomized to FF, 379 (62.2%) pregnancies were reported. The overall
FR comparing randomized vs. not randomized to FF was 0.95 (95% CI:
0.84-1.08). Among women who had been trying for 0-2, 3-4, and 5-6 cycles
at study entry, proportions conceiving among randomized and non-randomized
women were 66.4% vs. 69%, 58.7% vs. 57.3%, and 45.6% vs. 30.1%,
respectively. FRs for the association between randomization to FF and fecundability
were 0.90 (CI: 0.78-1.04), 0.97 (CI: 0.72-1.30), and 1.80 (CI:
1.09-2.99), respectively. Of those randomized to FF, 53.7% actually used
the software. The prevalence of FF use was similar across strata of attempt
time at entry (55.2%, 47.9% and 54.4%, respectively), but there were differences
in time to first use, number/type of features used, and intensity of use.
CONCLUSIONS: Overall, there was little evidence that randomization to
the FF menstrual cycle charting software program influenced fecundability
among pregnancy planners participating in an incentive-based internet study.
However, among those who had already been trying to conceive for 5-6 cycles
at enrollment, assignment to FF was associated with faster conception.
Supported by: This research was Supported by NICHD (R21-HD072326).
P-23 Tuesday, October 20, 2015
ARE PLACENTA SPECIFIC PROTEIN 1 LEVELS ASSOCIATED
WITH RECURRENT PREGNANCY LOSS OR IN VITRO FERTIL-
IZATION FAILURE. N. Yilmaz, a H. Timur, a S. Yilmaz, a
A. S. Erdinc, a S. Erkilinc, b H. A. Inal. c a Reproductive Endocrinology
Department, ZTB, Dr Zekai Tahir Burak Women’s Health Research and Education
Hospital, Ankara, Turkey; b Dr Zekai Tahir Burak Education and
Research Hospit, Ankara, Turkey; c Konya Education and Research Hospital,
Konya, Turkey.
OBJECTIVE: To investigate the relationship between the placenta specific
protein-1( PLAC 1) levels associated and recurrent pregnancy loss or in vitro
fertilization failure.
FERTILITY & STERILITY Ò
e113

DESIGN: A total of 28 patients with recurrent pregnancy loss (group 1), 30
unexplained infertile patients with IVF failure ( group 2), 29 fertil patients (
group 3) were included in this cross- sectional study, from January 2013 to
June 2014.
MATERIALS AND METHODS: Recruited patients were younger than 40
years and had no systemic disease. Antecubital venous samples were obtained
from each woman in the morning. Serum samples were separated
and Placenta Specific Protein-1 levels were determined with Human Placenta
Specific Protein-1 (Plac-1) (Cusabio) ELISA KIT method.
RESULTS: There was no significant difference in terms of age and BMI
(p¼ 0.93; 0.20). Serum PLAC 1 levels were significantly higher in group1
when compared to group 3 and 2 (Median of the serum PLAC1 levels:
10.5 vs 1; 10.5 vs 4 ng/ml, respectively) (p

69 minor surgeries and 130 IVF retrievals per year. Average practice size was 5
(range 1 - 50), with an average of 480 fresh IVF cycles (per program) per year.
28% worked with a fellow. 60% were salaried and 40% were equity partners.
Compensation was higher than listed by the American Medical Group Association
and was greatly skewed. Greater than 85% had a positive morale and view of
the future, 92% would choose REI as a career again and 79% would recommend
this career to their children. The most satisfying areas of reproductive medicine
were; patient interactions, intellectual stimulation, interactions with colleagues,
and work schedule. The least satisfying areas were work schedule and financial
compensation. Training was felt to be too focused on female infertility and basic
research with insufficient training on embryology, genetics, male infertility and
clinical research. In the next 5 years, 49% of respondents believed practice size
would increase, and 53% responded someone in their practice would retire. The
average age of planned retirement was 68. 55% suggested that the need for specialists
would stay the same (20% decrease, 18% increase). A total of 58% felt we
were training the right amount of fellows (36% felt we were training a surplus).
Compared to academia, those in private practice reported: higher compensation,
less major surgery, more IVF, less endocrinology and less research. Men worked
more hours, conducted more surgery (and IVF cycles), had higher compensation
and planned to retire later than women.
CONCLUSIONS: Our subspecialty has an extremely high morale. We are
a middle aged subspecialty with disparate compensation and a focused practice.
Some sense a need for a change in our training and most anticipate only
mild growth in our field.
Supported by: SREI
P-27 Tuesday, October 20, 2015
SHOULD ICSI BE RECOMMENDED ROUTINELY IN PATIENTS
WITH SINGLE OOCYTE RETRIEVED? L. Ma, a L. Cai, b J. Dong, b
M. Xia, c J. Liu, b R. Chian. d a Reproductive Medicine, The Center of Reproductive
Medicine, Nanjing, China; b The First Affiliated Hospital of Nanjing
Medical University, Nanjing, China; c Laboratory of Reproductive Medicine,
Nanjing, China; d McGill University, Nanjing, China.
OBJECTIVE: To compare clinical outcomes following in vitro fertilization
(IVF) and intracytoplamisc sperm injection (ICSI) when only a single
oocyte retrieved with non-male factor.
DESIGN: Retrospective study was performed in this study and included
2,133 patients with 3,310 completed treatment cycles from January 2007
to June 2014, who underwent either stimulated or spontaneous cycles that resulted
in only a single oocyte was retrieved.
MATERIALS AND METHODS: Fertilization, clinical pregnancy, miscarriage
and live birth rates per embryo transfer were compared with two insemination
methods and further analyzed whether the clinical outcomes were
affected by the patient’s diagnosis with primary infertility, secondary infertility
and age (38 years). Statistical analyses were performed using SPSS,
version 13.0 (SPSS, Inc), and the clinical outcomes between groups were
compared using the X2 test. The differences in means of demographic data
were calculated by t test, P38 years-old groups.
CONCLUSIONS: There were similar clinical outcomes with IVF or ICSI
as insemination method when the patients with only a single oocyte retrieved.
The choice for insemination method should be based primarily on sperm
quality. Therefore, the low number of eggs retrieved is not an indication
for performance of ICSI.
P-28 Tuesday, October 20, 2015
EXTERNAL VALIDATION AND CALIBRATION OF IVFPREDICT -
CONFIRMATION THAT PROGNOSTIC STRATIFICATION PRIOR
TO RCT ENTRY IS FEASIBLE. S. M. Nelson a A. D. Smith. b a School
of Medicine, University of Glasgow, Glasgow, United Kingdom; b MRC integrative
Epidemiology Unit, University of Bristol, Glasgow, United Kingdom.
OBJECTIVE: IVFpredict a model for prediction of live birth after in vitro
fertilization has been developed and externally validated in UK population
data. Whether IVFpredict is effective in predicting the outcome of multicenter
international randomized controlled trials (RCTs) or whether IVFpredict
could facilitate stratification of patient prognosis prior to RCT entry is
unknown.
DESIGN: Cohort study using participants from the ENGAGE, ENSURE
and PURSUE randomized controlled trials which compared corifollitropin
alpha and recombinant follicle-stimulating hormone.
MATERIALS AND METHODS: The IVFpredict tool uses patient age,
duration of infertility, cause of infertility, treatment, pregnancy and live birth
history, source of oocytes, and use of ICSI as predictors. The discriminatory
power (ability of the prediction tool to discriminate between cycles resulting
and not resulting in live birth) was assessed using the area under the receiveroperator
curve (AUROC). Calibration (how close the tool’s predictions match
observed live birth rates) was assessed by comparing predicted with observed
live birth rate in cycles with different patient and treatment characteristics.
Differential calibration was assessed using Pearson’s chi-square test.
RESULTS: 3292 fresh cycles of IVF were eligible for inclusion, of which
complete data was available for 2591 cycles. The AUROC was 0.599 (95%
confidence interval: 0.576 to 0.623). The predicted live birth rate in the validation
sample was 27.3% compared to an observed rate of 29.3%. There was
no evidence (all p R 0.25) of differential calibration with respect to different
patient ages, duration of infertility, causes of infertility, number of previous
cycles, or use of ICSI.
CONCLUSIONS: External validation, the assessment of a prediction tool
in a population that is different from that in which the tool was developed, is
an essential step towards analyzing a prediction tool’s impact. IVFpredict has
been validated in an international multicentre RCT population and demonstrates
acceptable discrimination and calibration. IVFpredict may be utilized
for stratification of patient prognosis prior to RCT entry.
P-29 Tuesday, October 20, 2015
EFFECT OF BODY MASS INDEX (BMI) ON OOCYTE QUALITY IN
IVF CYCLES. L. Aghajanova, M. Noel, C. Kao, M. Cedars. Obstetrics,
Gynecology and Reproductive Sciences, University of California, San Francisco,
San Francisco, CA.
OBJECTIVE: Data on the impact of BMI on IVF outcomes and human
oocyte quality are conflicting. We aimed to evaluate the effect of BMI on
measures of oocyte quality and treatment outcomes in women undergoing
IVF.
DESIGN: Retrospective cross-sectional study
MATERIALS AND METHODS: Clinical records were examined for all patients
undergoing their first autologous IVF cycle between January 2009-
October 2014 at a single academic institution. Cancelled and egg freezing cycles
were excluded. Patients were grouped by BMI into underweight (30). Basic demographic
characteristics, IVF cycle, oocyte and embryology parameters, and
pregnancy outcomes including implantation rate (IR), pregnancy rate (PR), clinical
pregnancy rate (CPR), miscarriage rate (MR) and live birth rate (LBR) were
evaluated. SAS v9.3 was used for statistical analysis. Data was analyzed with
either an ANCOVA or logistic regression controlling for age. Pairwise comparisons
were performed and adjusted with the Bonferroni method.
RESULTS: A total of 2309 patients met inclusion criteria. 2.8% were underweight
(n¼65), 69.1% normal weight (n¼1596), 18.7% overweight (n¼432)
and 9.4% obese (n¼216). When comparing cycle parameters, peak estradiol
levels were significantly lower in overweight patients compared to those with
normal BMI (difference 239.473.6, p

OBJECTIVE: Obstetricians often recommend waiting 3 months after a
pregnancy loss before attempting to conceive. However, little data supports
this recommendation and previous research has focused on answering the
question ‘‘When should couples achieve pregnancy after a loss?’’ instead
of the more relevant public health question: ‘‘When should couples start
trying to achieve pregnancy after a loss?’’ We compared time to pregnancy
(TTP) leading to a live birth among varying inter-trying intervals (ITI)
defined as time from last pregnancy loss to time starting conception attempt.
DESIGN: Secondary analysis of a multicenter, block-randomized, doubleblind,
placebo-controlled trial to evaluate the effect of preconceptioninitiated
daily low dose aspirin on reproductive outcomes in women with a
history of pregnancy loss.
MATERIALS AND METHODS: 1083 women, aged 18-40 and actively
trying to conceive with 1-2 prior losses and whose last pregnancy outcome
was a loss, were included. Participants were followed for up to 6 menstrual
cycles, and for women who became pregnant, until pregnancy outcome.
TTP was defined as time from starting to try to conceive until date of conception.
Cox models, accounting for left truncation and right censoring, estimated
fecundability odds ratios (FOR) adjusting for age, race, BMI,
education, and subfertility. Multiple imputation addressed missing data.
Sensitivity analyses assessed the potential biasing effects of additional confounding
factors or ITI exposure measurement error.
RESULTS: Couples with a 0-3 month versus >3 month ITI were more
likely to achieve a pregnancy leading to a live birth (53.2% versus 36.1%)
and have a shorter, albeit non-significant, TTP (FOR: 1.17 [95% CI: 0.86,
1.59]). Findings remained robust after adjustment for additional potential
confounding factors (Table). Sensitivity analyses testing the potential impact
of measurement error corroborated our findings.
CONCLUSIONS: Our findings suggest that the traditional recommendation
to wait at least 3 months after a pregnancy loss before attempting to
conceive may be unwarranted.
Sensitivity analyses assessing robustness of findings.
Inter-trying
Interval
Live Birth
n (%)
Sensitivity
Analysis 1
FOR (95% CI)
Sensitivity
Analysis 2
FOR (95% CI)
0-3 months 407 (53.2) 1.22 (0.87, 1.71) 1.16 (0.86, 1.58)
>3 months 84 (36.1) 1.0 1.0
1. Adjusted for age, race, BMI, education, subfertility, partner’s age, smoking,
alcohol intake, parity, previous number of losses, recency of loss, gestational
age of last loss, age of first intercourse, age of menarche, and dilation
and curettage performed for last loss.
2. Monte Carlo sampling applied to assess potential ITI exposure measurement
error. Models were adjusted for age, race, BMI, education, and subfertility.
The average FOR (95% CI) reported for 500 simulations.
Supported by: Intramural Research Program, DIPHR, PRAE, NICHD,
NIH
P-31 Tuesday, October 20, 2015
PERINATAL OUTCOME IN PREGNANT WITH SUBCLINICAL
HYPOTHYROIDISM. S. Agrawal. Obstetrics & Gynecology, Sudha
Hospital Med Research Center, Kota, India.
OBJECTIVE: Pregnancy is associated with major changes in the thyroid
function. The thyroid function is very closely related to the reproductive performance
in women. Hypothyroidism is one of the most common endocrinopathy
seen during pregnancy. Approximately 5% of all pregnancies are
affected by hypothyroidism. Majority of such cases have subclinical hypothyroidism,
which is defined by an elevated serum thyroid stimulating hormone
(TSH) concentration R 3.00 mU/L and a serum free thyroxine (FT4) in the
normal range, i.e. between 0.80 to 1.90 ng/dl. Hypothyroidism has been associated
with pregnancy complications such as preeclampsia, preterm labor,
low birth weight, placental abruption, recurrent abortions, perinatal death
and congenital hypothyroidism in the newborn. There has been a lot of debate
on the impact of subclinical hypothyroidism and pregnancy outcome. Therefore,
we did this large scale, multi-institutional prospective study to evaluate
the consequences of subclinical hypothyroidism on pregnancy outcome.
DESIGN: A multi-instituitional prospective study conducted in the Northern
part of India between April 2010 and December 2014.
MATERIALS AND METHODS: All pregnant women registered at our
antenatal clinics were screened for thyroid dysfunction by serum TSH and
free serum T4 using an immunometric TSH assay. Out of them, 489 women
with TSH concentration R 3.00 mU/L and a serum free thyroxine (FT4) in
the normal range, i.e. between 0.80 to 1.90 ng/dl were recruited as cases and
similar number of controls were recruited with normal TSH and free T4
values. These patients were followed up, delivered at the same hospitals
and their maternal and perinatal outcomes were compared.
RESULTS: Of all patients screened 489 subclinically hypothyroid pregnant
women fulfilled our inclusion criteria were compared with control group.
There was a significantly higher risk of spontaneous abortions, placental
abruption, preterm labor, abruption placentae, low birth weight and neonatal
hypothyroidism in pregnancies with subclinical hypothyroidism.
CONCLUSIONS: Pregnancies with subclinical hypothyroidism have
significantly higher number of adverse outcomes which can be potentially
be prevented by thyroxine supplementation. So routine screening for hypothyroidism
should be initiated for all pregnant women, however more such
studies are needed before making any conclusion.
References:
1. Leung AS, Millar LK, Koonings PP, Montoro M, Mestman JH (1993)
Perinatal outcome in hypothyroid pregnancies. Obstet Gynecol 81:
349-353.
2. Casey BM, Dashe JS, Wells CE, McIntire DD, Leveno KJ, Cunningham
FG 2006 Subclinical hyperthyroidism and pregnancy outcomes.
Obstet Gynecol 107:337-341
3. Abalovich M , Gutierrez S , Alcaraz G , Maccallini G , Garcia A , Levalle
O 2002 Overt and subclinical hypothyroidism complicating pregnancy.
Thyroid 12:63-68
P-32 Tuesday, October 20, 2015
FREQUENCY OF LOW SERUM LH LEVEL DURING OVARIAN
STIMULATION IS ASSOCIATED WITH EARLY PREGNANCY
LOSS IN IVF/ICSI CYCLES WITH A GNRH ANTAGONIST
PROTOCOL. S. Chen, a Y. Chiang, b G. Jou, a C. Chang, c C. Chen. a a Department
of Obstetrics and Gynecology, National Taiwan University Hospital,
Taipei, Taiwan;
b School of Medicine, National Taiwan University
Hospital, Taipei, Taiwan;
c Department of Medical Research, National
Taiwan University Hospital, Taipei, Taiwan.
OBJECTIVE: The influence of low serum LH level in the antagonist protocol
for controlled ovarian stimulation (COS) remained controversial. Previous
studies focused on the LH level on a certain day of COS. Our study is to
evaluate the association between frequencies of low serum LH levels and
pregnancy outcome in IVF/ICSI cycles with a GnRH antagonist protocol.
DESIGN: Retrospective cohort study
MATERIALS AND METHODS: A total of 695 patients underwent COS
for IVF/ICSI cycles using GnRH antagonists and recombinant FSH-only
from January 2011 to December 2012. Serial serum LH levels on day 2 of
menstruation, before and during the administration of GnRH antagonist were
measured until the day of hCG. We documented the lowest serum LH level
in the cycle for each patient, and a threshold value of 0.8 mIU/mL was determined
from logistic regression. Our control group were women with their
lowest serum LH above 0.8 mIU/mL. Women with one or more low serum
LH levels (LH ¼ 2 times) and early pregnancy loss were evaluated.
RESULTS: During COS, there were 535 patients with their lowest serum
LH above 0.8 mIU/mL. In the study group (n¼160), serum LH 0.8 mIU/mL, the incidence
of early pregnancy loss was increased by 1.6 folds in patients with
one episode of LH level

Incidence of early pregnancy loss in various episodes of low LH levels
during ovarian stimulation.
Episodes of LH
¼ 2 2.354 (1.076-5.150) .032
P-33 Tuesday, October 20, 2015
ACTIVIN-A PROMOTES LUTEAL REGRESSION BY DOWN-
REGULATING THE EXPRESSION OF STEROIDOGENIC EN-
ZYMES AND UP-REGULATING BMP-6 AND ACTIVIN-A SUBUNIT
IN HUMAN LUTEAL GRANULOSA CELLS. N. Akin, a G. Bildik, a
Y. Guzel, b A. Seyhan Ata, b B. Balaban, b B. Urman, c,b O. Oktem. c,b a Reproductive
Biology, Koc University Graduate School of Health Sciences, Istanbul,
Turkey;
b Women’s Health Center Assisted Reproduction Unit,
American Hospital, Istanbul, Turkey; c Obstetrics and Gynecology, Koc University
School of Medicine, Istanbul, Turkey.
OBJECTIVE: Activin-A up-regulates the expression of aromatase enzyme
and FSH receptor, and has mitogenic effect on the granulosa cells at early
stage of follicle growth. But its roles on corpus luteum are not clear.
DESIGN: A translational research study
MATERIALS AND METHODS: Human luteal granulosa cells (HLGCs)
retrieved from the patients undergoing natural (n¼10), GnRH antagonist
(n¼10) and agonist (n¼10) IVF cycles were incubated with recombinant activin-A
(60 ng/mL) for 72hrs. The expressions of activin-A beta-A subunit
(INHBA) and activin receptors (ACVR1B, ACVR2A and ACVR2B), steroidogenic
enzymes (stAR, Scc, 3-b HSD and aromatase), LH receptor
and bone morphogenetic proteins (BMPs) 4, 6, 7, 15, and growth and differentiation
factor- 9 (GDF-9) were quantitatively compared by qRT-PCR between
control and activin-A treated cells.
RESULTS: Activin-A and its receptors were expressed by HLGCs of natural
cycles at a significantly higher level than those of GnRH agonist and
antagonist cycles. Compared to control, activin-A significantly decreased
E2 and P production of the cells, reduced their expression of the steroidogenic
enzymes, LH receptor, BMP-4 and 15 and GDF-9; and increased activin-A
beta-A subunit and BMP-6 in both natural and GnRH agonist and
antagonist IVF cycles.
CONCLUSIONS: These results indicate that activin-A expedites luteal
regression in HLGCs. This action of activin-A may open a new avenue in
the treatment of ovarian hyperstimulation syndrome (OHSS).
Granulosa cells before and after activin-A treatment antagonist IVF cycle.
Gene Expression Control Activin-A P-Value
BMP-4 1.000.45 0.640.13

6.4% (3.7-10.9), and 9.8% (6.0-15.4) for AMH quartiles 1-4, respectively (ptrend¼0.52).
Cycle cancellation was also not associated with AMH quartiles
in a multivariate model (p¼0.91).
CONCLUSIONS: Our analysis suggests that AMH may not provide additional
benefit for predicting clinical pregnancy in both FSH and clomiphene
citrate IUI cycles among our patient population.
P-36 Tuesday, October 20, 2015
THE HETEROGENITY OF ANTI-MULLERIAN HORMONE
DEPLETION MODELS ACROSS ASSAYS IN 12917
WOMEN. K. Lukaszuk, a,b L. Plociennik, a A. Lukaszuk, a
D. Bialobrzeska. a a INVICTA Fertility and Reproductive Centre, Gdansk,
Poland; b Gynecology and Oncological Gynecology, Medical University of
Gdansk, Gdansk, Poland.
OBJECTIVE: A regular Anti-M€ullerian hormone diagnosis obtained with
different kits is still an open problem to opinion makers in the field of reproductive
medicine. As far now the literature points out that the profile of the
rate of AMH change varies for available assays. Thus, predominately important
is to measure the heterogeneity among kits.
DESIGN: The study was prospectively conducted on 12917 women (aged
between 20 and 50 years) examined with four AMH assays. AMH measurements
were performed during the time period between January 2007 and
December 2012 in Invicta Fertility Centre, Poland.
MATERIALS AND METHODS: 12917 women were examined with four
AMH assays: Immunotech I generation kit (ImI 4016 samples), Beckman
Coulter II generation kit RUO (BCII RUO 3430 samples), Beckman Coulter
II generation kit with IVD certificate (BCII IVD 830 samples) and Ansh Labs
I generation kit (AnshLabs 4641 samples).
RESULTS: The data revealed the lack of parallelism in AMH depletion
between assays at P< .05, when pairwise comparisons of assays were performed.
Also the distribution of AMH concentration in quartiles between
kits was statistically different (P< .05). Consequently, calculations proved
an interaction between age variable and assays.Based on evidences we
concluded that heterogeneity in the rate of AMH change exists and the central
tendency for AMH levels across assays is substantially different. Finally,
AMH diagnosis in women is dependent on an assay-age effect.
CONCLUSIONS: We claim future studies should endeavour to create a
profiles for heterogeneity and homogeneity of slopes between kits along
age periods. This will give a certain medical aid to decision makers (clinicians),
who perform opinions on infertility problem in patients. Those gynaecologists
concerned with programming options for fertility preservation
might benefit as well.
P-37 Tuesday, October 20, 2015
CREATION & VALIDATION OF THE FERTILITY AND INFER-
TILITY TREATMENT KNOWLEDGE SURVEY (FIT-
KS). R. Kudesia, a E. Chernyak, b B. McAvey. c a Department of Obstetrics
& Gynecology and Women’s Health, Albert Einstein College of Medicine,
New York, NY; b Albert Einstein College of Medicine, New York, NY; c Icahn
School of Medicine at Mount Sinai, Reproduct, New York, NY.
OBJECTIVE: Reproductive decision-making is complicated by a preponderance
of fertility misconceptions. Low fertility knowledge has been
demonstrated across educational levels, in the general population and even
among physicians. We therefore sought to validate a novel instrument assessing
fertility and infertility treatment knowledge for use in both patients and
providers.
DESIGN: Internet-based survey
MATERIALS AND METHODS: An internet-based survey regarding
fertility knowledge and treatment, along with relevant demographics, was
constructed by the research team and examined for clarity and thoroughness
by a panel of reproductive endocrinologists. The pilot survey was distributed
to female students and OB/GYN housestaff at two academic hospitals, as
well as to females in the general population via Amazon Mechanical Turk.
Item analysis and assessment for convergent and divergent validity was
then performed. The study was IRB-exempted.
RESULTS: 127 medical trainees (99 students, 28 housestaff) and 118
women participated. They represented a broad spectrum of geographic,
ethnic and faith backgrounds. The mean scores were 18.8/29 (64.9%) among
medical trainees (MT) and 16.2/29 (55.9%) among the general population
(GP). Among both groups, a majority underestimated the miscarriage rate
(50.4% MT, 59.3% GP), and over-estimated success in delayed-childbearing
scenarios relating to IVF for a 44-year old woman (73.2% MT, 68.6% GP)
and oocyte cryopreservation (96.1% MT, 91.5% GP). There were additional
inaccuracies relating to fertility-impacting lifestyle choices, such as the
impact of moderate alcohol consumption or lubricants. The instrument
demonstrated an appropriate profile in item analysis and validity testing.
CONCLUSIONS: This study demonstrated that the FIT-KS instrument is
an appropriate tool to evaluate understanding of fertility and infertility treatment
knowledge in both patients and providers. The validation process also
reaffirmed that significant gaps remain among medical trainees and within
the general population, and greater efforts to improve reproductive education
must be undertaken.
Supported by: Joan F Giambalvo Fund for the Advancement of Women,
Harold and Muriel Block Institute for Clinical and Translational Research
at Einstein and Montefiore (UL1 TR001073)
P-38 Tuesday, October 20, 2015
THE EFFECTS OF FORMOTEROL ON THE SERUM, PERITO-
NEAL VEGF, AND MDA AND VEGF LEVELS IN THE OVARIES
AND ENDOMETRIUM OF RATS WITH OHSS. H. A. Inal, a
Z. O. Inal, a N. Yilmaz, b H. Timur, c A. Sargin Oruc, d M. N. Kalem, e
O. Han. f a IVF Unit, Konya Education and Research Hospital, Konya, Turkey;
b Reproductive Endocrinology Department, ZTB, Ankara, Turkey; c IVF Unit,
Dr. Zekai Tahir Burak Women’s Health Research and Education Hospital,
Ankara, Turkey; d Zekai Tahir Burak Women’s Health Education and Res,
Ankara, Turkey; e Turgut Ozal University Hospital, Ankara, Turkey; f Guven
Hospital, Ankara, Turkey.
OBJECTIVE: To investigate the effects of formoterol (a beta2-adrenoreceptor
agonist) on serum and peritoneal fluid VEGF, and MDA levels and
on VEGF-stained cell counts in the ovaries and endometrium of rats with
OHSS within the framework of immunohistochemical analysis.
DESIGN: A total of 28 immature female Wistar rats were randomly
divided into four groups.
MATERIALS AND METHODS: Three groups were given 10 IU pregnant
mare serum gonadotropin/day on days 22-25 of life. They were administered
30 IU hCG on day 26 of life to mimic OHSS. On days 26 and 27 of life, 24
mcg/kg/day formoterol in group 3 and 48 mcg/kg formoterol in group 4 were
Comparison of the ovarian cortical and stromal VEGF, endometrial VEGF, serum and peritoneal VEGF lev.
Group
Group1
(Control)
Group 2
(OHSS-Placebo)
Group 3
(OHSS-Formoterol 24 mcg)
Group 4
(OHSS-Formoterol 48 mcg)
p value
Ovarian cortex VEGF* 99.14+8.07 a 156.57+47.80 a 142.86+30.39 121.43+26.24 0.013
Ovarian stromal VEGF 74.42+15.60 a,b,c 179.00+17.37 a 171.85+17.90 b 168.42+10.98 c 0.001
Endometrial VEGF 76.71+12.47 a 125.29+27.47a,d,e 82.86+21.10d 57.00+25.41e 0.001
Blood serum VEGF (mcg/mL) 19.53+6.47 25,83+7,22 25.30+6.00 24.67+6,69 0.281
Peritoneal VEGF (mcg/mL) 22.35+18.16 28.93+7.17 23.52+8.16 22.97+6.98 0.674
Blood serum MDA** (ng/mL) 11.94+2.66 13.37+2.57 13.31+1.88 12.30+1.58 0.543
Peritoneal MDA (ng/mL) 0.72+0.68 1.59+1.38 1.22+1.20 0.91+0.91 0.546
*VEGF: vascular endothelial growth factor; **MDA: malondialdehyde. a¼ The difference between groups 1 and 2 was statistically significant, b¼ the difference
between groups 1 and 3 was statistically significant, c¼ the difference between groups 1 and 4 was statistically significant, d¼ the difference between
groups 2 and 3 was statistically significant, and e¼ the difference between groups 2 and 4 was statistically significant.
e118 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

administered intraperitoneally per animal. The control group (group¼1) was
given the same dosage of 0.9% saline solution (ip) on days 22-26 day of life.-
The main outcomes were weight gain, ovarian volumes and weights, serum
and peritoneal fluid VEGF and MDA levels and VEGF-stained cell count
in the ovaries and endometrium were evaluated.
RESULTS: The weight gain and ovarian volumes were highest in the
OHSS-Placebo group (p¼0.001). The ovarian weights were lower in the control
and formoterol treatment groups than in the OHSS-Placebo group
(p¼0.001). Although, there were no statistically significant differences between
the groups in terms of serum and peritoneal fluid VEGF or MDA levels
(serum VEGF p¼ 0.281, peritoneal VEGF p¼ 0.674, serum MDA p¼ 0.543,
peritoneal MDA p¼ 0.506), there was a significant difference between the
control and the OHSS-Placebo groups (p¼0.013) regarding the VEGF in
the ovarian cortex. There was a significant difference between the control
and the other groups in terms of ovarian stroma (p¼0.001), and there was
also a statistically significant difference between the OHSS-Placebo and
the other groups regarding VEGF in the endometrium (OHSS-Placebo vs.
control group p¼0.002, OHSS-Placebo vs. the formoterol 24 mcg/kg group
p¼0.008, and OHSS-Placebo vs. the formoterol 48 mcg/kg group p¼0.001).
There were no significant differences between the control and formoterol
treatment groups (p¼0.955 and p¼0.372).
CONCLUSIONS: Formoterol represents a potential novel strategy for the
management of OHSS. Further studies, including those examining the
dosage of formoterol, are warranted.
P-39 Tuesday, October 20, 2015
ELEVATED ESTRADIOL LEVELS POST-TRIGGER ARE ASSOCI-
ATED WITH INCREASED ANEUPLOIDY RATES. A. P. Melnick,
R. Setton, E. M. Murphy, Z. Rosenwaks, S. D. Spandorfer. The Ronald
O. Perelman and Claudia Cohen Center for Reproductive Medicine, Weill
Cornell Medical College, New York, NY.
OBJECTIVE: To determine whether elevated estradiol levels post-hCG
trigger are associated with increased aneuploidy rates in patients undergoing
in vitro fertilization with preimplantation genetic screening (IVF-PGS).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients undergoing IVF-PGS with trophectoderm
biopsy and 24-chromosome screening over a three year period at
an academic fertility center were included. Patients were stratified into two
groups by estradiol level post-HCG per mature oocyte (MII) retrieved. The
primary outcome assessed was aneuploidy rate. Secondary outcomes
included total and mature oocyte yield, total days of stimulation, lead follicle
size at trigger, and estradiol level at trigger and one day post-trigger. Statistical
analysis included student’s t-test, chi-square test, and logistic regression.
P

P-42 Tuesday, October 20, 2015
OBESITY AND METABOLISM
OXIDATIVE STRESS AND QUALITY DEGRADATION IN MOUSE
OOCYTES WERE INDUCED BY MATERNAL
OBESITY. M. Kobayashi A. Yoshida. Kiba Park Clinic Research Center,
Tokyo, Japan.
OBJECTIVE: Obesity and overweight increase the risk of many health
problems. In reproductive outcomes of obese women, rates of infertility
and pregnancy loss are higher than normal women. Recently, the relationship
between obesity and oxidative stress is reported and such studies are progressing.
However, the mechanisms of the defects on oocytes from obese female
have not yet been cleared. The aim of this study was to estimate the
effect of maternal obesity on the function of organelles, embryonic development,
and redox status of oocyte.
DESIGN: The meiotic spindle morphology, mitochondrial membrane potential
(MMP), and developmental potency of ovulated oocytes derived from
obese mice were studied. Redox state in ovulated oocytes and sera were
compared between normal and obese mice.
MATERIALS AND METHODS: Female C57BL/6J mice were fed either a
control diet (control) or a high-fat diet (obesity) for 8 weeks. Then, matured oocytes
were retrieved from each group. Morphology of meiotic spindle was
examined immunohistochemically, and mitochondrial membrane potency
(MMP) was evaluated with JC-1 dye. For evaluation of the developmental ability,
oocytes were fertilized by ICSI and cultured in KSOM until to blastocyst
stage. Redox state of ovulated oocytes and sera derived from normal and obese
mice were assessed by dihydroethidium (DHE) staining and d-ROMs/BAP test,
respectively.Reactive oxygen species (ROS) in ovulated oocytes were assessed
by dihydroethidium (DHE) staining. Further, systemic redox states in normal
and obese mice were evaluated with d-ROMs and BAP test of sera.
RESULTS: After 8 weeks, mice fed high-fat diet were significantly heavier
than control (28.32.8g vs. 21.21.2g, p

attributes of a health promotion class (HPC) would be attractive to infertility
patients.
DESIGN: Cross-sectional survey study.
MATERIALS AND METHODS: Patients aged 21-35 presenting to an
infertility clinic were invited to complete an anonymous survey regarding demographics,
perceptions of health, weight status, the effect of excess preconception
weight and potential weight optimization interventional components.
Data was analyzed using regression models.
RESULTS: To date, 24 of the 50 eligible women completed the survey.
Women were typically young (30.1 6.5 years), married (87%), Caucasian
(78%), and college educated (61%) with average BMI of 32 kg/m2. On a 5-
point Likert scale, most consider themselves ‘‘somewhat healthy’’ (64%),
feel ‘‘very/completely confident’’ (64%) that they could perform moderate
physical activity (PA) for 30 minutes 5 times/week, correctly identify PA recommendations
(71%) but are aware that they do not meet these recommendations
(68%). In preparation for pregnancy, patients indicated they should
lose weight (65%) and increase PA (74%), and were aware that excess preconception
weight increases the risk of infertility (86%), pregnancy complications
(86%), and difficult childbirth (82%), however, only 50% were aware
of the increased risk of having an overweight/obese child. Of the 67% of patients
who indicated they would attend a health promotion class (HPC), many
would attend weekly or monthly meetings (58%) lasting 30-60 minutes
(63%) in a community room (42%), gym (42%), online (58%), or via email
(46%), without monetary incentive (85%). Most preferred a HPC that would
provide tips for cheap, healthy eating including recipes and progress
tracking.
CONCLUSIONS: Our study demonstrates that infertile patients are aware
of the risks of excess weight as well as PA recommendations, but do not meet
PA recommendations despite indicating they are physically capable of doing
so. A majority of patients would participate in a HPC without incentive. Preferences
for frequency, duration, location and type of class identified by this
study can be used to develop a HPC tailored to infertility patients.
Supported by: Study funded by Virginia Tech Fralin Life Sciences Institute
P-46 Tuesday, October 20, 2015
ADOLESCENT BODY MASS INDEX IS DECREASED IN CHIL-
DREN CONCEIVED BY INFERTILE COUPLES, REGARDLESS
OF FERTILITY TREATMENT. M. Link, a H. Hanson, b
J. M. Hotaling, c K. Smith, d K. Aston, e D. T. Carrell, f E. B. Johnstone. c a Obstetrics
and Gynecology, University of Utah, Salt Lake City, UT; b Family and
Preventive Medicine, University of Utah, Salt Lake City, UT; c University of
Utah, Salt Lake City, UT; d Population Sciences, University of Utah, Salt
Lake City, UT; e Andrology and IVF Laboratories, University of Utah, Salt
Lake City, UT; f University of Utah School of Medicine, Salt Lake City, UT.
OBJECTIVE: To determine whether birth weight and adolescent body
mass index (BMI) differ among offspring of infertile couples conceived
with and without the use of assisted reproductive technologies and whether
they differ from offspring of fertile controls.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Children of infertile Utah couples born
prior to 1999 were selected for study inclusion (n¼6,251). Data include 165 children
conceived through intrauterine insemination (IUI), 224 children conceived
through in vitro fertilization (IVF) and 5,862 natural conceptions (Infertile - NC).
Offspring of infertile couples were matched to offspring of fertile controls by sex,
birth year (+/- 1 year), year of BMI measurement (+/- 2 years), and multiplicity
(singleton, multiple birth). Primary outcomes of interest were birth weight and
BMI at first driver’s license record. Multivariate generalized linear models
were run adjusting for birth order, gestational age, maternal age, maternal/
paternal education, socioeconomic status, and maternal/paternal BMI.
RESULTS: Table 1 shows a summary of the results. For singleton births,
there were no significant differences in birth weight among IUI/IVF infants,
infertile-NC children, and children of fertile couples. BMI in adolescence
was lower in singletons conceived through IVF and Infertile-NC children
compared to children of fertile controls. For twin births, birth weight was
lower for IVF children but there were no other significant differences in birth
weight between children of infertile and fertile couples. In adolescence, all
twins of infertile couples, regardless of route of conception, had lower
BMIs than children of fertile controls.
CONCLUSIONS: Twins, and to a lesser degree, singletons born to infertile
couples have similar birth weights but slightly lower adolescent BMI than
children born to fertile couples, regardless of the route of conception. Factors
leading to infertility may contribute to in utero stress that is compounded by
twin gestation and may ultimately result in decreased BMI for offspring.
Table 1 - Birth Weight and Adolescent BMI of Children Born to Infertile and
Fertile Couples.
SINGLETONS
Birth Weight
(grams)
Adolescent
BMI
TWINS
Birth Weight
(grams)
Adolescent
BMI
Children of Fertile 3248 21.4 2701 20.0
Controls
Infertile - IUI 3255 21.0 2588 18.7*
Infertile - IVF 3245 20.6* 2467* 18.5*
Infertile - NC 3242 21.0* 2607 19.2*
P-47 Tuesday, October 20, 2015
OBESITY AND HIGH FAT DIET IMPAIR RESPONSE TO SUPER-
OVULATION BY SUPPRESSING GENES INVOLVED IN OVARIAN
FOLLICULOGENESIS. K. Thornton, a O. A. Asemota, b S. K. Jindal, a
M. Charron, a E. Buyuk. a a Obstetrics and Gynecology, Albert Einstein College
of Medicine/ Montefiore Medical Center, Bronx, NY; b Advanced
Fertility Center of Chicago, Gurnee, IL.
OBJECTIVE: Obese women undergoing controlled ovarian hyperstimulation
(COH) have higher cycle cancellation rates, fewer oocytes collected and
lower pregnancy rates. The exact mechanism of how obesity causes reproductive
dysfunction is not well understood. We hypothesize that obesity
and high fat diet are associated with altered ovarian gene expression, which
interferes with folliculogenesis and/or ovulation.
DESIGN: Prospective controlled study.
MATERIALS AND METHODS: Mice were subjected to dietary manipulation
starting at 6w to develop 2 female rodent models: Group 1: C57BL/6J normal
chow fed mice (NC) (N¼12) and Group 2: C57BL/6J high fat diet fed mice (HF)
(N¼12). At 20w of age, 6 mice in each group were sacrificed while another 6 underwent
superovulation (SO) with gonadotropins and then were sacrificed. At the
time of sacrifice, one ovary was snap frozen in liquid nitrogen and used for RNA
extraction and RT-PCR for detecting the following genes: Gdf-9, Bmp-15, Amh,
Amhr (Amh receptor), Foxl2, Gja1 (mouse homolog for human connexin 43),
Fshr and Lhcgr. Ribosomal protein 36b4 was used as loading control. Data given
as units of expression SEM and p

MATERIALS AND METHODS: We reviewed IVF cycles during the
period from 2010 through 2014 at a university IVF center. We included women
with PCOS under the age of 40 years who underwent autologous cleavage stage
(day 2 or 3) fresh embryo transfer. Women treated with metformin during the
IVF cycle were excluded. The included cycles were classified into 2 groups according
to their body mass index (BMI): Normal (BMI %25) and Overweight/
obese (BMI >25). The primary outcomes were clinical pregnancy and life birth
rates. Secondary outcomes included number of oocytes retrieved, number and
quality of embryos, and abortion rates. Good quality embryos included grade A
(cells 8 and 7, fragmentation 0-10% and symmetry perfect to moderate) and
grade B (cells 8 and 7, fragmentation up to 25% and symmetry perfect to moderate)
or (cells 6 and 5, fragmentation 0-10% and symmetry perfect to moderate).
The control group consisted of women without PCOS under the age of 40
undergoing IVF/ET for unexplained infertility.
RESULTS: 90 PCOS cycles were included in the analysis, 56 cycles in the
normal BMI group and 34 in the overweight and obese group. 152 non-PCOS
cycles were used as control. Overweight/obese women with PCOS compared
to normal weight women with PCOS were found to have an increased number
of poor quality embryos and decreased percentage of good quality embryos
(see table). Overweight/obese women with PCOS also had reduced clinical
pregnancy rate, increased abortion rate and a reduced live birth rate compared
to normal weight women with PCOS. In women without PCOS, overweight/
obesity did not have measurable adverse effects on IVF outcome.
CONCLUSIONS: Overweight and Obesity are associated with adverse IVF
outcomes in women with PCOS but not in those with unexplained infertility.
Outcomes by groups.
BMI%25
(n¼56)
PCOS infertility
BMI>25
(n¼34)
p value
Unexplained infertility
BMI%25
(n¼86)
BMI>25
(n¼66)
p value
BMI (Mean SD) 20.94 2.11 31.02 6.69

gas anesthesia. Isotopic tracers of 13C, D3-Met, 13C3-Ser and D2-Cys were
administered as a 4 hour prime-constant rate infusion. Blood samples were
obtained at 3, 3.5, 3.75 and 4 hours. After sacrifice, germinal vesicle stage
oocytes were isolated and denuded. The rate of appearance of each amino
acid being traced was calculated using the tracer dilution method.
RESULTS: Dietary protein restriction caused differential responses in
the kinetics and concentrations of plasma serine and glycine, and folate
supplementation did not ameliorate the observed responses. The total
flux of serine, a methyl group donor, was higher in the LP and LPF groups
(P

dopamine. It has been reported that dopamine negatively regulates follicle
stimulating hormone (FSH) and insulin secretion through the D2 receptor,
and this suggests that SLC18A2 might play a role in polycystic ovary syndrome
(PCOS). Until now, however, there have been no studies on the association
between SLC18A2 and PCOS. We investigated whether naturally
occurring genetic variations at the human SLC18A2 locus are associated
with the etiology of PCOS and/or with the FSH levels and insulin secretion
seen in PCOS pataints.
DESIGN: We undertook a systematic search for polymorphisms in
SLC18A2 by resequencing the gene and investigated whether the common
genetic variants are associated with PCOS in a large cohort
(n¼539). And we also investigated the relationship between genetic variants
and various kinds of metabolic traits in patients. Finally, we studied
function of corresponding genetic variant by luciferase expression experiment
in vitro.
MATERIALS AND METHODS: We genotyped common single-nucleotide
polymorphisms across the locus in 319 PCOS patients who were well
phenotyped for several metabolic traits to determine associations between
SLC18A2 variants and PCOS. We constructed wild-type and mutant plasmids
into HeLa cell and investigated functional of different allele by
measuring luciferase activity.
RESULTS: We found two common genetic variants in the 3’-untranslated
region(rs363282 and rs363238) that are strongly associated with serum FSH
concentration (P¼0.0004 and P¼0.0001, respectively), insulin level at 0.5h
after an oral glucose tolerance test (P¼0.032 and P¼0.009, respectively),
and insulin level at 1h after the test (P¼0.007 and P¼0.005, respectively).
Finally, we performed a functional study that showed that minor alleles of
the two variants decreased expression from a transfected luciferase reporter/SLC18A23’-UTR
expression plasmid.
CONCLUSIONS: Our results strongly suggest that common genetic variants
in SLC18A2 contribute to the phenotypic expression of PCOS and
suggest novel pathophysiological links between the SLC18A2 locus and
PCOS.
Table1.The association of the phenotype and genotype of rs363282 and
rs363238 in patients with PCOS.
Rs363282 Rs363282 Rs363282 Rs363238 Rs363238 Rs363238
AA+AG GG P-value CC+AC AA P-value
Number 245 72 240 67
Age (years) 26.02 0.40 24.66 0.68 0.135 25.94 0.41 24.95 0.74 0.236
BMI (kg/m2) 25.60 0.33 25.15 0.67 0.667 25.43 0.33 25.56 0.72 0.833
E2 (pmol/L) 188.39 9.10 190.89 17.98 0.930 195.25 19.33 195.25 19.33 0.784
T (nmol/L) 2.83 0.05 2.72 0.09 0.211 2.84 0.05 2.70 0.10 0.173
LH (IU/L) 8.91 0.35 10.31 0.87 0.102 8.91 0.35 10.41 0.95 0.074
FSH (IU/L) 7.14 0.16 8.17 0.25 0.0004 7.10 0.15 8.33 0.26 0.0001
FINS (mU/L) 16.57 0.88 17.62 1.53 0.294 16.52 0.89 17.97 1.65 0.288
0.5hINS (mU/L) 89.48 3.16 109.12 12.82 0.032 88.63 3.15 114.46 14.09 0.009
1hINS (mU/L) 110.97 3.70 135.74 13.84 0.007 109.95 3.44 139.92 15.22 0.005
Supported by: This work was Supported by the National Natural Science
Foundation of China (Grants 81270747) and the Shanghai City Board of
Education Scientific Research Innovation Key Projects (Grant 13ZZ001).
P-54 Tuesday, October 20, 2015
IMPACT OF OVARIAN VOLUME-BASED ADJUSTED THERMAL
DOSE VERSUS FIXED-PUNCTURE DOSAGE IN LAPAROSCOPIC
OVARIAN DRILLING ON OVARIAN RESERVE IN CLOMIPHENE
CITRATE-RESISTANT PCOS WOMEN. A. NASR. Obstetrics and Gynecology,
Women’s Health Center, Assiut University, Assiut, Egypt.
OBJECTIVE: To evaluate the impact of adjusted thermal dose on the basis
of ovarian volume versus fixed-puncture dosage in laparoscopic ovarian drilling
(LOD) on reproductive outcome and ovarian reserve assessed by serum
anti-Mullerian hormone (AMH) in women with clomiphene citrate(CC)-
resistant polycystic ovary syndrome (PCOS).
DESIGN: A randomized controlled clinical trial.
MATERIALS AND METHODS: Eighty women with CC-resistant PCOS
who underwent LOD were recruited. They were randomized into two groups:
group A [40 women; mean age: (SD) 27.7 (2.1) years], received an adjusted
thermal dose based on ovarian volume with use of a new model for dose
calculation (60 J/cm3 of ovarian tissue); group B [40 women; mean age:
(SD) 28.5 (1.9) years], received 600 J per ovary through four ovarian holes
regardless of size. A group of normally ovulating women; group C [40
women; mean age: (SD) 27.9 (2.2) years], served as controls. Basal serum
AMH levels were measured in all three groups and six months after LOD
in groups A and B. Women were followed-up for six months. Assuming a
20% difference between the groups, with an a of 5% and a b of 20%, it
was calculated that forty women are required in each arm of the study to
detect a true difference at the 95% confidence level with 80% power. Statistical
analysis was performed using SPSS software, version 17.0 (SPSS, Chicago,
IL, USA). P

of OHSS, especially in patients with PCO. IVM is a relatively new option for
ART promising significant benefits, but still controversial. However, we have
been using IVM as a first choice of ART mainly for PCO patients for more
than 10 years. The present study was conducted to validate if IVM was clinically
useful for infertility by analyzing all pregnant cases of the last 10 years.
DESIGN: Retrospective clinical study at private setting fertility clinic.
MATERIALS AND METHODS: Three hundred and thirty six cycles of
IVM performed from January 2004 to December 2013 were analyzed retrospectively.
Ultrasound monitoring was started from day 7 of menstrual cycle
and oocyte retrieval was performed when the smallest follicle reached 7 mm
in diameter. Regardless of the choice of priming, HCG 10,000 units was
administered and followed by oocyte retrieval 38 hours later. When endometrium
was thicker than 8mm at oocyte retrieval, fresh embryo transfer was
performed. Otherwise, all embryos were vitrified for frozen transfer cycle.
Frozen cycles were not included in the present study. Clinical outcomes
such as number of oocytes retrieved, maturation rate, embryo transfer rate,
and clinical pregnancy rate (GS positive) were analyzed. Moreover, appropriate
timing for oocyte retrieval was evaluated.
RESULTS: Average number of oocyte retrieved was 10.4 (3498 oocytes
per 336 retrievals) and maturation rate was 47.8 % (1683 metaphase two/
3498 retrieved oocytes). Fresh cycle embryo transfer rate was 47.8% (216
transfers per 336 retrievals). Oocyte retrievals on PCO patient rate was
66.9% (225 cycle per 336 cycles). Clinical pregnancy rate with gestational
sac was 31.5% (68 pregnancies per 216 transfers) and 53 babies were born
without any congenital abnormality. Appropriate cycle day for IVM oocyte
retrieval was day 15.4 (between day 9 and 28) for fresh cycle. However,
PCO patients have wide ranges of menstrual cycle days. Therefore, we determined
appropriate timing of retrieval by the percentage of individual menstrual
cycle length. Thirty three percent of cycle length is appropriate for
retrievals in fresh cycle. Average estradiol value of pregnant patients on
the day of HCG was 192 pg/ml.
CONCLUSIONS: Average clinical pregnancy rate of IVM was 31.5% and
it was considered acceptable for clinical validity not only because of less
physical and financial burdens, but also absolute no risk of OHSS. Overall,
IVM is a valid treatment of ART especially for PCO patients when appropriately
performed.
References: Validation of clinical IVM.
P-57 Tuesday, October 20, 2015
CLINICAL OUTCOME OF PCOS PATIENTS UNDERGOING ASSIS-
TED REPRODUCTIVE TECHNOLOGY: THE ROLE OF IN VITRO
MATURATION. T. Takeuchi, a N. Aono, a N. Oka, a R. Obata, a
N. Okuyama, a S. Yanagihori, a T. Okuda, a K. Kyono. a,b a Kyono ART Clinic
Takanawa, Minatoku, Tokyo, Japan; b Kyono ART Clinic, Sendai, Miyagi,
Japan.
OBJECTIVE: Assisted reproductive technology in PCOS patients is often
associated with ovarian hyperstimulation syndrome (OHSS). The severe
form of OHSS is an iatrogenic complication and possibly a life-threatening
condition resulting from ovarian stimulation. In vitro maturation (IVM) techniques
have been developed and employed in clinical settings to circumvent
OHSS by minimizing or eliminating gonadotropin administration. The aim
of this study was to assess the efficacy of IVM in PCOS patients by
comparing its clinical outcome with that of conventional ovarian stimulation
protocols.
DESIGN: Retrospective review of clinical outcome in PCOS patients.
MATERIALS AND METHODS: The average maternal age of the patients
involved was 32.6 3 years. In IVM cycles, no gonadotropin was administered,
but 10,000 IU of hCG 36 hours prior to oocyte retrieval. Oocytes were
cultured in an IVM medium for 24 hours and were inseminated by ICSI. The
endometrium was prepared by administering estradiol from the day of oocyte
retrieval, followed by luteal support. When IVM cycles were not successful,
same patients underwent controlled ovarian stimulation (COS) for harvesting
mature oocytes. Embryological parameters, clinical outcome and complications
were compared between IVM and COS cycles.
RESULTS: A total of 20 consenting PCOS patients underwent 22 IVM
treatment cycles. Following unsuccessful IVM treatment 14 patients underwent
24 COS cycles. The average number of retrieved oocytes in IVM cycles
was 16.9 7, with the maturation and fertilization rate being 41.7% and
49.0%, respectively, while these were 10.4 3, 80.8% and 52.7%, respectively
in COS cycles. IVM yielded more oocytes; however, the number of
fertilized oocytes was similar between the two. Only mild or moderate
form of OHSS was observed in 15 out of 24 (62.5%) COS cycles, while
none developed in IVM cycles. Following fresh and frozen transfers, 8 out
of 20 (40.0%) became pregnant in IVM; similarly, 26 transfers resulted in
10 (41.7%) pregnancies in COS. Pregnancy rate per patient was 33.3% (6/
20) in IVM, and 57.1% (8/14) in COS, cumulatively 70% (14/20).
CONCLUSIONS: Overall clinical pregnancy rate of PCOS patients
following ART was 70%. IVM completely eliminated OHSS and reduced
costs while maintaining a satisfactory pregnancy rate. Thus, IVM can be
offered to PCOS patients undergoing ART as a first line treatment.
P-58 Tuesday, October 20, 2015
ANTI MULLERIAN HORMONE (AMH) LEVELS PREDICT CAR-
DIOVASCULAR RISK ASSESSMENT IN YOUNG WOMEN WITH
PCOS. R. Feldman, a,b S. Butts, c A. Dokras. a,b a University of Pennsylvania,
Philadelphia, PA; b Obstetrics and Gynecology, University of Pennsylvania,
Philadelphia, PA; c Perelman School of Medicine, Philadelphia, PA.
OBJECTIVE: Serum levels of anti-mullerian hormone (AMH), a member
of the TGFb family, are elevated in women with PCOS. There is conflicting
information in women with and without PCOS regarding the association between
low AMH levels and increased cardiometabolic risk. Young women
with PCOS have an increased risk of metabolic syndrome and we hypothesized
that serum AMH levels would predict risk of metabolic syndrome in
this population.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Women seen at the Penn PCOS center
from 2010-2014 with complete metabolic work up were included in this
study (n¼469). Metabolic syndrome (Met Syn) was defined by modified
NCEP-ATP III criteria (BMIR30, sBP/dBPR135/85mmHg or taking antihypertensives
medications, fasting glucose R100mg/ml or taking medications
for diabetes, TGR150mg/ml, HDL-C %50mg/ml). AMH was
measured using the Gen II ELISA, lipids were measured by standard enzymatic
methods and total testosterone was measured by radioimmunoassay.
Spearman correlation coefficients were used to determine relationships between
continuous variables. Linear regression was used to model associations
between AMH and lipids adjusting for confounders. AMH was
dichotomized based on tertiles and univariate tests and logistic regression
modeling were used to evaluate associations of selected variables with tertiles
of AMH.
RESULTS: The median AMH level in the entire cohort was 5.06ng/ml
(IQR 3.08-7.97) . The mean age of the group was 27.66years, 42.1%
were non-white and 8.1% were smokers. The overall prevalence of Met
Syn was 19%. AMH levels positively correlated with total testosterone
(p

ovarian response to gonadotropin stimulation, cannot be a definitive marker
for OHSS. In addition to the desired cohort of leading follicles, gonadotropins
also tend to recruit several AMH producing secondary upcoming follicles
in the mid-follicular phase (around day7) of the stimulation cycle.
Therefore, it would be more prudent to estimate serum AMH level on day
7 of stimulation to get a forewarning of the risk of OHSS in that particular
cycle.
DESIGN: Prospective study of PCOS women (n¼240) undergoing IVF/
ICSI procedure using standard protocol involving COH with 225 IU daily
of recombinant FSH and antagonist.
MATERIALS AND METHODS: Day of commencement of stimulation
protocol was considered day 1. After six days of ovarian stimulation, overnight
fasting serum AMH on day seven (D7) was measured. Antral follicle
count and follicular development was continuously monitored by TVS
from day 7 until at least R 3 lead follicles reached 18 mm diameter size
on attainment of which injection hCG (5000IU) was administered. Serum
Estradiol levels were measured on day of hCG administration. Oocyte
retrieval was done 34 - 36 hours of injection hCG. PCOS women were classified
on the basis of dhCG serum E2 level. group A: no OHSS: E2 8000pg/
ml) was observed in this study. Embryo transfer (ET) was done in all women
except 12 patients of group C.
RESULTS: There was parity in all women w.r.t. age, infertility period,
body mass index, and waist: hip ratio. No significant difference was found
in baseline serum AMH (p¼0.09), and AFC on day1 of stimulation (p¼
0.11) respectively. However, compared to group A, D7 AMH level was
significantly higher in group B (p ¼ 0.0024) and group C (p¼0.0021). There
was also a highly significant difference in D7 AMH levels between group B
and group C (p¼0.0002). Interestingly, the 12 patients in this study where no
ET was done had significantly elevated D7 AMH level compared to the other
30 women of the same group i.e. group C (p

aimed to investigate the diagnostic value of the levels of Progranulin in the
clinical setting of PCOS, and its metabolic effects.
DESIGN: Prospective case-control study.
MATERIALS AND METHODS: Forty-one adolescent patients with
PCOS and 39 adolescent patients as a control group were recruited for
this study in a tertiary referral center in the capital of Turkey. The diagnosis
of PCOS was dependent upon the recent Amsterdam ESHRE/ASRM proposal
and the presence of all three of the Rotterdam criteria for diagnosing
PCOS in adolescents was required (1). Progranulin levels and individual
characteristics of the two groups were compared. Indices of insulin sensitivity,
metabolic variables, circulating androgen levels, lipidemic markers
were measured and blood pressures (BP) were also assessed. To diagnose
the cases with metabolic syndrome (MetS), Cook/Ford modified criteria
were used and patients who had at least 3 of the 5 criteria were diagnosed
with MetS (2).
RESULTS: Progranulin levels in patients with PCOS (7.481.93 ng/mL)
were significantly higher than in the age-BMI matched controls (6.251.98
ng/mL) (p ¼ 0.006). Luteinizing hormone (LH) levels, LH / Follicle stimulating
hormone (FSH) ratios, free testosterone, dehydroepiandrosterone
sulfate (DHEA-S), C-reactive protein (CRP) levels in patients with PCOS
were significantly higher than in the control group (p 0.05). The MetS
was present in 8 (19.5%) of the patients in the study group and in 1
(2.3%) of the patients in the control group (p ¼ 0.029). There was significant
inverse correlation between serum High-density lipoprotein cholesterole
(HDL-C) and Progranulin levels in patients diagnosed with PCOS (p ¼
0.008).
CONCLUSIONS: The MetS is more common among the adolescent patients
with PCOS. There was a significant association between Progranulin
and HDL-C levels. Progranulin may be a novel biomarker for cardiovascular
risk in patients with PCOS, thus these cases should be directed to close
follow-up for possible cardiovascular diseases. Future larger studies should
focus on this entity, thus providing a further vantage point into the role of Progranulin
in the pathogenesis of PCOS.
Demographic and laboratorial features of the subjects.
Variables
PCOS group
(n:41)
Control
group (n:39)
p value
Age (years) 18.92.3 18.72.5 0.629
BMI(kg/m2) 22.73.1 22.22.3 0.440
Waist to Hip
0.800.08 0.790.07 0.436
Circumference Ratio
Progranulin (ng/mL) 7.481.93 6.251.98 0.006
LH/FSH 1.30.6 1.00.6 0.040
Free Testosterone (pg/mL) 1.5 (0.8-4.0) 1.2 (0.6-2.7) 0.014
DHEA-S (mg/dL) 329.811.3 259.0101.8 0.004
CRP (mg/L) 2.2 (0.5-9.9) 1.9 (0.6-6.9) 0.025
natural killer cells and they have been shown to play important roles in cardiovascular
diseases (CVD) and diabetes. We aimed to determine serum perforin
and granzyme B levels in adolescent polycystic ovary syndrome
(PCOS) patients, and to investigate whether they are associated with insulin
sensitivity, obesity and CVD risk markers.
DESIGN: Case-control study.
MATERIALS AND METHODS: A total of 172 adolescents (83 PCOS patients
and 89 healthy controls) were recruited consecutively. Homeostasis
model assessment (HOMA-IR), lipid parameters, anthropometric measurements
were determined. Serum perforin and granzyme B levels were
measured by commercially available ELISA kits. HOMA-IR>2.5 was
considered to indicate the presence of insulin resistance. Multiple Logistic
Regression Analyses were applied for each clinical condition after adjustment
for body mass index.
RESULTS: Adolescents with PCOS had significantly higher levels of fasting
glucose, insulin, HOMA-IR, and total cholesterol/HDL ratio when
compared with controls. Granzyme B levels were also significantly higher
in PCOS patients (median 3.09pg/mL vs 2.21 pg/mL, P¼0.002). No statistically
significant differences were determined among serum perforin levels.
According to the ROC analysis performed for the diagnostic performance
of granzyme B levels in PCOS patients for body mass index (BMI) of >
25 kg/m2, waist circumference (WC) >80 cm, waist to hip ratio (WHR)>
0.8 and HOMA-IR >2.5, the AUC values were all statistically significant.
The best granzyme B cut-off values for distinguishing the groups, and also
the sensitivity, specifity, positive and negative predictive values are given
in Table. Multivariate logistic regression analysis was then used to determine
if a relationship between groups and the defined cut-off levels of granzyme B
was present. Granzyme B levels were found to be independently associated
with increased WHR (OR¼4.256, 95%CI: 1.550-11.687, p¼0.005) and
HOMA-IR (OR¼46.748, 95%CI: 12.431-175.801, p2.06 85.4% 50.0% 59.3% 80.0%
Waist circumference >2.06 80.4% 52.6% 69.5% 66.7%
R80 cm
Waist-to-hip ratio >1.865 84.3% 47.4% 68.3% 69.2%
R0.80
HOMA-IR >2.5 >3.595 87.5% 89.5% 82.4% 92.7%
References:
1. Fauser BC, Tarlatzis BC, Rebar RW, Legro RS, Balen AH, Lobo R, et
al. Consensus on women’s health aspects of polycystic ovary syndrome
(PCOS): the Amsterdam ESHRE/ASRM-Sponsored 3rd PCOS
Consensus Workshop Group. Fertil Steril 2012;97:28-38.
2. Cook S, Auinger P, Li C, Ford ES. Metabolic syndrome rates in United
States adolescents, from the National Health and Nutrition Examination
Survey, 1999-2002. J Pediatr. 2008;152(2):165-70.
P-63 Tuesday, October 20, 2015
INCREASED GRANZYME B LEVELS ARE ASSOCIATED WITH IN-
SULIN RESISTANCE IN ADOLESCENT POLYCYSTIC OVARY
PATIENTS. E. Oztas, a S. Ozler, b A. Tokmak, a N. Yilmaz, c
N. Danisman, a M. Ergin, d H. I. Yakut. a a Zekai Tahir Burak Women’s Health
Education and Res, Ankara, Turkey; b Obstetrician, Perinatolog, Ankara,
Turkey; c Reproductive Endocrinology Department, ZTB, Ankara, Turkey;
d Ataturk Training and Research Hospital, Ankara, Turkey.
OBJECTIVE: Perforin, pore-forming protein and granzyme B, a serin protease
are stored in secretory granules inside the cytotoxic T lymphocytes and
P-64 Tuesday, October 20, 2015
CHOLESTEROL HAS ASSOCIATION WITH BIRTH WEIGHT IN
POLYCYSTIC OVARIAN SYNDROME. A. K. Ahmad, a C. Kao, b
M. Cedars, b H. Huddleston. c a Obstetrics, Gynecology & Reproductive Sciences,
University of California San Francisco, San Francisco, CA; b University
of California San Francisco, San Francisco, CA; c UCSF, Woodside, CA.
OBJECTIVE: Low birth weight (LBW) has been reported to associate
with insulin resistance and increased cardiovascular risk later in life. We assessed
whether birth weight had an association with the development of insulin
resistance and cardiovascular risk in a polycystic ovarian syndrome
(PCOS) population.
DESIGN: Cross sectional.
MATERIALS AND METHODS: Women seen in a multidisciplinary
PCOS clinic from 2006 to 2014 who were diagnosed with PCOS by 2003
Rotterdam criteria were consented and considered for inclusion. PCOS patients
who were born pre-term were excluded from the study population.
Data were systematically collected and included the following metabolic
FERTILITY & STERILITY Ò
e127

and cardiovascular parameters: body mass index (BMI), waist circumference,
cholesterol, low density lipoprotein (LDL), high density lipoprotein
(HDL), triglycerides (TG), aspartate aminotransferase (AST), alanine
aminotransferase (ALT), C-reactive protein, fasting & two-hour insulin, fasting
& two-hour glucose, homeostasis model assessment of insulin resistance
(HOMA-IR), systolic and diastolic blood pressure (SBP, DBP). Birth weight
was analyzed as a predictor of metabolic and cardiovascular parameters as a
continuous variable using multiple and simple regression. Logistic regression
was used to assess the relationship between biochemical hyperandrogenism
(yes or no) and BW as a continuous variable. P-value was set to 0.05. BMI
and age were controlled for as confounding factors.
RESULTS: 230 full-term PCOS patients were identified with mean birth
weight of 3367g (SD 497.5) and mean age at evaluation of 28.43years
(SD 5.64). Of metabolic parameters reviewed, BW was a significantly associated
with cholesterol (-0.0148, p¼0.0043), meaning for each 100g decrease
in BW, cholesterol would increase by 1.48. There was a trend for an association
of BW on hyperandrogenism (OR 0.9995, p¼0.0698).
CONCLUSIONS: There was a significant association between low birth
weight and higher cholesterol levels. In a population already at increased cardiovascular
risk, PCOS patients with LBW should be followed closely with
more frequent screening.
BW as a predictor of metabolic & cardiovascular parameters.
Metabolic &
Cardiovascular
Parameters
Simple
Regression
Coefficient
P-value
Multiple
Regression
Coefficient
(Controlling
for BMI & Age)
P-value
BMI -0.0014 0.2190 — —
Cholesterol -0.0183 0.0005 -0.0148 0.0043
LDL -0.0085 0.0564 -0.0062 0.1523
HDL -0.0024 0.2998 -0.0028 0.1873
TG -0.0130 0.1645 -0.0067 0.4475
CRP -0.0001 0.8994 0.0006 0.5751
Fasting Insulin -0.0007 0.8664 0.0010 0.7764
Fasting Glucose -0.0027 0.1677 -0.0016 0.4092
Two-hour Glucose -0.0122 0.0314 -0.0082 0.1218
HOMA-IR -0.0004 0.7100 0.0003 0.7681
DBP 0.0004 0.8077 0.0014 0.3291
P-65 Tuesday, October 20, 2015
PREDICTIVE MARKERS OF ABNORMAL GLUCOSE INTOLER-
ANCE IN POLYCYSTIC OVARY SYNDROME. D. Lee, J. Jeon,
S. Park, H. Chung, K. Jeong. Department of Obstetrics and Gynecology,
School of Medicine, Ewha Womans University, Seoul, Korea, Republic of.
OBJECTIVE: To identify predictive markers of abnormal glucose metabolism
(IGT/NIDDM) in Korean women with polycystic ovary syndrome(P-
COS).
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: A total of 312 PCOS women were evaluated.
All patients underwent 75g-oral glucose tolerance tests (OGTTs). The
2 hour plasma glucose level was used to categorize subjects as IGT or
NIDDM. Areas under the curve (AUC) of the receiver operating characteristic
curves (ROC) were used to compare the power of serum markers. Multiple
linear regression analysis was used to evaluate the contribution of each
confounding factor for 2hr post-load glucose value.
RESULTS: Two hundred eighty five (91.3%) PCOS women with normal
glucose tolerance and 27 (8.7%) PCOS women with abnormal glucose metabolism
(IGT/NIDDM) were evaluated in this study. AUC of HbA1c, high
sensitivity CRP, lipid accumulation product (LAP) index, and triglycerides
were 0.780, 0.772, 0.762, and 0.758 respectively. ROC analysis suggested
a threshold value of 5.45 in HbA1c (71.4% sensitivity and 70.0% specificity),
a value of 1.16 in high sensitivity CRP (70.3% sensitivity and 80.1% specificity),
a value of 12.98 in LAP index (88.5% sensitivity and 52.3% specificity)
and a value of 88.0 in triglycerides (77.8% sensitivity and 63.5%
specificity) for prediction of abnormal glucose metabolism. However,
glucose to insulin ratio (G:I ratio) and quantitative insulin sensitivity check
index (QUICKI) could not replace the role of OGTT in screening of IGT
and NIDDM.
CONCLUSIONS: HbA1C, high sensitivity CRP, LAP index, and triglycerides
can be useful predictive markers of abnormal glucose metabolism
(IGT/NIDDM) in Korean PCOS women.
Comparison of areas under the ROC curves(95% CI) for potential markers
for normal and abnormal(IGT/T).
Areas under
ROC(95%-CI)
Supported by: Korea Centers for Disease Control and Prevention.
P-66 Tuesday, October 20, 2015
ABERRANT EXPRESSION OF MICRORNAS IN CUMULUS CELLS
ISOLATED FROM PCOS PATIENTS. X. Huang C. Hao. Reproductive
Medicine Centre, Affiliated Hospital of Qingdao Medical University, Yantai
Yuhuangding Hospital, Yantai, China.
OBJECTIVE: Recent data has pointed to the importance of miRNAs in
disease and embryo development. The developmental competence of oocytes
and embryos in PCOS patients is reduced to a certain extent during the in vitro
fertilization (IVF) process. Cross-talk between the oocyte and cumulus
cells is critical for oocyte maturation and embryo competence. The aim of
this study is to determine if microRNAs are differentially expressed in the
cumulus cells of PCOS patients compared to non-PCOS patients and identify
associated altered gene expression.
DESIGN: Descriptive study.
MATERIALS AND METHODS: MicroRNA analysis was performed using
Human miRNA Microarray Release 18.0 microarray between cumulus
cells isolated from PCOS (n¼5) and non-PCOS (n¼6) patients. Several miR-
NAs were selected to validate the microarray results using quantitative RT-
PCR on additional samples. Potential gene targets were identified and their
expression analyzed by quantitative RT-PCR.
RESULTS: The microarray profiling of cumulus cells revealed expression
of 386 miRNAs, 17 of them were differentially expressed between the two
groups. Annotation of predicted gene targets for these differentially expressed
miRNAs included genes involved in several pathways as focal adhesion,
regulation of actin cytoskeleton, MAPK signaling pathway, and Wnt
signaling pathway. Several predicted miRNA target genes were selected
for analysis and demonstrated significant altered expression consistent
with aberrant miRNA profiles.
CONCLUSIONS: This study describes for the first time that aberration of
cumulus cells microRNA expression is associated with PCOS. With growing
evidence indicating the importance of miRNAs during oocyte development
of PCOS patients, new molecular biomarkers or a new basis for the diagnosis
of PCOS would be provided and the etiology of PCOS might be improved.
Supported by: This study was Supported by the National Natural Science
Foundation of China (Grant 81401172 and 81170622) and the Natural Science
Foundation of Shandong Province (Grant ZR2013HQ004).
P-67 Tuesday, October 20, 2015
p-value
(asymptotic
significance
Cut-off
(calculated by Sensitivity Specificity
of area¼0.5) Youden’s index) (at cut-off) (at cut-off)
HbA1C 0.780(0.66-0.89)

OBJECTIVE: The benefit of choosing GnRH agonist versus antagonist in
women with PCOS has yet to be determined. The existing literature does not
reach a firm consensus. The objective of this study was to compare reproductive
outcomes between agonist and antagonist protocols in PCOS patients undergoing
IVF.
DESIGN: Retrospective cohort study in an academic setting.
MATERIALS AND METHODS: Women undergoing IVF from 2007 to
2015 and meeting Rotterdam PCOS criteria were included. Data regarding
baseline demographics, clinical parameters, and treatment outcomes was
collected. Provider choice determined participants’ IVF protocol. Descriptive
statistics were computed. Bivariate analysis was performed using Student’s
t, Mann-Whitney U, Pearson’s chi-squared, and Fisher’s exact tests
as appropriate. Multivariate analysis was completed by stratifying outcomes
by demographic and baseline clinical parameters when p

explore the relationship between sRAGE and PCOS and the relationship between
sRAGE and IVF-ET Results in PCOS.
DESIGN: Prospective study.
MATERIALS AND METHODS: A total of 80 patients of childbearing age
were observed and 30 PCOS and 28 non-PCOS women were enrolled into the
final analyses for parameters of IVF-ET results. Measurements of sRAGE,
VEGF in plasma at early follicular phase and leading follicular fluid aspirated
without blood at the time of egg collection by ELISA to examine the
difference and the correlation. Parameters of IVF-ET results including the
number of the oocyte retrieved, the number of the mature follicles, the normal
fertilization rates, and the high-quality embryos.
RESULTS: 1 Compared with the non PCOS group, the concentration of
sRAGE in plasma and follicular fluid in the PCOS group both were significantly
higher (plasma, 1351.79290.62 vs 1185.00205.71, t¼2.148,
P¼0.046. follicle fluid, 1650.23239.711 vs 1034.18156.010, t¼11.510,
P¼0.000). In PCOS group, the follicular fluid concentration of sRAGE
was positively related to the VEGF (r¼0.988, P¼0.000). 2 In PCOS, the concentration
of sRAGE in follicular fluid has no relationship with the number of
the oocyte retrieved(r¼0.375 ,p¼0.103) and the number of the mature follicles
(r¼0.409,P¼0.074), but it was positively related to the IVF-ET parameters
including the normal fertilization rates(r¼0.540,P¼0.014), normal
cleavage rates(r¼0.489,P¼0.029)and the high-quality embryos(r¼0.539,
P¼0.014 ) when the age,BMI,T and LH/FSH were adjusted.
CONCLUSIONS: sRAGE may affect the development of inflammatory
factor generated in PCOS. The concentration of sRAGE in the follicle fluid
was positively related to the IVF-ET parameters,and could be a biomarker to
predicate IVF-ET results with PCOS.
P-71 Tuesday, October 20, 2015
THE CORRELATION BETWEEN AMH AND LABORATORY PA-
RAMETERS IN PCOS WOMEN ACCORDING TO SUBTYPE: A PI-
LOT-STUDY. Y. Kim, S. Lee, K. Yi, H. Park, J. Shin, T. Kim,
J. Hur. Department of Obstetrics and Gynecology, Korea University Guro
Hospital, SEOUL, Korea, Republic of.
OBJECTIVE: Do AMH level correlate with hormonal and metabolic parameters
in women with PCOS according to subtype?
DESIGN: A university hospital-based observational study.
MATERIALS AND METHODS: The observational study of women with
PCOS between January 2013 and November 2014 in University-based medical
center. Seventy of women with PCOS were classified according to insulin
resistance(IR) by the homeostasis model assessment-estimated insulin
resistance(HOMA-IR) or androgen excess(AE) by free androgen index(FAI).
The following parameters were analyzed; age, BMI, basal and 2 hour-post
prandial(PP2) insulin level, basal and PP2 glucose, HbA1c, testosterone,
DHEA, SHBG, AMH, cholesterol, triglyceride, LDL, HDL, and CRP. Between
IR(-, n¼33) and IR(+, n¼37), and AE(-, n¼25) and AE(+, n¼45),
the correlation between AMH and analyzed parameters were investigated.
RESULTS: In overall analysis, AMH had significant positive correlation
with SHBG (r¼0.385, P¼ .001), testosterone (r¼0.349, P¼ .004), and
HDL (r¼0.340, P¼ .006). In IR (+) type, AMH showed correlation positive
with testosterone (r¼0.337, P¼ .041), HDL (r¼0.486, P¼.003) and negative
with CRP (r¼-0.376, P¼ .024), basal and PP2 glucose (r¼-0.343, P¼ .037;
r¼-0.347, P¼.035) whereas not in IR (-) type. In AE (+) type, AMH showed
correlation positive with HDL (r¼0.372, P¼ .014) and negative with PP2
glucose (r¼-0.303, P¼ .038) although not in AE (-) type.
CONCLUSIONS: AMH may show positive correlation with SHBG and
testosterone regardless of PCOS subtype, and have different correlation
with metabolic parameters according to IR and AE. To clarification of its
clinical meanings, further study with large scale could be necessary.
DESIGN: Cross-sectional study with control group.
MATERIALS AND METHODS: 242 patients with PCOS (Rotterdam
criteria, 2003) aged 16-34 years and 166 women (control group) aged 16-
35 years were studied between 01.01.12 and 07.31.14.Prior institutional
ethical approval and informed written consent (from all participants/ legal
guardians) were obtained. Body mass index (BMI; Kg/M2), abdominal
circumference (AC, cm), waist-hip ratio (WHR), Ferriman Galwey score,
presence of acne (%) and acanthosis nigricans (AN, %) and blood pressure
were assessed. Serum testosterone (T), sex hormone binding globulin
(SHBG), post-prandial serum glucose (PPG) and insulin (PPI) levels 2 hours
after 75 glucose intake were measured. FAI was calculated as [T (ng/ml) x
100 x 3.47] / SHBG (nmol/l). PPG R 140 mg% was considered as AGT.
PPG: PPI % 1.0 was considered as indicative of IR 1.
RESULTS: Prevalence of AGT was 11.9% (95% CI ¼ 7.9%, 16.1%)
among PCOS patients and 12.6% (95% CI ¼ 7.6%, 17.7%) in the control
group (not significantly different). PCOS group was younger and had higher
BMI and AC. In regression analysis, after controlling for the covariates,
PCOS status was not associated with AGT in our study population; BMI
(p¼ 0.011) and WHR (p¼0.027) were the only significant positive predictors
(for AGT). Among the PCOS patient population (n¼242), univariate analysis
demonstrated that those with IR were significantly younger (p¼0.003) with
higher BMI (p

P-74 Tuesday, October 20, 2015
EXCESSIVE BODILY RETENTION OF ORGANOCHLORINE
PESTICIDE IS ASSOCIATED WITH ENERGY IMBALANCE AND
INFLAMMATION IN WOMEN WITH PCOS: A CASE CONTROL
STUDY. Y. Zhao. Peking University Third Hospital, Beijing, China.
OBJECTIVE: Our aim was to investigate the relationship of the serum
levels of organochlorine pesticides (OCPs) and energy homeostasis factor
adropin with the risk of polycystic ovary syndrome (PCOS), as well as
explore the association between bodily retention of OCPs and metabolic homeostasis
and inflammation of PCOS patients.
DESIGN: This was a preliminary case-control study undertaken at the Division
of Reproductive Center, Peking University Third Hospital. A total of
50 participants affected by PCOS and 30 normal controls were recruited between
August and October 2012 from Northern China. All participants were
Han women.
MATERIALS AND METHODS: PCOS participants were diagnosed according
to the 2003 Rotterdam criteria. The control participants were nonpregnant
females unable to conceive solely due to male azoospermia. Serum
levels of three typical OCPs were analyzed using gas chromatographic mass
spectrometry (GC-MS), including p,p’-dichlorodiphenyldichloroethylene
(p,p’-DDE), hexachlorobenzene (HCB), and b-hexachlorocyclohexane (b-
HCH). Serum levels of adropin and C-reactive protein (CRP) were measured
by ELISA kits (R&D System).
RESULTS: Serum levels of three OCPs were significantly higher in the
PCOS group than the control group. After adjustment for age and BMI, concentrations
of p,p’-DDE and the sum of all OCPs ( P OCPs) in serum above
median levels remained significantly associated with PCOS, and ORs were
2.94 [95% confidence interval (CI), 1.11-7.78] and 5.03 [95% confidence interval
(CI), 1.79-14.11], respectively. Partial least-squares-discriminant analysis
(PLS-DA) confirmed that serum levels of OCPs were risk factors of
PCOS, especially for p,p’-DDE. Moreover, dramatically reduced serum level
of adropin and enhanced serum level of CRP was detected in PCOS group
compared with the control group, indicating the energy metabolism imbalance
and inflammatory status in PCOS women. Furthermore, serum levels
of adropin correlated negatively with p,p’-DDE (R¼0.219, P¼0.035), b-
HCH (R¼0.450,P

similar cycle outcomes can be achieved in comparison to their normal baseline
E2 counterparts.
DESIGN: Retrospective analysis.
MATERIALS AND METHODS: 110 patients undergoing oocyte cryopreservation
or IVF at our center from 2013-2014 were included. Estrogen
prime patients were excluded. Patients were separated into two groups.
Group 1 included 31 patients with E2 levels greater than 70 pg/ml on day
2 with subsequent ganirelix acetate given for 2-3 days and repeat E2 levels
of less than 70 pg/ml. Group2 included 79 patients with standard day 2 E2
levels of less than 70 pg/ml whom initiated their cycles immediately without
the use of ganirelix acetate. Groups were compared with t-tests of the means
as well as chi-squared analysis for cycle outcomes.
RESULTS: Group 1 was composed of 12 egg freeze patients and 19 IVF
patients. Group 2 was composed of 1 egg freeze patient and 78 IVF patients.
Table 1 shows the difference in baseline characteristics between group 1 and
group 2. Specifically, there were no significant differences seen in age, FSH
levels, as well as historic high FSH levels and AMH levels. Day 2 E2 levels
were significantly different between the groups, as seen in table 1. There was
also no difference in cycle stimulation parameters. Group 1 had an average
egg yield of 9.2, as opposed to 14.6 in group 2. This was statistically significant
with a p-value of 0.01. Of patients who underwent trophoectoderm (TE)
biopsy (53 patients in both groups), group 1 had 33% (8/24 total biopsied embryos)
euploid embryos as compared to group 2 which had 36.6% euploid
embryos (91/249 total biopsied embryos). This was not significant, p-value
of 0.75. A total of 6 transfers in group 1 and 25 transfers in group 2 were performed.
66.67% of patients in group 1 had a positive beta HCG after transfer
as opposed to 76% in group 2 which was not significant, p-value of 0.49.
Clinical pregnancy rates in group 1 were 66.67% in group 1 and 72% in group
2, also not significant, p-value of 0.58.
CONCLUSIONS: Patients with high baseline E2 levels on day 2 of cycle
demonstrate lower egg yields despite adequate suppression of E2 with ganirelix
acetate however, this did not translate to lower clinical pregnancy rates or
lower euploid embryo rates. Given the lower egg yields in the high day 2 E2
group this may be of clinical importance in oocyte cryopreservation cycles.
Cycle characteristics and outcomes.
Group 1
(E2>70 pg/ml)
Group 2
(E2)
p-value (*all t-tests
of the mean,
**chi-square)
Age (years) 36 (+/- 6.2) 37 (+/- 4.9) 0.22*
Day 2 E2 (pg/ml) 106 (+/- 45) 44 (+/- 14.3)

ulation but is a relatively poor test for prediction of pregnancy and live birth.
Patients with low levels of AMH still can achieve reasonable treatment outcomes
and should not be precluded from attempting IVF solely on the basis
of an AMH value.
Supported by: Natural Science Foundation of Guangdong Province (Grant
No. S2013040013849).
P-80 Tuesday, October 20, 2015
THE EFFECT OF POSTPARTUM PROGESTOGEN ON CARDIAC
FUNCTION, NT-PROBNP, AND INFLAMMATORY BIOMARKERS
IN WOMEN WITH GESTATIONAL HYPERTENSION. J. S. Yeh, a
M. Daubert, b M. Kuchibhatla, c F. Z. Stanczyk, d T. M. Price. a a Department
of Obstetrics & Gynecology, Division of Reproductive Endocrinology &
Infertility, Duke University Medical Center, Durham, NC; b Department of
Medicine, Division of Cardiology, Duke University Medical Center,
Durham, NC; c Department of Biostatistics & Bioinformatics, Duke University
Medical Center, Durham, NC; d Department of Obstetrics and Gynecology,
University of Southern California, Los Angeles, CA.
OBJECTIVE: The discovery of a mitochondrial progesterone receptor
(PR-M) established a mechanism by which progesterone increases cardiac
cellular respiration. Because human myocardium contains the most mitochondria
per volume than any other tissue, progesterone may have a role
in increasing energy production to meet the greater metabolic demands of
pregnancy. Furthermore, transgenic mice over-expressing PR-M show
higher levels of cardiac cellular respiration and are less susceptible to
heart failure after aortic constriction. Therefore, our primary objective
was to investigate if a progestogen can affect human heart function by
determining if postpartum (PP) administration of Depot medroxyprogesterone
acetate (DMPA) has an effect on cardiac performance, NT-ProBNP,
and/or inflammatory biomarkers in women with gestational hypertension
(GH).
DESIGN: Prospective longitudinal cohort pilot study.
MATERIALS AND METHODS: Ten patients with GH participated in
this study. The DMPA group (DG) consisted of five subjects given
DMPA for contraception on PP day 1, while the control group (CG)
received no exogenous hormones. Subjects underwent echocardiograms
and serum collection for IL-6, TNFa, and NT-ProBNP at three testing sessions
(TS): 2 weeks prior to delivery, 1 day PP, and 2 weeks PP. Absolute
values, net change and percent change were calculated for and between
each TS. Data were analyzed using Student’s t-test, Mann-Whitney U
test, and Spearman’s correlations.
RESULTS: Subjects completed the three TS at an average of 37.5 weeks,
1.0 day PP, and 15.0 days PP. All patients showed evidence of at least mild
left ventricular (LV) hypertrophy by the first TS. By the third TS, the DG
had significantly better diastolic function than the CG, with function positively
correlated to serum levels of medroxyprogesterone acetate (Spearman’s
r(3)¼1.000, p

Comparison of groups related to cycle outcomes.
Group A (P4)
Group B
(P4>1.5 ng/ml)
p-value
(*t-test,
**chi-sqaure)
Age (years) 38.4 +/- 4.1 38.4 +/-3.7 0.89*
Total Gonadotropins (IU) 3795.34 +/- 1539 4395.2 +/- 1521.7 0.02*
Estradiol at trigger (pg/ml) 2483.8 +/- 1048.1 3034.2 +/- 1495.3 0.02*
# Eggs retrieved 14.5 +/- 7.9 15.7 +/-7.0 0.34*
# Embryos biopsied 5.0 +/- 3.97 5.2 +/-3.5 0.73*
% Euploid Embryos 32.7 +/-31.5 29.4 +/- 29.2 0.51*
% No diagnosis embryos 10.4 +/- 26.4 11.2 +/-26.4 0.86*
% No normal embryos 32.1 (26-38.9) 25.5 (15.1-39.6) 0.19**
Clinical pregnancy rate (%) 72 (62.1-79.5) 66.7 (47.7-81.5) 0.31**
Implantation rate (%) 59.8 (50.6-68.4) 66.7 (47.7-81.5) 0.43**
Ongoing/Live birth rate (%) 65.6 (56-74.2) 66.7 (47.7-81.5) 0.46**
P-83 Tuesday, October 20, 2015
OVARIAN FUNCTION
EXPRESSION OF HUMAN TELOMERASE REVERSE TRAN-
SCRIPTASE (TERT) IS STAGE SPECIFIC IN THE OVARIAN
FOLLICLE. J. A. Dundee, a N. Esfandiari, b M. M. Blanchette Porter, c
E. McGee. d a ObGyn, University of Vermont, Burlington, VT; b ObGyn, Dartmouth
Hitchcock Medical Center, Lebanon, NH; c Dartmouth Hitchcock
Medical Center, Lebanon, NH; d University of Vermont, Burlington, VT.
OBJECTIVE: The TERT gene encodes telomerase, the activity of which is
required to maintain telomere length. Shorter telomeres and decreased telomerase
activity are associated with cellular dysfunction, ovarian insufficiency
and poorer ART outcomes. In this study TERT expression was
evaluated in human mural granulosa cells and follicles at varying stages in
ovarian tissue.
DESIGN: basic science research using human cells and tissues.
MATERIALS AND METHODS: Mural granulosa cells were isolated
from follicular fluid pooled from women undergoing IVF. Samples
were used to determine expression of mRNA encoding TERT and c-
Myc in both fresh and cultured granulosa cells. Immunohistochemistry
was performed using a TERT antibody on paraffin sections from four
normal ovaries from a pathology tissue bank. TERT protein was also
localized in cultured human granulosa cells. Telomerase activity
(TRAPeze assay; Millipore) was measured in DNA extracted from fresh
mural granulosa cells.
RESULTS: Unexpectedly, TERT mRNA was not detected in human mural
granulosa cells by PCR. To verify the samples and the technique, control
studies were performed. The same technique amplified TERT from other tissues.
C-Myc, which controls TERT gene transcription in breast cancer cells,
did amplify in the same samples for which no TERT was amplified. To
further characterize the timing and nature of TERT expression in granulosa
cells, we performed IHC for TERT protein in ovarian sections from 4
different women. TERT protein is expressed in the nucleus of granulosa cells
from the primary stage onward, in a stage specific pattern. Expression of
TERT protein is fairly uniform in granulosa cell nuclei in pre-antral and
small antral follicles. Expression in large antral follicles is strongest in the
granulosa cell layer adjacent to the antrum. In cultured granulosa cells,
TERT protein is distributed in the cytoplasm. However, in freshly isolated
granulosa cells functional TERT protein was demonstrated by measurable
telomerase activity.
CONCLUSIONS: The TERT message ceases to be expressed in luteal
granulosa cells, though the protein is still present. This is consistent with
the role of TERT to preserve DNA telomere length and the nature of granulosa
cells to undergo controlled senescence. Healthy follicle growth and
development early in folliculogenesis requires the elaboration of numerous
generations of granulosa cells, but continued survival of granulosa cells
beyond the necessary cell divisions could result in pathologic follicle cysts
or tumors. On the other hand, early suppression of TERT could result in
inadequate follicle growth and fewer cells able to express aromatase and
produce estrogen needed for normal folliculogenesis and fertility. Further
study of TERT regulation could provide important clues for conditions
with aberrant folliculogenesis such as PCOS and diminished ovarian
reserve.
Supported by: New England Fertility Society/ Ferring REI Fellow grant.
P-84 Tuesday, October 20, 2015
AKAP13 IS REQUIRED FOR NORMAL MURINE OVARIAN
DEVELOPMENT. X. Wu, a,b K. Devine, c,b C. Quaglieri, b P. Driggers, a
J. Segars. a a Department of Gyn/OB, Johns Hopkins School of Medicine, Baltimore,
MD; b Program in Reproductive and Adult Endocrinology, NICHD,
NIH, Bethesda, MD; c Shady Grove Fertility Center, Washington, DC.
OBJECTIVE: Induction of aromatase by FSH is a key step in ovarian folliculogenesis.
We previously reported that AKAP13 siRNA transfection was
accompanied by a significant reduction in aromatase transcripts in COV434
cells, suggesting a role for AKAP13 downstream of FSH. Here we sought to
test whether AKAP13 might be involved in induction of aromatase and
ovarian growth in vivo.
DESIGN: Murine Cre/loxP conditional tissue-specific knockout model.
MATERIALS AND METHODS: To examine the role of AKAP13 in
ovary we used a Cre/LoxP tissue-specific knockout approach with Cre driven
by Amhr2 and a floxed Akap13 gene (CKO¼Akap13Flox/Flox/Cre+). Quantitative
real-time polymerase chain reaction (qRT-PCR) was used to measure
akap13 mRNA in CKO ovaries. qRT-PCR was also used to quantify the aromatase
transcripts and 18S rRNA was used for normalization. To assess effects
on ovary size, CKO and wild type (WT) murine ovaries were freshly
collected, fixed, sectioned and stained with Weigert’s iron/picric blue. Images
of stained sections were imported to ImageJ and section surface area
was measured. Differences between groups was tested with a student t-test
or ANOVA and p

PGRMC1, PGRMC2, and PAQR7 attenuated the P4’s ability to suppress cell
cycle entry. However, this was not seen with siRNA treatment for PGR.
Further, it was found that PGRMC1 interacts with PGRMC2 and PAQR7,
in addition to PGR in the cytoplasm; however, there was limited interaction
between PGR and PGRMC1 in the nucleus, despite having a nuclear localization
on immunocytochemistry. Finally, PGR was not found to interact with
PGRMC2 or PAQR7.
CONCLUSIONS: The present experiments are the first to reveal the
complex interaction that occurs among PGRMC1, PGRMC2, and
PAQR7 as well as their involvement in P4’s ability to regulate cell cycle
entry. These mediators may be acting, at least in part, as a complex
through a common mechanism of action; that does not involve PGR. In
addition, PGR and PGRMC1 were found to interact in the cytoplasm,
although what role if any this interaction plays in regulating granulosa/
luteal cell function is currently unknown. Further studies are now needed
to assess the downstream processes of these P4 mediators, for a more
complete understanding of the potentially implicated pathways that
mediate their action.
P-86 Tuesday, October 20, 2015
KDM4A AND KDM4B EXPRESSION IN OVARIAN CUMULUS
CELLS IN WOMEN UNDERGOING IN VITRO
FERTILIZATION. F. Grimstad, a S. Krieg, b K. Roby, c K. Foster, a
A. Krieg, a K. Marquis, a S. Mullinax. a a Department of Obstetrics and Gynecology,
University of Kansas, Kansas City, KS; b University of Kansas Medical
Center, Kansas City, KS; c Department of Anatomy and Cell Biology,
University of Kansas, Kansas City, KS.
OBJECTIVE: In vitro fertilization (IVF) requires superovulation to
create adequate gametes for transfer. Several groups have suggested that
this process may cause epigenetic changes that could have negative health
outcomes for offspring. However, studies have also demonstrated epigenetic
modifications are important to the process of normal gametogenesis.
The study objectives were to assess expression of the histone demethylases
KDM4A and KDM4B in granulosa collected from women undergoing
oocyte retrieval and to determine if expression correlated with pregnancy
outcome.
DESIGN: Prospective Laboratory Study.
MATERIALS AND METHODS: Patients undergoing IVF by a single provider
were stimulated using microdose flare or antagonist protocols and
monitored by serial ultrasound and estradiol levels. At time of egg retrieval
mural and cumulus granulosa were collected and prepared for RNA isolation.
Total RNAwas isolated using TRIzol reagent, reverse transcribed, and cDNA
subjected to quantitative real-time PCR using SyBR Green with specific
primers and the ABI Prism 7500 System. The comparative DCT method
was used for relative quantitation of specific mRNA normalized to 18S
RNA. The mean DCT was determined with relative expression (2DDCt)
calculated for each patient sample compared to the overall mean. The
average relative expression for each gene and cell type was compared with
pregnancy outcome, significance was tested by Mann-Whitney. KDM4B protein
was measured by immunostaining in sections from normal ovaries of
reproductive age women. Tissues were obtained from the KUCC Biospecimen
Facility.
RESULTS: KDM4A and KDM4B mRNA expression was overall higher
in cumulus compared to mural granulosa. When comparing cumulus granulosa
demethylase gene expression, KDM4B mRNA expression was 1.8-
fold higher in granulosa from not-pregnant patients compared to pregnant
patients (P¼0.05). KDM4A displayed a similar trend (P¼ 0.057). In
contrast, there was no significant difference in expression of KDM4A
and KDM4B in mural granulosa from the same patient populations. Immunohistochemistry
detected KDM4B protein in granulosa cells at all stages
of follicle development with more robust expression observed in less
mature follicles.
CONCLUSIONS: The present studies are the first to demonstrate
expression of the histone demethylases KDM4A and KDM4B in granulosa
cells. Data reveal a differential expression where mRNA expression
was higher in cumulus compared to mural granulosa. Expression of
both KDM4A and KDM4B was lower in cumulus granulosa from
pregnant compared to not-pregnant patients. Ongoing studies are
exploring specific genes affected by KDM4A and KDM4B in cumulus
and mural granulosa. These findings suggest that altered expression of
histone demethylases may impact epigenetic changes associated with
pregnancy.
P-87 Tuesday, October 20, 2015
CHARACTERIZATION OF THE HUMAN OVULATORY NON-COD-
ING TRANSCRIPTOME REVEALS MIRNAS AS NEW REGULA-
TORS OF THE OVULATORY CASCADE. A. Hourvitz, a L. Ophir, b
Y. Yung, c R. Orvieto, d G. M. Yerushalmi. e a IVF Unit, Sheba Medical Center,
Ramt-Gan, Israel; b Sheba Medical Center, Petah Tiqva, Israel; c Embryology,
Sheba Medical Center, Ramat Gan, Israel; d Sheba Medical Center, Ramat Gan,
Israel; e Sackler Faculty of Medicine, Tel Aviv University, Tel Hashomer, Israel.
OBJECTIVE: Ovarian follicular development and ovulation in mammals
is a highly complex and tightly regulated process. Using whole transcriptome
sequencing, we previously identified mRNAs that are differentially expressed
between immature early antral follicles and mature preovulatory follicles.
To complete the identification of factors involved in the ovulatory
process, we aim to generate a library of global miRNAs involved in this process.
Using advanced bioinformatics tools, this library will enable us to identify
the ovulatory miRNA-regulated gene and networks and link them to our
previously described library of mRNA ovulatory genes.
DESIGN: Experimental study.
MATERIALS AND METHODS: Cumulus granulosa cells (CGCs) obtained
from GV cumulus-oocyte complex (COC) aspirated from IVM, MII
COC aspirated from IVF and MII COC obtained from 30 h in vitro matured
oocytes. Nanostring nCounter miRNA expression assay was performed in order
to generate regulated miRNA library. Cross analysis between mRNA and
miRNA libraries was performed using miRTrail bioinformatics tool.
RESULTS: Altogether, we found 124 miRNAs expressed in at least one of the
three groups tested. When comparing miRNA from CGCs of MII IVF COC
(mature oocytes) and GV IVM COC (immature oocytes), we found 16 upregulated
and 1 down-regulated miRNAs. Using miRTrail software, we found 1724
potential miRNA regulated target genes by the 17 differentially expressed
miRNA. One hundred and forty one of these potential target genes were also
found in our previously generated ovulatory gene library. Pathway analysis found
4 significant KEGG pathways categories including Cell cycle, ECM-receptor
interaction, Hedgehog signaling pathway and Focal adhesion. These pathways
are probably of major importance for ovulation and corpus luteum formation.
When comparing the miRNA expression pattern between the three groups we
observed that the MII IVM COC miRNA profile was close to the GV COC
miRNA profile, but differ from the MII IVF COC transcriptome. This may correspond
to the observed dysregulated coding gene expression reported in IVM
matured COC and may help explain the lower efficacy of IVM treatments.
CONCLUSIONS: The linkage bioinformatics analysis between the libraries
of the coding genes from our preliminary study with the newly generated
library of regulatory miRNAs enable us to better understand the
regulation of coding ovulatory genes by non-coding transcripts and their
function from the single molecule level to the whole pathways.
P-88 Tuesday, October 20, 2015
THE DETECTION AND CLINICAL UTILIZATION OF ANTI-
MULLERIAN HORMONE (AMH) IN POSTMENOPAUSALWOMEN
USING THE NEW HIGHLY SENSITIVE ELISA. J. Buckbinder, a
S. Mucowski, b H. Burks, c F. Z. Stanczyk, c H. N. Hodis, c W. Mack, d
K. A. Bendikson, c D. Shoupe. c a LAC-USC Medical Center, Los Angeles,
CA; b Dallas Infertility, Frisco, TX; c USC Keck School of Medicine, Los Angeles,
CA; d University of Southern California, Los Angeles, CA.
OBJECTIVE: To determinewhether Anti-M€ullerian Hormone (AMH) levels
as measured by a highly sensitive AMH assay have clinical utility in postmenopausal
women as a marker of low levels of ovarian function that may relate to
other markers of health status in aging women, such as assessments of cardiovascular
and bone health. AMH has been used as a biomarker of ovarian aging
but it has not been useful after onset of menopause as levels are unmeasurably
low. Given that postmenopausal ovaries often contain primordial follicles histologically,
use of a new assay with a very low threshold of detection (3 pg/mL) [a
picoAMH ELISA from AnshLabs] may allow measurement of very low levels
of AMH in menopausal women that are undetectable using earlier versions of
the AMH assay (threshold of detection¼73 pg/mL).
DESIGN: Secondary analysis of a prospective randomized controlled trial,
the Women’s Isoflavone Soy Health Study (WISH) and measurement of
AMH in serum samples of participants with intact ovaries.
MATERIALS AND METHODS: Healthy postmenopausal women participating
in the WISH trial without history of oophorectomy and not using hormone
replacement therapy were included in the study. All participants
underwent baseline venipuncture upon enrollment, and serum was stored
FERTILITY & STERILITY Ò
e135

at -80 C and available for analysis. The picoAMH ELISA from Ansh Labs
was used to measure serum levels of AMH using these stored samples.
RESULTS: Of the 133 samples, the mean age of the postmenopausal
women was 57 6.9 years with a range of 45-78 yrs. The mean time from
the onset of menopause was 7.9 years (range 1-31 years) with 9 of the participants
only being 1 year out from menopause. The mean BMI was 26.1 5.1
kg/m 2 . The ethnicity of the patients was as follows: 55.6% Caucasian, 19.6%
Hispanic, 18.1% Asian or Pacific Islander, and 6.8% African American. In all
133 serum samples, AMH levels were < 3 pg/ml, below the detection limit of
the assay.
CONCLUSIONS: Despite several participants’ recent menopausal transition,
serum AMH was undetectable by a highly sensitive AMH assay in the
entire cohort of postmenopausal women. Measurement of AMH in postmenopausal
women is not clinically useful as levels are unmeasureable, even using
a highly sensitive assay.
Supported by: AnshLabs provided the assays at no cost to investigators.
P-89 Tuesday, October 20, 2015
THE PROSTAGLANDIN TRANSPORTER (PGT): A NOVEL INDIS-
PENSABLE MEDIATOR OF OVULATION. G. M. Yerushalmi, a
Y. Yung, a R. Orvieto, a E. Y. Adashi, b A. Hourvitz. c a Department of Obstetrics
and Gynecology, Sheba Medical Center, Ramat Gan, Israel; b The Warren
Alpert Medical School Brown University, Providence, RI; c Sheba Medical
Center, Ramt-Gan, Israel.
OBJECTIVE: To investigate the physiological ovarian role of the prostaglandin
transporter (PGT).
DESIGN: Experimental study.
MATERIALS AND METHODS: PGT expression in vivo and in vitro in
human mural granulosa cells (MGC) and in the mouse ovary was determined
by qPCR, WB and IHC. MGCs were treated with human chorionic gonadotropin
(hCG) and different activators and inhibitors of the LH pathway and
PGT expression was analyzed by qPCR. in vitro transport assays of PGT in
MGCs using EIA and radiolabeled PGE2. in vivo PGT function studies
were performed in female mice, subjected to a superovulation protocol.
The effect of PGT inhibitors, DIDS (4,4 0 -Diisothiocyanatostilbene-2,2 0 -disulfonic
acid) and Bromocrestol Green (BCG), on ovulation was examined
by histological evaluation, oocyte number and maturation, serum progesterone
levels and gene expression.
RESULTS: The expression of PGT, a transmembrane PG carrier protein,
is markedly upregulated in preovulatory MGCs. Treatment with hCG, an
ovulatory trigger, significantly increases the expression of PGT mRNA
and protein both in vivo and in vitro. The hCG-induced increase in PGT
expression is mediated via protein kinase A and protein kinase C by way
of the extracellular signal-regulated kinase (ERK) pathway. PGT also mediates
the uptake of PGE2 in cultured MGCs, thereby regulating its extracellular
concentration. in vivo treatment with PGT inhibitors effectively
inhibits ovulation in mice. Inhibition of PGT was also associated with inhibition
of luteal function and a marked downregulation of key ovulatory
genes including AREG, EREG, PR, TNFAIP6, StAR, ADAM-TS1, and
CSTL.
CONCLUSIONS: We hypothesize that the inhibition of PGT in GCs increases
the extracellular concentrations of PGE2, the ability of which to exert
its ovulatory effect is compromised by the desensitization of its cognate receptors.
These findings suggest a new mechanism for the regulation of PGE2
during ovulation, and prove PGT as an indispensable mediator of the ovulatory
process, the inhibitors of which constitute potential novel candidates for
non-hormonal contraception.
P-90 Tuesday, October 20, 2015
THE MRNA EXPRESSION OF MT-ENCODED GENES OF OXPHS
IN OVARIAN GCS OF INFERTILE WOMEN AND THE EFFECT
OF NMN ON EXPRESSION OF MT-ENCODED GENES IN SVOG
CELLS. Y. Li X. Liang. Reproductive Medicine Research Center, Sixth
Affiliated Hospital, Sun Yat-sen University, Guangzhou, China.
OBJECTIVE: To detect the mRNA expression of mitochondria(mt)-encoded
genes in oxidative phosphorylation system(OXPHS) in ovarian granulosa
cells(GCs) of infertile women with advanced age or poor ovarian
response(POR), and to explore the relationship between mRNA expression
with in vitro fertilization(IVF) outcomes. In addition, effect of nicotinamide
mononucleotide(NMN)-a NAD+ precursor on mRNA expression of mt-encoded
genes in OXPHS is to be detected.
DESIGN: Retrospective and experimental study.
MATERIALS AND METHODS: 45 infertile women were divided into the
old group(>¼ 37 years old), the young group(< 37 years old) with POR and
the young group with normal ovarian response(NOR). On the day of oocyte
retrieval, GCs were purified from follicular fluid in all the patients for use. In
separate experiments, immortalized human granulosa(SVOG) cells were
treated with nicotinamide mononucleotide(NMN) or NMN adenylyltransferase-1(NMNAT-1)
small interfering RNA(siRNA) to see effect of NMN on
expression of mt-encoded genes in OXPHS. Real-time quantitative PCR
was conducted to detect the mRNA expression of mt-encoded genes CYB,
ND1, ATP6, CO1 and nuclear(nu)-encoded genes ATP5A1, SDHB,
UQCRC1, NDUFS8 in GCs or SVOG cells. mRNA expression in different
groups and the relationship with IVF outcomes, such as number of retrieved
oocytes, achievement of good quality embryos, clinical pregnancy or live
birth were evaluated by Kruskal-Wallis H test, Mann-Whitney or one-way
ANOVA as appropriate.
RESULTS: The mRNA expression of the mt-encoded genes in young
POR group and old group were significantly lower than young NOR group,
besides patients who retrieved more than 3 oocytes or had high quality embryos
showed higher mt-encoded genes expression in GCs. In SVOG cells,
the expression of mt-encoded genes was significantly decreased in the
NMNAT-1 gene knockdown group compared with control and control+NMN
group. Additionally, there were no significant differences in
the mRNA expression of nu-encoded genes among these groups in GCs
or SVOG cells.
CONCLUSIONS: Young POR and old infertile patients may have
impaired mitochondrial function in GCs. Higher expression of mt-encoded
genes in GCs implies good ovarian response and high quality embryos. Besides,
supplementation of NAD+ precursors such as NMN may restore mitochondrial
function and prevent ovarian cell aging.
P-91 Tuesday, October 20, 2015
ANTI-MULLERIAN HORMONE (AMH) MAY INHIBIT OOCYTE
MATURATION AND FOLLICULAR VASCULARIZATION IN HU-
MAN OVARIAN CORTEX. L. Detti, a L. J. Williams, a
N. M. Fletcher, b G. M. Saed. b a Obstetrics and Gynecology, University of
Tennessee, Memphis, Memphis, TN; b Wayne State University, Detroit, MI.
OBJECTIVE: AMH inhibits follicle recruitment [1, 2]. In fact, in the
absence of AMH, primordial follicles are recruited at a faster pace, a common
complication after transplantation of ovarian cortex blamed on a defective
re-vascularization. VEGF stimulates neovascularization and GDF9 is
essential for oocyte-dependent ovarian follicle development [3]. We tested
the hypothesis that administration of recombinant AMH to ovarian cortex
fragments would inhibit follicular development by modulating GDF9 and
VEGF expression.
DESIGN: Pilot experimental study with ovarian cortex obtained from 3
patients.
MATERIALS AND METHODS: Immediately after explant the ovarian
cortex specimens were divided into 5 equal fragments. One fragment was
flash-frozen (untreated) and four were incubated for 48 hours 37 Cina
pH-adjusted gamete buffer media with increasing AMH concentrations of
0-5-25-50 ng/ml. After incubation, all specimens were rinsed and flashfrozen
for PCR analyses, which were executed in triplicates. We utilized
real time RT-PCR to determine mRNA levels for VEGF and GDF9 in ovarian
cortex tissue . We performed ANOVA with Tukey post hoc tests to evaluate
changes in mRNA levels among the five different fragments. A p

TABLE. VEGF and GDF9 mRNA expression in the different treatment groups.
Treatment
VEGF espression
(pg/mcg)
GDF9 espression
(pg/mcg)
Untreated 8.71.3 18.21.9
Media Control
826.08.3
45.27.5
(no AMH)
5 ng AMH 706.256.2 49.41.0
25 ng AMH 422.234.0 18.40.6
50 ng AMH 194.314.8 14.92.7
References:
1. Durlinger ALL, Kramer P, Karels B, et al. Control of primordial follicle
recruitment by anti-Mullerian hormone in the mouse ovary. Endocrinol
1999;140:5789-96.
2. Durlinger ALL, Visser JA, Themmen APN. Regulation of ovarian
function: the role of anti-Mullerian hormone. Reprod
2002;124:601- 9.
3. Dong J, Albertini DF, Nishimori K, Kumar TR, Lu N, Matzuk MM.
Growth differentiation factor-9 is required during early ovarian folliculogenesis.
Nature 1996; 383: 531-535.
in the pups born from natural breeding may indicate that further investigation
is warranted. If appropriately powered the results may reveal a benefit
IVF/ICSI.
CONCLUSIONS: A single nonablative dose of CYC does not immediately
alter semen parameters. IVF/ICSI with CYC exposed sperm decreases
embryo development but does not affect the quality of
surviving embryos. CYC exposure resulted in decreased litter size but
not birth rate following IVF/ICSI-ET or NB. IVF/ICSI with CYC exposed
sperm does not lead to increased genetic variants in the resulting 8 cell
embryos compared with embryos generated using PreCYC or VEH
treated sperm.
Supported by: Eunice Kennedy Shriver National Institutes of Health and
Childhood Development grants HD055475, HD076412, HD075795, Magee-Womens
Research Institute and Foundation and gift funds from Sylvia
Bernassoli.
P-93 Tuesday, October 20, 2015
WITHDRAWN
Supported by: UTHSC Departmental Funds.
ENVIRONMENT AND TOXICOLOGY
P-92 Tuesday, October 20, 2015
EXPOSURE OF TERMINALLY DIFFERENTIATED SPERM TO A
SINGLE NONABLATIVE DOSE OF CYCLOPHOSPHAMIDE
DOES NOT INCREASE THE NUMBER OF GENETIC VARIANTS
IN EMBRYOS OR LIVE BORN OFFSPRING PRODUCED BY IVF/
ICSI. M. D. Johnson, a,b M. Sukhwani, b K. Peters, b S. Malik, a,b
J. S. Sanfilippo, a K. E. Orwig. a,b a Department of Obstetrics, Gynecology
& Reproductive Sciences, University of Pittsburgh School of Medicine, Pittsburgh,
PA; b Magee-Womens Research Institute, Pittsburgh, PA.
OBJECTIVE: Determine the effect of cyclophosphamide on sperm, embryo
development after in vitro fertilization (IVF)/intracytoplasmic sperm injection
(ICSI), birth rate after IVF/ICSI-embryo transfer (ET) or natural
breeding, and genetic integrity of embryos and offspring.
DESIGN: Prospective Laboratory Study.
MATERIALS AND METHODS: Sperm was collected from the epididymis
of male B6D2 mice. 7 days later intraperitoneal cyclophosphamide
(CYC, 300 mg/kg) (n¼10) or vehicle (VEH) (n¼10) were administered.
Sperm were recovered from the contralateral epididymis 7 days later. A
post thaw semen analysis was performed on one straw from each Pre
and Post treatment sample. Pre and Post treatment sperm from CYC
and VEH treated animals were thawed and morphologically normal,
motile sperm were selected to fertilize eggs from B6D2 females by
ICSI. Embryos were monitored for in-vitro development for two days
then collected for exome sequencing. In a separate experiment, birth
rate and genetic integrity of live born offspring was examined after ET
of day two embryos produced by IVF/ICSI with CYC exposed and pre
exposed sperm. These results were compared with offspring produced
by natural breeding (NB) of males before and after exposure to CYC. Genetic
integrity was assessed by exome sequencing of tail DNA from IVF/
ICSI-ET and NB offspring.
RESULTS: CYC did not affect sperm count (P¼.15), motility (P¼.86) or
morphology (P¼.98). 983 embryos from 22 IVF/ICSI cycles were monitored
for preimplantation development. The number of 8 cell embryos produced
using PostCYC sperm was reduced compared to those using PreCYC
sperm (47% vs 18%, P92% of 8 cell embryos in all groups
were Grade A (P>.50). Birth rates after ET were similar for the PreCYC
and PostCYC groups (100% vs 60%, P¼.7) but litter size (3.8 vs 0.8,
P

P-94 Tuesday, October 20, 2015
DIETARY BISPHENOL-A EXPOSURE ALTERS THE METHYL-
OME OF RAT TROPECTODERM AS DETERMINED BY REDUCED
REPRESENTATION BISULFITE SEQUENCING. A. E. Batcheller, a
G. L. Christensen, a Y. Leung, b J. Biesiada, b X. Zhang, b
M. Medvedovic, b M. A. Thomas, a S. Ho. b a Division of Reproductive Endocrinology
and Infertility, Department of Obstetrics and Gynecology, University
of Cincinnati, West Chester, OH; b Department of Environmental Health,
University of Cincinnati, Cincinnati, OH.
OBJECTIVE: To determine if maternal dietary bisphenol-A (BPA) exposure
alters the rat trophectoderm methylome as determined by reduced representation
bisulfite sequencing (RRBS).
DESIGN: Basic research comparative animal study.
MATERIALS AND METHODS: Female Sprague Dawley rats were fed
either AIN-93G control (CTL n¼7) or 250 mg/kg body weight/day BPA
(BPA n¼8) diets during the periconceptional time period. Dams were sacrificed
on gestational day 5, and embryos were flushed from the uterine
horns. Blastocyst trophectoderm (TE) biopsies were obtained using an embryo
splitter (Bioniche). TE biopsies (CTL n¼42, BPA n¼48) were pooled
to make 5 samples in the CTL group, and 6 samples in the BPA group, then
lysed. Genomic DNA was digested with Msp I, size selected and bisulfite
modified. The library was enriched and amplified, then sequenced using Illumina
HiSeq1000. Data were trimmed with Trim Galore. Overall read
quality was determined with FastQC. Read alignment, methylation, and
CpG information was extracted with Bismark. CTL and BPA samples
were collapsed into two groups for analysis. For sites with at least 10
fold coverage, the methylation change was calculated as bBPA(methylated
reads/total reads)-bCTL(methylated reads/total reads). Differences greater
than 60% were considered important. Subsequent statistical analyses
were conducted.
RESULTS: 20464 sequence reads exhibited at least 10 reads per
sequence in both groups, of which 1963 reads demonstrated a difference
in methylation of more than 60%. These reads were located in 230 introns,
five 3’untranslated regions, 16 coding sequences, 625 intragenic regions,
and 16 promoters. Five genes showed more than one differentially methylated
CpG site in the promoter: ERRFI1,Gak, Vom1r90, Rn5-8s, and
Tmem175.
CONCLUSIONS: RRBS can be used to identify differentially methylated
genes in rat trophectoderm. We noted hypomethylation of ERRFI1 promoter
CpG sites resulting from dietary BPA exposure. ERRFI1 is involved in the
embryo/maternal interaction and represents an interesting direction for future
research.
Supported by: Center for Environmental Genetics Innovator Award NIH/
NIEHS P30ES006096. Center for Environmental Genetics New Investigator
Scholar Award NIH/NIEHS P30ES006096.
region as a determinant of LBR after adjusting for recipient’s age, BMI,
race, day and number of donor egg embryos transferred in fresh and FET cycles.
RESULTS: Of 76,296 donor egg IVF cycles reported to SART, regional
information was available for 71,182. Specified regions differed significantly
in: number of donor egg IVF cycles (with highest number of cycles being undertaken
in NE at 20,351, followed by the W (n¼9,051), S(n¼8,054), and
MW (least at 12,241), recipient’s age (p< 0.001), BMI (p< 0.001) and
race (p< 0.001). Regional differences in LBR following fresh and FET
were noted, with lowest LBR’s reported for cycles undertaken in the NE
(46.12% for fresh ET and 26.02% for FET) whereas highest LBR’s were
noted for cycles undertaken in the W (54.23% for fresh ET and 30.12% for
FET, p< 0.001). On adjusted analyses, compared to the NE, donor egg
IVF-ET cycles undertaken in the W region were significantly more likely
to yield LB following fresh ET (OR1.32, 95% CI 1.24-1.41) and FET
(1.18, 95% CI 1.06-1.30) (Table 1).
CONCLUSIONS: Accounting for regional differences in the demographics
of the donor egg recipients, our current analyses (undertaken on
an extended dataset of donor egg IVF cycles) reaffirms our earlier observation.
There are significant regional differences in LBR’s for both fresh and
FET donor egg IVF cycles. Consistent with our earlier observation, highest
LBR’s were observed for donor egg IVF cycles undertaken in the West region
of the USA. Potential influences of ecological nuances on IVF outcome are
suggested.
Association between region and live birth outcome for frozen and fresh embryo
transfer cycles.
Frozen Embryo Transfer
Region Unadjusted OR p Adjusted OR p
(95% CI)
(95% CI)
Northeast Reference — Reference —
South 1.14 (1.05, 1.23) 0.003 1.15 (1.03, 1.27) 0.010
Midwest 1.08 (0.99, 1.18) 0.093 1.08 (0.96, 1.21) 0.189
West 1.23 (1.14, 1.32)

were whether or not a state had mandated coverage for either IVF or some
aspect of infertility care.
RESULTS: Our findings were that classical economic drivers relating to
wealth or economic well-being were not predictive of IVF demand. Rather,
IVF demand was best predicted by general medical insurance coverage,
specific IVF coverage, and the proportion of the population with a selfidentified
Catholic background in a state. The model constructed was:
Log(IVF Cases/Pop.) ¼ -2.96 + 2.41 Catholic + 2.88 Insurance Coverage
+ 0.43 IVF Coverage (se) (1.05) (0.72) (1.33) (0.17) (p-value) (0.007)
(0.0018) (.036) (0.015) The F-statistic was 16.2; and the R-squared was
0.53.
CONCLUSIONS: Independent variables associated with various measures
of income and economic outlook were not helpful in explaining
the variability of IVF utilization among states. The number of IVF cases
done in a state was not related to the effectiveness of IVF in that state nor
the convenience of having an increased number of IVF programs. The
most unexpected finding was that a 1% increase in the proportion of
self-identified Catholics in a state was associated with a 2.4% increase
in the number of IVF cases per 1000 reproductive aged women. Given
the Donum Vitae, one might have expected the impact of an increased
proportion of Catholics to have a negative impact on IVF utilization.
We hypothesized that the variable of being a self-identified Catholic encompassed
microeconomic factors that played a major role in the decision
to undertake IVF. These might include love of family and an increased
desire to have children.
P-97 Tuesday, October 20, 2015
ASSOCIATION OF PREPUBERTAL SERUM DIOXIN CONCEN-
TRATIONS WITH SEMEN QUALITY IN A PROSPECTIVE
COHORT OF YOUNG RUSSIAN MEN. L. Minguez-Alarcon, a
O. Sergeyev, b,c J. S. Burns, a P. L. Williams, a M. M. Lee, d
S. A. Korrick, e L. Smigulina, c B. Revich, b R. Hauser. a a Harvard T H
Chan School of Public Health, Boston, MA; b Russian Academy of Science,
Moscow, Russian Federation; c Chapaevsk Medical Association, Moscow,
Russian Federation;
d University of Massachusetts Medical School,
Worcester, MA; e Brigham and Women’s Hospital, Harvard Medical School,
Boston, MA.
OBJECTIVE: To explore associations of prepubertal serum concentrations
of dioxins, furans and PCBs with semen parameters.
DESIGN: A prospective cohort study conducted in Chapaevsk,
Russia.
MATERIALS AND METHODS: From 2003 to 2005, 499 boys were
enrolled at age 8-9 years and their growth and development was assessed
annually for ten years. Serum dioxins, furans and PCBs were measured
by the U.S. CDC using samples collected at enrollment. At ages 18-19
years, 133 young men provided up to two semen samples collected
one week apart (257 samples) which were analyzed for volume, sperm
concentration and motility according to NAFA and ESHRE-SIGA
manual. Linear mixed models with random intercepts were used to
examine the relation between quartiles of serum concentrations of dioxins,
dioxin toxic equivalents (TEQs), furans and PCBs with semen parameters.
Models were adjusted for BMI, season, and abstinence time,
with further adjustment by time from ejaculation to start of semen analysis
for sperm motility.
RESULTS: Men had a median sperm concentration and motility of 52
mill/mL and 64%, respectively. Higher prepubertal serum TCDD levels
were associated with lower semen parameters. The adjusted difference
(95% CI) in percent change in sperm concentration, total sperm count, and
total motile sperm count between the lowest and highest quartile was
-23.8% (-33.6, -10.5) (p trend¼ .008), -41.5% (-68.9, 1.9) (p trend¼.06)
and -27.6% (-45.2, -1.2) (p trend¼ .06), respectively. In addition, there was
a suggested negative association of PCDD TEQs and P PCDD concentrations
with sperm concentration (p trend¼ .06) and motility (p trend¼.10),
respectively.
CONCLUSIONS: Our results were consistent with the Seveso study that
showed an association of early life exposure to TCDD with poorer semen
quality, indicating that the prepubertal period is a sensitive window of exposure.
Supported by: NIH grants R01ES0014370, P30ES000002, and Russian
Science Foundation grant 14-45-0065.
P-98 Tuesday, October 20, 2015
NEW METHODS IN QUALITY CONTROL: DETECTION OF SUB-
LETHAL TOXICITY. A. Ainsworth, a J. Fredrickson, a D. Morbeck. b
a Obstetrics and Gynecology, Mayo Clinic, Rochester, MN; b Mayo Clinic,
Rochester, MN.
OBJECTIVE: Success of in vitro fertilization (IVF) depends on sound laboratory
methods, culture conditions, and quality supplies, which must be
monitored using sensitive quality control (QC) testing. Current QC testing
using mouse embryo assays (MEA) detects overt toxicity but fails to detect
sublethal toxicity. Sublethal toxicity has been associated with suboptimal
embryo development and occasional manufacturer recalls. This study aimed
to improve the sensitivity of routine MEA in detection of sublethal mineral
oil toxicity.
DESIGN: Two experiments were conducted to examine the detection of
toxicity with sublethal levels of peroxide. Each experiment used fresh 1-
cell mouse embryos, cultured individually, in previously recalled IVF-grade
mineral oil. In experiment 1, an extended MEA (144 h) was compared to
routine MEA (96 h) and cell counts. In experiment 2, sensitivity of the
extended MEA was compared to morphokinetics using serial dilutions of
oil high in peroxides.
MATERIALS AND METHODS: In experiment 1, embryos were cultured
under mineral oil with known toxicity (Oil 31). After 96 hours, half of the
embryos were removed for cell counts. The remaining embryos continued
in culture until 144 hours. In experiment 2, embryos were cultured in serial
dilutions of mineral oil with known toxicity (Oil 44). Morphokinetic data
from the Embryoscope was compared to blastocyst rates at 96 and 144
hours. The results were compared to control oil outcomes using Dunnett’s
t-test.
RESULTS: Extended MEA and cell counts, from experiment 1, identified
sublethal peroxide toxicity not detected by evaluation of blastocyst rates at 96
hours. Blastocyst rates were similar between control and toxic oil at 96 hours,
86% vs. 90%, but significantly different at 144 hours, 30% vs. 0% (p < 0.01).
Cell counts at 96 hours also reflected statistically significant differences, 61
cells in control oil vs. 52 cells in toxic oil (p < 0.01). Morphokinetic data,
from experiment 2, showed delayed cell division with small dilutions of toxic
oil (Table 1). Extended MEA demonstrated increased sensitivity in detection
of trace toxicity when compared to blastocyst rates at 96 hours (Table 1).
CONCLUSIONS: Current methods of QC testing are not sensitive enough
to detect sublethal embryo toxicity. Extended culture to 144 hours, cell
counts at 96 hours, and time-lapse morphokinetic assessment are more effective
than current methods in detection of peroxide toxicity and should be used
to improve reproductive outcomes.
Table 1: Blastocyst Rates and Morphokinetics in Extended MEA.
Oil Blastocyst
Dilutions Rate at 96h
Blastocyst
Rate at 120h
Blastocyst
Rate at 144h
Morphokinetics
i3(h)
Morphokinetics Morphokinetics
cc2(h) tB(h)
1:20 21.88%
(p

MATERIALS AND METHODS: This study included 20 adult men who
reported smoking only cigarettes, referred to the Human Reproduction Section
of the Sao Paulo Federal University for andrological evaluation. Twenty
adult men without semen alterations were included as controls. Men with
confounding factors of male infertility and those reporting fever in the 90
days preceding semen collection were excluded. Semen was collected and
analyzed as per the WHO 2010 guidelines. DNA fragmentation was assessed
using an alkaline Comet assay, classified as grade I (no DNA fragmentation)
to class IV (high DNA fragmentation); mitochondrial activity was analyzed
by a reagent which stains only active mitochondria (3,3’-diaminobenzidine
[DAB]), and classified as DAB I (all mitochondria active) to DAB IV (all
inactive); and acrosome integrity was evaluated by a fluorescent stain
(PNA-FITC). The seminal plasma was used to assess oxidative stress by
TBARS quantification and for label-free quantitative proteomics, performed
by liquid chromatography - tandem mass spectrometry (LC-MS/MS).
Groups were compared using a Student’s T-test (p

increase was maintained with increasing concentrations of up to 1000 ng/ml.
A time-dependent increase in sperm motility was observed in Ritalin-treated
samples until 240 minutes at all concentrations. Thereafter, a decrease was
observed at 300 minutes, even though motility remained significantly higher
compared to basal levels. No significant changes were seen in sperm motion
kinetic parameters.
CONCLUSIONS: Ritalin increased sperm motility at a concentration
of 1 ng/mL. It had no toxic effect on sperm motility even at a concentration
of 1000ng/mL. More research is needed to evaluate whether Ritalin
can be used to improve sperm quality in asthenozoospermic specimens
from patients undergoing assisted reproductive procedures. Further
investigation is ongoing to evaluate the effect of Ritalin on other sperm
parameters such as acrosome reaction, DNA fragmentation and membrane
integrity.
In vitro effect of Ritalin on motility of spermatozoa (%, MeanSEM, n¼10).
Time (min)/
Ritalin (ng/ml)Y 0’ 60’ 150’ 240’ 300’
Control 58.33.79 a 58.44.18 a 59.14.06 a 58.24.14 a 56.34.52 a
1 57.04.25 a 63.93.59* b 66.33.64* b 68.93.83* b 60.53.65* b
10 58.03.69 a 63.23.87* b 66.53.82* b 68.84.21* b 63.23.98* b
100 59.03.55 a 63.44.07* c 66.43.40* c 68.73.90* c 62.93.69* c
1000 59.24.18 a 65.04.02* b 66.53.66* b 68.53.83* b 63.13.61* b
OBJECTIVE: Among Endocrine Disruptors, Bisphenol A (BPA), a
monomer widely used as plasticizer, showed adverse effects on reproductive
system. BPA it was previously foundinovarianfollicularfluid
at approximately 1-2 ng/ml but the direct effects on ovarian somatic
cells are still poor elucidated. In this light, the aim of this study was
to evaluate by FT-IR microspectroscopy and qPCR how BPA affects
metabolism and steroidogenic activity of living human Granulosa cells
(GCs).
DESIGN: This study has been conducted between June and Dicember
2014 on women undergoing a COH for an IVF treatment (n ¼ 20), referring
male factor infertility, (age¼364). GCs have been in vitro exposed
for 48h to different concentrations of BPA (1ng/ml, 10ng/ml and 100ng/
ml).
MATERIALS AND METHODS: FT-IR analysis will be carried out on
living GCs. At this regard, a specific microfluidic device was designed and
used. Q-PCR was utilised to detect gene expression modulation induced by
BPA.
RESULTS: Results from both FTIR and QPCR analises are showed in the
following table.
CONCLUSIONS: The results obtained clearly evidenced a detrimental
effect of BPA on GCs seriously compromising metabolism and AMH
synthesis activity and inducing lipid peroxidation and cell death via
apoptosis and autophagy process. Particularly, the lowest dose of BPA,
the environmentally relevant one, induced the highest negative impact
on GCs.
FTIR RESULTS Control BPA (1ng/ml) BPA (10ng/ml) BPA (100ng/ml)
P-102 Tuesday, October 20, 2015
GYNECOLOGIC DISEASE RISK OF EXPOSURE TO BISPHENOL
A AND PHTHALATE. J. Jeon, S. Park, D. Lee, K. Jeong, H. Chung.
Obstetrics & Gynecology, Ewha Womans University College of Medicine,
Seoul, Korea, Republic of.
OBJECTIVE: To investigate the risk of gynecologic disease is related to
endocrine disrupting chemicals exposure in Korean reproductive women.
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: A total of 307 women aged 30 to 49,
who have lived in the same area for more than 18 months were evaluated.
Anthropometric measurements, laboratory tests with urine and blood sampling
and pelvic ultrasound examinations were performed. Logistic regression
analysis was used to evaluate the contribution of each endocrine
disrupting chemicals exposure for myoma, infertility, endometriosis, obesity,
and metabolic syndrome.
RESULTS: The mean age was 36.764.39 years old and the highest proportion
(45.6%) was between 30 and 35 years old. The mean body mass index
(BMI) was 22.43.1 kg/m2 and 50 women (16.3%) were diagnosed with
obesity (BMI>25.0 kg/m 2 ). Bisphenol A was significantly higher
(1.992.02 ug/g crea) in women with overweight (BMI>23.0 kg/m 2 ) and
obesity. Logistic regression analysis suggested infertility was increased in
high bisphenol A exposure group and the oods ratio (4.248) was significant
statistically after adjusted for age, birth control pills, age of menarche, parity,
and waist circumference. High phthalate exposure was associated to endometrial
polyp after adjustment (OR: 2.742). However, exposure to these endocrine
disrupting chemicals may not lead to myoma, endometriosis, and
metabolic syndrome.
CONCLUSIONS: Bisphenol A exposure is associated with overweight
and infertility and endometrial polyp is increased in Korean reproductive
women with high phthalate exposure.
P-103 Tuesday, October 20, 2015
HUMAN GRANULOSA CELLS IN VITRO EXPOSURE TO BISPHE-
NOL A: FTIR MICROSPECTROSCOPY AND MOLECULAR
EVIDENCES. G. Gioacchini, a,b E. Giorgini, a L. Vaccari, c E. Sereni, b
C. Zaca, b O. Carnevali, a A. Borini. b a Universita Politecnica delle Marche,
Ancona, Italy; b Tecnobios Procreazione, Bologna, Italy; c Elettra Synchrotron,
Trieste, Italy.
LIPIDS/CELL(%) 14.661.25% 25.072.97%*** 22.122.64%* 21.332.85%
PHOSPHOLIPIDS/LIPIDS(%) 15.990.69% 21.080.12%*** 19.560.16% 19.260.16%
LIPIDS PEROXIDATION(%) 1.250.01% 2.040.01%** 1.460.01% 1.630.02%*
PROTEIN/CELL(%) 30.540.83% 20.120.28%** 22.610.36%** 19,870.31%**
UNFOLDED PROTEIN (%) 15.213.65% 43.094.71%*** 35.545.01%** 39.734.13%***
RNA/DNA (%) 10.512.46% 4.600.36%*** 8.981.68% 9.582.01%
QPCR RESULTS Control BPA (1ng/ml) BPA (10ng/ml) BPA (100ng/ml)
Peroxisome proliferator 1.260.27 10.190.86*** 1.230.39 3.330.78*
activated receptorg
(lipid metabolism)
CASPASE3 (apoptosis marker) 1.230.29 8.700.97** 3.301.31* 1.370.45*
BECLIN1 (autophagy marker 1.130.40 5.000.88* 4.170.83* 4.500.90*
Anti Mullerian Hormone 6.231.29 1.400.11 ** 2.130.12** 1.600.48**
*¼P

mean cumulative pre-test scores. At curriculum completion 89% reported
they would adopt or modify strategies for managing patients’ fish intake;
at follow-up 79% had adopted new practices such as providing lists of fish
to avoid and information about adding fish to diet; improving screening for
at-risk groups based on ethnicity and fish consumption levels; adding questions
to electronic medical record screens; and providing patients with magnets
and placing posters on the walls of their waiting rooms.
CONCLUSIONS: Prenatal providers completing the Healthy Fish
Choices CME curriculum can help patients maximize the benefits of fish consumption
while minimizing their exposure to contaminants in fish.
Supported by: This project was Supported by grant #GL-00E00536-0 from
the Great Lakes Restoration Initiative of the USEPA.
P-105 Tuesday, October 20, 2015
GESTATIONAL HYPERTENSION COULD LED TO ADULT
OFFSPRING VASCULAR DYSFUNCTION IN RATS. M. Guo a
H. Huang. b a The Key Laboratory of Reproductive Genetics, Ministry of Education,
Zhejiang University School of Medicine, Hangzhou, China; b Shanghai
Jiao Tong University School of Medicine, Shanghai, China.
OBJECTIVE: To evaluate if offsprings from gestational hypertension
(GH) has vascular dysfunction long term in life.
DESIGN: Fetal origins adult disease has become a lively topic of reproductive
science recently and epidemiology research has proved that intrauterine
environment could impact growth and health of offspring. Our previous
proteomics study on umbilical artery detected several altered proteins
expression during GH related to cardiovascular genesis. As the umbilical artery
could appropriately represent fetal vascular, we assumed that the general
cardiovascular system of offspring could also been affected during this disorder
both short term and long term. To investigate long term effect of offsprings
vascular function changes, we built an animal model use Sprague-
Dawley rats. Body weight and blood pressure were measured continuously.
Futhermore, mesentric arteries were isolated from both groups for contraction
and relaxation function test and morphology analysis were also carried
out use same arteries.
MATERIALS AND METHODS: Animal model were built through L-
NAME application. Maternal blood pressure were measured 2 days before
mate till 2 days after delivery using tail clamp. Body weight and blood pressure
of offsprings were continuously measured from birth to one and half
year-old. 6 offsprings from each group were sacrificed at 8 week-old and 1
year-old for the wire myography (model 620A, DMT). Excess mesentric arteries
from same area were collected for the HE staining. The independentsamples
t test were used to evaluate the statistical significance between two
groups.
RESULTS: Offspring birth weight from GH were lower but become identical
after 3 weeks compared with the control group. GH offspring blood
pressure increase begin at 6 month-old and mesentric artery contraction reaction
is greater than control through same stimulation at 8 week-old. At 1
year-old, not only contraction, the relaxation ability of GH offspring has
also became obtuse to the vasodilator. Through HE staining, mesentric
wall from GH group were much thicker than the control.
CONCLUSIONS: GH is one of the major cause of maternal and neonatal
mortality and morbidity. Besides the negative effects we observe short term
at birth, it can also led to long term increasing risk of cardiovascular diseases
in offspring. In this research, we revealed that the resistance arteries
has both contraction and relaxation ability dysfunction from adult offsprings
of GH and these arteries has a thicker artery wall. Blood pressure
was also increased accompany with the function loss. Further investigation
should be proceed to find out what the mechanism underling these phenomenon.
Supported by: This work was Supported by the National Basic Research
Program of China (No.2012CB944900 to H.F.H); the National Natural Science
Foundation of China (No.81270708 to J.Z.S.).
P-106 Tuesday, October 20, 2015
WITHDRAWN
P-107 Tuesday, October 20, 2015
HEALTH DISPARITIES
IVF OUTCOMES DISPARITIES IN THE UNITED STATES: DO
WOMEN OF MIDDLE EASTERN/NORTH AFRICAN ETHNICITY
EXPERIENCE IN VITRO FERTILIZATION (IVF) DISPARITIES
COMPARED TO A CAUCASIAN COHORT?. W. H. Salem, a
F. Sharara, b O. Abuzeid, c T. I. Abozaid, c M. Ashraf, c M. I. Abuzeid. c
a OB/GYN and Reproductive Sciences, University of California, San Francisco,
San Francisco, CA; b Virginia Center for Reproductive Medicine, Reston,
VA; c IVF Michigan, Rochester Hills, MI.
OBJECTIVE: Health disparities have been demonstrated for ethnic groups
undergoing IVF in the United States however outcomes remain contradictory
(1-3). There are no studies to our knowledge that address the Middle Eastern/
North African (MENA) ethnic group in the U.S. Under SART reporting data,
this group is generally categorized under the Caucasian ethnic group. This
study investigates IVF outcomes and demographic metrics of MENA women
in comparison to a control group of Caucasian women.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: A group of 50 randomly selected
MENA women were compared to a control group of 100 randomly
selected Caucasian women undergoing IVF at a private IVF clinic in
Michigan over the period 2011-2013. Inclusion criteria were met if the
subjects were undergoing an IVF cycle with a fresh day 5 blastocyst,
morula or mixed transfer. Patients were excluded if they had incomplete
data, underwent preimplantation genetic screening or utilized donor gametes.
49 MENA patients and 99 Caucasian patients were included in the
analysis. All continuous variables were analyzed with a two sample t
test while categorical information was analyzed with a chi squared analysis.
RESULTS: This pilot study suggests that there is no difference in IVF
pregnancy or live birth rates between MENA and Caucasian women (67%
vs 72% p¼0.5 and 57% vs 59% p¼0.87). There was a trend toward a higher
miscarriage rate among MENAwomen (10% vs 4% p¼0.14). MENAwomen
also had a trend toward a lower BMI than Caucasian women (25.0 vs 27.1,
p¼0.06). This may be reflective of a lower prevalence of polycystic ovarian
syndrome in the MENA group (26.5% vs 41.4%, p¼0.13). There were no differences
in age, etiology of infertility and the rate of primary infertility.
Embryological data demonstrated no significant differences in the number
of fertilized oocytes, day 5 morulae, blastocysts or total embryos transferred.
e142 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

MENA women required a lower number of stimulation days (8.7 vs 9.4,
p¼0.01).
CONCLUSIONS: This preliminary study suggests that there are no differences
in pregnancy outcomes between MENA and Caucasian women in a
sample U.S. population. This is the first study to support the categorization
of MENA women under the Caucasian ethnic group for SART outcomes.
A more robust analysis of MENA women with regards to IVF outcomes disparities
is forthcoming.
Demographics, Embryological and IVF Outcome Data of MENA vs Caucasian
Women.
MENAN¼49 Caucasian N¼99 P Value
Age (years) 31.8 32.7 0.14
BMI (kg/m^2) 25.0 27.1 0.06
Male Factor 57.1% N¼28 54.5%N¼54 0.09
PCOS/Ovulatory 28.6%N¼14 41.4%N¼41 0.13
Tubal Factor 26.5%N¼13 28.3%N¼28 0.82
Uterine 34.7%N¼17 36.4%N¼36 0.84
Unexplained Infertility 4.1%N¼2 0%N¼0 0.04
Antral Follicle Count 11.8 15.1 0.10
Total Embryo Transfer 2.18 2.17 0.92
Miscarriage 10.2%N¼5 4.0%N¼4 0.14
Live Birth 57.1%N¼28 58.6%N¼58 0.87
rate was higher in the Caucasian group but not statistically significant likely
secondary to inadequate power (46% vs. 31%; OR 1.95; CI [0.87,4.36]). Live
birth rate after a clinical pregnancy was documented was similar between
groups (86% vs. 81%; OR 0.72; CI [0.14,3.81]).
CONCLUSIONS: Asian ethnicity is associated with a lower clinical pregnancy
rate even when donor oocytes are used. However, they have similar
live birthrates after clinical pregnancy is documented. Differences in IVF
pregnancy outcomes between Asian and Caucasian patients, even with
oocyte donation, are likely secondary to impaired implantation in women
who self declare Asian ethnicity.
Asian (N¼39)
Median
(interquartile range)
Caucasian (N¼84)
Median
(interquartile range)
P value
Donor Age 25 (6) 25 (6) 0.31
Recipient Age 44 (8) 43 (5) 0.75
Donor BMI 20 (2) 21 (3) 0.01
Recipient BMI 22 (3) 22 (4) 0.52
Total Gonadotropin (ampules) 30 (12) 30 (29) 0.53
Peak E2 (pg/mL) 3815 (2476) 3650 (1854) 0.62
Oocytes retrieved 21 (13) 24 (13) 0.06
Mature oocytes 17 (11) 18 (10) 0.39
Oocytes fertilized 15 (12) 16 (10) 0.38
E2/oocyte retrieved (pg/mL) 174 (89) 150 (70) 0.03
References:
1. Dayal MB, Gindoff P, Dubey A, Spitzer TL, Bergin A, Peak D, et al.
Does ethnicity influence in vitro fertilization (IVF) birth outcomes?
Fertil Steril. 2009;91:2414-8.
2. Sharara FI, McClamrock HD. Differences in in vitro fertilization (IVF)
outcome between white and black women in an inner-city, university
based IVF program. Fertil Steril 2000;73:1170.
3. Wellons MF, Fujimoto VY, Baker VL, et al. Race Matters: A Systematic
Review of Racial/Ethnic Disparity in Society for Assisted Reproductive
Technology(SART) Reported Outcomes Fertil Steril. 2012
Aug;98(2):406-409.
P-108 Tuesday, October 20, 2015
WHY ARE IVF PREGNANCY RATES LOWER IN WOMEN OF
ASIAN ETHNICITY: AN ANALYSIS OF ETHNICITY-MATCHED
OOCYTE DONOR CYCLES. K. Louie, a L. Ross, a T. Jones, a
B. Rudick, b K. Chung, c K. Bendikson. d a Division of Reproductive Endocrinology
and Infertility, University of Southern California, Los Angeles, CA;
b Division of Reproductive Endocrinology and Infertility, Columbia University,
New York, NY; c USC Keck School of Medicine, Los Angeles, CA;
d USC Fertility, Los Angeles, CA.
OBJECTIVE: To resolve controversies regarding decreased pregnancy
rates in women of Asian ethnicity in comparison to Caucasian women, we
sought to evaluate clinical pregnancy rates between ethnically matched Asian
and Caucasian egg donation cycles.
DESIGN: A retrospective cohort study.
MATERIALS AND METHODS: First-time anonymous oocyte donors
of Asian or Caucasian ethnicity who underwent oocyte stimulation between2006and2013thatresultedinafreshembryotransferwereidentified.
Only the first stimulation of egg donors and first transfers of
recipients were considered. Ethnicity was self-reported by both the
donor and the intended recipient. Only ethnicity matched donor-recipient
pairs were included. Fisher’s exact, Wilcoxon rank-sum, binomial
and multivariate logistic regression were used as appropriate for analysis.
RESULTS: 39 Asian and 123 Caucasian ethnically matched oocyte donor
and recipient pairs meeting inclusion criteria were identified. Baseline characteristics
were similar between Asian and Caucasian donors and recipients
except for BMI in donors. There were no differences in stimulation parameters.
E2 levels per oocyte retrieved were higher in Asian donors. The number
of embryos transferred was the same between groups (2 1 vs. 2 1;
p¼0.64). The Caucasian group had significantly higher implantation rates
(42% 41% vs. 21% 29%; p¼0.01) and clinical pregnancy rates (57%
vs. 36%; OR 2.38; CI [1.09,5.22]) compared to the Asian group. Live birth
P-109 Tuesday, October 20, 2015
OVERVIEW OF 2012 U.S. ASSISTED REPRODUCTIVE TECHNOL-
OGY (ART) TREATMENT OUTCOMES AND CONTRIBUTION TO
MULTIPLE BIRTH AND PRETERM INFANTS. S. Sunderam, a
D. M. Kissin, b S. Crawford, a S. Folger, a D. Jamieson, a L. Warner, a
W. Barfield. a a Division of Reproductive Health, Centers for Disease Control
and Prevention, Chamblee, GA; b Centers for Disease Control and Prevention,
Atlanta, GA.
OBJECTIVE: To report U.S. ART statistics and compare ART infant outcomes
to all U.S. infant outcomes.
DESIGN: Population-based retrospective analysis.
MATERIALS AND METHODS: Data were obtained from CDC’s National
ART Surveillance System and National Vital Statistics System. The
following were calculated for each state, the District of Columbia, and Puerto
Rico: number of ART procedures performed per million women of reproductive
age (ART utilization), average number of embryos transferred, rates of
elective single embryo transfers (eSET), as well as rates of ART-conceived
multiple-birth, low birth weight, and preterm infants. The proportion of
ART infants among all infants, multiple-birth, low birth weight and preterm
infants was also calculated.
RESULTS: Among 3,991,741 infants born in 2012 in the U.S. and Puerto
Rico, 1.5 % (61,432) were conceived with ART (range: 0.2% in Puerto Rico
to 5% in Massachusetts). ART utilization ranged from 323 to 7,221 procedures
per million women aged 15-44 years, and was higher than the national
average of 2,483 in 13 reporting areas, many of which were located in the
northeast. The national eSET rate among women

P-110 Tuesday, October 20, 2015
OXIDATIVE STRESS
FOLLICULAR FLUID FROM INFERTILE WOMEN WITH MILD
ENDOMETRIOSIS IMPAIRS IN VITRO BOVINE EMBRYO DEVEL-
OPMENT AND N-ACETYL-CYSTEINE STIMULATES THE EM-
BRYO HATCHING. V. S. Giorgi, a B. T. Jianini, a M. G. Da Broi, a
C. C. De Paz, b R. Ferriani, a P. A. Navarro. a a Department of Obstetrics
and Gynecology, Faculty of Medicine of Ribeirao Preto, University of S~ao
Paulo, Ribeirao Preto, Brazil; b Department of Genetics, Faculty of Medicine
of Ribeirao Preto, University of S~ao Paulo, Ribeirao Preto, Brazil.
OBJECTIVE: To assess the effect of the addition of follicular fluid (FF)
from infertile women with and without mild endometriosis (ME) and of
the antioxidant N-acetyl-cysteine (NAC) on embryo development, using
bovine model.
DESIGN: Experimental study.
MATERIALS AND METHODS: FF samples were obtained from 22 infertile
women undergoing ovarian stimulation for intracytoplasmic sperm injection
[11 with ME (EFF) and 11 with tubal and/or male factor of infertility
(CFF)], pooled, and utilized in 5 in vitro maturation (IVM) experiments
with immature bovine oocytes (IBO). IBO were submitted to IVM divided
in 5 groups: without FF (NF), with 1% of FF from ME patients (EFF) or control
patients (CFF), EFF + 1.5mM of NAC (ENAC), CFF + 1.5mM of NAC
(CNAC). Then, in vitro fertilization (IVF) was performed and embryos were
in vitro cultured. We analyzed cleavage rate, blastocyst production, and
hatched blastocysts rate. Data were analyzed by gamma distribution.
RESULTS: The cleavage rate was similar comparing NF with CFF
(p¼0.606), CNAC (p¼0.206) and ENAC (p¼0.179); and lower comparing
EFF group with NF (p¼0.007) and CFF (p¼0.029) groups. The blastocyst production
rate was similar between NF and ENAC (p¼0.544) groups, and lower
in CFF (p¼0.019), CNAC (p¼0.003) and EFF (p¼0.004) compared to NF. The
hatched blastocyst rate was similar between NF and CFF (p¼0.837) groups; the
EFF had the lowest hatched blastocyst rate compared to all other groups
(p

Mitochondrial thioredoxin dependent peroxide reductase and thioredoxin
related transmembrane protein 4 were uniquely expressed in high ROS
group providing additional defense. Overexpression of Apolipoprotein D
is suggestive of better androgen response despite oxidative stress in this
group.
CONCLUSIONS: Based on the comparative proteome profiles, we
conclude that fertility in men with high seminal ROS may be related to better
stress response and enhanced antioxidant defense.
P-113 Tuesday, October 20, 2015
MYELOPEROXIDASE SERVES AS A REDOX SWITCH THAT REG-
ULATES APOPTOSIS IN HUMAN LEIOMYOMAS. M. S. Abusamaan, a
N. M. Fletcher, a M. G. Saed, a A. Al-Hendy, b M. P. Diamond, c J. M. Berman, a
G. M. Saed. a a Obstetrics and Gynecology, Wayne State University, Detroit, MI;
b Medical College of Georgia, Georgia Regents Univer, Augusta, GA; c Georgia
Regents University, Augusta, GA.
OBJECTIVE: To determine the expression of myeloperoxidase (MPO)
and nitric oxide (NO), modulators of oxidative stress, and their impact on
apoptosis of leiomyoma cells under normal and hypoxic conditions.
DESIGN: Prospective experimental study.
MATERIALS AND METHODS: We have utilized an immortalized human
leiomyoma and a matched myometrial cell line. Cells were exposed
to normal (20% O2) and hypoxic (2% O2) conditions for 24 hours in Smooth
Muscle Medium-2. Bax, Bcl-2 and caspase-3, markers of apoptosis, and
MPO were measured by real-time RT-PCR. The Nitrate/Nitrite Colorimetric
Assay was used to measure NO levels. Data were analyzed by Student’s t-
test.
RESULTS: MPO and NO levels were higher in leiomyoma (0.043 0.006
fg/ug RNA and 7.0 0.09 mM) than myometrial cells (0.020 0.006 fg/mg
mRNA and 6.3 0.18 mM) (p¼0.07 and p

P-116 Tuesday, October 20, 2015
ESTABLISHING THE OXIDATION-REDUCTION POTENTIAL IN
SEMEN AND SEMINAL PLASMA. A. Agarwal, a S. S. Du Plessis, a,b
R. Sharma, a L. Samanta, a,c A. Harlev, a,d G. Ahmad, a,e S. Gupta, a
E. S. Sabanegh. f a Center For Reproductive Medicine, Cleveland Clinic,
Cleveland, OH; b Medical Physiology, Stellenbosch University, Tygerberg,
South Africa; c Redox Biology Laboratory, School of Life Sciences, Ravenshaw
University, Orissa, India; d Soroka Medical Center, Ben-Gurion University,
Beer Sheva, Israel; e Physiology and Cell Biology, University of Health
Sciences, Lahore, Pakistan; f Urology, Cleveland Clinic, Cleveland, OH.
OBJECTIVE: Oxidation-reduction potential (ORP) is a novel measure of
oxidative stress or redox imbalance in biological samples. Static ORP
(sORP) provides an integrated measure of the balance between total oxidants
and reductants in a biological system, wheras capacity ORP (cORP) equates
to the amount of antioxidant reserves. sORP has been shown to correlate well
with illness and injury severity that accompanies the state of oxidative stress;
cORP correlates with the ability to respond to illness or injury. Our objectives
were to evaluate whether 1) ORP can be measured in semen and seminal
plasma samples and 2) ORP levels correlate with sperm motility.
DESIGN: Prospective study measuring ORP in both semen and seminal
plasma.
MATERIALS AND METHODS: Semen samples (n¼18) from normal
control subjects were divided into two fractions and the seminal plasma
was isolated from one fraction (300 x g, 7min). Sperm count and motility
were assessed manually. sORP (mV/10 6 sperm) and cORP (mC/10 6 sperm)
were measured in both fractions (RedoxSYSÒ, Aytu BioScience). Values
are reported as Mean SEM. Spearman correlation and Receiver Operating
Characteristic curves (ROC) were used for statistical analysis.
RESULTS: sORP and cORP levels in semen correlated significantly with
the levels in seminal plasma. A significant negative correlation existed between
sperm motility and sORP in both semen (r¼-0.609; p¼0.004) and seminal
plasma (r¼-0.690; p¼0.002). Furthermore, a sORP cutoff of 4.73mV/
10 6 sperm in semen (sensitivity ¼ 100%, specificity ¼ 89.5%,
AUC¼0.947) and 4.65mV/10 6 sperm in seminal plasma (sensitivity ¼
100%, specificity ¼ 93.8%, AUC ¼ 0.969) was highly predictive of abnormal
sperm motility.
CONCLUSIONS: RedoxSYSÒ accurately measured sORP and cORP in
both semen and seminal plasma samples. Based on high sensitivity as assessed
by ROC analysis, sORP levels can be used to screen infertile men
with oxidative stress. These results are being validated in a larger cohort of
infertile men.
ORP levels in semen and seminal plasma.
sORP
(mV/10 6 sperm)
cORP
(mC/10 6 sperm)
Semen 2.790.66 0.200.08
Seminal Plasma 2.670.71 0.310.16
Correlation (r) 0.994 0.959
P

iogenesis (WT, 3.420.54 x 10 5 vs Tg 2.480.64 x 10 5 copies/blastocyst;
p

showed that there was no DNA methylation in SOD1 CpGs in promoter and
intron regions, as well as SOD2 promoter, while SOD2 intron 2 displayed
moderate methylation. Moreover, the methylation level of SOD2-intron in
preeclampsia placentae increased significantly compared with that in normal
third trimester (P < 0.05), which was confirmed by BSP. The methylation
level of SOD1 was inhibited dose-dependently by ADC, and the mRNA
expression of SOD2 elevated following the decrease of methylation.
CONCLUSIONS: The present study demonstrates that the reduced
expression of both SOD1 and SOD2 may be engaged in the increased oxidative
stress in preeclampsia, which is a crucial pathogenesis in the progress of
preeclampsia. In addition, SOD2 expression was down-regulated by elevated
DNA methylation in intron 2 region. This helps us to understand the etiology
of preeclampsia better and to develop new agents to restrain the progress of
preeclampsia.
Supported by: National Basic Research Program of China
(2012CB944900), the Natural Science Foundation of China (81200446,
31171444 and 30973209), the National Science and Technology Support
Program (2012BAI32B01).
P-122 Tuesday, October 20, 2015
REVISITING AN OLD SEMINAL BIOMARKER TO GAUGE
GAMATE MICROENVIRONMENT AND PREGNANCY
OUTCOME. L. Park, Q. V. Neri, L. Reisman, T. Paniza,
Z. Rosenwaks, G. D. Palermo. Reproductive Medicine, Weill Cornell Medical
College, New York, NY.
OBJECTIVE: To assess the efficacy of additional seminal biomarkers in
predicting pregnancy outcome with IUI.
DESIGN: In a time interval of 12 months, we assessed gamete microenvironment
by measuring total antioxidant capacity (TAC) and seminal fructose
(FRU) on 134 men. To account for fructose consumption by spermatozoa, the
fructose value was corrected (cFRU) by multiplying it with the logarithm of
sperm concentration. Correlation between TAC and cFRU as well as with
clinical outcome were assessed.
MATERIALS AND METHODS: FRU (mg/ml) and TAC (nmol/ml) was
assessed using a colormetric assay on an automated microplate reader
(DOD405/570). cFRU was calculated by multiplying the seminal fructose
by the logarithm of the sperm concentration (for men with >1 million
sperm/ml). IUI pregnancy outcomes were assessed.
RESULTS: A total of 135 men (38.5 8 years old) had a sperm concentration
of 26.8 29.4million/ml with a motility of 39.4 14.2% and
morphology of 2.3 1.0%. The average TAC was 1834.5 278.4nmol/ml
and significantly correlated with fructose (1.97 1.13mg/ml, P <
0.000005). Once fructose was corrected for sperm concentration, the average
cFRU was 2.531.61mg/ml and also correlated with TAC (P < 0.0000001),
as well as with male age (P ¼ 0.03) and sperm motility (P ¼ 0.009). A group
of 41 men were treated in 100 IUI cycles with their female partner (35.9 5
years) and reported a clinical pregnancy rate of 15.0%. The average TAC was
1862.6193.6mM and the average cFRU was 3.02 1.58mg/ml. There were
no significant differences among male and female ages as well as sperm parameters
between couples that achieved a clinical pregnancy (n ¼ 15) and
those who did not (n ¼ 26). However, TAC and cFRU in the successful
outcome group was both remarkably higher than those that failed (P ¼
0.002 and P ¼ 0.02 respectively), even after controlling for an eventual female
factor (%35 years; P ¼ 0.02).
CONCLUSIONS: Information acquired from standard semen analysis has
a limited predictability on the fertile status of men with adequate semen parameters.
Additional seminal vesicle biomarkers such as TAC and corrected
fructose level provide information on the male genital tract microenvironment
and male gamates’ ability to achieve pregnancy.
Supported by: WCMC.
OBJECTIVE: Does Fertilix, a novel antioxidant formulation designed to
alleviate oxidative stress in the male reproductive tract, reduce Sperm
DNA Damage (SDD) and increase pregnancy rates in mouse models of
Sperm Oxidative Stress (SOS)?
DESIGN: Fertilix efficacy was evaluated in two, well-established mouse
models of SOS; Glutathione Peroxidase 5 knockout (GPX-5 KO) mice and
Scrotal Heat Shock (SHS) treatment, each with n¼12, by independent laboratories.
Mice were given Fertilix in their drinking water for 2 months (GPX-
5 KO) and 2 weeks (SHS) and then compared with control groups for SDD
and pregnancy rates.
MATERIALS AND METHODS: In GPX-5 KO mice, SDD was evaluated
by immunocytochemical detection of 8-Hydroxy-deoxy Guanosine (8-
OHdG) residues, a biomarker of DNA oxidation. In the SHS model, each
male’s fertility was tested by partnering with three females for 5 days. The
percentage of pregnant females, number of vaginal plugs, resorptions per
litter, and litter size were recorded.
RESULTS: The average background levels of 8-OHdG in WT mice is
around 30%. This level doubles up to about 60% in transgenic male mice
deficient in the antioxidant enzyme GPX-5. Our results indicate that a 2
month pretreatment of GPX-5 KO mice with Fertilix provides complete protection
of sperm DNA against oxidation. In male mice exposed to the SHS
model, only 35% (19/54) of partnered female mice got pregnant resulting
in 169 fetuses. In contrast, male mice pretreated with Fertilix for 2 weeks
yielded 74% (42/57) of partnered female mice becoming pregnant, resulting
in 427 fetuses. The role of chance in obtaining supporting results for the efficacy
of Fertilix in both models is minimal although it was not possible to
ensure that every mouse took 100% of the product for the treatment period.
CONCLUSIONS: Oral administration of Fertilix significantly reduces
SDD in GPX-5 knockout mice and restores pregnancy rates almost back to
normal levels in mice subjected to SHS. These results, if confirmed in humans,
will impact clinical fertility practice. Current clinical studies confirm
moderate to severe SDD in about 60% of all men visiting IVF centers and in
about 80% of men diagnosed with idiopathic male infertility, figures which
are gravely concerning. Antioxidant supplementation will be an adjuvant
therapy prior to undertaking ART procedures to improve fertilization rates,
maintain a healthy pregnancy, and reduce de novo sporadic mutations being
passed onto children.
Supported by: The study was funded by the University of Clermont and the
University of Madrid.
P-124 Tuesday, October 20, 2015
WITHDRAWN
REPRODUCTIVE IMMUNOLOGY
P-123 Tuesday, October 20, 2015
FERTILIX, A NOVEL ANTIOXIDANT FORMULATION DESIGNED
TO TREAT MALE INFERTILITY EMANATING FROM SPERM
OXIDATIVE DNA DAMAGE: PROMISING PRECLINICAL
EVIDENCE FROM MOUSE MODELS. A. Moazamian, a
P. Gharagozloo, a J. Drevet, b A. Gutierrez-Adan, c A. Kocer, b A. Calle, c
E. Pericuesta, c A. M. Polhemus, a R. J. Aitken. d,a a CellOxess LLC, Princeton,
NJ; b University Blaise Pascal, Clermont-Ferrand, France; c INIA Animal
Reproduction, Madrid, Spain; d The University of Newcastle, Callaghan,
NSW, Australia.
e148 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

4. Meuchel LW, Stewart A, Smelter DF, Abcejo AJ, Thompson MA, Zaidi
SL, et al. Neurokinin-neurotrophin interactions in airway smooth muscle.
Am J PHysiol Lung Cell Mol Physiol 2011;301:L91-8.
5. Argawal A, Saleh Ra, Bedaiwy A. Role of reactive oxygen species in
the pathophysiology of human reproduction. Fertil Steril
2003;79:829-43.
P-126 Tuesday, October 20, 2015
CONCENTRATIONS OF INSULIN-LIKE GROWTH FACTOR (IGF)-
1 AND 2 AT THE BEGINNING OF A MATCHED DONOR CYCLE
BEFORE STIMULATION PREDICT OUTCOME IN
RECIPIENTS. T. Kanninen, a I. Ramer, a G. Sisti, a S. S. Witkin, a
S. D. Spandorfer. b a Weill Cornell Medical College, New York, NY; b Cornell
University Medical Center, New York, NY.
P-125 Tuesday, October 20, 2015
SERUM LEVELS OF BRAIN-DERIVED NEUROTROPHIC FACTOR
PRIOR TO INITIATION OFAN IN VITRO FERTILIZATION CYCLE
PREDICT OUTCOME. I. Ramer, a T. T. Kanninen, a G. Sisti, a
S. S. Witkin, a S. D. Spandorfer. b a Cornell University Medical College,
New York City, NY; b Cornell University Medical Center, New York City, NY.
OBJECTIVE: Prior research has shown that brain-derived neurotrophic
factor (BDNF) levels in sera from women on day 28 of an in vitro fertilization
(IVF) cycle were highly predictive of a live birth. Most BDNF that is present
in sera is derived from activated platelets and recent data has shown that
platelet activation is essential to the initiation of innate and acquired immune
system activation in response to non-physiological conditions. Thus, the
release of BDNF by platelet activation may serve as a sensitive signal for
the presence of physiologic stress. The goal of this study was to determine
if the concentration of circulating BDNF prior to cycle initiation (day 2) predicts
outcome in women undergoing IVF.
DESIGN: This is a cross-sectional, retrospective study of women undergoing
IVF at Weill Cornell Medical College.
MATERIALS AND METHODS: Sera was obtained on day 2 of an IVF
cycle from 54 (23.9%) women with a live birth, 45 (19.9%) with a spontaneous
abortion, 38 (16.8%) with a biochemical pregnancy, 54 (23.9%) who
did not become pregnant, and 35 (15.5%) with an ectopic pregnancy.
BDNF concentration was determined by ELISA. Associations with clinical
parameters were assessed by the Kruskall-Wallis and Mann-Whitney tests.
The ability to predict an ectopic pregnancy was determined by receiver operator
curve analysis.
RESULTS: Median serum concentrations of BDNF were lowest in women
who had a subsequent live birth (3.6 ng/ml), compared to women with an
ectopic pregnancy (7.3 ng/ml), those who did not become pregnant (5.5
ng/ml), women with a biochemical pregnancy (3.8 ng/ml) or a spontaneous
abortion (4.2 ng/ml) (p37 weeks) in
women who were recipients.
DESIGN: Sera from 59 oocyte recipients who had a liveborn, collected at
the initiation of their recipient cycle and frozen, were thawed and assayed for
concentrations of mediators reported to be involved in determining pregnancy
outcome. All of the patients underwent fresh anonymous oocyte donation.
After down regulation with OCPs and leuprolide acetate, serum was
collected. This was before any therapy with estradiol in the egg donation cycle.
MATERIALS AND METHODS: IGF-1, IGF-2, IGF binding protein-1,
pro-inflammatory cytokines (Interleukin-1beta, interleukin-6, tumor necrosis
factor alpha) and anti-inflammatory cytokines (interleukin-13, interleukin-
17) in sera were assayed in duplicate by commercial ELISA kits. Values
were converted to ng/ml or pg/ml by reference to a standard curve generated
in parallel to each assay. Differences in biomarker levels between women
who delivered at term (>37 weeks) or preterm were analyzed by the
Mann-Whitney. Associations between gestational age at delivery and either
biomarker concentration or maternal age were analyzed by the Spearman
rank correlation test.
RESULTS: Ten women delivered preterm (32-36.5 weeks) while the remaining
49 had a term birth (37-41.6 weeks). The median concentration of
IGF-1 was 17.1 ng/ml in women who delivered at term as compared to <
0.03 ng/ml in women with a preterm birth (p ¼ 0.0014). IGF-1 levels were
positively associated with gestational age at delivery (p ¼ 0.0345) but not
associated with maternal age. Conversely, median IGF-2 levels were higher
in women with a preterm delivery (452 pg/ml) than in those with a term birth
(365 pg/ml) (p ¼ 0.0090). IGF-2 levels were inversely correlated with gestational
age at delivery (p ¼ 0.0055) but not with maternal age.
CONCLUSIONS: Determination of IGF-1 and IGF-2 levels in sera of
oocyte donor recipients at the start of their IVF cycle can predict who will
most likely have a preterm birth and who will deliver at term.
P-127 Tuesday, October 20, 2015
INTRAUTERINE HCG INFUSION AFFECTS THE DISTRIBUTION
OF NATURAL KILLER CELLS IN THE ENDOMETRIUM OF
FERTILE OOCYTE DONORS. E. Giuliani, a M. Olson, b M. Strug, b
J. Young, c V. Shavell, c W. Dodds, c R. Leach, b A. Fazleabas. b a Department
of Obstetrics and Gynecology, Grand Rapids Medical Education Partners/
Michigan State University, Grand Rapids, MI; b Department of Obstetrics,
Gynecology and Reproductive Biology, Michigan State University, Grand
Rapids, MI; c The Fertility Center, Grand Rapids, MI.
OBJECTIVE: Reports on intrauterine (IU) human chorionic gonadotropin
(hCG) administration prior to embryo transfer suggest a potential positive effect
on pregnancy rates possibly by promoting the expression of genes and
proteins that are critical for implantation (1,2,3). A prior study by our group
demonstrated that patients with infertility and endometriosis have dysregulated
expression of endometrial CD16 and CD56 natural killer cells (NK)
(4). NK cells play a key role in implantation since they can produce angiogenic
factors that promote spiral artery remodeling and cytokines (LIF, leukemia
inhibitory factor) that direct the migration of trophoblasts (4). The goal
of this study was to investigate if a single IU hCG infusion during the period
FERTILITY & STERILITY Ò
e149

corresponding to embryo transfer could induce histological changes in the
distribution of NK cells in the uterine endometrium.
DESIGN: Case-control study.
MATERIALS AND METHODS: Three days following controlled ovarian
hyperstimulation and oocyte retrieval, fertile oocyte donors were randomly
assigned to receive an IU infusion of either 500 IU hCG (n¼7) or Vehicle
(n¼8). Endometrial biopsies were performed 48h post infusion. Subsequent
analysis included blinded histological staining and quantification (Image J
software) of CD56+ and CD16+ stroma NK cells in the endometrial biopsies.
Paired T-test was used to analyze the results between the two treated groups.
RESULTS: No statistically significant differences in the age, parity, oocytes
retrieved and estradiol levels before retrieval were found between the
two study groups. The percentage of CD16+ stroma cells was higher after
the infusion of hCG (1.320.05 vs 1.180.13, p4 fold; P

OBJECTIVE: Advanced ovarian age is often implicated in implantation
failure and early pregnancy loss in women aged 40 and over. The role of
the uterus in reproductive aging has been investigated, but with conflicting
conclusions. We seek to investigate the impact of uterine age on reproductive
capacity while controlling for oocyte quality by using Comprehensive Chromosomal
Screened Embryos.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Women undergoing in vitro fertilization
(IVF) with preimplantation genetic screening (PGS) from September 2010 to
March 2015, who used either donor or autologous oocytes, were included.
Fresh or thawed euploid blastocysts, confirmed by PGS, were selected for
embryo transfer (ET). Student’s t-test calculated mean values and a Poisson
regression analysis examined the effect of uterine and oocyte age on IVF outcomes.
RESULTS: Eight hundred and sixty nine women’s (range of 23-54 years
(36.75+/-4.3)) cycles were analyzed. Autologous and donor oocytes ranged
from 23-39 years (35.88+/-4.46). Per each additional year of embryo recipient
age, the endometrial thickness decreased by 0.028mm (p¼0.086). Per
each additional year of oocyte age, the number of eggs retrieved significantly
decreased by a factor of 2.5% (p

P-133 Tuesday, October 20, 2015
THE PREVALENCE AND MANAGEMENT OF SMALL TESTIC-
ULAR MASSES INCIDENTALLY DISCOVERED ON ULTRASOUND
EVALUATION OF MALE INFERTILITY. J. M. Bieniek, a T. Juvet, a
M. Margolis, b E. D. Grober, a K. C. Lo, a K. A. Jarvi. a a Urology, University
of Toronto, Toronto, ON, Canada; b Radiology, University of Toronto, Toronto,
ON, Canada.
OBJECTIVE: Scrotal ultrasonography (US) is easily employed in the
evaluation of men in infertile couples but increases the prevalence of incidental
testicular lesions, therefore, this study was performed to assess the natural
history of these small testicular masses (STMs) and possible indications
for intervention.
DESIGN: Retrospective review of patients presenting for male infertility
evaluation who were identified to have an incidentally-found small testicular
mass with analysis of patient variables and management strategies.
MATERIALS AND METHODS: After obtaining institutional review
board approval, patients presenting to a high-volume infertility clinic were
reviewed to identify those who had tumor markers drawn or completed
more than one scrotal US between 2001 and July 2014. Charts were reviewed
to identify patients with one or more incidentally discovered small (10mm or
less) hypoechoic masses. All other testicular lesions were excluded. Patient
demographics, further imaging, laboratory evaluation, and treatment were
collected and statistical analysis performed with Student’s t tests and chi
square tests.
RESULTS: Of 4088 men completing a scrotal US for evaluation of infertility
during the study time period, 120 (2.9%) were found to have small
testicular masses. The mean age of men with a STM finding was 36.7 (range
23-62) years. Patients were followed with serial imaging for these lesions unless
concerns existed regarding the US appearance, prior history of testis cancer,
or due to patient concerns. Average follow-up duration was 1.3 years
(0.1-16.9) with 18 men (15.0%) having more than two years of follow-up.
Mean initial maximum mass dimension was 4.14mm (2.0) with vascular
flow noted in 38 patients (42.2%). Tumor markers were obtained in 54
(45.0%) with only one positive result (1.9%). Among patients followed at
least one month, interval mass growth averaged 0.1mm/yr. Eighteen patients
(15.0%) underwent extirpative surgery with 6 (33.3%) having malignant lesions
and the remaining 12 (66.7%) benign pathologies. All cases of malignant
disease were pure seminomas, measured greater than 5.2mm on US, and
demonstrated vascularity. Significant differences were found between the
surgery and surveillance groups in initial max lesion size (5.38 vs 3.92mm,
p¼ .004) and presence of vascularity (81.3% vs 33.8%, p¼ .001). Due to relatively
small numbers, no significant differences were found between men
with benign and malignant lesions on pathology.
CONCLUSIONS: This series represents the largest of its kind with nearly
3% of men undergoing scrotal US for infertility evaluation being diagnosed
with a STM. While the majority of incidental small testicular masses did not
demonstrate significant growth, lesions presenting at greater than 5mm in
size with vascularity were more likely to undergo surgery and potentially harbor
a malignancy necessitating urologic evaluation in this population.
P-134 Tuesday, October 20, 2015
NON-INVASIVE TECHNOLOGY COMBINING TIME-LAPSE IM-
AGING AND STATISTICAL MODELING: BRINGING AUTOMA-
TION INTO THE LAB TO IMPROVE BLASTOCYST
SELECTION. B. Behr, a L. Tan, b J. Conaghan, c J. Liebermann, d
A. Bartolucci, e A. A. Chen. b a Stanford Fertility and Reproductive Medicine
Center, Palo Alto, CA; b Progyny, Menlo Park, CA; c PFC, San Francisco, CA;
d Fertility Centers of Illinois, Chicago, IL; e CARS, Farmington, CT.
OBJECTIVE: The Eeva Test combines automated time-lapse analysis
with statistical modeling to predict continued embryo development. It automatically
captures quantitative image features that the human eye cannot
detect, and incorporates statistical modeling of dynamic parameters to maximize
the test’s predictive power. A new predictive algorithm includes parameters
of patient prognosis, morphology, early cleavage timings and
quantitative image features to generate a 5-category score for developmental
potential. The objective of this multi-center study was to validate this new
algorithm against embryo implantation following blastocyst transfer.
DESIGN: Retrospective multi-center study
MATERIALS AND METHODS: The study included a total of 151 patients
from 9 centers who consented to use the Eeva Test. Embryos were
transferred on Day 5, and embryos with known implantation data were
analyzed. Images and clinical data were processed by the Eeva Test’s new
Xtend algorithm, which utilizes automated image analysis software to classify
embryos into five categories by incorporating key cell division timing
parameters, Day 3 cell#, age and quantitative image features reflective of activity
during the third cell cycle.
RESULTS: The overall known implantation rate (IR) for the study population
was 39% (84/216). The 5-category output of the extended algorithm
correlated positively with blastocyst IR: 51% (36/71), 45% (33/74), 25%
(10/40), 24% (5/21), 0% (0/10). In patients younger than 35yo, the overall
known IR was 50% (62/123); however, the IR for ‘‘Category 1’’ embryos
was 60% (32/53). In the same patient group, IR for the good morphology
blastocysts was 56% (54/97); however, embryos with good morphology
and Category 1 achieved a 61% (31/51) IR.
CONCLUSIONS: The novel aspect of the new algorithm is including
computer-extracted quantitative image attributes, patient characteristics
and traditional morphology parameters in the predictive model. Although
the model was built using blastocyst as outcome variables, in current study
we were able to demonstrate that 5-category output in the new algorithm
correlated positively with blastocyst implantation. Specifically, in young patients
who are more likely in need of embryo selection, Category 1 identified
a subset of embryos with 60% implantation rate, which approaches the implantation
rate of euploid blastocysts (Forman et al, 2013). The algorithm
also identified embryos with better implantation potential among good
morphology blastocysts from young patients. Combining extended algorithm
and traditional morphology may present a non-invasive approach assisting
embryologists to select the best blastocyst(s) for transfer, and encourage
the practice of elective SET.
Supported by: Progyny (formerly Auxogyn).
P-135 Tuesday, October 20, 2015
IMPACT OF SALINE INFUSION SONOGRAM PERFORMED DUR-
ING CONTROLLED OVARIAN STIMULATION ON IN VITRO
FERTILIZATION-EMBRYO TRANSFER OUTCOMES OF THE
SAME CYCLE. N. Pereira, a A. P. Hutchinson, b J. Lekovich, a
R. T. Elias, a Z. Rosenwaks. b a The Ronald O. Perelman and Claudia Cohen
Center for Reproductive Medicine, New York, NY; b Weill Cornell Medical
College, New York, NY.
OBJECTIVE: To compare the impact of saline infusion sonogram (SIS)
performed after initiation of gonadotropin stimulation during an ongoing
in vitro fertilization (IVF) cycle to SIS performed in the follicular phase of
an earlier menstrual cycle.
DESIGN: Retrospective single-center cohort study.
MATERIALS AND METHODS: All patients undergoing fresh in vitro
fertilization (IVF) - embryo transfer (ET) cycles between January 2008
and June 2013. Patients were stratified into 2 groups based on whether
they underwent SIS during gonadotropin stimulation of an ongoing IVF cycle
or during the follicular phase of an earlier menstrual cycle. Demographic and
baseline characteristics were extracted from patient charts and included age,
gravidity, parity, body mass index (kg/m 2 ), total days of ovarian stimulation,
total dosage of gonadotropins administered (IU), number of oocytes
retrieved, and number of embryos transferred. Clinical pregnancy, biochemical
pregnancy, spontaneous miscarriage and live birth rates were compared
between the two groups. Tests of equivalence i.e., student’s t-tests and Chisquare
(c2) tests were used as indicated. Statistical significance was set at
P

Group 1: SIS performed during same IVF cycle; Group 2: SIS performed in a
previous menstrual cycle.
Parameter Group 1 (n¼152) Group 2 (n¼7648) P
Age (years) 35.7 (4.99) 35.9 (4.95) 0.62
BMI (kg/m2) 23.2 (5.99) 23.1 (5.89) 0.84
Total stimulation days 9.28 (1.90) 9.13 (1.54) 0.24
Total gonadotropins (IU) 3161.7 (1433.4) 3118.6 (1498.1) 0.72
Number of oocytes retrieved 12.1 (7.61) 11.7 (6.42) 0.45
Number of embryos transferred 2.34 (1.16) 2.41 (1.19) 0.47
Clinical pregnancy rate 64 (42.1%) 2967 (38.8%) 0.41
Biochemical pregnancy rate 12 (7.89%) 578 (7.56%) 0.88
Spontaneous miscarriage rate 8 (5.26%) 275 (3.60%) 0.28
Live birth rate 55 (36.2%) 2687 (35.1%) 0.79
P-136 Tuesday, October 20, 2015
ARE MORPHOKINETIC PARAMETERS AFTER THAWING
RELATED TO IMPLANTATION IN DAY 3 CRYOPRESERVED
EMBRYOS?. E. Fernandez Gallardo, C. Spiessens, T. D’Hooghe,
S. Debrock. Leuven University Fertility Centre, Leuven University Hospital,
Leuven, Belgium.
OBJECTIVE: To study, for the first time, the relation between morphokinetic
parameters after thawing and implantation in intact and non-intact vitrified/warmed
and slow frozen/thawed embryos.
DESIGN: Presence of mitosis, presence of compaction, time to mitosis
and time to compaction were analyzed retrospectively for 82 embryos (35
slow frozen and 47 vitrified). After survival, embryos were cultured overnight
in ASTECÒ time lapse incubator (TLI) and transferred. Implantation
was defined as the presence of fetal sac (intra- or extrauterine) at 6-8 weeks
pregnancy after transfer. All transfers included in the study had either 0% or
100% embryos implanted.
MATERIALS AND METHODS: Embryos were vitrified (EmbryoStore,
GynemedÒ) or slow frozen (Vitr Kit Freeze, Irvine ScientificÒ) on day 3
if R6 cells and

concerns (75%) in comparison to other women in the study- 15% Caucasian,
5% Hispanic and 5% Asian.
CONCLUSIONS: Most of the women in this qualitative study raised financial
concerns relating to their fibroids in their interviews. African-American
women, however, brought up financial issues substantially more than White,
Hispanic or Asian women. This is particularly concerning given that fibroids
disproportionately affect African-American women. For the women who expressed
financial challenges, they were mainly related to an inability to pay
for diagnostics and treatment needed to alleviate symptoms or for definitive
treatment. More work needs to be done to improve insurance coverage for
fibroid diagnosis and treatment as well as to quantitatively define the financial
impact of these prevalent tumors on individual women.
Supported by: NIH WRHR Program K12HD050121; RWJ Foundation;
NMH; Evergreen Foundation.
P-139 Tuesday, October 20, 2015
ROLE OF DOPAMINE AGONISTS IN UTERINE
MYOMA. B. Bhagavath, a H. Prizant, b S. R. Hammes. c a OBGYN, Strong
Fertility Center, University of Rochester, Rochester, NY; b OBGYN, University
of Rochester, Rochester, NY; c University of Rochester, Rochester, NY.
OBJECTIVE: 1. Study the expression of dopamine receptors, prolactin
and prolactin receptors in leiomyoma in an animal model for uterine fibroids
(Uterine-specific Tsc2 knockout (Tsc2 KO) mice)2. Study the effect of dopamine
agonists on the growth of leiomyoma in Tsc2 KO mice
DESIGN: Interventional study using a mouse model for leiomyoma
MATERIALS AND METHODS: Tsc2 KO mice are known to develop
leiomyoma spontaneously. Previously performed experiments have established
that these tumors are estrogen sensitive and respond well to treatment
with aromatase inhibitor, Letrozole, which decreases circulating levels of estrogen.
Real time quantitative polymerase chain reaction (qPCR) was performed
on mRNA from the uterine tissue to compare prolactin (PRL),
prolactin receptor (PRLR) and dopamine receptor (DRD2) expression with
that of wild type (wt) mice. Serum PRL levels were compared between wt
mice and Tsc2 KO mice. Ovaries were surgically removed (ovariectomy)
from 18 week wt and TSC2 KO mice and mice were treated with estradiol
for 1 week. Uterine PRL, PRLR and DRD2 expressions were studied in these
mice. To study the effect of dopamine, four knockout animals were injected
with cabergoline (dopamine agonist) subcutaneously three times a week for
12 weeks from 18 weeks age. At 30 weeks age, these animals were sacrificed
and their uteri subjected to gross examination. In addition, the uteri were
sectioned, fixed and stained with hematoxylin and eosin, and compared
with uteri of wt mice.
RESULTS: In Tsc2 KO mice at 18 weeks age, PRLR but not DRD2
expression markedly and significantly increased after administration of estradiol
for one week. Local PRL levels are undetectable in the uteri of wt mice
and detected in small quantities in Tsc2 KO mice at 18 and 30 weeks age.
Ovariectomy at 18 weeks age for a period of 12 weeks significantly reduced
PRLR expression and PRL is no longer detectable in the uteri of uterine-specific
Tsc2 KO mice at 30 week age. Serum prolactin levels were significantly
suppressed in Cabergoline treated mice. Gross pathological examination did
not show any improvement in the leiomyoma in Tsc2 KO mice treated with
Cabergoline. Examination of fixed and stained sections of uteri are indistinguishable
from those of untreated Tsc2 KO mice.
CONCLUSIONS: Presence of local uterine PRL production in Tsc2 KO
mice along with PRLR suggests a complex, estrogen dependent role in leiomyoma
development. Although DRD2 agonist did not show a significant
clinical effect on leiomyoma development, it is possible that a PRLR antagonist
might have a clinical effect on leiomyoma development.
Supported by: This work was Supported by a grant from the Richard W.
Goode and Mae Stone Goode Foundation.
P-140 Tuesday, October 20, 2015
HISTORY OF UTERINE LEIOMYOMATA AND TIME-TO-PREG-
NANCY IN BLACK WOMEN. L. A. Wise, a R. Radin, b
L. Rosenberg. c a Department of Epidemiology, Boston University School of
Public Health, Boston, MA; b NIH, Rockville, MD; c Slone Epidemiology
Center, Boston University, Boston, MA.
OBJECTIVE: To evaluate the extent to which a history of uterine leiomyomata
(UL) and its characteristics (tumor location, size, and number) are associated
with delayed time-to-pregnancy (TTP).
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: We assessed the association between UL
and fecundability among participants aged 21-40 years from the Black
Women’s Health Study. In 2008, women with incident diagnoses of UL
(1995-2008) reported detailed data on their date of UL diagnosis, symptoms,
tumor characteristics and treatment via supplemental questionnaire. In 2011,
women reported the time-to-pregnancy (TTP) in months for each of their
planned pregnancies resulting in a live birth during their reproductive life.
The incident period for the analysis was restricted to 1995-2011 and only
women with UL diagnoses prior to a given pregnancy attempt were considered
‘‘exposed.’’ Proportional probabilities regression models were used to
estimate fecundability ratios (FRs) and 95% confidence intervals (CI), with
adjustment for age and calendar year at pregnancy attempt, prepregnancy
BMI, physical activity, education, and smoking history.
RESULTS: During 1995-2011, 1,353 women contributed 1,572 births and
8,250 months of attempt time. A history of UL was associated with a slight
delay in conception: the FR for a history of UL diagnosis was 0.92 (95% CI:
0.81-1.05) relative to no UL diagnosis (referent for all comparisons). The FR
for submucosal UL was 0.71 (95% CI: 0.45-1.12) and 0.91 (95% CI: 0.63,
1.31) for intramural UL. FRs for women with 1, 2, and R3 UL tumors
were 0.89 (95% CI: 0.63-1.24), 0.89 (95% CI: 0.65-1.21), and 0.87 (95%
CI: 0.68-1.10), respectively. FRs for women with UL tumors

References:
1. Kho K, Nezhat C. Evaluating the Risks of Electric Uterine Morcellation.
JAMA 2014 (311) 9: 905-906.
2. Seidman MA, Oduyebo T, Muto MG, Crum CP, Nucci MR, Quade BJ.
Peritoneal dissemination complicating morcellation of uterine mesenchymal
neoplasms. PLoS One. 2012;7(11):e50058.
3. Steiner RA, Wight E, Tadir Y, et al. Electrical cutting device for laparosopic
removal of tissue from the abdominal cavity. Obstet Gynecol.
1993;81:471-474.
4. Milad MP, Milad EA. Laparoscopic morcellator-related complications.
J Minim Invasive Gynecol. 2014;21(3):486-491.
5. American Association of Gynecologic Laparoscopists-Tissue Extraction
Task Force. Morcellation during uterine tissue extraction. Available at:
www. Aagl.org/wp-content/uploads/2014/05/Tissue_Extraction_TFR.pdf.
P-142 Tuesday, October 20, 2015
PARADOXIC EFFECT OF MIFEPRISTONE ON LEIOMYOMA
EXTRACELLULAR MATRIX COMPONENT DERMATOPONTIN
EXPLAINED. A. Patel, a M. Malik, a J. L. Britten, a J. Cox, b
W. H. Catherino. a a Obstetrics and Gynecology, Uniformed Services University
of the Health Sciences, Bethesda, MD; b Program in Reproductive and
Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child
Health and Human Development, Bethesda, MD.
OBJECTIVE: Progesterone stimulates leiomyoma growth, but the impact of
this hormone on the extra cellular matrix (ECM) genes remains unknown. Dermatopontin
is a unique ECM gene that is known to be decreased in leiomyomas
compared to patient-matched myometrium, in contradistinction to other
biomarker ECM genes which are elevated. If progesterone promotes the leiomyoma
phenotype of excessive and aberrant ECM, then treatment should result
in decreased dermatopontin, while treatment with the anti-progestin mifepristone
should increase expression. Our objective is to characterize the mechanism
of progesterone on the expression of the ECM protein dermatopontin.
DESIGN: Laboratory setting
MATERIALS AND METHODS: Immortalized leiomyoma cells were
treated with the progestin R5020 and the anti-progestin mifepristone at clinically-relevant
concentrations and various time-points up to 120 hours. Dermatopontin
protein concentration was quantified using Western blot.
RESULTS: At 1 and 6 hours exposure, paradoxically both progestin and
mifepristone-treated cultures resulted in a reduction of dermatopontin protein
concentrations (0.50.23 fold and 0.7 0.08 fold at 1 hour and
0.40.19 fold and 0.6 0.14 fold at 6 hours) respectively. At 72 and 120
hours, dermatopontin expression remained decreased (0.17 0.2 fold) with
mifepristone treatment. However, in the same time-frame, progestin impact
on dermatopontin expression was lost (1.0 0.25 fold at 72 hours) and (0.6
0.34 fold at 120 hours ).
CONCLUSIONS: Our results demonstrate that mifepristone has a progestational
effect on dermatopontin protein concentration, and this effect is
maintained despite a diminution of effect by a standard progesterone receptor
agonist. These findings suggest the production of specific signaling pathway
proteins by progestins that regulate dermatopontin expression. Better understanding
of this pathway would result in improved therapeutic options for
women suffering with symptomatic uterine leiomyomas.
Supported by: This research was Supported by Intramural grant from Uniformed
Services University of the Health Sciences, QP85GF13 and R21,
R085193713.The research was also Supported, in part, by the intramural
research Program in Reproductive and Adult Endocrinology, NIH.
P-143 Tuesday, October 20, 2015
SEEKING THE TRUTH: A QUALITATIVE ASSESSMENT OF
WOMEN’S EXPERIENCES SEEKING AND OBTAINING KNOWL-
EDGE OF UTERINE FIBROIDS. M. Ghant, a K. Sengoba, b
G. Mendoza, b E. E. Marsh. b a University of Illinois at Chicago College of
Medicine, Chicago, IL; b Northwestern University Feinberg School of Medicine,
Chicago, IL.
OBJECTIVE: To identify thematic content on health information-seeking
amongst women with fibroids.
DESIGN: Qualitative semi-structured interviews and demographic surveys
MATERIALS AND METHODS: Women between the ages of 25-55, who
either currently had symptomatic uterine fibroids or received treatment for
their uterine fibroids within the past 12 months, were recruited from an urban
academic medical center and community-based organizations. Women
completed in-depth, one-on-one recorded interviews, a demographic survey
and a health literacy assessment tool. Interviews were transcribed verbatim
and uploaded to NVivo version 10 for data management and thematic coding.
Three coders used a grounded theory approach to identify major themes and
subthemes.
RESULTS: Sixty women completed the study, resulting in a total of 35
hours of interviews yielding 1,357 transcribed pages. The k across coders
was 0.94. The mean age of participants was 43.0 6.8 (mean SD).
61.7% of participants self-identified as African-American, 25.0% as Caucasian,
8.3% as Hispanic and 5.0% as Asian. 68.3% of the participants had at
least a 4-year college degree. The average health literacy score was 6.8 on
a 7-point scale. Four themes were identified regarding information seeking
and sharing amongst participants. Theme 1 focused on sources of information,
with many women reporting that they used the Internet, family and
friends, physicians, books, the radio and church to obtain information on
uterine fibroids. Theme 2 focused on women’s dissatisfaction with available
information. Despite utilization of multiple resources, the majority of women
(78.3%) were dissatisfied with either the clarity, quality, or quantity of available
information on fibroids for patients. Theme 3 focused on hindsight, as
many women expressed that if they had more knowledge regarding fibroids,
they would have made different decisions regarding their treatment. A fourth
and final theme highlighted what subjects felt that women with fibroids need
to know about the tumors, and centered on the promotion of self-education
and the importance of finding a trustworthy physician.
CONCLUSIONS: A diverse cohort of well educated, health literate
women with a history of symptomatic uterine fibroids felt that there is not
an existing source of high-quality patient-centered information available.
Despite utilizing multiple resources and venues to educate themselves,
women felt that more needed to be done to educate patients and ensure
that they had access to unbiased counseling and trustworthy providers.
Increasing women’s knowledge on fibroids empowers them in the face of
their diagnosis, allowing them to make informed decisions regarding their
treatment options without regret.
Supported by: NIH WRHR Program K12HD050121; RWJ Foundation;
NMH; Evergreen Foundation (EEM).
P-144 Tuesday, October 20, 2015
TISSUE-SPECIFIC EXPRESSION OF ESTROGEN RECEPTOR 1 IS
REGULATED BY DNA METHYLATION IN A T-DMR (TISSUE-
DEPENDENT AND DIFFERENTIALLY METHYLATED
REGION). H. Asada, M. Okada, H. Tamura, N. Sugino. Obstetrics
and Gynecology, Yamaguchi University School of Medicine, Ube, Japan.
OBJECTIVE: The mechanism controlling tissue-specific expression of
ESR1 is still unclear. It has been reported that DNA methylation of a specific
region of the gene has an important role in determining tissue- and cell-specific
gene expression. This region is called the T-DMR (tissue-dependent and
differentially methylated region). We previously found a possible link between
the mRNA expression of ESR1 and the DNA methylation status of
ESR1 promoter, suggesting that the region includes the T-DMR for tissuespecific
ESR1 expression. It is known that ESR1 has several transcription
starts sites (TSS) and corresponding upstream exons (upstream Exon-A to
-E1). The transcription of ESR1 starts from any of these upstream exons,
and the upstream exons are used in a tissue-dependent manner. Three upstream
exons, upstream Exon-A, -B and -C, are often used in the tissues
with high ESR1 expression. In the present study, we investigated whether human
ESR1 has a T-DMR and whether DNA methylation of the T-DMR regulates
its expression. We also investigated whether T-DMR is present in each
upstream exon of ESR1.
DESIGN: basic research
MATERIALS AND METHODS: We obtained informed consent from patients
and approval by Yamaguchi University. DNA methylation profiles and
mRNA expression profiles of ESR1 were analyzed in the endometrium,
mammary gland, placenta, skin, and breast cancer tissues by sodium bisulfite
sequencing. T-DMR-methylated reporter assay was performed to examine
whether DNA methylation at the T-DMR actually suppresses transcription
of ESR1.
RESULTS: ESR1 expression was tissue-specific, being high in the endometrium
and mammary gland and low/nil in the placenta and skin. In all
of the tissues, the proximal promoter regions were unmethylated. On the
other hand, the distal regions were unmethylated in the endometrium and
mammary gland, but were hypermethylated in the placenta and skin,
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indicating this region is a T-DMR. T-DMR-methylated reporter assay revealed
that DNA methylation of the T-DMR suppressed ESR1 transcription.
DNA methylation analysis around upstream Exon-A, -B and -C showed each
upstream exon has its own T-DMR and DNA methylation of the region is
associated with transcriptional regulation in each upstream exon. In some
breast cancer cases, T-DMRs regulate ESR1 transcription via DNA methylation
in a manner similar to normal tissues, while in other cases, ESR1 transcriptional
regulation deviates from the regulation seen in normal tissues.
CONCLUSIONS: This is the first report to demonstrate that ESR1 has T-
DMRs, and that the T-DMRs regulate tissue-specific ESR1 expression via
DNA methylation in normal tissues. We also found that each upstream
exon has a corresponding T-DMR, of which DNA methylation status is
involved in regulating transcription of the upstream exon. Furthermore, our
results show some breast cancer cases deviate from the normal regulatory
mechanism of the transcription regulation of ESR1.
P-145 Tuesday, October 20, 2015
THE ROLE OF TYPE 2 DIABETES IN MODIFYING THE RISK FOR
FIBROIDS. D. R. Velez Edwards, a M. Wellons, b K. Hartmann, a
T. L. Edwards. c a Obstetrics and Gynecology, Vanderbilt University, Nashville,
TN; b Division of Diabetes, Endocrinology, and Metabolism, Department
of Medicine, Vanderbilt University, Nashville, TN;
c Medicine,
Vanderbilt University, Nashville, TN.
OBJECTIVE: Uterine fibroids (UF) affect 77% of women by menopause,
and account for $9.4 billion in annual healthcare costs. Type-2-diabetes
(T2D) diagnosis has inconsistently associated with UF risk in prior studies.
Differences in results across studies may be due to differences in T2D
severity and treatment. To further evaluate the relationship between T2D
and UFs we tested for association between T2D and UF risk before and after
adjustment for T2D severity and stratifying by T2D treatments.
DESIGN: This nested case-control study of a clinical cohort limited to
women a UF diagnosis after T2D diagnosis and who had a pelvic imaging
both prior to T2D diagnosis and after T2D diagnosis.
MATERIALS AND METHODS: UF cases were those diagnosed with a
UF using pelvic imaging after T2D diagnosis. T2D outcome and treatments
were assessed using inpatient records and natural language processing of
clinical notes implemented in MedEx software. Logistic regression, adjusted
for covariates, was used to test for association between T2D outcome and UF
risk. Secondary analyses were also performed adjusting models for HgBA1C
levels and evaluating diabetics on specific treatments (metformin, thiazolidinedione,
or insulin) for association with UF risk.
RESULTS: We identified 3,789 subjects with T2D outcome, 714 were diabetics
and 3,075 were T2D controls. Among these 16% (N¼608) had UFs.
In adjusted models we observed that T2D decreased risk for UFs (adjusted
odds ratio [OR] 0.61, 95% confidence interval [CI] 0.47 to 0.80). However,
when models were further adjusted for HgbA1C effects were inversed
(adjusted OR¼1.55, 95% CI 0.96 to 2.49), suggesting the models were
confounded by T2D severity. Additional analyses stratifying by diabetics
with HgbA1C

increasingly in cancer genomics research. We use the NGS to investigate the
process of malignant transformation of endometriosis.
DESIGN: We collected four parts of pathological specimen, consisting of
eutopic endometrium, non-atypical endometriosis tissue, atypical endometriosis
and ovarian carcinoma tissue from EAOC patients. NGS was applied to
investigate the gene mutations during the transformation process.
MATERIALS AND METHODS: In this project, we use the Ion AmpliseqTM
Comprehensive Cancer Panel (CCP) which targets the exons of
409 tumor suppressor genes and oncogenes in order to identify pathogenic
mutations associated with EAOC. Ten cases of EAOC were enrolled in this
study. For each case, macrodissection was performed to separate different
four types of cells from formalin-fixed paraffin-embedded (FFPE) sections.
RESULTS: The DNA of endometriosis patients was extracted from
formalin-fixed paraffin embedded (FFPE) tissue. Samples include four transformation
processes that are normal eutopic endometrium, nonatypical endometriosis,
atypical endometriosis and carcinoma tissue from the same
patients of endometriosis associated with ovarian cancer. The NGS direct
sequencing found that coding sequence of exon 1 in ARID1A gene and exons
20 in PIK3CA gene were screened for mutations. Our results showed that
ARID1A and PIK3CA mutation (c.6488delG and c.3140A>G) were identified
in the carcinoma tissue, but not in endometrium, non-atypical endometriosis
and atypical endometriosis.
CONCLUSIONS: Our study aims to develop and validate NGS platform
for identifying the critical factor in the early event of the malignant transformation
process of endometriosis. ARID1A and PIK3CA mutations
contribute to the transformation process in our study. We believe this study
will shed new light on fundamental aspects in the understanding the molecular
pathogenesis of malignant transformation of ovarian endometriosis.
Supported by: This work is Supported by MST Taiwan [grant number 102-
2628-B-037-011-MY3 and 102-2632-B-037-001-MY3].
P-148 Tuesday, October 20, 2015
COMPARATIVE ANALYSIS OF MEMBRANE MICROPARTICLES
(MP) EXPRESSION IN WOMEN WITH EPITHELIAL OVARIAN
CANCER (EOC) AND ENDOMETRIOMAS. J. A. Falcao, Jr., a
F. F. Nunes, b J. Marinuzzi, c O. A. Martins-Filho, b A. T. Carvalho, b
R. M. Lamaita, a M. M. Carneiro, a A. L. Silva-filho. a a Obstetrics and Gynecology,
Federal University of Minas Gerais, Belo Horizonte, Brazil; b Centro
de Pesquisas Rene Rachou - Fiocruz, Belo Horizonte, Brazil; c UNESP Botucatu,
Botucatu, Brazil.
OBJECTIVE: Microparticles (MP) are small vesicles derived from cell
membranes which have been recognized as important mediators of cell activity
and may associated with pathological and physiological processes such as
immune response, cell differentiation, vascular disorders and cancer. The aim
of this study is to compare circulating MP according to their specific cellular
origin in women with endometrioma, EOC and controls.
DESIGN: Prospective evaluation of 60 women from March 2010 to
October 2013 divided into three groups: EOC (n¼26); endometriomas
(n¼18); control (n¼16; women operated for benign gynecologic disease
with normal ovaries).
MATERIALS AND METHODS: A convenience sample was used due to
the low prevalence of EOC and the strict inclusion and exclusion criteria.
Surgical staging was performed according to the FIGO classification or
ASRM endometriosis staging as appropriate. Exclusion criteria were: previous
chemotherapy and/or radiotherapy; immune system diseases diagnosis
and / or use of immunosuppressive drugs within the past 6 months, acute infections,
identification of distinct malignancy from EOC in the histopathological
exam. Serum levels of CD66 +/neutrophils, CD45 +/leukocytes,
CD14 +/monocytes, CD235 +/erythrocytes, CD51 +/endothelium, CD41
+/platelets, CD3 +/lymphocytes were performed by flow cytometry. The differences
between groups were evaluated by Mann-Whitney or Kruskal-
Wallis tests. P < 0,05 was considered statistically significant.
RESULTS: Mean patient age was: EOC (62 14.06), endometrioma
(3710.31) and control (408,8) years. Ten cases were identified as FIGO
stage I and II, and 16 as III/IV. Fifteen women were classified as ASRM stage
III and 3 as stage IV. Women with endometrioma were associated with higher
circulating levels of CD45 +/leukocytes (p¼ 0.0292), CD14 +/monocytes
(p¼0.012), CD235 +/erythrocytes (p¼0.0341), CD51 +/endothelium
(p¼0.0228) and CD41 +/platelets (p¼0.0464) compared to patients with
EOC and control group. No association was found between age of the patients
of the 3 groups and the MP assay neither with maximum diameter of
the lesion in the endometrioma group.
CONCLUSIONS: Our data shows for the first time that women with endometrioma
present with higher MP circulating levels compared EOC patients.
MP have potential use in understanding the cellular microenvironments associated
with these diseases and may contribute to the improvement in
screening, diagnosis and therapeutic strategies.
P-149 Tuesday, October 20, 2015
THE LONG TERM RECURRENCE RATE AFTER CONSERVATIVE
SURGICAL TREATMENT OF ENDOMETRIOSIS IN
ADOLESCENTS. Y. Cho, a S. Lee, b M. Kim, c J. Bae, d M. Han, d
J. Park. d a Department of Obstetrics and Gynecology, Dong-A University
Medical Center, Dong-A University, College of Medicine, Busan, Korea, Republic
of; b Dankook University, School of Medicine, Cheil Gene, Seoul, Korea,
Republic of; c Department of Obstetrics and Gynecology, CHA Gangnam
Medical Center, CHA University, Seoul, Korea, Republic of; d Dong-A University
Hospital, Busan, Korea, Republic of.
OBJECTIVE: Endometriosis, while generally considered as a disease that
affects adult women, has become increasingly recognized as a chronic illness
that can begin during adolescent and young adulthood. This patient group
presents particular challenges in terms of differential diagnosis, variable presentation
and symptoms, and choice of treatment. There is very limited
research in adolescents with endometriosis and long term studies about the
recurrence or progression are not well understood. We aimed to evaluate
the long term recurrence rate of ovarian endometriomas in adolescents
following the first conservative surgical treatments.
DESIGN: Multicenter retrospective cohort study
MATERIALS AND METHODS: Patients % 20 years of age who were
surgically treated with laparoscopic enucleation of ovarian endometrioma
were selected. We included patients only who were followed up more than
36 months. We excluded patients who had reproductive tract anomalies,
those who underwent non-conservative procedures, such as oophorectomy,
those who underwent cyst aspiration. Recurrence of the endometrioma was
established on the basis of transvaginal or transrectal sonography documenting
the presence of a cystic mass with a diameter of R 20 mm. Baseline surgical
characteristics were analyzed.
RESULTS: We recruited 51 adolescent patients who were followed up
more than 36 months. The mean age of patients was 19.0 1.1 years
(range, 16-20 years). According to our definition of recurrence, 15 patients
(29.4%) experienced recurrence of ovarian endometrioma after first
laparoscopic cyst enucleation. The overall cumulative recurrence rates of
ovarian endometrioma per patient at 24, 36, 60, and 96 months after
first-line surgery as 8.2%, 10.2%, 20.8%, and 37.4%, respectively.
Surgical characteristics, such as the diameter of the cyst, disease stage,
unilateral or bilateral involvement, and co-existence of deep endometriosis,
and postoperative medical therapy were not associated with recurrence in adolescents.
CONCLUSIONS: The long-term recurrence rate in adolescent after the
conservative surgery was dependent on the months that had elapsed since
treatment, and this value increased over time. Long-term and continuous
follow up is needed who have undergone surgical treatment for endometriosis
in adolescent period.
P-150 Tuesday, October 20, 2015
EXPRESSION OF HOXB4 IN ENDOMETRIAL TISSUES FROM
WOMEN WITH OR WITHOUT ENDOMETRIO
SIS. G. M. Alkusayer, a,b B. Peng, c C. Klausen, d S. Lisonkova, d
M. Kinloch, e P. Yong, f M. A. Bedaiwy. g a Department of Clinical Sciences,
College of Medicine, Princess Nourah Bint Abdulrahman University, Riyadh,
Saudi Arabia; b Department of Obstetrics and Gynaecology, University of British
Columbia, Vancouver, BC, Canada; c University of British Columbia, CFRI,
Vancouver, BC, Canada; d University of British Columbia, Vancouver, BC, Canada;
e Vancouver General Hospital, Vancouver, BC, Canada; f Assistant Professor,
Vancouver, BC, Canada; g BC Women’s Hospital, Vancouver, BC, Canada.
OBJECTIVE: HOX genes play important roles in the functional differentiation
of adult tissues by regulating proliferation, angiogenesis, adhesion
and motility. The aim of our study was to examine the expression and localization
of HOXB4 in normal human endometrial tissues as well as eutopic
endometrium and ectopic implants from women with endometriosis
throughout the menstrual cycle.
DESIGN: Case-control study.
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MATERIALS AND METHODS: Normal endometrial tissues were
collected from 20 pre-menopausal women with no history of endometriosis.
Additionally, matched eutopic endometrium and ectopic endometrial implants
were obtained from 40 women with endometriosis and no recent history
of hormonal therapy [deep infiltrating endometriosis (DIE), n¼20;
endometrioma, n¼20]. Using the Histoscore method, a pilot immunohistochemical
analysis of HOXB4 was performed on 9 normal endometrial tissues
and 12 eutopic/ectopic endometrial tissue pairs. Kruskal-Wallis non-parametric
test was used to compare multiple categories, Wilcoxon test was
used for pair-wise comparisons.
RESULTS: HOXB4 was immunolocalized in the nuclei of glandular
epithelial cells in normal endometrium and eutopic/ectopic endometrial tissues
from women with endometriosis. HOXB4 immunoreactivity was absent
from endometrial stromal cells in all tissues examined. No significant difference
in HOXB4 immunostaining intensity was found between controls and
eutopic endometrium from endometriosis. Interestingly, HOXB4 immunoreactivity
was significantly reduced in ectopic DIE compared to eutopic endometrium
[P¼ 0.041] and endometrioma [P¼ 0.05] in women with
endometriosis. Moreover, HOXB4 immunoreactivity in eutopic endometrium
of disease and control groups combined was significantly higher in
the proliferative phase than in the secretory phase [P¼ 0.026]. Conversely,
HOXB4 immunoreactivity in ectopic implants was lower in proliferative
phase than in the secretory phase [P¼ 0.021].
CONCLUSIONS: During the menstrual cycle the expression of HOXB4 in
endometrial glandular epithelial cells is higher in the proliferative phase than
the secretory phase. On the whole, HOXB4 expression does not appear to
differ between normal endometrium and eutopic endometrium from endometriosis.
However, its expression is reduced in DIE, but not endometrioma, and
may be dysregulated in ectopic implants depending on the phase of the menstrual
cycle.
Supported by: Internal fund, Department of Obstetrics and Gynecology,
University of British Columbia, Vancouver, BC, Canada. Graduate Sponsorship
Program,Princess Nourah bint Abdulrahman University,Riyadh,
Kingdom of Saudi Arabia.
P-151 Tuesday, October 20, 2015
COULD THE UNILATERAL OVARIAN ENDOMETRIOSIS
AFFECT THE CONTRALATERAL OVARY? NEW INSIGHTS
FROM FOURIER TRANSFORM INFRARED (FTIR)
SPECTROSCOPY. G. Gioacchini, a E. Sereni, b C. Zaca’, b
E. Giorgini, c V. Notarstefano, a L. Vaccari, d O. Carnevali, a A. Borini. b
a Universita Politecnica delle Marche, Ancona, Italy; b Tecnobios Procreazione,
Bologna, Italy; c Polytechnic University of Marche, Ancona, Italy; d E-
lettra-Sincrotrone Trieste, Trieste, Italy.
OBJECTIVE: Up to date, endometriosis’ possible impact on follicle cells
metabolism has never been highlighted. In this light, the aim of this study was
to evaluate, by FT-IR microspectroscopy, metabolic changes on granulosa
cells (GCs) isolated from endometriosis affected ovaries and contralateral
healthy ones
DESIGN: This prospective non-randomized study has been conducted
from January to September 2014 on women undergoing a controlled ovarian
hyperstimulation for an IVF treatment. In particular, GCs were collected
from both ovaries of 10 women with a diagnosis of unilateral ovarian endometrioma
at the time of oocytes retrieval. 9 women with male, idiopathic or
tubal-factor infertility diagnosis were selected for the control group. The
three experimental groups matched for female age (36.24.1 vs 35.42.6)
MATERIALS AND METHODS: GCs obtained from follicles aspirates
were isolated from red blood cells and follicular fluid by density gradient
centrifugation. FTIR analysis was performed by using a Bruker Vertex 70
Interferometer with a Hyperion 3000 Vis-IR microscope. QPCR analysis
Supported FTIR results. . Data are presented as mean S.D. Two-Way AN-
OVA followed by Tukey test as Multiple comparisons test, was used for comparison
among experimental groups. All statistical analyses were performed
using the statistical software package Prism5 (Graphpad Software, Inc. USA)
with significance accepted at P

Cornell University, New York, NY; c The Ronald O. Perelman and Claudia
Cohen Center, New York, NY.
OBJECTIVE: Mechanism of infertility in endometriosis potentially involves
oocyte/embryo defects and lower fertilization and implantation rates.
Cumulus complex in endometriosis patients demonstrates mitochondrial
dysfunction and higher oxidative stress. We sought to investigate if ICSI,
which diminishes oxidative stress when compared to insemination, improves
IVF outcomes in patients with endometriosis.
DESIGN: Retrospective cohort
MATERIALS AND METHODS: 221 patients who had surgically or sonographically
confirmed endometriosis were compared with 150 patients with
unexplained infertility. Patients with endometriosis as a sole cause of infertility
were included. Exclusion criteria [1. > 41yo; 2. Male factor (sperm
count < 15 x 106/cc, motility less than 40%, older than 55yo]. Primary outcomes:
1. Number of retrieved eggs; 2. Number of 2 pronuclear zygote (2PN)
stage embryos; 3. Proportion of day 5 transfers; 4. Number of frozen blastocysts.
Secondary outcomes: 1. Clinical intrauterine pregnancy [IUP] rate; 2.
Live birth rate. Student’s t-test and ANOVA were used to continuous and
Fisher’s exact and Chi-square tests for categorical variables.
RESULTS: Within the endometriosis group, 124 patients utilized standard
insemination, whereas 97 utilized ICSI. Endometriosis-insemination, endometriosis-ICSI
and the control groups were matched for age, BMI, anti-mullerian
hormone (AMH) and the number of eggs retrieved [11.95.3 vs. 13.13.1 vs.
13.1 5.9, P¼0.07]. The control and endometriosis-ICSI groups did not differ
in terms of number of 2PNs, number of day 5 transfers, IUP and live birth rates
(Table). Endometriosis-ICSI group demonstrated significantly higher percentage
of 2PN embryos, percentage of day 5 transfers, number/proportion of clinical
IUP as well as live birth rates when compared with the endometriosisinsemination
group. Additionally, endometriosis-ICSI group had a lower proportion
of cycles with no normal fertilization [0 (0%) vs 4 (3.2%), but this difference
did not reach statistical significance.
CONCLUSIONS: Our findings suggest that endometriosis might be an
indication for ICSI, as patients with endometriosis who utilized ICSI
achieved better outcomes (higher proportion of normal fertilization, higher
number of day 5 transfers, clinical pregnancy and live birth rates) when
compared to standard insemination cycles.
Table 1
Endometriosis- Endometriosis-
Insemination(124) ICSI(97) P 1 Controls(150) P 2 P 3
Mean # eggs retrieved 11.95.3 13.13.1 0.5 13.15.9 0.2 0.8
Total# of retrieved eggs(%) 1301 1269 1954
Mean # of 2PN 6.93.9 8.44.9 0.01 8.85.1

Obstetrics and Gynecology, West China Second University Hospital of Sichuan
University, Chengdu, Sichuan, China.
OBJECTIVE: Endometriosis-related infertility is common in women of
reproductive age. Progesterone resistance, especially aberrantly expressed
progesterone receptor in the eutopic endometrium, is considered as a key
causal factor. Our objective is to explore microRNA-mediated mechanism
controlling aberrant progesterone receptor expression in infertile women
with minimal or mild endometriosis.
DESIGN: microRNA Array, combined with Human Gene Expression microarrays,
was used to screen eutopic endometrium of infertile women with
minimal or mild endometriosis. Bioinformatics analysis predicted that microRNA-196a
target the PGR 3 0 UTR. We studied the relationship between
microRNA-196a level and PGR expression in endometrial stromal cells
(ESCs). According to mRNAs microarray result, MAPK pathway is activated
in eutopic mid-secretory endometrium. The role of MAPK pathway involved
regulation of miR-196a on PGR was also investigated.
MATERIALS AND METHODS: The study was conducted at West China
Second University Hospital of Sichuan University. 22 infertile women with r-
AFSI-IIendometriosis and 20 disease-free control subjects were enrolled.
MiRCURY LNATM microRNA Array(v.18.0)(Exiqon), Human 444K
Gene Expression Microarrays v2(Agilent), qRT-PCR, cell culture, transfections,
luciferase reporter assays, and western bolt were used in this study.
RESULTS: 66 differently expressed microRNAs (fold changeR2.00 and
p12wks, FSH > 12). The number of oocytes retrieved, fertilization
rate, clinical pregnancy rate and miscarriage rate were compared. Statistical
comparisons were done with Stata10.0 software.
RESULTS: The clinical pregnancy rate was 114/355(32.1%) in women
with only endometriosis, 17/88(19.3%) in endometriosis with adenomyosis
and 12/64(20.3%) in only adenomyosis. The odds ratio for clinical pregnancy
showed that the adenomyosis groups (Group B & C) had significantly lower
pregnancy rate (Avs B; OR: 0.51, 95% CI : 0.28-0.89, p< 0.02; A vs. C; OR:
0.49, 95% CI:0.25-0.94, p< 0.03).Miscarriage was observed in 26/355 in
Group A,13/88 in Group B and 10/64 in Group C. Miscarriage rate was
significantly higher in pregnancy associated with adenomyosis (Avs.B;
OR: 0.45, 95% CI : 0.22-0.92, p< 0.03;A vs.C; OR: 0.42, 95% CI:0.19-
0.93, P< 0.03).Number of oocytes retrieved and fertilization rates were comparable
in all the three groups.
CONCLUSIONS: Presence of adenomyosis appeared to have adverse effects
on IVF/ICSI outcomes in terms of lower pregnancy rate and higher
miscarriage rate in women pretreated with long-term GnRH-agonist.
Screening for adenomyosis before going for IVF/ICSI may be considered,
so that counseling can be done regarding decreased probability of viable
pregnancy in women having adenomyosis.
P-158 Tuesday, October 20, 2015
ANGIOGENIC ACTIVITY OF PGF2A IN WOMEN WITH
ENDOMETRIOSIS. H. Rakhila, a M. Bergeron, b M. Daris, b
M. Leboeuf, b M. Lemyre, b C. Rheaume, b M. Pouliot. a a Department of
Microbiology and Immunology, Centre Hospitalier Universitaire de Quebec,
Faculty of Medicine, Universite Laval, Quebec, QC, Canada; b Department of
Obstetrics and Gynecology, Centre Hospitalier Universitaire de Quebec, Faculty
of Medicine, Universite Laval, Quebec, QC, Canada.
OBJECTIVE: To investigate interleukin-8 (CXCL8) and vascular endothelial
growth factor (VEGF) expressions (potent angiogenic factors) in
endometrial stromal cells of women with endometriosis compared with controls
in response to Prostaglandin F2a (PGF2a), an important regulators of
female reproductive function.
DESIGN: Primary cultures of eutopic and ectopic endometrial stromal
cells isolated from endometrial biopsies of endometriosis patients (n ¼ 9),
as well as eutopic endometrium of control patients (n¼5) in proliferative
and secretory cycle phases exposed to PGF2a, with assessment of CXCL8
and VEGF expression.
MATERIALS AND METHODS: Cells were cultured in DMEM-F12 medium
with 10% FBS, 1% antibiotics, 5 mg/mL insulin and 5mg/mL transferrin.
Cells grown to confluence were preincubated for 1 h with PGF2a-
FP receptor antagonist (AL8810) (50ug/mL) or cyclo-oxygenase-2 (cox-2)
inhibitor (NS398) (50 ug/mL) and stimulated with different concentrations
of PGF2a (0-1000ng/mL) or PGF2a analog (Fluprostenol) (0-1000 ng/mL)
for different periods of time (0-48 h). PGF2a biosynthetic pathways were
analyzed by RT-qPCR and Western blot. While, CXCL8 and VEGF secretions
were analyzed by ELISAs. An unpaired t test and one-way ANOVA followed
by the Dunnett’s test were performed for this study.
RESULTS: Our study showed that PGF2a stimulated CXCL8 and VEGF
expressions in ectopic endometrial stromal cells of women with endometriosis
in the proliferative and the secretory phases of the menstrual cycle,
without noticeable change in eutopic endometrial stromal cells of women
with or without endometriosis. Cell exposure to PGF2a analog (Fluprostenol)
confirmed these previous observations. After 24h of stimulation, diminutions
of CXCL8 and VEGF were significant when PGF2a receptor (FP) antagonist
was added with either PGF2a or Fluprostenol. Meanwhile, we observed that
PGF2a increased significantly cox-2 expression while its other biosynthetic
enzymes showed no statistical difference. We, therefore, used cox-2 inhibitor
(NS398) which correlated a statistical decrease for both VEGF and IL-8 secretions.
CONCLUSIONS: This study showed that angiogenic pathways are
involved in PGF2a-mediated activation through FP and Cox-2 in ectopic
endometrial stromal cells of women with endometriosis. This is in keeping
with our previous data showing multiple abnormalities in PGF2a pathways
in ectopic endometrial tissue, leading to an abnormal behavior. Expression
of CXCL8 and VEGF in ectopic endometrial cells throughout the menstrual
cycle of women with endometriosis may, considering the role of these cytokines
in cell growth and angiogenesis, play an important role in the capability
of endometrial cells to develop and to survive in ectopic locations.
Supported by: grant MOP-123259 to MP from the Canadian Institutes for
Health Research.
e160 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

P-159 Tuesday, October 20, 2015
ENDOCERVIX MICRORNA EXPRESSION IN PATIENTS WITH
DEEP ENDOMETRIOSIS. C. V. de Carvalho, P. Rozenchan,
T. Bonetti, A. Kopelman, M. B. Girao, E. Schor. Gynecology, Universidade
Federal de S~ao Paulo - UNIFESP/EPM, S~ao Paulo, Brazil.
OBJECTIVE: The identification of molecular differences in eutopic endometrium
of women with endometriosis is an important step toward understanding
it pathogenesis and developing novel strategies for the treatment
of associated infertility and pain. A number of differences in gene expression
were found in patients with endometriosis. miRNA are small molecules and
may act as post-transcriptional gene expression regulator. Many miRNAs are
tissue specific and play a significant role in oncogenes, metastasis and tumor
invasion. It has also been found that most miRNAs have dual nature, targeting
both oncogenes and tumor suppressor genes. Endocervix could represent a
less-invasive approach to detect patients with endometriosis, so the aim of
this study was to perform miRNA expression screening in endocervix of patients
with deep endometriosis compared to healthy controls.
DESIGN: Experimental transversal study
MATERIALS AND METHODS: It were included six patients with deep
endometriosis and six control patients without endometriosis or other previous
gynecological illness. Patients underwent an endocervical brush during
the proliferative phase of menstrual cycle. The samples were submitted to
RNA purification and were analyzed by miRNA PCR-array (miScript
miRNA PCR Array, Qiagen), containing 86 different miRNAs.
RESULTS: We were able to found 7 miRNAs significantly upregulated:
Has-miR210-3p (fold: 3.4, p: 0.04), Has-miR23b-3p (fold: 3.5, p: 0.01),
Has-miR151a-5p (fold: 8.2, p: 0.01), Has-miR222-3p (fold: 2.6, p: 0.04)
Has-let7e-5p (fold: 3.4, p 0.05), Has-let7b-5p (fold: 4.6, p: 0.02), Haslet7c-5p
(fold: 3,0, p: 0.04) and one miRNA downregulated in endometrium:
Has-miR142-3p (fold: -8.8, p: 0.004), in patients with deep endometriosis
when compared to control group.
CONCLUSIONS: We found eight microRNAs differentially expressed in
endometrium of patients with deep endometriosis when compared to control
group, highlightening those miRNAs as biomarkers candidates for endometriosis.
This study is a screening of miRNA expression in endocervix of patients
with deep endometriosis and our next step is the validation of these
miRNAs in a different set of patients.
References: Pelvic Pain Unit - UNIFESP-EPM
Supported by: Fundaç~ao de Amparo a Pesquisa do Estado de S~ao Paulo
(FAPESP) Coordenaç~ao de Aperfeiçoamento de Pessoal de Nıvel Superior
(CAPES)
P-160 Tuesday, October 20, 2015
EFFECT OF LUPRON VS NORETHINDRONE TREATMENT ON
LIPID PROFILE OF WOMEN WITH SYMPTOMATIC
ENDOMETRIOSIS. C. Charles, a O. Muneyyirci-Delale, a,b N. Sinaii, c
M. Dalloul, a P. Stratton. d a OB/GYN, SUNY Downstate Medical Center,
Brooklyn, NY; b Ob/Gyn, Kings County Hospital Center, Brooklyn, NY; c National
Institutes of Health, Bethesda, MD; d NICHD, NIH, Bethesda, MD.
OBJECTIVE: To assess changes in the lipid profiles of women with symptomatic
endometriosis who underwent treatment with Leuprolide Acetate
Depot form (LD) vs Norethindrone Acetate (NA).
DESIGN: Prospective, randomized, double-masked clinical trial.
MATERIALS AND METHODS: 62 women with endometriosis-associated
pain were randomized to receive LD 11.25mg every 3 months or NA
5mg daily for 24 weeks in Phase I, then all were given NA for another 28
weeks in Phase II. After 52 weeks of treatment, women were followed for
up to one year (PTFU). Lipid testing included Total Cholesterol (CHOL), Triglycerides
(TRIG), High Density Lipoprotein (HDL) and Low Density Lipoprotein
(LDL). All parameters were assessed at entry, end of Phases I and II
as well as during PTFU. Data were analyzed using paired or two-sample t-
tests, or Wilcoxon signed rank and Wilcoxon rank-sum tests. Regression
modeling adjusted for continued NA use in PTFU.
RESULTS: Women were predominantly Black (82%), between age 21 and
47 years with 31 NA and 31 LD. A decrease in HDL and an increase in LDL
values were noted during Phase I in NA (p

Medical School, Boston, MA; d Brigham and Women’s Hospital and Harvard
Medical School, Boston, MA.
OBJECTIVE: Develop an in vitro model of human disease for endometriosis
DESIGN: Basic science
MATERIALS AND METHODS: G4 mouse embryonic stem cells
(mESCs) were cultured andallowed to differentiate for 21 days as colonies
or as suspended embryoid bodies (EBs). Differentiation of G4 ESCs was
induced by transferring the cells to mouse embryonic feeder cell culture media
(Dulbecco’s modified Eagle’s Medium (DMEM) with 10% fetal bovine
serum (FBS)) for 3 weeks. G4 ESCs were also differentiated with differentiation
media (DMEM, FBS, Bone morphogenetic protein (BMP)-4 50ng/ul
and Activin-A 20ng/ul). Post-differentiation analysis profiles of the mESCs
and EBs were assessed for the endometrium and endometriosis markers
CD9 / CD13 by immunocytochemistry and transcriptional analysis using
reverse and quantitative PCR. Differentiated CD9 + CD13 + were then prepared
for sorting using FACS for further characterization.
RESULTS: Colonies of mESCs showed progressive differentiation of CD9
immunoreactive cells over a 3 week period (26.6%, 48.9%,and 76.4%, weeks
1-3, respectively), while populations of differentiated CD13 cells appeared
relatively constant over this same duration (40.6%, 47.0% and 47.4%).
40% of EBs displayed CD9 immunoreactivity with 61.2% also immunolableing
for CD13. These observations were confirmed using quantitative PCR analyses.
Additional markers of endometrium (CD146, PGDF-b) were also
identified in these differentiating cultures.
CONCLUSIONS: The results from this study demonstrate the generation
of presumptive endometrial precursors in differentiating mouse ESCs. The
ability to produce CD9 + CD13 + cells in vitro, isolate the same by FACS and
continue their in vitro growth shows the utility of ESCs for preclinical
modeling of a human disease, endometriosis. Furthermore this system affords
the unique opportunity to investigate novel therapeutic approaches for the
management of this debilitating disease using a stem cell disease platform.
CONCLUSIONS: We observed that the levels of serum anti-endometrial
antibodies to SLP2, TMOD3 and TPM3 did not appear to elevate further
in advanced stages (Stage III-IV) of endometriosis.
Supported by: Department of Biotechnology, Government of India (BT/
PR4589/MED/30/731/2012) and ICMR (NIRRH/MS/RA/184/09-2014).
P-164 Tuesday, October 20, 2015
WITHDRAWN
P-163 Tuesday, October 20, 2015
STAGE-WISE COMPARISON OF ANTI-ENDOMETRIAL-ANTI-
BODIES AGAINST PEPTIDES OF SLP2, TMOD3 AND TPM3 IN
DIAGNOSIS OF ENDOMETRIOSIS. T. Bendigeri, a A. Ghuge, a
K. Bhusane, a S. Begum, b N. Warty, c R. Sawant, d P. Dasmahapatra, e
K. Padte, f A. C. Humane, g A. Chauhan, h R. Gajbhiye. a a Department of Clinical
Research, National Institute for Research in Reproductive Health, Mumbai,
India; b Department of Biostatistics, National Institute for Research in
Reproductive Health, Mumbai, India; c Sanjivani Diagnostic Centre and General
Maternity, Mumbai, India; d Sanjivani Diagnostic Centre and General
Maternity Home, Mumbai, India; e Spectrum clinic & Endoscopy Research
Institute, Kolkata, India; f Dr Kedar’s Maternity, Infertility and Surgical Hospital,
Endoscopy and IVF Center, Panjim, India; g Goverment Medical College,
Nagpur, India; h Department of Obstetrics and Gynecology, Seth GS
Medical College and King Edward Memorial (KEM) Hospital, Mumbai, India.
OBJECTIVE: The lack of non-invasive diagnostic test for early diagnosis
of endometriosis results in 8-11 years delay in diagnosis leads to deterioration
of quality of life. Previously, we identified anti-endometrial-antibodies
against peptides of Stomatin like protein 2 (SLP2), Tropomodulin 3
(TMOD3) and Tropomyosin 3 (TPM3), and further proposed their utility
as non-invasive biomarkers for early diagnosis of endometriosis.The aim
of the present study was to investigate whether the levels of biomarkers
vary with disease progression and to compare the sensitivity, specificity
and diagnostic accuracy of the biomarkers in diagnosis of endometriosis in
early stages (Stage I-II) versus advanced stages (Stage III-IV).
DESIGN: Multi-centre, cross sectional study
MATERIALS AND METHODS: Women with endometriosis (Stage I-II
n¼133, Stage III-IV n¼133)and healthy controls (n¼104) were screenedfor
eleven novel autoimmune markers (anti-endometrial-antibodies of SLP2a,
SLP2b, SLP2c, TMOD3a, TMOD3b, TMOD3c, TMOD3d, TPM3a,
TPM3b, TPM3c and TPM3d) using the peptide ELISA.The statistical analysis
was performed using STATA software (version 8.2, Texas, USA).
RESULTS: The mean serum levels of anti-endometrial-antibodies of
SLP2a, SLP2b, SLP2c, TMOD3a, TMOD3b, TMOD3c, TMOD3d,
TPM3a, TPM3b, TPM3c and TPM3d) in early stages (Stage I-II) were significantly
higher than that of advanced stages (Stage III-IV) of endometriosis
(p

P-167 Tuesday, October 20, 2015
DOES THE RECURRENCE OF OVARIAN ENDOMETRIOMA
AFFECT THE PREGNANCY RATES IN IVF?. K. Aslan, a
I. Kasapoglu, b B. Avci, c B. Ata, d G. Uncu. e a Uludag University School
of Medicine Depart, Gynecology, Bursa, Turkey; b Obstetrics and Gynecology,
Uludag University School of Medicine, Bursa, Turkey; c Histology/
Embryology, Uludag University Faculty of Medicine, Bursa, Turkey; d Reproductive
Endocrinology and Infertility, _Istanbul, Turkey; e Uludag University,
Bursa, Turkey.
P-166 Tuesday, October 20, 2015
DOWN REGULATION OF BIOMARKERS AND INCREASED PREG-
NANCY RATE FOLLOWING GNRH AGONIST TREATMENT IN
PATIENTS WITH ENDOMETRIOSIS. E. Rahmawati, a
P. K. Maurya, b A. Kao, c H. Chen, d C. Tzeng. e a Clinical Medicine, Taipei
Medical University, Taipei, Taiwan; b Amity Institute of Biotechnology, Amity
University, Uttar Pradesh, India; c Obstetrics and Gynecology, Taipei
Medical University, Taipei, Taiwan; d Graduate Institute of Toxicology, College
of Medicine, National Taiwan University, Taipei, Taiwan; e Taipei Medical
University, Taipei, Taiwan.
OBJECTIVE: The present study was undertaken to evaluate the effect of
GnRHa in endometriosis patients undergoing IVF. We further investigated
the effect of GnRHa on the genes expression and proteins level in endometriotic
tissues. We hypothesized that some protein related with pathogenesis of
endometriosis are down regulated in the patients following GnRHa treatment.
DESIGN: Retrospective case control clinical study combined with analysis
of gene expression profile and the protein level in patients with endometriosis
after GnRHa treatment.
MATERIALS AND METHODS: This study recruited 282 endometriosis
women without GnRHa treatment and 96 endometriosis patients with
GnRHa treatment. IVF outcome was measured by evaluation on oocyte number,
embryo transfer, pregnancy rate, implantation rate and abortion rate. Biopsy
of endometriotic tissues was done to evaluate distinction of genes
expression with and without GnRHa treatment using affymetrix microarray
and independently validated by q-RT PCR, western blot and IHC. The candidate
biomarkers from serum were measured by ELISA. Primary culture of
endometriotic stroma cells to identify the role of estrogen in the expression
of candidate proteins. Statistical analysis was performed using Mann-Whitney
U test. P < 0.05 is considered significant.
RESULTS: IVF outcome shown that implantation rate increased significantly
in endometriosis patients with GnRHa treatment (25.4%) compared
with untreated group (18.7%) (p¼0.001) and the pregnancy rate increased
significantly in treated patient (39.3%) compared with untreated group
(19.7%) (p¼0.003). This data was Supported by microarray data that approximately
65 genes exhibited alterations in expression following GnRHa treatment.
Validation using q-RT PCR, WB, IHC and ELISA indicated that TNS1,
MMP 14, CAV 2, NRN1 and ATP2A3 were significantly lower in patients
with GnRHa treatment. P values are in the following manner: Tensin 1
(p¼ 0.0100), MMP 14 (p¼ 0.013), Caveolin 2 (p¼ 0.023), Neuritin 1 (p¼
0.040) and ATP2A3 (p¼ 0.044). Immunohostochemistry indicated that stromal
cells expressed the five candidates proteins. Treatment with estradiol on
stroma endometriotic cells increased the expression of those proteins. Tensin
1 and MMP 14 play important role in cell migration. The role of Caveolin 2 is
poorly understood. Neuritin 1 is associated with pain. ATP2A3 is involved in
calcium sequestration associated with muscular excitation and contraction.
CONCLUSIONS: GnRHa treatment in endometriosis patients increased
implantation and pregnancy rate in women undergoing IVF and down regulated
the promising prognostic biomarkers: Tensin 1, MMP 14, Caveolin 2,
Neuritin 1 and ATP2A3.
Supported by: This work was Supported by the funding from the
Academia Sinica (BM104010117) and MOST(103-2314-B-038-054-MY2)
of Taiwan
OBJECTIVE: To determine whether recurrence of endometrioma affects
ongoing pregnancy rates in IVF cycles.
DESIGN: Retrospective Study
MATERIALS AND METHODS: This is a retrospective study conducted
at Uludag University Faculty of Medicine, Department of Obstetrics and Gynecology.
Electronic data of the years between 2011-2015 was screened and
IVF patients with endometriosis were selected. Patiens were classified into
three subgroups; patients with recurrence of endometrioma (Group 1), patients
with primary endometrioma (Group-2) and patients without recurrence
of endometrioma after surgery(Group-3) . Baseline characteristics, embroloygy
laboratory parameters and pregnancy outcomes were analyzed and
compared.
RESULTS: Total 62 infertile patients with endometriosis were selected
from electronic database. Baseline characteristics were similar in each group.
Median duration after surgery was 2 years for IVF. 24 of these 42 patients
with surgical history have had postoperative recurrence of endometrioma
during ovarian stimulation. There have been 42 patients with endometrioma
during ovarian stimulation. Mean endometrioma size was 3,48 cm (+/- 2,1).
24 of 42 patients had recurrence of endometrioma and 18 of 42 patients had
primary endometrioma. The rest of 62 patients had been surgery before and
there were no recurrence of endometrioma in these patients. Embryologic
laboratory parameters and ongoing pregnancy rates (OPR) were similar in
three groups. Number of picked up oocytes were 7,8 (+/-4,8), 12,3(+/-8,7)
and 10,2(+/-6,6), p:0,258, respectively group 1,2,3. Number of metaphase-
2 oocytes were 5,6(+/-3,7), 7,89(+/-5,1) and 7,9(+/-6,1), p:0.164, respectively
group 1,2,3. Pregnancy outcomes were analyzed in two parameters;
positive B-hCG and ongoing pregnancy rate. Positive B-hCG rates were %
58.3, %33.3, %55, respectively group 1,2,3, p:0.244. Ongoing pregnancy
rates were %33.3, %22.2, %35, respectively group 1,2,3, p:0.655.
CONCLUSIONS: Presence of endometrioma during ovarian stimulation
in infertile patients does not affect embryology laboratory parameters and
pregnancy outcomes. After surgical removal, presence of recurrence does
not decrease pregnancy outcomes. Thus in conclusion, there is no need to
re-operate the patients for removal of recurrence for better IVF outcomes.
Embryology and Pregnancy Outcomes.
Group 1
Surgery (+)
Endometrioma (+)
Group 2
Surgery (-)
Endometrioma (+)
Group 3
Surgery (+)
Endometrioma (-)
p value
Number of oocyte (sd) 7,8 (+/-4,8) 12,3(+/-8,7) 10,2(+/-6,6) 0,164
Number of metaphase 5,6(+/-3,7) 7,89(+/-5,1) 7,9(+/-6,1) 0.258
2 oocyte (sd)
Number of 2PN (sd) 4,17(+/-2,8) 5,4(+/-4) 5,2(+/-3,9) 0.518
Positive b-hCG (%) 14/24 (%58.3) 6/18 (%33.3) 11/20 (%55) 0.244
Ongoing Pregnancy
Rate (%)
8/24 (%33.3) 4/18 (%22.2) 7/20 (%35) 0.655
P-168 Tuesday, October 20, 2015
IMPROVEMENT IN ENDOMETRIOSIS-RELATED PELVIC PAIN
WITH LEUPROLIDE OR NORETHINDRONE TREAT-
MENT. O. Muneyyirci-Delale, a,b C. Charles, a N. Sinaii, c M. Dalloul, a
P. Stratton. d a OB/GYN, SUNY Downstate Medical Center, Brooklyn, NY;
b Ob/gyn, Kings County Hospital Center, Brooklyn, MD; c National Institutes
of Health, Bethesda, MD; d NICHD, NIH, Bethesda, MD.
OBJECTIVE: To compare the long-term effectiveness of 24 weeks Leuprolide
Acetate Depot form 11.25mg (LD) vs Norethindrone Acetate 5mg
(NA) followed by NA alone for 28 weeks in relieving pain from endometriosis.
DESIGN: Prospective double-masked randomized clinical trial
FERTILITY & STERILITY Ò
e163

MATERIALS AND METHODS: 62 women with symptomatic endometriosis
were randomized to LD (N¼31) or NA (N¼31) for 24 weeks in Phase
I; then all were given NA for 28 weeks in Phase II. After 52 weeks of treatment,
women were followed for up to one year (PTFU) with the option to restart
NA. Visual analog scale (VAS) was used to assess menstrual (MP),
coital (CP), and non-menstrual pelvic pain (NMP), evaluated at entry and
compared to Phase I, Phase II and PTFU to assess improvement. Data
were analyzed as intent-to-treat; VAS deltas were compared within phases
using paired tests and between groups by two-sample tests. PTFU data
considered NA use.
RESULTS: Women were predominantly Black (81%) and single (66%),
had a mean age of 34.1 6.7 yr and reported pelvic pain for 14.3 8.0 yr.
Groups did not differ in these characteristics, stage of endometriosis,
gravidity, BMI, or pain scores upon entry.In Phase I, women using NA had
a significant reduction in VAS for MP (p

compared to controls. DNA methylation arrays showed heightened methylation
of this gene’s promoter region in tissues from endometriosis patients
with pain compared to controls.
CONCLUSIONS: Our results thus far suggest that the upregulation of polycomb
proteins in endometriotic tissues modulates DNA methylation of the
forkhead protein resulting in its decreased expression. This likely leads to
increased growth of the endometriotic tissue in the peritoneal environment,
as well as increased pain. Our results have uncovered a network of proteins
that may aid in the progression of endometriotic lesions. This underlying
mechanism has yet to be exploited for therapeutic value in endometriosis.
Supported by: Marshall University/University of Kentucky CCTS (NIHfunded).
P-173 Tuesday, October 20, 2015
EFFICACY OF NATURAL CYCLE ENDOMETRIAL PREPARA-
TION FOR FROZEN-THAWED EMBRYO TRANSFER IN PATIENTS
WITH ENDOMETRIOSIS. H. Guo. Shanghai Ninth People’s Hospital,
Shanghai, China.
P-172 Tuesday, October 20, 2015
THE EPIGENETIC ROLE OF EZH2-FOXP3 CROSSTALK IN ENDO-
METRIOSIS AND ITS ASSOCIATED PAIN. K. Wright N. Santanam.
Pharmacology, Physiology, and Toxicology, Marshall University, Huntington,
WV.
OBJECTIVE: Determine the epigenetic mechanisms involved with polycomb
group proteins and associated genes in the development and progression
of endometriosis and its associated pain.
DESIGN: Patient tissues classified by location (eutopic or ectopic) and the
presence of pain were studied to determine trends in genetic expression,
interaction, and methylation.
MATERIALS AND METHODS: This study involved tissues from control
(non-endometriosis) subjects without pain (n¼4), as well as eutopic (nonlesion,
n¼7) and endometriotic (ectopic, n¼4) tissues from patients with
endometriosis and pain. All specimens were obtained from IRB-approved
and consented subjects. Real-time PCR was used to analyze gene expression
of polycomb repressor complex 2 (PRC2) components and Western blot was
used to determine the relative protein levels. Interaction of tumor suppressor
Forkhead box P3 (FoxP3) as a potential target for the PRC2 catalytic subunit,
Enhancer of zeste 2 (EZH2), was also measured using chromatin immunoprecipitation
and immunoblotting. Lastly, Qiagen EpiTect Methyl II Array
was also used to identify methylation patterns of genes associated with
inflammation and autoimmunity in endometriotic fluids compared to controls.
RESULTS: Expression of EZH2 was elevated in ectopic tissues compared
to controls, while lowered in eutopic tissue in endometriosis patients (fold
change ¼ 3.54, 0.11, respectively). Western blots also revealed increased
levels of the tri-methylation of lysine 27 on histone 3 (H3K27me3). FoxP3
gene expression was much lower in both ectopic and eutopic tissues
OBJECTIVE: To assess the efficiency of natural cycle(NC) endometrial
preparation for frozen-thawed embryo transfer (FET) in women with endometriosis(EMS).
DESIGN: Retrospective study
MATERIALS AND METHODS: This was a retrospective study of the use
of the natural cycle for endometrial preparation following FET cycles in patients
with endometriosis from March, 2011 to August, 2013. The study
groups included 97 patients with stage I-II endometriosis who underwent a
total of 120 FET cycles(group A, included cases of mild and minimal endometriosis)
and 179 patients with stage III-IVendometriosis who underwent a
total of 233 FET cycles(group B, included cases of moderate and severe
endometriosis). The control group included 258 patients with tubal factor
infertility who underwent a total of 300 FET cycles(group C). In stage III-
IV endometriosis , 40 patients (47 FET cycles,groupB1) had endometrial
cyst recurrence after laparoscopic or laparotomy treatment. All patients
enrolled in the study met the following inclusion criteria: (i) normal uterine
cavity as assessed by ultrasonography, hysterosalpingography or hysteroscopy;
(ii) high-quality frozen embryos were transplanted. (iii)normal male
semen. The exclusion criteria comprised: (i) presence of hydrosalpinges;
(ii) presence of adenomyosis and (iii) past history of myomectomy;(iv) polycystic
ovary syndrome(PCOS); (v)repeated failed intrauterine insemination
(IUI ). For group A , B and C ,we used the natural cycle for endometrium
preparation. The patients’ characteristics, including age, duration of infertility,
body mass index, number of embryos transferred, average endometrial
thickness of ET (mm), basal FSH, serum E2 and P of transplantation day, No.
of previous failed embryo transfer cycle, the rates of basal FSH>10 were recorded.
Secondary measures included clinical pregnancy rate, live birth rate,
ongoing pregnancy rate, implantation rate, and pregnancy complication rate.
RESULTS: There were comparable clinical pregnancy rate (43.33% stage
I-II, 50.21% stageIII-IV and 45.33% tubal factor )and live birth rate(36.67%
stage I-II, 41.63% stageIII-IV and 40.00% tubal factor ) among the three
groups, not depending on severity of endometriosis . No differences were
found in other pregnancy parameters , in terms of ongoing pregnancy
rate(36.67% stage I-II, 41.63% stageIII-IVand 40.00% tubal factor ) , miscarriage
rate(14.89% stage I-II, 16.24% stageIII-IV and 10.29% tubal factor ),
pregnancy complication rate(7.69% stage I-II, 10.26% stageIII-IV and
12.50% tubal factor ). No congenital birth defects were found in the endometriosis
groups. In addition, when high-quality embryos are transferred, the
pregnancy results are not affected by endometriomas.
CONCLUSIONS: Natural cycle endometrial preparation for FET obtains
similar pregnancy outcomes in patients with endometriosis compared with
tubal infertility, and don’t increase the risk of birth defects and other complications.
P-174 Tuesday, October 20, 2015
SURGICAL MANAGEMENT OF ENDOMETRIOSIS DOES NOT
AFFECT OXIDATIVE DAMAGE IN ENDOMETRIOTIC TISSUE
OVER TIME. I. Al-Aref, L. R. Goodman, R. Flyckt, A. Goyal,
T. Falcone. Cleveland Clinic, Cleveland, OH.
OBJECTIVE: Oxidative stress has been considered as a potential factor
involved in the pathology and progression of endometriosis. The objective
of this study was to evaluate the natural progression of endometriosis by
FERTILITY & STERILITY Ò
e165

comparing oxidative damage in endometriotic tissue at two distinct time
points.
DESIGN: Retrospective cohort trial
MATERIALS AND METHODS: Women who had presented between
1999-2009 with pelvic pain and/or infertility between the ages of 18-45 years
with a history of two distinct surgeries with biopsy proven endometriosis
were included if there was banked blood and tissue samples available for review.
Histopathochemistry was used to evaluate oxidative damage by
measuring hydroxy deoxyguaninine (8OHdG) in endometriotic cells and to
evaluate the cellular ability for DNA repair with 8-oxoguanine glycosylase
1 (OGG1). The markers were measured in a biopsy sample of pathology
proven endometriosis for each patient at each surgery and compared to
each other using Wilcoxan rank-sum tests. The pathologist was blinded to
whether the samples were from the primary or secondary surgery. All surgeries
were performed by the same surgeon.
RESULTS: There were seven patients who met criteria and were included
in the study. The average age at first surgery was 32.4 +/- 4.4 years. The
average time between surgeries was 2.0 +/- 1.1 years. Stage of disease and
marker intensity and presence in percentage of cells at first and second surgery
are shown in table 1.
Table 1: Stage of endometriosis and markers at first and second surgery.
Stage
8-OHdG
Intensity
8-OHdG
% of cells
OGG1
Intensity
OGG1
% of cells
1 IV/IV 0/1 0/10 2/2 90/80
2 IV/IV 2/3 80/90 3/2 90/30
3 IV/IV 3/3 100/80 2/3 10/80
4 IV/IV 2/2 80/60 2/2 50/80
5 IV/IV 3/2 75/75 3/3 95/95
6 IV/IV 2/2 40/5 3/3 90/90
7 III/III 2/3 60/80 3/3 95/90
Despite complete excision of all visible endometriosis at the time of first
surgery, all patients had return of the disease to the same stage at their second
surgery. Temporally, there was no significant difference between percentage
of cells that expressed 8-OHdG (p ¼ 0.79) or OGG1 (p ¼ 0.81) between the
two surgeries. There was a correlation of 8-OHdG percent of cells and intensity
(r ¼ 0.80, p < 0.01) and OGG1 percent of cells and intensity (r ¼ 0.65, p
< 0.01), but no temporal relationship between surgeries.
CONCLUSIONS: Overall, There was no relationship of progression of
oxidative damage over time. Based on these results, it appears that baseline
oxidative damage in endometriotic cells is predictive of future cell damage
and that surgical excision of endometriosis has no benefit in changing the underlying
pathophysiology leading to oxidative damage.
P-175 Tuesday, October 20, 2015
WITHDRAWN
P-176 Tuesday, October 20, 2015
THE RELATIONSHIP OF CASPASE-3, CASPASE-9, MMP-9
EXPRESSION AND C-1562T MMP-9 GENE POLYMORPHISM IN
MENSTRUAL BLOOD AS THE ETIOPATHOGENESIS MARKER
TO CLINICAL ENDOMETRIOSIS MANIFESTATION. T. Madjid.
Faculty of Medicine, University of Padjadjaran, Bandung, Indonesia.
OBJECTIVE: The aim of this study is to discover non-invasive diagnostic
method of endo-metriosis using menstrual blood, and also to reveal clearer
understanding in patho-genesis of endometriosis
DESIGN: A case control study involving 149 women who visited the
Reproductive Endocrinology and Fertility Clinic, FKUP/RSHS and RSHS
network hospitals was performed, from February 2007 to February 2008.
MATERIALS AND METHODS: Screening for suspected endometriosis
was performed by history taking, physical examination, and additional examination.
Diagnostic laparoscopy or laparotomy and biopsy was performed afterward.
The immunocytochemical examination on caspase-3, caspase-9,
MMP-9, and mmp-9 gene polymorphism of menstrual blood were performed.
Based on the microscopic confir-mation of histopathological result,
the relationship of endometriotic and non-endometriotic clinical manifestation
with menstrual blood biomolecular marker was assessed.
RESULTS: Sixty-three (42.28%) endometriosis cases and 86 (57.78%)
non-endometriosis cases were found. From those subjects, 34 endometriosis
cases and 48 non-endometriosis cases with complete data were enrolled in
this study. The endometrial cells were successfully isolated using preservative
solution, and with immunocytochemical assay, all samples from 34
endometriosis subjects could be analyzed for the expression of caspase-3,
caspase-9, and MMP-9. The decreased expression of caspase-3 and caspase-9,
and increased expression of MMP-9 in endometriosis group were
higher than those in non-endometriosis group (82.4% vs 77.1%, p¼0.562;
97.1% vs 87.5%, pE-F¼0.129; and 85.3% vs 85.4%, p¼0.988 respectively).
The frequency of allele C in -1562T region of mmp-9 gene was significantly
increased in endometriosis group (p

P-177 Tuesday, October 20, 2015
INCREASED HIPPO SIGNALLING PATHWAY PROMOTES CELL
PROLIFERATION AND ANTI-APOPTOSIS IN
ENDOMETRIOSIS. Y. Song, a W. Huang, b M. Zhou, a L. Xiao, a
X. Feng. a a Obstetrics and Gynecology, West China Second University Hospital
of SCU, Chengdu, China; b West China Second University Hospital of
Sichuan University, Chengdu, China.
OBJECTIVE: Endometriosis is one of the common reasons that results in
female infertility and pelvic pain in reproductive women. However, the pathogenesis
is unclear. The imbalance in proliferation and apoptosis of endometrial
stromal cells(ESCs) is considered as an important role in pathogenesis of endometriosis.
Several signalling pathways involve in these processes and exact
regulation need to be confirmed in endometriosis. A newly established signalling
pathway–Hippo signalling pathway participates in organ development and
tumorigenesis which indicates that it plays a critical role in proliferation and
apoptosis processes. However, the study about Hippo pathway and endometriosis
is lack. Our objective is to explore the mechanism of Hippo signalling
pathway controlling ESCs proliferation and apoptosis in endometriosis.
DESIGN: We investigated expression levels of Hippo signaling components
in the endometrium of women with or without endometriosis. We identified
YAP regulated proliferation and apoptosis ability of endometrial
stromal cells by cell culture and transfections. And endometriosis animal
model of nude mice and verteporfn treatments were used to identify the intervention
effect of Hippo signaling on endometriotic lesions.
MATERIALS AND METHODS: Study population, Quantitative real-time
RT-PCR, Western blotting, immunohistochemistry , cell culture, transfections,
BrdU labeling, TUNEL assay, immunoprecipitation-quantitative
PCR and animal model, verteporfn treatments were used in this study.
RESULTS: Our data showed that increased expression of YAP and
decreased expression of p-YAP in ectopic and eutopic endometrium, compared
with normal endometrium. Further, we demonstrated that knockdown of YAP
in eutopic ESCs could decrease proliferation and enhance apoptosis companied
with decreased expression of CTGF and increased BAX/BCL-2 ratio. In addition,
overexpression of YAP could make normal ESCs cells get higher proliferation
and lower apoptosis ability, with increased CTGF and decreased
BAX/BCL-2 ratio. Furthermore, We identified, by chromatin immunoprecipitation-quantitative
PCR, that CTGF, Bcl-2 were direct downstream target genes
of YAP in the regulation of proliferation and apoptosis, and the expression of
CTGF, Bcl-2 were through the YAP-TEAD complex. At last, endometriosis animal
model of nude mice were treatment with verteporfin–specific inhibitor of
Hippo pathway, which disrupts the formation of the YAP-TEAD complex,
significantly reduced endometric lesions and cell proliferation.
CONCLUSIONS: These data indicate that hippo signalling pathway play
a critical role in the pathogenesis of endometriosis and could represent as a
novel direction for specific diagnosis and therapeutic method.
References:
1. Nasu K, Yuge A, Tsuno A, Nishida M, Narahara H.Involvement of
resistance to apoptosis in the pathogenesis of endometriosis. Histol Histopathol.
2009 ;24:1181-92.
2. Zhao B, Li L, Guan KL.Hippo signaling at a glance. J Cell Sci.
2010;123:4001-6.
P-178 Tuesday, October 20, 2015
THE USE OF LEVONORGESTREL-RELEASING INTRAUTERINE
SYSTEM FOR LONG-TERM MAINTENANCE AFTER CONSERVA-
TIVE LAPAROSCOPIC SURGERY FOR ENDOMETRIOSIS: IN THE
RESPECT OF RECURRENT OVARIAN ENDOMETRIOMA. M. Kim, a
Y. Cho, b S. Seong. a a Department of Obstetrics and Gynecology, CHA Gangnam
Medical Center, CHA University, Seoul, Korea, Republic of; b Department of Obstetrics
and Gynecology, Dong-A University Medical Center, Dong-A University,
College of Medicine, Busan, Korea, Republic of.
OBJECTIVE: A levonorgestrel-releasing intrauterine system (LNG-IUS)
can be used for the secondary prevention of endometriosis-associated dysmenorrhea
after surgery for endometriosis. Although the LNG-IUS effectively reduces
endometriosis-associated pain, its efficacy in preventing ovarian
endometrioma recurrence is questionable.In this study, we evaluated recurrence
rate of ovarian endometrioma and surgical characteristics in patients who used
LNG-IUS after the conservative laparoscopic surgery for endometriosis.
DESIGN: Multicenter retrospective cohort study
MATERIALS AND METHODS: We performed a retrospective review of
women who underwent conservative laparoscopic enucleation of ovarian endometrioma
and were treated with LNG-IUS at the CHA Gangnam Medical
Center and Dong-A University Medical Center between January 2007 and
September 2014. The inclusion criteria were as follows: (1) pathologically
proven ovarian endometrioma; (2) undergone conservative laparoscopic surgery;
(3) premenopausal status; (4) no residual ovarian lesions, as confirmed
by transvaginal ultrasonography (TVS) before insertion of the LNG-IUS. Patients
were excluded if they had undergone laparotomy or laparoscopic hysterectomy
with or without oophorectomy. Endometrioma recurrence was
defined as a cystic mass with a diameter of R20 mm on TVS.
RESULTS: A total 61 patients were included in this study. The mean age of
the patients was 36.2 5.9 years (range, 23-48 years), and the mean follow-up
duration was 41.6 21.4 months (range, 8-98 months). Ovarian endometrioma
recurrence was detected on TVS in 7 of the 61 (11.5%) patients. The cumulative
recurrence rates of ovarian endometrioma per patient at 24, 36, and 60 months
were 5.7%, 11.9%, and 23.1%, respectively. Surgical characteristics, such as the
diameter of the cyst, disease stage, unilateral or bilateral involvement, and coexistence
of deep endometriosis, and postoperative medical therapy were not
associated with recurrence. Nulliparity at surgery was a significant risk factor
for endometrioma recurrence during the treatment with LNG-IUS. When we anlayzed
the recurrence rate according to the age, there was no recurrence of
ovarian endometrioma in patients over 35 years old during the follow up period
CONCLUSIONS: Postoperative long-term maintenance therapy with
LNG-IUS could be a treatment option for prevention of recurrent endometrioma
after conservative laparoscopic surgery for endometriosis, especially
for women in older age.
P-179 Tuesday, October 20, 2015
EVALUATION OF FSHR GENE POLYMORPHISMS IN INFERTILE
WOMEN WITH AND WITHOUT ENDOMETRIOSIS AND ITS COR-
RELATION WITH HUMAN ASSISTED REPRODUCTION
OUTCOMES. B. Bianco, a C. M. Trevisan, b R. Fernandes, a
R. Oliveira, a C. Peluso, a D. M. Christofolini, b C. P. Barbosa. a a Faculdade
de Medicina do ABC, Santo Andre, Brazil; b FMABC, Santo Andre, Brazil.
OBJECTIVE: We aimed to characterize the current relationship between
FSHR gene polymorphisms (Ala307Thr and Asn680Ser) with FSH serum
levels and assisted reproduction outcomes [controlled ovarian hyperstimulation
response, antral follicle counting (AFC), MII (metaphase II oocytes) and
embryos] in Brazilian women that underwent assisted reproduction treatments
with and without endometriosis.
DESIGN: Cross-sectional study comprising 277 infertile women (136
with endometriosis and 141 controls) submitted to assisted reproduction
treatment.
MATERIALS AND METHODS: All patients (cases and controls) were
%38 years old, had the presence of both ovaries without morphological abnormalities,
regular ovulatory cycle, body mass index %30, and no evidence
of endocrine disorders. The control group was formed by patients with tubal
obstruction or women with male factor involved in infertility causes. The
endometriosis group included women with diagnosis of endometriosis
confirmed by laparoscopy and/or laparotomy, with the disease staging according
to American Society for Reproductive Medicine (ASRM, 1996)
and histological evidence of disease. Genotyping of FSHR polymorphisms
were performed using TaqMan methodology by real time PCR. FSH was
measured by Enzyme-linked fluorescence assay. The data was analyzed statistically
(p-value

P-180 Tuesday, October 20, 2015
ENDOMETRIUM
THE MIXTURE OF RECOMBINANT HUMAN EPIDERMAL
GROWTH FACTOR, POLOXAMER AND SODIUM ALGINATE
PROMOTES ENDOMETRIAL GROWTH IN INFERTILE WOMEN
WITH THIN ENDOMETRIUM. C. Kim, a J. Moon, a B. Kang, a
W. Cho, b I. Park, b H. Kim. c a Obstetrics and Gynecology, College of Medicine,
University of Ulsan, Asan Med, Seoul, Korea, Republic of; b R&D
Center, Genewel, Co., Seongnam, Korea, Republic of; c R & D Center, Genewel,
Co., Seoul, Korea, Republic of.
OBJECTIVE: To evaluate the effect of the mixture of recombinant human
epidermal growth factor (rhEGF), poloxamer and sodium alginate on endometrial
growth and subendometrial artery (SEA) blood flow in infertile
women with poorly developed thin endometrium.
DESIGN: Prospective observational study.
MATERIALS AND METHODS: A total of 68 infertile patients with thin
endometrium less than 6mm were recruited for this study between March
2013 and September 2014. All subjects aged 28-41 years underwent intrauterine
treatment with either the mixture of rhEGF, poloxamer and sodium
alginate (study group, n¼38) or rhEGF alone solution (control group,
n¼30) every 2-3 days in the follicular phase. Six hundred IU of vitamin E
was orally administered daily to all subjects throughout the whole treatment
period.Mean values were expressed as meanstandard deviation (SD). Student’s
t-test was used to compare mean values. Chi-square test and Fisher’s
exact test were used to compare fraction. Statistical significance was defined
as P

P-183 Tuesday, October 20, 2015
EMBRYO-ENDOMETRIUM ASYNCHRONY MAY AFFECT
THAWED EMBRYO CYCLES USING GENETICALLY SCREENED
BLASTOCYSTS. B. S. Shapiro, S. Daneshmand, F. Garner,
M. Aguirre, C. Hudson. Fertility Center of Las Vegas, Las Vegas, NV.
OBJECTIVE: Compare implantation rates of day 5 blastocysts transferred
on day 6 and day 6 blastocysts transferred on day 7 after embryos were held
overnight in order to obtain results of genetic screening.
DESIGN: IRB-approved retrospective analysis.
MATERIALS AND METHODS: This study included cycles in which
thawed 2pn oocytes were cultured to the blastocyst stage before trophectoderm
biopsy, genetic testing, and transfer in a 3-year time period (2012-
2014). All cycles had artificial endometrial preparation with exogenous estradiol.
Progesterone injections were initiated the evening before 2pn oocyte
thaw. Embryos were cultured to the blastocyst stage, biopsied, and genetically
screened for aneuploidy by array CGH. Transfers occurred the day after
blastocysts formed and were biopsied, as genetic test results arrived the
following morning, so that blastocysts that formed on day 5 of embryo development
were transferred on day 6 and blastocysts that formed on day 6 were
transferred on day 7 of embryo development. These corresponded with transfers
on days 6 and 7 of progesterone exposure, respectively. Implantation and
clinical pregnancy calculations were based on fetal hearts observed on ultrasound
at 6-7 weeks gestation.
RESULTS: The study included 39 day 6 transfers and 28 day 7 transfers
following biopsy and genetic screening. The two groups did not differ significantly
in age at retrieval or number of embryos transferred. The number of
hours of pre-transfer progesterone exposure was significantly lower in the
day 6 transfer group when compared to the day 7 transfer group (mean
137 hours vs 160 hours, P

P-186 Tuesday, October 20, 2015
USE OF CLOMIPHENE CITRATE IN MINIMAL OVARIAN STIMU-
LATION CYCLES NEGATIVELY IMPACTS ENDOMETRIAL
THICKNESS, INDEPENDENT OF PEAK ESTRADIOL LEVELS:
AN ARGUMENT FOR A FREEZE-ALL APPROACH. B. G. Reed,
M. Ezzati, S. N. Babayev, V. Libby, B. Carr, O. Bukulmez. UT Southwestern
Medical Center, Dallas, TX.
OBJECTIVE: Minimal ovarian stimulation cycles use daily clomiphene
citrate (CC) and a small amount of gonadotropin on days 5, 7, and 9 of the
ovarian stimulation. Extrapolating from data on intrauterine insemination cycles
that indicate CC may have a detrimental effect on the endometrial stripe
(ES), we avoid fresh embryo transfer in patients undergoing minimal stimulation
despite the lack of data from IVF cycles to support this freeze-all practice.
We sought to determine if ES is negatively affected in minimal
stimulation as compared to other types of ovarian stimulation that do not utilize
CC. In addition, we sought to compare each patient’s ES during her minimal
stimulation cycle(s) with her ES during her subsequent frozen embryo
transfer (FET) cycle.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: A total cohort of 141 cycles (75 patients)
were analyzed: 78 minimal ovarian stimulation cycles and their 31 subsequent
FETs, 13 mild stimulation cycles, and 19 high dose gonadotropin
GnRH antagonist cycles. One way ANOVA and Tukey’s test were used for
continuous variables. Chi square test was used to compare the cumulative
live birth rate per patient. Paired t-test was used to compare the ES in the
same patient during the minimal stimulation and her subsequent FET cycle.
RESULTS: Therewas no statistically significant difference in age, body mass
index (BMI), antimullerian hormone (AMH) level, or cumulative live birth rates
between the study groups (Table 1). Maximum ES in the minimal stimulation
group was significantly lower than the other two groups (8.1 mm versus 13.6
mm and 13.9 mm). Peak estradiol level was significantly higher in the high
dose gonadotropin group as compared to both the minimal or mild stimulation
groups. Despite a significant difference in peak estradiol levels, there was no difference
in ES between the mild and high-dose stimulation groups. In patients
who underwent minimal stimulation IVF followed by FET, significantly thicker
ES was observed during their FET cycles (7.9 mm vs 10.3 mm).
CONCLUSIONS: Based on our data, endometrial thickness is negatively
impacted during minimal stimulation IVF cycles, independent of peak estradiol
levels. The negative effects on the endometrial thickness are not
observed in the subsequent FET cycles and, hence, a freeze-all approach
can be adopted to mitigate potential adverse endometrial effects in minimal
stimulation IVF cycles.
Table 1. Demographics and outcomes.
Minimal
stimulation
(Included use
of CC)
Mild
stimulation
(Did not
include
the use of CC)
High dose
gonadotropin
GnRH antag
stimulation
(Did not include
the use of CC)
Frozen embryo
transfer following
minimal
stimulation
(Did not include
the use of CC)
p-value
Age (y) 384 38.83.5 37.64.5 - NS
BMI 264.8 25.65.7 25.24.1 - NS
AMH (ng/ml) 0.60.7 0.50.5 1.00.7 - NS
Maximum ES (mm) 8.12 13.63.5 13.93.8 -

P-189 Tuesday, October 20, 2015
GENE EXPRESSIONS OF L-SELECTIN LIGANDS ARE
DECREASED AT MID-SECRETORY PHASE IN WOMEN WITH
ADENOMYOSIS. T. Lai, a,b,c F. Lee, a,b Q. Ling. c,d a Obstetrics and Gynecology,
Cathay General Hospital, Taipei, Taiwan; b School of Medicine, Fu
Jen Catholic University, New Taipei, Taiwan; c Institute of Systems Biology
and Bioinformatics, National Central University, Taoyuan, Taiwan; d Cathay
Medical Research Institute, Cathay General Hospital, Taipei, Taiwan.
OBJECTIVE: To evaluate the gene expression of L-selectin ligands’ subtypes
during different phases of the menstrual cycles in women with adenomyosis.
DESIGN: A prospective study on endometrial samples from reproductiveaged
women with adenomyosis at various phases in the normal menstrual cycles.
MATERIALS AND METHODS: The Institutional Review Board of Cathay
General Hospital approved this study. Tissue samples of endometrium
were collected from women (age 35-49 y) with adenomyosis undergoing hysterectomy.
Forty-two endometrial biopsies included 12 at proliferative (day7-
14), 10 at early secretory (day15-19), 9 at mid-secretory (day20 to 24) and 11
at late secretory phase (dayS 25). Immunohistochemistry was performed using
the antibody, MECA-79, which recognizes the ligands for the L-selectin
adhesion molecule. The mRNA expression profiles of L-selectin ligand’s
genes were analyzed using RT-PCR for the 4 L-selectin ligand’s subtypes: podocalyxin,
endomucin, GlyCAM-1 and CD34. Western blot were used to
quantify protein levels. Kruskal-Wallis (K-W) analysis with multiple comparisons
was performed to examine differences between different phases in the
menstrual cycle. Findings with a two-sided P value < 0.05 were considered
to indicate statistically significant differences between the phases.
RESULTS: Mean age and BMI of the patients among 4 phases were no
different (P>0.05). Immunohistochemistry showed L-selectin ligands were expressed
in the luminal and glandular epithelium of the endometrium at all the
phases. The intensity of L-selectin ligands was greatest at mid-secretory phase
which was at the stage of implantation window. However, the gene expression
profiles of L-selectin ligands including endomucin, GlyCAM-1 and CD34 revealed
gradually decreased from proliferative to mid-secretory phase. Endomucin,
GlyCAM-1 and CD34 had the lowest expression during mid-secretory
phase. Interestingly, the highest gene expression of podocalyxin, endomucin,
GlyCAM-1 and CD34 were found at late secretory phase (P

OBJECTIVE: It has been reported that exogenous hCG from a trigger injection
impairs embryo implantation and endometrial receptivity. This study
was performed to determine if a relationship exists between exogenous hCG
exposure (dose, serum level) and clinical pregnancy in fresh autologous embryo
transfer cycles, while controlling for confounding variables.
DESIGN: IRB-approved retrospective study.
MATERIALS AND METHODS: Cycles in which both hCG and GnRH
agonist were used in combination for oocyte maturation were selected so
that (1) a broad range of hCG doses would be available, and (2) the effect
of hCG in oocyte maturation would be mitigated by the redundant LH
response to the agonist. This study included all such cycles (agonist+hCG)
that had fresh autologous embryo transfer. hCG doses were varied among patients
and were steadily decreased as the protocol matured over the 9-year
study period (2006-2013). GnRH agonist dose was 4mg leuprolide acetate
(off-label use) throughout the study period. Serum levels of hCG were
measured approximately 12 hours after trigger. Fresh embryos were transferred
at the blastocyst stage. Stepwise logistic regression was used to identify
significant predictors of clinical pregnancy. Chi-square analysis was used
for univariate comparison of clinical pregnancy rates.
RESULTS: There were 349 included cycles. hCG doses varied from 500IU
to 10,000IU (17721501 IU), and post-trigger hCG serum levels varied from
11 to 354 IU/L (51.539.7 IU/L). Logistic regression identified day of transfer
(P

P-195 Tuesday, October 20, 2015
DONOR TREATMENT WITH MYCOPHENOLATE MOFETIL
ATTENUATES UTERINE ISCHEMIA/REPERFUSION
INJURY. G. Sahin Ersoy, a M. Eken, b O. Cevik, c O. T. Cilingir. d a Obstetrics
and Gynecology, Kartal Dr Lutfi Kirdar Research and Education Hospital,
Istanbul, Turkey; b Zeynep Kamil Education and Research Hospital,
Istanbul, Turkey;
c Cumhuriyet University, Faculty of Pharmacy, Sivas,
Turkey; d Marmara University School of Medicine, Istanbul, Turkey.
OBJECTIVE: Uterine transplantation is a potential option to treat uterine
factor infertility. Mycophenolate mofetil (MMF) is a powerful immunosuppressive
agent that is currently used in uterine transplantation. However
ischemia/reperfusion (I/R) injury has a detrimental effect on early allograft
dysfunction in organ transplanted patients. Therefore we aimed to investigate
whether MMF could attenuate uterine I/R injury.
DESIGN: Experimental study.
MATERIALS AND METHODS: 28 female Wistar rats used for all experiments.
Group I (M): underwent sham surgery and received 20 mg/kg/day
MMF (0.5% sodium carboxymethylcellulose), Group 2 (C): underwent
sham surgery and received vehicle control, Group 3 (IR+V): underwent uterine
I/R procedure and received vehicle control, Group 4 (IR+M): underwent
uterine I/R procedure and received 20 mg/kg/day MMF. All treatments were
conducted by gavage and started 5 days before surgery and maintained until
uteri were removed. Ischemia reperfusion was conducted for 30 minutes.
Specifically ischemia was created by clamping the distal abdominal aorta followed
by 4 hours reperfusion. Uterine tissue ischemia modified albumin
(IMA), malondialdehyde (MDA), glutathione (GSH), superoxide dismutase
(SOD) and miyeloperoxidase (MPO) levels were determined as markers of
oxidative injury. 8 hydroxydeoxyguanosine (8-OHdG) was investigated to
evaluate the levels of oxidative DNA injury. Histopathologic examination
were conducted following hematoxylin-eosin staining.
RESULTS: Histologic examination of IR+M group had more regular
endometrial contours, glands and less polymorphonuclear cell infiltration
than the IR+V group.
CONCLUSIONS: Pretreatment with MMF attenuates I/R injury. This protection
may have occurred by an anti-inflammatory and antioxidant effect.
Therefore MMF used as part of an immunosuppressant regimen may also
provide a cytoprotective effect against uterine warm ischemia injury.
Table 1. The levels of oxidative injury and DNA injury markers.
Groups C IR+V IR+M
IMA (U/mL) 0.23 0.05 0.56 0.08* 0.41 0.07*#
GSH (umol/mg protein) 2.31 0.35 1.74 0.23* 1.88 0.20
MDA (mmol/mg protein) 10.5 1.50 15.96 1.21**12.13 1.14##
MPO (U/mg protein) 1.60 0.43 6.27 2.26* 3.05 0.70*#
SOD (U/mg protein) 4.95 1.39 1.98 0.45** 3.76 1.30
8OHdG (ng/ug DNA) 1.06 0.12 1.82 0.40* 1.22 0.12#
Data are shown mean SD. * P

P-198 Tuesday, October 20, 2015
EVALUATION OF OVARIAN RESERVE BEFORE AND AFTER
SURGERY IN WOMEN WITH OVARIAN ENDOMETRIOMA
COMPARED TO PELVIC ENDOMETRIOSIS AND NEGATIVE
CONTROLS: A PROSPECTIVE COHORT TRIAL. L. R. Goodman,
J. M. Goldberg, R. Flyckt, M. Gupta, N. Gueye, k. J. Holoch,
T. Falcone. Cleveland Clinic, Cleveland, OH.
OBJECTIVE: To determine the impact of surgical excision of endometriosis
and endometrioma compared with controls on ovarian reserve.
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: Women aged 18-43 years presenting
with pelvic pain and/or infertility undergoing surgical management of suspected
endometriosis or endometrioma between October 2013 and April
2015. Patients were excluded if they had prior ovarian surgery. Statistical
analysis included student t-tests, paired t-tests and ANOVA where appropriate.
RESULTS: A total of 116 patients were included with 58 suspected endometriomas
and 58 controls with suspected pelvic endometriosis but no evidence
of ovarian involvement on pelvic imaging. All surgeries were
performed laparoscopically. All of the suspected endometriomas were
completely removed by cystectomy and confirmed by pathology. Of the 58
controls, 31 had biopsy confirmed pelvic endometriosis and 27 patients
had no evidence of endometriosis on laparoscopy with pathology negative
for endometriosis on biopsy. We prospectively evaluated ovarian reserve
measured by Anti-Mullerian Hormone (AMH) prior to surgery, at one month
and six months post-operatively. Age, body mass index and proportion of patients
with pain and/or infertility did not differ significantly between groups.
Table 1: Means (95% Confidence Intervals), { } are p-values compared to preoperative
value.
Endometrioma
(n ¼ 56)
Pelvic
Endometriosis
(n ¼ 31)
Negative
Laparoscopy
(n ¼ 27) P-Value
Age (years) 31.5 (30.1 - 33.3) 31.1 (28.9 - 33.3) 30.9 (28.8 - 33.1) 0.55
Length of 119.8 (105.8 - 128.1) 87.7 (72.6 - 102.7) 69.7 (53.6 - 85.9)

P-201 Tuesday, October 20, 2015
ASSOCIATION BETWEEN STATE-MANDATE INSURANCE
COVERAGE FOR INFERTILITY TREATMENT AND SURGICAL
APPROACH FOR MANAGEMENT OF ECTOPIC
PREGNANCY. R. Schickler, a E. Mikhail, b J. Salemi, c A. Imudia, d
S. Plosker, d H. Salihu. c a Department of Obstetrics and Gynecology, University
of South Florida Morsani College of Medicine, Tampa, FL; b Department
of Minimally Invasive Surgery and Department of Obstetrics and Gynecology,
University of South Florida Morsani College of Medicine, Tampa,
FL; c Department of Family and Community Medicine, Baylor College of
Medicine, Houston, TX; d Department of Reproductive Endocrinology and
Department of Obstetrics and Gynecology, University of South Florida Morsani
College of Medicine, Tampa, FL.
OBJECTIVE: To estimate the association between trends of surgical management
of ectopic pregnancy and the availability of state-mandated infertility
insurance coverage (SIC) including in-vitro fertilization.
DESIGN: A cross-sectional analysis of the Healthcare Cost and Utilization
Project’s Nationwide Inpatient Sample (HCUP-NIS), the largest allpayer
inpatient database in the United States, was conducted for all patients
who underwent surgical management of ectopic pregnancy between 1998
and 2011.
MATERIALS AND METHODS: All community hospitals in the US
participating in HCUP, excluding rehabilitation and long-term acute care
hospitals were included. The study includes all inpatient hospitalizations
for women receiving treatment for ectopic pregnancy as identified using
ICD-9-CM diagnosis codes. Joinpoint regression was used to estimate temporal
trends (including the annual percent change [APC]) in salpingectomy
versus salpingostomy during the study period. Stratified analyses were performed,
investigating trends in states with and without SIC (including invitro
fertilization). Those states with SIC were Connecticut, Illinois, Massachusetts,
New Jersey, and Rhode Island.
RESULTS: Between 1998 and 2011, there were over 419,000 inpatient
hospitalizations for ectopic pregnancy, approximately 14% of which
occurred in states with SIC. We observed a gradual increase in the rate of salpingectomy
in states with SIC from 69.3% in 1998 to 80.9% in 2011 (APC:
1.5%; 95% CI: 1.2, 1.8). Conversely, among ectopic pregnancies in states
without SIC, there were no statistically significant changes in temporal trends
of salpingectomy. Overall, there were significant decreasing temporal trends
in both groups when it comes to salpingostomy. The APC for salpingostomy
during the study period in states with SIC was -7.7% (95% CI: -9.0, -6.3). In
non-mandated states, the APC for salpingostomy changed from -2.9% (95%
CI: -5.7, -0.1) from 1998-2004 to -9.8% (95% CI: -12.9, -6.7) from 2004-
2011.
CONCLUSIONS: The availability of state mandated insurance coverage
for infertility treatment is correlated with the choice of surgical management
approach for ectopic pregnancy. In states with SIC, the rate of salpingectomy
increased significantly over time compared to states without such mandate.
Irrespective of infertility treatment mandate status, there was a decline in
the rate of salpingostomy as a surgical management approach for ectopic
pregnancy.
P-202 Tuesday, October 20, 2015
REVIEW AND OUTCOMES OF POWER MORCELLATION USING
AN INNOVATIVE CONTAINED BAG SYSTEM. C. E. Miller, a
C. Steller, b K. Sasaki, c A. Cholkeri-Singh. d a The Advanced IVF Institute,
Naperville, IL; b Advocate Lutheran General Hospital, Park Ridge, IL; c Advanced
Gynecologic Surgery Institute, Naperville, IL; d The Advanced Gynecologic
Surgery Institute, Naperville, IL.
OBJECTIVE: To assess feasibility of an innovative method of power morcellation
within a specimen bag.
DESIGN: Retrospective Chart Review from a specialty practice in suburban
Chicago.
MATERIALS AND METHODS: The study included patients who underwent
power morcellation during a laparoscopic or robotic-assisted hysterectomy
or myomectomy from May 2014 through March 2015. Exclusion
criteria were known uterine malignancy or successful specimen removal
without morcellation.The procedure was performed using the Espiner Eco-
Sac 230. The bag was inserted through the umbilical port and the specimen
placed inside. The cinched bag edge was pulled out through a 15mm umbilical
incision. The bag was insufflated to 25mm Hg allowing a lateral trocar to
be placed through the bag for laparoscope access. Morcellation was performed
through the umbilical incision.
RESULTS: Of the 93 procedures performed, 72% were myomectomies
and 27% were hysterectomies. The patients’ mean age was 39 years old
and mean BMI was 28.5 (range 17.2-49.9). The average specimen weight
was 288 grams, with the largest weighing 2,134 grams. Estimated blood
loss (EBL) averaged 77 milliliters. The average operating time was 2.6 hours.
The postoperative admission rate was 11%, majority of which were due to
nausea. Four patients (4.6%) had minor postoperative complications and 1
patient was readmitted to an outside hospital with vomiting and constipation
which resolved with conservative treatment. There were no bag failures or
complications that were due to use of the specimen bag or due to morcellation.
CONCLUSIONS: Using the Espiner EcoSac 230 specimen bag was successfully
performed in 93 patients with minimal complications. This is a
feasible, reliable and reproducible method of contained power morcellation,
even for a large specimen.
P-203 Tuesday, October 20, 2015
SURGICAL OUTCOMES AND COST ANALYSIS OF ABDOMINAL,
MINI-LAPAROTOMY, AND TRADITIONAL AND ROBOTIC-ASSIS-
TED LAPARASCOPY WITH AND WITHOUT TANDEM MINI-
LAPAROTOMY: A COMPARISON OF MYOMECTOMY
TECHNIQUES. J. Stanhiser, B. Mouille, J. M. Goldberg, T. Falcone,
L. R. Goodman. Women’s Health Institute, Cleveland Clinic, Cleveland, OH.
OBJECTIVE: With the recent FDA warning against laparoscopic power
morcellation necessitating larger incisions for leiomyoma extraction at the
time of myomectomy, the objective of this study was to compare the surgical
outcomes and cost of myomectomy by abdominal, mini-laparotomy, and
traditional and robotic-assisted laparoscopy with and without tandem minilaparotomy
techniques.
DESIGN: Retrospective cohort study of patients undergoing myomectomy
January 2010 to February 2015 in a tertiary care center.
MATERIALS AND METHODS: Surgical outcomes included myomectomy
type, operative time, estimated blood loss (EBL), myoma weight,
length of stay (LOS) and postoperative complications. Hospital costs were
assessed. Statistical analysis was by Student’s t-test and Wilcoxon rank
sum test.
RESULTS: 274 myomectomies were included. The average age was 38.7
5.7 years and did not differ between groups. Body mass index (BMI) and
outcomes are listed in Table 1. In laparoscopic cases, the addition of a minilaparotomy
significantly lengthened operative time (p < 0.01) and increased
EBL (p < 0.01), but the myoma weight was significantly higher (p < 0.01).
These differences were not observed in robotic-assisted cases (p¼0.69,
p¼0.07, p¼0.57 respectively), however, robotic cases were significantly
longer and had a higher EBL than laparoscopic cases (p

procedures; however when combined, LOS significantly increased (p <
0.01). The average robotic myomectomy spent one night in the hospital
and the addition of a mini-laparotomy did not significantly increase LOS
(p¼0.41). Complications and blood transfusions were significantly more
common in the abdominal and traditional laparoscopic with tandem minilaparotomy
techniques (p < 0.01). Cost was lowest with solo mini-laparotomy,
higher in the laparoscopic and highest with the robotic group. The
cost of traditional laparoscopy significantly increased 40% when a tandem
mini-laparotomy was performed (p¼0.04), however cost was not significantly
increased in robotic-assisted cases (p¼0.88).
CONCLUSIONS: The addition of tandem mini-laparotomy to laparoscopic
myomectomy increases operative time, EBL, hospital length of
stay, postoperative complications, and cost. This study supports the need
for further investigation of minimally invasive myoma extraction techniques.
P-204 Tuesday, October 20, 2015
VALUE OF HERBAL MEDICINE IN PREVENTION OF POSTOPER-
ATIVE INTRAUTERINE ADHESIONS (MISURATA
EXPERIENCE). A. M. Elbareg. Obstetrics & Gynaecology, Misurata
University, Misurata, Libyan Arab Jamahiriya.
OBJECTIVE: Intrauterine adhesions (IUAs) are the major long-term complications
of operative hysteroscopy(OH). It may result in poor reproductive
outcomes, menstrual abnormalities and pelvic pain. Several methods have
been tried to prevent post-surgical IUAs but all without significant benefits.
Moist exposed burn ointment (MEBO) is a herbal product, containing phytosterols
with anti-inflammatory, anti-bacterial and analgesic effects mainly
used for wound healing and management of burns and ulcers, such properties
might be useful to avoid IUAs, therefore, aim of this study is to assess the
efficacy and safety of MEBO in prevention of de novo IUAs following OH.
DESIGN: A randomized controlled University hospitals-based study.
MATERIALS AND METHODS: Selected 72 patients during a period of
12 months were enrolled in this study. Inclusion criteria: hysteroscopic diagnosis
of submucous myomas, polyps, uterine septa or DUB requiring endometrial
resection. Exclusion criteria were: age > 50, obesity, pregnancy,
prolapse, malignancy, and presence of IUAs. Patients were randomized
into 2 groups, and without significant differences in age, weight, uterine
size and party. Group (A) of 40 patients underwent hysteroscopic surgery(HS)
plus intrauterine application of MEBO. Group (B), the remaining 32
patients, underwent (HS) only (control). Follow-up diagnostic hysteroscopy
was performed for each patient, 6 weeks later. Adhesion score assessment according
to the American Fertility Society (AFS) classification system. Statistical
analysis performed using SPSS package. P-value considered to be
significant if (< 0.05).
RESULTS: Eleven women ( 4 from group A & 7 from group B) did not
attend for follow-up. At 6 weeks follow-up, a highly significant lower rate
of post-surgical IUAs was found in group (A): only one out of 36 women
compared with group (B): 8 out of 25 patients {2.8 % versus 32 % ; P <
0.05}. Evaluation of IUAs staging by AFS system, showed a significant
decrease in adhesions severity in the patient from group A ( stage I ) when
compared with women from group B: 6 patients (stage II) & 2 patients (stage
III ){P < 0.05}. No complications or adverse MEBO-related effects were detected
in group A patients.
CONCLUSIONS: (MEBO), is safe, non-expensive and with a highly significant
reduction rate in the incidence and severity of de-novo formation of
IUAs after HS.
P-205 Tuesday, October 20, 2015
THE IMPACT OF THE TECHNIQUES OF HYSTEROSCOPIC SEP-
TOPLASTY ON THE REPRODUCTIVE OUTCOMES _IN PATIENTS
WITH SEPTATE UTERUS. O. Dural, C. Yasa, E. Bastu, F. Gungor
Ugurlucan, S. Can, G. Yilmaz, F. Buyru. Department of Obstetrics and Gynecology,
Istanbul University School of Medicine, Istanbul, Turkey.
OBJECTIVE: The study was aimed to determine the reproductive outcomes
of two different techniques of hysteroscopic septoplasty.
DESIGN: We retrospectively studied 49 patients who underwent hysteroscopic
septoplasty (HS) for symptomatic septate uteri between January 2010
and April 2014.
MATERIALS AND METHODS: We divided the patients into two groups
on the basis of the technique of HS. Group I involved 27 patients with a history
of HS which was performed by monopolar hook using the operating hysteroscope
8 mm in diameter. Group II involved 22 patients with a history of
HS which was performed by scissors using office hysteroscopy 5mm in diameter.
We evaluated pregnancy outcomes within first year after HS for Group I
and II.
RESULTS: Reproductive outcomes were obtained from 44 patients who
tried to get pregnant after HS within first year. Grup I and II include 25
and 19 patients with median age 27,5 (19-38) and 28 (21-39), respectively.
There were a total of 23 pregnancies in Group I, of which 15 (65.2%) were
carried to term, 3 (13%) resulted in preterm live birth, 5 (21.7%) resulted
in pregnancy loss in the first or second trimester. There were a total of 17
pregnancies in Group II, of which 11 (64.7%) were carried to term, 2
(11.7%) resulted in preterm live birth, 4 (23.5%) resulted in pregnancy
loss in the first or second trimester. The overall life birth rate was 78.2 %
in Group I and 76.4% in Group II (p¼0.85). One case of uterine rupture occured
at 37 week gestation in a subsequent pregnancy in Group I.
CONCLUSIONS: There is a scarcity of data regarding whether using of a
specific instrument during HS may improve reproductive outcome. Our data
support that the rates of pregnancy to term and live birth are similar between
two techniques of HS. In accordance with a few previous case reports , uterin
rupture occured after HS with monopolar diathermy. Additional studies are
needed to evaluate the impact of the techniques of HS on the reproductive
outcomes.
P-206 Tuesday, October 20, 2015
LAPAROSCOPIC AND ROBOTIC MYOMECTOMY: COMPARISON
OF COST AND PERIOPERATIVE OUTCOMES FOR COMPLEX
CASES. M. V. Vargas. Department of Obstetrics and Gynecology, Division
of Gynecology, George Washington University Medical Center, Washington,
DC.
OBJECTIVE: To compare cost and outcomes between laparoscopy and robotic-assistance
in women undergoing complex myomectomy.
DESIGN: Retrospective chart review (Canadian Task Force Classification
II-2).
MATERIALS AND METHODS: We reviewed charts of women laparoscopic
and robotic myomectomy by two expert surgeons from January
2014 to December 2014. Each surgeon utilized exclusively one modality.
Characteristics of patients with largest myoma size > 8cm and/or specimen
weight > 250g and/or > 5 myomas removed were compared by surgical modality.
RESULTS: The cohort consisted of 70 patients with 54% laparoscopic and
46% robotic myomectomies. Aside from higher rates of Medicaid insurance
in the robotic group (28% versus 8%, P¼.0017), there were no differences in
demographics and medical/surgical history. The robotic group had significantly
lower rates of concomitant operative hysteroscopy (32% versus 2%,
P¼.014), mean (SD) specimen weights in grams (351.5 (417.6) versus
574.0 (525.5), P¼.014), and mean (SD) operative time in minutes (150
(62.9) versus 216.7 (84.8), P¼.0006), but no difference in number of myomas
removed (median (range) 4 (1-16) versus 4 (1-20), P¼.057) or size of dominant
myoma (median (range) 8 (2-20 versus 10 (2-20)), P¼.18). There was no
difference in estimated blood loss or rate of complications. The mean (SD)
direct in dollars for robotic cases was lower (5861.3 (2273.9) versus
7081.7 (3373.5), P¼.0402) but indirect costs did not significantly differ
9833.5 (3749.7) versus 12048.6 (6396.3), P¼.058). Cost was significantly
increased by operative time, specimen weight, estimated blood loss, prior
laparotomy, complications, and length of stay.
CONCLUSIONS: In this pilot study of expert surgeons using their
preferred modality for complex myomectomy, outcomes and costs were
comparable. Costs were most influenced by factors that increased surgical
complexity and were contained in the robotics group. This preliminary
data demonstrates a potential capacity for cost containment with the use of
robotic surgery.
P-207 Tuesday, October 20, 2015
WITHDRAWN
e176 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

P-209 Tuesday, October 20, 2015
REPRODUCTIVE OUTCOMES AFTER LAPAROSCOPIC MYO-
MECTOMY (LM) AND LAPAROSCOPIC RADIOFREQUENCY
VOLUMETRIC THERMAL ABLATION UTERINE
LEIOMYOMAS. K. Isaacson. Gynecology, Harvard Medical School,
Newton, MA.
P-208 Tuesday, October 20, 2015
WITHDRAWN
OBJECTIVE: To evaluate reproductive outcomes in women who were
randomized to laparoscopic ultrasound-guided LM and RFVTA for symptomatic
myomas.
DESIGN: Single-center, prospective, randomized controlled trial.
MATERIALS AND METHODS: Fifty women with self-reported heavy
menstrual bleeding and/or bulk symptoms consented to intraoperative
randomization (1:1) to LM or RFVTA for treatment of their symptomatic
myomas. All participants desired uterine and reproductive conservation.
RESULTS: Eight women ages 28 to 35 years (median, 33 years) conceived
9 times between 3.2 to 23.5 months after excision or ablation of intramural
and subserosal myomas. Of the RFVTA cohort (n ¼ 25), 3 women, ages
31 to 35 years old, underwent ablation of 1 or 2 fibroids (intramural and/or
subserosal) measuring 0.5 to 5.7 cm in greatest diameter. No intraoperative
or postoperative complications were reported, and the patients conceived
at 7 to 23.5 months post procedure. Two of the pregnancies resulted in
full-term spontaneous vaginal births of healthy infants, and the third pregnancy
is expected to result in a full-term vaginal birth in August 2015 for
a pregnancy rate of 12% (3/25). Five women in the LM cohort (n ¼ 25)
conceived 6 times for a pregnancy rate of 24% (6/25). One 28-year-old patient
conceived at 3.2 months post myomectomy, but the pregnancy ended
in a therapeutic abortion. She conceived again at 13.8 months post myomectomy
and delivered a healthy infant by C-section. Three myomectomy patients
conceived and delivered healthy infants by vaginal (n ¼ 2) and
Cesarean delivery (n ¼ 1). The fifth myomectomy patient’s pregnancy is
ongoing.
CONCLUSIONS: Viable, full-term pregnancies culminating in vaginal
births of healthy infants are possible after either RFVTA of symptomatic
uterine myomas or laparoscopic myomectomy. Patients in this study will
be followed for 5 years.
P-210 Tuesday, October 20, 2015
PERINATAL RISK IN MICROTUBAL REANASTOMOSIS VERSUS
IN VITRO FERTILIZATION IN PATIENTS WITH TUBAL FACTOR
INFERTILITY. S. A. Greenberg, a S. O’Gradney, a L. Davis, b
E. A. Zbella, a M. Sanchez. a a Bayfront Health St Petersburg, St Petersburg,
FL; b OBGYN, Bayfront Health St Petersburg, St Petersburg, FL.
OBJECTIVE: To assess the perinatal outcome and evaluate the risk of preterm
delivery, low birth weight, multiple gestation, spontaneous abortion,
and ectopic pregnancy in women with tubal factor infertility who underwent
either Microtubal Reanastomosis (MTR) versus In Vitro Fertilization (IVF).
DESIGN: Retrospective Cohort.
FERTILITY & STERILITY Ò
e177

MATERIALS AND METHODS: We analyzed the perinatal outcome of 74
IVF and 39 MTR patients who had tubal factor infertility with documented
fetal cardiac activity between 2009 and 2013. Preterm delivery, low birth
weight, multiple gestation, spontaneous abortion and ectopic pregnancy
were then analyzed between IVF and MTR. Preterm delivery and low birth
weight were only compared amongst cohorts with singleton pregnancies.
We performed multivariable analyses using Fisher’s exact and calculated
P-value and adjusted risk ratio at a 95% confidence intervals.
RESULTS: IVF had an increased risk of preterm delivery (27.7%
compared with 3.33%, p-value 0.0153, adjusted RR 1.92, 95% CI), spontaneous
abortion (24.3% compared with 7.63%, p-value .0485, adjusted RR
1.4, 95% CI) and multiple gestation (35.7% compared with 3.22%, p-value
.0067, adjusted RR 1.74, 95% CI). Although there was an increased risk of
low birth weight, this was not statistically significant (22.2% compared
with 6.66%, p-value .2863, adjusted RR 1.42, 95% CI). MTR had an
increased risk of ectopic pregnancy (15.3% compared with 0%, p-value
0.0031, adjusted RR 3.18, 95% CI).
CONCLUSIONS: Patients who have undergone IVF have an increased
risk for preterm birth, multiple gestation, and spontaneous abortion when
compared to those who have undergone MTR for tubal factor infertility.
P-211 Tuesday, October 20, 2015
EFFECT OF HYSTEROSCOPIC SURGERY FOR UTERI WITH
CAVITARY LESIONS ON CLINICAL PREGNANCY RATE IN
WOMEN SCHEDULED FOR IVF. A. M. Abdelmagied, a,b
M. A. Kamel, b A. M. Abuelhasan, b I. Elnashar, b A. A. Abdelaleem, b
T. A. Farghaly, b A. A. Nassr, a,b M. H. Makarem. b a Obstetrics and Gynecology,
Mayo Clinic, Rochester, MN; b Obstetrics and Gynecology, Women
Health Hospital, Assiut University, Assiut, Egypt.
OBJECTIVE: Adoption of routine office hysteroscopy (OH) before In-Vitro
Fertilization (IVF) to diagnose and treat cavitary lesions (CL) is still
debatable both from evidence and cost perspectives. Our objective was to
evaluate the role of OH in detecting CL and the effect of correction of these
lesions before IVF on clinical pregnancy rate (CPR).
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: Women with primary infertility, before
their enrollment in In-Vitro Fertilization (IVF), were examined by 2 diagnostic
modalities to detect possible CL: trans-vaginal ultrasound (TVUS)
and OH, as a gold standard,. All CL diagnosed by OH were treated before
IVF. TVUS and OH findings, IVF related data and CPR were reported for
all women. The t-test, Wilcoxon rank sum test, Chi-square test, Fisher exact
test and logistic regression were used for comparisons.
RESULTS: Sixty six women were included. In all, 20 women (30.3 %) had
CL, 8 (40 %) of them were missed by TVU. The overall implantation rate
(IR) and CPR were 16% and 36.4% respectively. Women with (n¼20) and
without (n¼46) CL were similar in age, BMI, duration of infertility, antral follicle
count, AMH, number of oocytes retrieved, number and quality of embryos transferred
and IR. However, women with CL were more likely to have previous
failed IVF attempts (55% vs. 26%, P¼0.02), and yet higher CPR (55% vs 28,
P¼0.04) than women without CL. Univariate analysis was conducted for potential
confounders that may influence CPR. CL treated uteri, adjusted for these confounders,
were comparable to uteri with no CL in terms of CPR (aOR ¼ 1.79,
95% CI¼0.52-6.17, P>0.05). A similar trend for CPR (58% vs 50%, P>0.05)
was observed when comparing TVUS suspected CL group (n¼12) to missed
CL group (n¼8). Of the overall CPR, 16.7% was attributed to the OH diagnosis
and treatment of the missed lesions. Accordingly, 14 women with normal TVUS
were needed to be screened by OH to achieve one clinical pregnancy.
CONCLUSIONS: Cavitary lesions were common among women scheduled
for IVF and a considerable percentage of these lesions were missed
on TVUS. Women with treated CL and those with no CL had comparable
CPR. The role of OH in recognizing TVUS missed CL should not be underestimated
as considerable increase in CPR could be achieved by their correction.
Supported by: Research support in university hospitals.
P-212 Tuesday, October 20, 2015
THE SURGICAL INDICATIONS FOR SEPTATE UTERUS IN INFER-
TILE POPULATION BEFORE IN VITRO FERTILIZATION. Y. Yi, a
X. Li, b Y. Ouyang, c G. Lu. a a Institute of Reproduction & Stem Cell Engineering,
Central South University, Chang-sha, China; b Reproductive and Genetic
hospital of CITIC-Xiangya, Chang-sha, China;
c Central South
University, Changsha, China.
OBJECTIVE: To investigate whether the surgical indications are reasonable
for infertile patients with septate uterus before IVF-ET (in vitro fertilization-embryo
transfer) and whether there are differences about pregnancy
outcome between infertile patients with septate uterus and infertile patients
with normal uterus after IVF-ET.
DESIGN: Retrospectively analyzed pregnancy outcome in patients with
normal uterus and patients with septate uterus after IVF-ET.
MATERIALS AND METHODS: 2637 infertile patients who got clinical
pregnancy after IVF-ET from April 2012 to March 2014 were identified.
There were 2100 patients (A group) with normal uterus and 537 patients
with septate uterus. Hysteroscopic metroplasty was performed before IVF-
ET in 231 patients (B group) with septate uterus who met surgical indications
that the depth of septum >10mm or depth of septum ranges from 5 to 10mm
accompanied by history of unexplained recurrent miscarriage. 306 patients
(C group) with septate uterus were managed expectantly because the depth
of septum < 10mm and without history of unexplained recurrent miscarriage.
Pregnancy outcome including miscarriage rate in 1st and 2nd trimester, preterm
delivery rate, term delivery rate and ectopic pregnancy rate were
compared among above 3 groups by Chi-Square Test. The difference was statistically
significant when P < 0.05.
RESULTS: Though the miscarriage rate in 1st trimester (14.1%) in C
group (Managed expectantly) was higher than that in other two groups, the
difference was not significant (P¼0.130). And miscarriage rate in 2nd
trimester, preterm delivery rate, term delivery rate and ectopic pregnancy
rate indicate no significant difference in these three groups (Table).
Pregnancy outcome in 3 groups (P¼0.130).
Pregnancy outcome Total
A group (Normal) 2100
(100%)
B group 231
(Hysteroscopic (100%)
metroplasty)
C group (Managed
expectanly)
306
(100%)
Miscarriage
in first
trimester
214
(10.2%)
27
(11.7%)
43
(14.1%)
Miscarriage
in second
trimester
91
(4.3%)
15
(6.5%)
12
(3.9%)
Preterm
delivery
Full term
delivery
276 1467
(13.1%) (69.9%)
37 150
(16.0%) (64.9%)
32 212
(10.5%) (69.3%)
Ectopic
pregnancy
52
(2.5%)
2
(0.9%)
7
(2.3%)
CONCLUSIONS: For infertile patients with septate uterus, the indication
of surgery that the depth of septum>10mm or depth of septum ranges
from 5 to 10mm accompanied by history of unexplained recurrent miscarriage
were reasonable before IVF-ET. According to above surgical
indications, the infertile patients with septate uterus could get similar
pregnancy outcome compared with infertile patients with normal uterus
after IVE-ET.
P-213 Tuesday, October 20, 2015
SMALLER ENDOMETRIOTIC CYST REMOVAL ALONG WITH
LONGER DURATION OF THE CYST HAS HIGHER IMPACT ON
OVARIAN RESERVE-ASSESSMENT OF ANTIMULLERIAN
HORMONE AND ANTRAL FOLLICULAR COUNTS, POST
CYSTECTOMY - A PROSPECTIVE OBSERVATIONAL
STUDY. N. K. Pratap, R. Sandya. Obstetrics and Gynecology, Kasturba
Medical College, Manipal, India.
OBJECTIVE: The damage inflicted to the ovary by cystectomy is a major
concern in balancing the fertility risks and benefits. This study
compared the cyst of less than 5 cms to more than 5 cms post laparoscopic
cystectomy with Antimullerian hormone (AMH) and Antral Follicular
Counts (AFC).
DESIGN: Prospective observational study was done following endometrioma
cystectomy and compared post operatively the cyst size with AMH
and AFC for ovarian reserve after one month.
MATERIALS AND METHODS: Group A less than 5 cms cyst and Group
B with more than 5 cms cyst were compared. Statistical Mauchly’s test of
sphericity was done. AMH was grouped as I, II and III depending on the
values of less than 1,1-3.5, 3.5ng/ml and further analyzed comparing the
cyst size by Pearson Chi square test.Ovarian sections were taken from the
thickest areas of the cyst wall to analyze histologically the amount of primodial,
primary and secondary follicles.
RESULTS: There was an overall drop of AMH from 3.83.01ng/ml to
2.671.92ng/ml (p¼ less than 0.001). AMH was clearly low in Group III
following surgery (46.4% to 26.0%). AMH dropped from 4.53.4ng/ml to
e178 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

3.192.18 with less than 5 cms cyst (Group A), compared to 2.41.2 to
1.70.85 with more than 5 cms cyst (Group B). A mean drop of 1.31 and
0.7ng/ml in Group A and B respectively (p value less than 0.01) was seen.
Overall AFC was 5.171.44 and 3.611.61 pre and post operatively in
the ovary operated. By the Statistical test - Paired t test it was significant.
Drop in AFC was 1.2 and 2.2 in Group A and B. With unilateral cyst, 5 of
the 15 in Group A had very low AMH post operatively (less than 1ng/ml)
with the duration of the cyst being more than four years. Histopathological
analysis of cyst wall based on semi quantitative scale which had grades 0-
3 depending on the loss of follicles were analyzed. Irrespective of the loss
of follicles or not, there was decline in both AMH and AFC. Of the total
cyst wall analyzed 76% did not have loss of follicles. However, with more
loss of follicles, there was more decline of AFC and AMH.
CONCLUSIONS: Smaller the endometriotic cyst and longer the duration,
more is the damage to the ovarian reserve. Hence, with smaller cysts meticulous
surgical techniques are needed for better prognosis compared to the
larger cyst.
P-214 Tuesday, October 20, 2015
PREDICTORS OF HEMORRHAGE AND TRANSFUSION FOR
WOMEN UNDERGOING LAPAROSCOPIC AND ROBOTIC-ASSIS-
TED MYOMECTOMY. M. V. Vargas. Department of Obstetrics and Gynecology,
Division of Gynecology, George Washington University Medical
Center, Washington, DC.
OBJECTIVE: To identify predictors of hemorrhage and/or transfusion for
women undergoing minimally invasive myomectomy.
DESIGN: Retrospective chart review (Canadian Task Force Classification
II-2).
MATERIALS AND METHODS: We reviewed the charts of women who
underwent minimally invasive myomectomy by three expert surgeons from
April 2011 to December 2014. Characteristics of patients with estimated
blood loss (EBL) > 1000mL and/or transfusion were compared to characteristics
of all other patients in the cohort.
RESULTS: The cohort consisted of 220 patients with 100 laparoscopic and
120 robotic myomectomies. The median (range) number of myomas
removed was 3 (1-20) and dominant myoma size in cm was 8 (4-20). The
mean (SD) specimen weight in grams was 409 (396) and ranged from 4 to
2894 grams. The mean (SD) operative room time (ORT) in minutes was
190 (80) and EBL in mL was 352 (494). The complication rate was
13.6%. Twenty-four patients (11%) experienced hemorrhage and/or transfusion,
accounting for 80% of complications. These patients experienced
longer ORT (mean in minutes (SD), 272 (100) versus 180 (73), P¼.0007)
and length of stay (median days (range), 2 (0-6) versus 1 (0-3), P¼.0001),
as well as higher rates of conversion (13% versus 0%, P¼0.0025) and ICU
admission (13% versus 0%, P¼0.0025). The probability of hemorrhage/
transfusion rose significantly with dominant myoma > 10cm (OR 2.83;
95% CI 1.34 to 5.87), removal of > 5 myomas (OR 3.33; 95% CI 1.1 to
10.5), transmural location 2.94 (95% CI 1.27 to 6.81), and specimen weight
> 500g (OR 5.84; 95% CI 2.70 to 12.67).
CONCLUSIONS: In this surgically complex cohort, minimally invasive
myomectomy had a complication rate that was comparable to other reports
of high volume surgeons. Hemorrhage and transfusion were the most common
complications and were associated with case complexity as well as
increased surgical morbidity. Anticipating these complications can facilitate
preoperative planning, patient counseling, and the implementation of preventative
interventions.
P-215 Tuesday, October 20, 2015
SHIFTING ANAEROBIC TO AEROBIC METABOLISM STIMU-
LATES APOPTOSIS IN ADHESION FIBROBLASTS THROUGH
THE MODULATION THE CELLULAR REDOX
HOMEOSTASIS. N. M. Fletcher, a M. G. Saed, a B. R. Neubauer, a
H. Abu-Soud, a A. O. Awonuga, b M. P. Diamond, c G. M. Saed. a a Obstetrics
and Gynecology, Wayne State University, Detroit, MI; b Wayne State University,
Southfield, MI; c Georgia Regents University, Augusta, GA.
OBJECTIVE: To compare the effect of shifting anaerobic to aerobic metabolism
on key regulators of oxidative stress, including extracellular superoxide
dismutase (SOD3), inducible nitric oxide synthase (iNOS) and its
product, nitric oxide (NO), as well as mitochondrial potential (Djm) and
apoptosis in normal peritoneal and adhesion fibroblasts.
DESIGN: Prospective experimental study.
MATERIALS AND METHODS: The normal peritoneal and adhesion fibroblasts
established from normal peritoneal and adhesion tissues from the
same patient(s) were treated with dichloroacetate (DCA 0, 20, 40, 80 mg/
ml, 24 hrs). Additionally, cells were treated with hypoxia (2% O2, 24 hrs)
in the presence of DCA (80 mg/ml). Expression of SOD3 and iNOS and
NO levels were determined by a combination of real-time RT-PCR and
Greiss assay. The Djm was evaluated by the JC-1 Mitochondrial Membrane
Potential Assay. Apoptosis was determined by caspases-3 activity and TU-
NEL assays. Data were analyzed using SPSS 22.0 for Windows. Mixed
model repeated measures ANOVA was used with treatment as the within factor
and cell type as the between factor. Paired comparisons with a Bonferroni
correction were used to compare pairs of treatments. Significant interactions
between treatment and cell type were analyzed with independent sample t-
tests by cell type on each treatment.
RESULTS: Levels of iNOS, SOD3, NO (nitrate/nitrite), and caspase-3 activity
are summarized in Table 1. There was a marked increase in TUNEL
staining in both normal peritoneal and adhesion fibroblasts in response to
DCA (80 mg/ml). There was also enhanced Djm in adhesion fibroblasts,
indicative of high oxidative stress, as compared to normal peritoneal fibroblasts.
Further enhancing oxidative stress by hypoxia treatment markedly
increased Djm in normal peritoneal to levels observed in adhesion fibroblasts.
Treatment with DCA (80 mg/ml) was found to protect against the effects
of hypoxia.
CONCLUSIONS: DCA modulates the redox homeostasis protecting cells
against anaerobic metabolism and the associated oxidative stress. These findings
may provide targets for therapeutic intervention for the reduction of profibrotic
disorders, including postoperative adhesions.
Levels of iNOS, SOD3, NO, and activity of caspase-3 in normal peritoneal and
adhesion fibroblasts.
iNOS
(fg/mg RNA)
SOD3
(fg/mg RNA)
Nitrate/
Nitrite (mM)
Caspase-3
Activity (mM)
Normal fibroblasts - 1.8 0.3 81.4 9.0 7.0 0.4 12.6 0.6
Untreated
Normal fibroblasts - 2.3 0.6 111.7 2.6* 7.0 0.1 15.5 0.9*
20 mg/ml DCA
Normal fibroblasts - 4.5 0.8* 97.9 2.1 9.4 0.04* 18.3 1.2*
40 mg/ml DCA
Normal fibroblasts - 16.8 0.1* 91.8 5.8 10.3 0.5* 21.6 0.7*
80 mg/ml DCA
Adhesion Fibroblasts - 2.0 0.2 27.9 0.6* 4.4 0.5** 8.1 0.8*
Untreated
Adhesion Fibroblasts - 2.4 0.3 116.2 4.8** 4.2 0.1 11.5 0.6**
20 mg/ml DCA
Adhesion Fibroblasts - 4.4 0.5** 65.9 2.3** 5.7 0.4** 17.5 0.8**
40 mg/ml DCA
Adhesion Fibroblasts - 21.1 0.3** 82.9 0.2** 13.2 0.4** 21.4 1.1**
80 mg/ml DCA
*p
P-216 Tuesday, October 20, 2015
30-DAY POSTOPERATIVE OUTCOMES OF MINIMALLY INVA-
SIVE VERSUS ABDOMINAL MYOMECTOMY: AN ANALYSIS OF
THE NATIONAL SURGICAL QUALITY IMPROVEMENT PRO-
GRAM (NSQIP) DATABASE 2005-2013. S. Moustafa, a C. M. Duke, b
A. Sood, c E. C. Dun, d A. Dabaja. c a Department of Obstetrics, Gynecology,
& Reproductive Sciences, Yale School of Medicine, New Haven, CT; b Yale
University School of Medicine, New Haven, CT; c Vattikuti Urology Institute,
Henry Ford Health System, Southfield, MI; d Yale School of Medicine, New
Haven, CT.
OBJECTIVE: To compare complication/adverse event rates between
minimally invasive (MI; laparoscopic and robotic-assisted) myomectomy
and abdominal myomectomy. Further, to assess the independent effect of
MI myomectomy on complications/adverse events in these patients.
DESIGN: Retrospective cohort study using the National Surgical Quality
Improvement Project Database (NSQIP).
MATERIALS AND METHODS: Patients undergoing either MI or abdominal
myomectomy, from 2005-2013, were identified using their CPT codes
[58140, 58146, 58545, 58546]. The primary endpoint was postoperative
complications. Secondary endpoints included operative time, length of stay
FERTILITY & STERILITY Ò
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(LOS), re-intervention, and readmission rates. Multivariable logistic regression
models assessed the independent effect of MI on postoperative outcomes.
RESULTS: 5338 patients were identified: 2261 (42.4%) underwent MI
myomectomy and 3074 (57.6%) underwent abdominal myomectomy. The
mean age of the patients in the MI group and abdominal myomectomy group
were 38.4 (SD¼7.3) years and 37.4 (SD¼6.4) years (p

References:
1. Committee on Adolescent Health C. Committee opinion: no. 562:
mullerian agenesis: diagnosis, management, and treatment. Obstetrics
and gynecology. May 2013;121(5):1134-1137.
2. Smith NA, Laufer MR. Obstructed hemivagina and ipsilateral renal
anomaly (OHVIRA) syndrome: management and follow-up. Fertility
and sterility. Apr 2007;87(4):918-922.
3. Cooper AR, Merritt DF. Novel use of a tracheobronchial stent in a patient
with uterine didelphys and obstructed hemivagina. Fertility and
sterility. Feb 2010;93(3):900-903.
P-219 Tuesday, October 20, 2015
ASSOCIATION BETWEEN BODY MASS INDEX (BMI), UTERINE
SIZE, AND OPERATIVE MORBIDITY IN WOMEN UNDERGOING
MINIMALLY INVASIVE HYSTERECTOMY. D. K. Shah, a B. Van
Voorhis, a A. Vitonis, b S. A. Missmer. b,c,d a Division of Reproductive Endocrinology
and Infertility, University of Iowa, Iowa City, IA; b Department of
Obstetrics, Gynecology, and Reproductive Biology, Brigham and Women’s
Hospital, Boston, MA; c Harvard Medical School, Boston, MA; d Harvard
School of Public Health, Boston, MA.
OBJECTIVE: To examine the association of surgical approach and operative
morbidity after minimally invasive hysterectomy and determine
whether the association varies with uterine size and patient BMI.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: Data abstracted from the American College
of Surgeons National Safety and Quality Improvement Project registry
included 36,855 women who underwent vaginal (VH), laparoscopic-assisted
vaginal (LAVH), or total laparoscopic (TLH) hysterectomy between January
2005-December 2012. Associations between surgical approach, BMI, and
operative morbidity were examined, stratifying based on uterine size and adjusting
for age, race, ethnicity, year of surgery, smoking, diabetes, and American
Society for Anesthesiology physical classification. Adjusted means, rate
ratios, or odds ratios (OR) with 95% confidence intervals (CI) were calculated
using linear, Poisson, or logistic regression.
RESULTS: Operative times were uniformly shorter in women undergoing
VH as compared to LAVH or TLH (p 40 kg/m 2 had 76% lower
odds of blood transfusion (CI¼0.1-0.6; unadjusted rates 0.8% vs. 2.2%)
and 39% lower odds of surgical site infections (CI 0.4-1.0; unadjusted rates
2.4% vs. 3.9%) after TLH as compared to VH. The association between
wound infection and BMI was attenuated or absent in women with large uteri
or in those that underwent LAVH.
CONCLUSIONS: When comparing minimally invasive approaches to
hysterectomy, operative times are shortest with VH, particularly in obese
women with small uteri. Rates of blood transfusion and wound infection
are lowest after TLH but absolute risks are low regardless of surgical
approach. Laparoscopic assisted vaginal hysterectomy appears to confer no
specific advantage over VH or TLH.
Supported by: Departmental funding, University of Iowa.
P-220 Tuesday, October 20, 2015
ART - CLINICAL
A RANDOMIZED CONTROLLED TRIAL OF ORAL ACETAMINO-
PHEN FOR ANALGESIC CONTROL AFTER TRANSVAGINAL
OOCYTE RETRIEVAL. S. Kassira, a G. Bates, b M. Powell, c
J. M. Bouknight, b M. McLean. b a School of Medicine, UAB, Birmingham,
AL; b Division of Reproductive Endocrinology and Infertility, UAB, Birmingham,
AL; c Dept of Anesthesiology, UAB, Birmingham, AL.
OBJECTIVE: To evaluate the efficacy of 1000mg of oral acetaminophen
onpost-procedure pain after transvaginal cyst puncture (TVCP).
DESIGN: Double-blind randomized controlled trial of acetaminophen vs
placebo in women undergoing IVF at an academic infertility clinic. Women
with an allergy to acetaminophen, narcotic dependence or use of acetaminophen
within 24 hours were excluded.
MATERIALS AND METHODS: All eligible patients presenting for
TVCP were approached for enrollment. Subjects were randomized at the
time of TVCP to acetaminophen vs placebo using a permuted block
schedule. Pills were taken under supervision of study personnel 1 hour prior
to TVCP. Demographics, IVF cycle characteristics and a baseline pain
score using the visual analogue scale (VAS) were collected. Conscious
sedation was administered by anesthesiology per routine. Procedure time,
intraoperative medications, and number of oocytes retrieved were recorded.
PACU pain scores (15, 30, 45, 60 minutes) and pain medication use in recovery
was collected by the nurse. At 24 hours, post-procedure pain scores
and use of pain medication was assessed. Total pain score was a sum of
PACU pain scores and 24 hour pain score. Wilcoxon rank-sum test was
used to compare pain scores between groups. Chi squared and t-tests
were used to assess factors associated with pain scores. For multivariable
analysis ‘‘moderate pain’’ was considered a cumulative pain score >8 for
PACU pain scores and >10 for 24-hr pain scores. Logistic regression was
used to compare the incidence of moderate pain while controlling for confounders.
Planned enrollment of 100 subjects will provide 80% power to
detect a 2-point difference in pain on the VAS.
RESULTS: To date 32 subjects have been enrolled with mean age
32.8+/- 5.0 years. Mean duration of oocyte retrieval was 23 +/- 10min.
9 subjects (28%) required pain medication in the PACU. 23 subjects
(72%) used an opioid in the 24 hours post-procedure. 16 subjects
(50%) did not undergo fresh embryo transfer, including 5 oocyte donors.
There was no difference in the total pain score (median[IQR]) between
placebo and acetaminophen groups (5.0 [1-13] vs 5.5 [0-10.5] p ¼0.7)
nor in PACU pain scores (6.5 [1.5-12] vs 7.0 [4-15] p ¼0.7). There was
no difference in the use of pain medication post procedure. Baseline
pain score, BMI, or diagnosis of endometriosis was not associated with
pain scores. Oocyte donors had similar pain scores to those undergoing
autologous IVF (8.0 vs 7.0; p ¼ 0.87). Procedure time and ibuprofen
use prior to TVCP was associated with moderate pain scores. There
was no difference in PACU or total pain scores between groups after controlling
for these factors(p¼0.89; 0.55).
CONCLUSIONS: Preoperative administration of oral acetaminophen
does not reduce PACU or 24-hour cumulative pain scores after TVCP. Procedure
time and ibuprofen use prior to TVCP are associated with post-operative
pain after TVCP.
P-221 Tuesday, October 20, 2015
BLASTOCYST VERSUS CLEAVAGE-STAGE ELECTIVE SINGLE
EMBRYO TRANSFER - A COMPARATIVE RETROSPECTIVE
STUDY. Z. Veleva, a S. Boulet, b S. Makinen, c H. Martikainen, d
A. Tiitinen, a J. Tapanainen, a D. M. Kissin. b a Ob/Gyn, University of Helsinki
and Helsinki University Central Hospital, Helsinki, Finland; b Centers for
Disease Control and Prevention, Atlanta, GA; c Family Federation of Finland,
Helsinki and Oulu Infertility Clinics, Helsinki, Finland; d Ob/Gyn, University
of Oulu and Oulu University Hospital, Oulu, Finland.
OBJECTIVE: To compare cumulative term live birth rates (ctLBR) after
elective single embryo transfer (eSET), using data from two countries –
Finland where cleavage stage (day 2/3) eSET is almost exclusively used,
and the U.S. where blastocyst (day 5/6) eSET is more common.
DESIGN: Retrospective cohort study comparing data from Finland’s
largest infertility clinics (LUMI database, cleavage stage eSET performed
in 50.5% of all fresh cycles) with data from the U.S. National Assisted Reproductive
Technology (ART) Surveillance System (blastocyst eSET performed
in 3.9% of all fresh cycles).
MATERIALS AND METHODS: The analysis included all fresh, nondonor
eSET (n¼16,276) and subsequent frozen embryo transfer cycles
(n¼11,625) performed during 2008-2012 in the U.S. and Finland among
women aged %40 years with no prior use of ART. U.S. clinics were classified
into those with high (R52 cases/year) and low (

OR 1.91, 95% CI 1.68-2.16). Gonadotropin dose per oocyte was higher in
cleavage stage Finnish cycles (184.3141.0 IU), compared with U.S. blastocyst
cycles (low eSET use: 154.2129.5, P

We observed that the overweight sample showed despite their efforts, an increase
in weight compared to their reported weight one year earlier (mean ¼
0.3kg SD 8.75 kg).
CONCLUSIONS: The majority of the women had made one or more
health promoting changes, but a considerable proportion of patients kept lifestyle
behaviors not optimal for conception, such as obesity and use of tobacco.
Therefore women, who have problems in conceiving, should be
encouraged to seek help earlier than did women in this sample. Fertility
care should include, as a routine, both assessments for lifestyle factors and
interventions to lifestyle changes.
Reference: Key Words: infertility, lifestyle, obesity, alcohol, diet.
Supported by: Funding of the study was received from the Foundation
Family Planning Fund in Uppsala, the College of Medicine at Uppsala University,
Sweden, V€astmanland county council, Sweden and the Uppsala-
€Orebro Regional Research Council, Sweden. None of the authors have any
conflict of interest to declare.
P-225 Tuesday, October 20, 2015
PROGESTERONE LEVEL ON THE DAY OF HCG TRIGGERING: A
NEW TOOL FOR A SUCCESSFUL MODIFIED NATURAL IN VITRO
FERTILIZATION CYCLE? C. Kamga-Ngande, a,b I. Kadoch, a,b
L. Lapensee. a,b a Reproductive Endocrinology and Infertility, Centre Hospitalier
Universitaire de Montreal, Montreal, QC, Canada; b Ovo Fertility
Clinic, Montreal, QC, Canada.
OBJECTIVE: Premature progesterone elevation before hCG triggering
is known to occur occasionally in stimulated IVF although causes for
this elevation are still unclear. The negative effect of this rise on the
endometrium therefore affecting IVF outcomes is acknowledged. However,
this is not documented in mnIVF condition. The aim of this study
is to determine whether PG levels on the day of hCG could predict no
oocyte at egg collection and both biochemical and clinical pregnancy
rates (PR).
DESIGN: We conducted a quality control (from 01-01-2012 to 30-06-
2013) involving 581 women and 1074 mnIVF cycles.
MATERIALS AND METHODS: Progesterone (PG) and Estradiol (E2)
values on the day of hCG triggering were available. Multivariate analyses
were performed tacking into account diagnosis, days of GnRH antagonist,
antimullerian hormone (AMH) level, and age >35 years.
RESULTS: Female were aged 32.43.5years,withAMH,E2andprogesterone
levels at 1.71.9 ng/mL, 916303 pmol/L, and 1.6 0.8 nmol/
L, respectively. The ‘‘no oocyte retrieved at egg collection per cycle’’ rate
was 9.5%. Per egg collection, 94% of oocytes were mature. Biochemical
and clinical PR were respectively 17.8% and 16.2% per cycle and 35.9%
and 32.8% per embryo transfer. Associations were found between PG
level and no oocyte retrieved at egg collection (OR ¼ 1.61 (1.2-2.16);
p¼0.001), biochemical PR (OR ¼ 0.62 (0.41-0.93); p¼0.02), and clinical
PR (OR ¼ 0.63 (0.42-0.95); p¼0.02). Each increase in PG significantly
increased the risk of not finding oocytes at egg collection by 61%, and
decreased chance of biochemical and clinical PR by 38% and 37%,
respectively.
CONCLUSIONS: PG appears to have a detrimental effect on mnIVF
outcomes. It has been proposed that PG increase in the late follicular
phase influences endometrial maturation. This may lead to decreased
endometrial receptivity and asynchrony between endometrium and the
embryo. More research is needed to determine PG threshold allowing
clinical decisions: cancel the cycle or freeze embryo and transfer in
another cycle.
P-226 Tuesday, October 20, 2015
CLINICAL OUTCOMES OF INTRAFALLOPIAN TRANSFER OF
CRYOPRESERVED-THAWED EMBRYOS IN A NATURAL MEN-
STRUAL CYCLE IN OLDER FEMALES. A. Tanaka, M. Nagayoshi,
I. Tanaka, S. Ikuma, T. Miki, T. Yamaguchi. Saint Mother Hospital, Kitakyusyu,
Japan.
OBJECTIVE: Assisted reproductive technology (ART) in females aged 40
years or older is characterized by low pregnancy and high miscarriage rates.
The impaired quality of the oocytes is thought to be the main cause, but the
environment of the in vitro culture may be another factor. Considering this,
we have carried out thawed embryos intrafallopian transfer (EIFT) in a natural
cycle to expect embryogenesis under the most natural environment. In
this study, we aimed to investigate whether intrafallopian transfer of thawed
embryos in a natural menstrual cycle would improve the implantation rate in
older females.
DESIGN: Retrospective cohort study of intrafallopian transfer of cryopreserved-thawed
embryos in a natural menstrual cycle in older females.
MATERIALS AND METHODS: The subjects were females aged 40 years
or older that had received in vitro fertilization (IVF) and embryo transfer
(ET), three or more times unsuccessfully, but had no passage obstruction
in fallopian tubes. When there were 3 or more antral follicles on day 3 of
the menstrual cycle, a GnRH agonist (the short method) was given. Meanwhile,
when there were 2 or less, the regimen of clomiphene citrate (SerofeneÒ)
plus recFSH every other day was employed. A 4 - 6-cell embryo 2
days after egg collection was frozen by the slow freezing method (1.8M
ethylene glycol + 0.2M sucrose + 20% fetal bovine serum-supplemented
D-PBS). In the group with a regular menstrual cycle, natural ovulation was
first confirmed. Then, the frozen embryo was thawed two days after the
ovulation and transferred into the fallopian tube laparoscopically under the
intravenous anesthesia with Propofol and Ketalar. In principle, the fallopian
tube on the ovulation side was selected for the transfer. For supporting luteal
function, a progesterone vaginal suppository (1000mg) was administered for
two weeks.
RESULTS: ET was carried out in 195 cycles from January 2010 to
November 2013. 30 females became pregnant (pregnancy rate per cycle,
15.4%) and 3 had a miscarriage (miscarriage rate, 10.0%). As a control,
the pregnancy rate in the group of over 40 years and miscarriage rate by intracytoplasmic
sperm injection and ETwere 13.0% (198/1527) and 60.6% (120/
198), respectively.
CONCLUSIONS: In older females cases when in vitro fertilization repeatedly
failed or had miscarriages, the results showed that intrafallopian transfer
of embryos in a natural menstrual cycle was useful.
P-227 Tuesday, October 20, 2015
EVALUATING OOCYTE YIELD, QUALITY & FERTILIZATION
FOLLOWING A CHANGE IN ANESTHESIA PROTOCOL. E. Li, a
B. M. Hanson, a J. Gordon, b M. DiMattina, b G. Celia, b M. Payson. b a Inova
Fairfax Hospital, Falls Church, VA; b Dominion Fertility, Arlington, VA.
OBJECTIVE: Compare oocyte yield, quality, and fertilization rates between
two anesthesia protocols (Demerol/Versed vs. Propofol) for transvaginal
oocyte retrieval in stimulated IVF (SIVF) and natural cycle IVF (NCIVF).
Previous studies have shown that Propofol accumulates in follicular fluid and
may impact fertilization rates (Piroli 2012, Christiaens 1999).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: SIVF and NCIVF patients at a private
fertility clinic who underwent transvaginal oocyte retrieval with Demerol/
Versed from 2012 to 2013 were compared to a similar cohort of patients
from 2013 to 2014 who had the same procedure using Propofol. Number
of oocytes retrieved, percent of MII oocytes, and fertilization rates were
compared.
RESULTS: 216 SIVF transvaginal oocyte retrievals were performed under
conscious sedation with Versed/Demerol. 2,677 total oocytes were retrieved
in this group. After a change in anesthesia protocol, 259 SIVF patients underwent
the same procedure with general anesthesia using Propofol, yielding
3,053 oocytes. No significant difference was noted in number of oocytes
retrieved per patient (12.39 in the Versed/Demerol group; 11.79 in the Propofol
group). A similar percentage of oocytes were noted to be in the MII stage
in both anesthesia groups (78.1% in the Versed/Demerol group; 79.2% in the
Propofol group). The number of oocytes transferred per patient was 1.7 in
both groups. Fertilization rates were similar in both protocols (74.6% in
the Versed/Demerol group; 77.1% in the Propofol group). 543 NCIVF patients
underwent transvaginal oocyte retrieval with Versed/Demerol. 488 total
oocytes were retrieved. 549 NCIVF patients underwent the same
procedure with Propofol, resulting in 477 oocytes. Similar to findings in
the SIVF cohort, no significant difference was noted in number of oocytes
retrieved per patient (0.90 in the Versed/Demerol group; 0.87 in the Propofol
group). A similar percentage of oocytes were noted to be in the MII stage in
both groups as well (82.2% in the Versed/Demerol group; 83.9% in the Propofol
group). Number of oocytes transferred per patient was 1.0 in both
groups. Fertilization rates were similar in both protocols (84.5% in the
Versed/Demerol group; 84.8% in the Propofol group).
FERTILITY & STERILITY Ò
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CONCLUSIONS: There was no significant difference observed in number
or quality of oocytes retrieved after the use of Versed/Demerol versus Propofol.
There was also no difference noted in fertilization rates. The use of Propofol
was not found to be detrimental to oocyte quality when compared to the
use of Versed/Demerol for retrieval anesthesia.
References:
1. Piroli A, Marci R, Marinangeli F, et al. Comparison of different anaesthetic
methodologies for sedation during in vitro fertilization procedures:
effects on patient physiology and oocyte competence. Gynecol
Endocrinol. 2012;28(10):796-9.
2. Christiaens F, Janssenswillen C, Verborgh C, et al. Propofol concentrations
in follicular fluid during general anaesthesia for transvaginal
oocyte retrieval. Hum Reprod. 1999;14(2):345-8.
P-228 Tuesday, October 20, 2015
ENDOMETRIAL BIOPSY PRIOR TO IVF STIMULATION SHOWS
IMPROVED RATES OF IMPLANTATION. H. Ahmed. Princeton
IVF, Lawrenceville, NJ.
OBJECTIVE: To evaluate the benefit of endometrial injury in an IVF cycle
and to study in a set patient population the implantation rates, biochemical
pregnancy rates, first trimester pregnancy loss, second/third trimester loss,
and live birth rates.
DESIGN: Retrospective data analysis.
MATERIALS AND METHODS: Local endometrial injury, by way of
endometrial biopsy prior to initiation of gonadotropins for controlled ovarian
hyperstimulation for IVF in 120 patients that were randomly selected from
234 patients.
RESULTS: Local endometrial injury in cycles with the transfer of either 2-
3 embryos (experimental group ¼ 120 and control group ¼ 114) resulted in a
significantly increased implantation rate (53.3% vs. 38.6%, p 0.025) and clinical
pregnancy rate (43.3% vs. 29.0%, p 0.022) compared to controls. There
was no statistical significance found in any of the negative outcomes which
included overall pregnancy loss, overall spontaneous miscarriage, biochemical
pregnancy, first trimester pregnancy loss, second/third trimester pregnancy
loss.
CONCLUSIONS: There is a clinically relevant improvement in implantation
and clinical pregnancy rates when local endometrial injury by way of
endometrial biopsy was performed prior to COH for IVF-ET, although this
did not result in a significant increase in live birth rates. There was also no
significant increase found in the rates of spontaneous miscarriage after local
injury to the endometrium by way of endometrial biopsy prior to COH for
IVF-ET. Further investigation is needed to determine what patients may
best benefit from this treatment modality.
P-229 Tuesday, October 20, 2015
IS LUTEAL PHASE SUPPORT FOR IUI CYCLES WITH GONADO-
TROPIN REALLY NECESSARY TO IMPROVE
OUTCOME? N. Karaca, a P. Ozcan, b E. E. Kovalak, c G. Batmaz, d
S. Ates. e a Bezmi Alem University Hospital, Istanbul, Turkey; b Yeditepe University
Hospital, Istanbul, Turkey; c Bagcilar Research and Training Hospital
Obstetrics, Istanbul, Turkey; d MD, _Istanbul, Turkey; e Bezmialem Vakif University,
Faculty of Medicine, Istanbul, Turkey.
OBJECTIVE: to evaluate the effectiveness of vaginal micronized progesterone
for luteal phase support in improving clinical PRs in IUI cycles with
OS using gonadotropin and to compare the results of progesterone supplementation
between recombinant follicle stimulating hormone (rFSH) and human
menopausal gonadotropin (hMG) for IUI cycles.
DESIGN: retrospective case-control study.
MATERIALS AND METHODS: A total of 579 women underwent OS
with gonadotropins who had admitted at IVF Center and Training and
Research Hospital between January 2011 and December 2014 were included
in this retrospective case-control study. Of 579 patients, 284 cycles receive
luteal phase support in the form of vaginal micronized progesterone capsules
(progesterone group), or not to receive luteal support (no progesterone
group).
RESULTS: Of IUI cycles, 132 (22,7%) resulted in a clinical pregnancy.
Between no progesterone group (n¼59 (20%) and progesterone group
(n¼73 (25.7%)) there was no significant difference in terms of clinical
PRs (p ¼ 0.055). Clinical PRs were similar to in women underwent OS
with rFSH or hMG (p ¼ 0.41 and 0,06, respectively) in progesterone group
and no progesterone group .
CONCLUSIONS: The results of study do not support to efficacy of routine
supplementation of the luteal phase with vaginal progesterone in IUI cycles
with OS to improve PRs. Further randomized trials are needed to evaluate if
luteal support is necessary in selected patients.
P-230 Tuesday, October 20, 2015
DOES FOLLICLE FLUSHING DURING OOCYTE RETRIEVAL
IMPROVE FERTILITY PRESERVATION CYCLE
OUTCOMES? V. Emirdar, a V. Turan, a F. Pacheco, a K. H. Oktay. a,b
a New York Medical College, Valhalla, NY; b Innovation Institute for Fertility
Preservation, New York, NY.
OBJECTIVE: Given that cancer patients are at risk for losing their ovarian
reserve post-chemotherapy, maximizing oocyte recovery is crucial in fertility
preservation (FP) cycles. Our aim was to determine the usefullness of follicular
flushing in maximization of oocyte recovery and embryo yield in women
undergoing controlled ovarian stimulation to preserve oocytes or embryos
with letrozole plus gonadotropins (LG) for FP.
DESIGN: Retrospective controlled cohort study of women with breast
cancer who underwent fertility preservation.
MATERIALS AND METHODS: Eleven cycles where oocytes were aspirated
using a double lumen flushing approach (DLF) were compared to 31
age-matched cycles where oocytes were retrieved via single lumen aspiration
(SLA) . All follicles that were visible by ultrasound, including those that are
10-mm on the pre-trigger ultrasound,
a similar number of follicles was detected (DLF vs. SLA: 16.21.2
vs. 12.51.3, p¼0.16). A significantly higher total number of oocytes (17.4
2.3 vs. 10.9 1.2, respectively; p ¼ 0.015) and a significantly higher number
of immature oocytes were recovered with DLF (8.36 1.12 vs. 3.53
0.45, p < 0.001). However the difference in the number of cryopreserved embryos
did not reach statistical significance (DLF vs. SLA: 7.2 1.94 vs. 5.2
0.79, p ¼ 0.279).
CONCLUSIONS: Utilization of DLF in FP cycles may facilitate the recovery
of immature oocytes from small follicles and increase the initial
oocyte yield. The value of DLF in enhancing final embryo yield should be
studied in larger prospective studies among women undergoing fertility preservation.
Cycle characteristics and fertility preservation outcomes.
DLF (n¼11) SLA (n¼31) P value
Age (years) 39.8 0.48 37.9 0.74 0.163
AMH (ng/ml) 1.6 0.37 1.4 0.25 0.635
Total FSH dose (IU) 5345 344 5017 108 0.236
Total Letrozole dose (mg) 59.09 2.84 57.90 1.08 0.633
Peak E2 level (pg/mL) 1022.46 205.45715.51 33.69 0.026
Number of follicles on 16.2 1.2 12.5 1.4 0.166
trigger day
No. of oocytes retrieved 17.4 2.34 10.9 1.26 0.015
No. of immature oocytes 8.36 1.12 3.53 0.45 p < 0.001
No. of embryos frozen 7.2 1.94 5.2 0.79 0.279
References:
1. Mok-Lin E, Brauer AA, Schattman G, Zaninovic N, Rosenwaks Z,
Spandorfer S.Follicular flushing and in vitro fertilization outcomes in
the poorest responders: a randomized controlled trial.Hum Reprod.
2013;28:2990-5.
e184 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

2. Levy G, Hill MJ, Ramirez CI, Correa L, Ryan ME, DeCherney AH,
Levens ED, Whitcomb BW.The use of follicle flushing during oocyte
retrieval in assisted reproductive technologies: a systematic review
and meta-analysis. Hum Reprod. 2012;27:2373-9.
P-231 Tuesday, October 20, 2015
CERVICAL MUCUS SCORE AT THE TIME OF INTRAUTERINE
INSEMINATION AND ITS ASSOCIATION WITH CONCEPTION
PROBABILITY. E. A. Evans-Hoeker, a,b A. Liberty, c A. Z. Steiner. b a Reproductive
Medicine and Fertility, Carilion Clinic, Virginia Tech Carilion
School of Medicine, Roanoke, VA; b Obstetrics and Gynecology, University
of North Carolina, Raleigh, NC; c University of North Carolina School of
Medicine, Chapel Hill, NC.
OBJECTIVE: Studies demonstrate that cervical mucus scores at time of
ovulation predict fecundability among couples attempting to conceive naturally.
It is unknown if this is due to the cervical mucus (CM) itself or associated
factors. The aims of this study were to determine 1) the prevalence of
fertile type CM, 2) factors associated with fertile type CM, and 3) whether
fertile type CM predicts pregnancy following intrauterine insemination (IUI).
DESIGN: Clinical cohort study.
MATERIALS AND METHODS: All clinical providers were trained on
CM scoring. Providers recorded the CM score at time of insemination. CM
was classified as absent, type 3 or type 4. Fertile type CM was defined as
types 3 and 4. This analysis includes all cycles with CM type documented
at the time of IUI performed at our academic center between 8/2012-7/
2013. Data was collected regarding patient demographics, medical history,
treatment types and outcome, and were analyzed using regression models
with a cluster term.
RESULTS: One hundred and seventy nine women contributed 321 IUI cycles
during the study time period. One hundred eighty cycles (56%) had
documented CM type and were included in our analysis. There were no differences
between cycles with and those without documented CM. Women
(n¼125) were typically Caucasian (76%), with an average age of 34.4 years,
body mass index (BMI) of 25.4 kg/m2 and a diagnosis of unexplained (29%)
or male factor (22%) infertility. A majority of cycles utilized oral ovulation
induction agents (73%) and ovulation prediction kit monitoring for IUI
timing (67%). Gonadotropins were used in 18% of cycles and 9% were
un-medicated. Fertile type CM was present in 91% of cycles. There were
no differences between fertile versus non-fertile type CM in regards to patient
age, race, BMI, AMH, number of follicles on midcycle ultrasound, endometrial
thickness, method used for IUI timing, fertility medications, or infertility
diagnosis. Cycle pregnancy rate did not differ between fertile type CM (17%)
and non-fertile type CM (19%), p¼0.75.
CONCLUSIONS: Most patients undergoing IUI demonstrate fertile type
cervical mucus on the day of IUI. Type of cervical mucus does not predict
probability of conceiving following IUI. Therefore, it is likely the cervical
mucus itself that facilitates fertilization during procreative intercourse.
P-232 Tuesday, October 20, 2015
TO WHAT DEGREE SHOULD THE ZONA PELLUCIDA BE CUT
OPEN IN ASSISTED HATCHING FOR BEST CLINICAL
RESULTS? H. Watanabe, R. Suzuki, M. Kobayashi, H. Hasegawa,
K. Tsukamoto, S. Saitou, J. Kobayashi. Kanagawa Ladies Clinic, Yokohama,
Japan.
OBJECTIVE: Assisted hatching is widely used to improve implantation
by opening or thinning of Zona Pellucida artificially. However, there is no
consensus to what degree the Zona Pellucida should be cut open for best clinical
results. We compared the size of the opening of the Zona Pellucida with
clinical results, using Saturn 5 ActiveTM (Research Instruments Ltd. UK) for
lazar assisted hatching (LAH) and EmbryoScopeBR(Vitrolife AB. Sweden)
for continuous observation.
DESIGN: prospective randmaized study.
MATERIALS AND METHODS: Cryopreserved blastocysts (SBB
grade) which were donated by patients for research were used for this
study. LAH was performed on warmed blastocysts after one hour of
culturing. The blastocysts were divided into 5 groups by the size of the
opening for assisted hatching: Group1 has been performed 50% opening
(n¼9) and Group2 has been performed 30% opening (n¼12). Group3 has
been conducted 12 micrometers opening (n¼12) so as Group4 has been
done the thinning (n¼12). For Group5 no LAH has been performed
(n¼12).We observed the start timing of hatching, time of hatching,
hatched rate and frequency of contraction until blastocysts finished hatching
by Time Lapse after LAH.
RESULTS: The time until the initiating of hatching was significantly
longer for groups 4 and 5 when compared to other groups. In fact, blastocysts
of groups 4 and 5 needed 1-3 times of contraction to start hatching as found
by Time Lapse observation. Time of hatching for Group 3 was significantly
slower than for other groups. In that group, embryos were stuck by the small
hole of the Zona Pellucida and repeated contraction occurred during hatching.
The hatched rates were significantly lower in groups 3, 4 and 5 when
compared to groups 1 and 2. Frequency of contraction until the blastocysts
finished hatching was higher for groups 3, 4 and 5. There was no significant
difference between groups 1 and 2.
CONCLUSIONS: From this study, we found that adequate LAH improve
subsequent hatching rates. In fact, the Zona Pellucida should not merely be
thinned by rather cut open, at least 30%.
P-233 Tuesday, October 20, 2015
ART - IN VITRO FERTILIZATION
NOVEL APPROACH FOR MANAGING DECREASED FERTILIZA-
TION WITH SEQUENTIAL ARTIFICIAL OOCYTE ACTIVATION:
PROSPECTIVE RANDOMIZED CLINICAL TRIAL. M. Fawzy, a
H. Abdelghaffar, b A. Alaboudy, a M. Sabry, b H. Kasem, a
M. Y. Abdel-Rahman, b M. Gad, a A. Metwallley, c E. Othman, d
A. Mahran, d S. M. Rasheed. b a Ibnsina IVF Center, Sohag, Egypt; b Sohag
Faculty of Medicine, Sohag, Egypt; c AlBarka IVF Center, Manama, Bahrain;
d Banon IVF Center, Assiut, Egypt.
OBJECTIVE: To compare outcomes between two different strategies of
artificial Oocyte activation: Calcium Ionophore activation protocol versus
a novel sequential protocol using Ca Ionophore followed by Strontium Chloride,
In cases of previously decreased fertilization ICSI cycles.
DESIGN: Prospective Randomized Clinical Trial.
MATERIALS AND METHODS: Patients with previous low fertilization
who presented to Ibnsina IVF Center, Sohag, Egypt (private center), between
January 2015 to March 2015 were recruited to undergo ICSI into
this study. Mature Oocytes (840) collected from 69 patients injected and
randomly assigned to two groups (420 each). Group I Oocytes activated artificially
for 15 min in ready made Ca Ionophore (GM508 Cult-Active, GY-
NEMED, Germany) immediately after ICSI followed by culture in
(GLOBAL TOTALÒ, LIFEGLOBALÒ, Canada). Group II Oocytes underwent
sequential activation using Ca ionophore (GM508 Cult-Active, GY-
NEMED, Germany) same protocol followed by another activation step
by washing and incubation for 60 min in Strontium Chloride (Sigma
69042). The Strontium Chloride Prepared in our laboratory at a concentration
of 10 micromol/ml in culture media (GLOBAL TOTALÒ, LIFEGLO-
BALÒ, Canada). The activated oocytes cultured traditionally according to
our protocol. All patients underwent Day 5 extended culture, and the embryos
selected had been randomized either from group I or II to be transferred
using closed envelope method.Outcome measures included:
Fertilization rate, top quality embryos day at 3, blastocyst formation rate
and finally pregnancy rate for each group. We compare the data of our
two group with the Chi-square test.
RESULTS: Study groups patients were similar regarding mean age, BMI,
the dose of FSH/HMG used, number of oocytes collected and number of cycle
days. There were a statistically significant increase in the rate of fertilization
in group II versus group I (group II 69% versus group I 46% P value <
0.01). We documented higher blastocyst formation rate in group II versus
group I (group II 59% compared to 38% group I P

assessment of clinical efficacy regarding multiple other clinical parameters
in a larger study in our center (Clinical Trial Registration:
NCT02418416).
P-234 Tuesday, October 20, 2015
DUAL TROPHECTODERM BIOPSY ON THE SAME BLASTOCYST
DOES NOT IMPAIR CLINICAL OUTCOMES. J. E. Swain, a,b
W. B. Schoolcraft, c M. Katz-Jaffe. c a Fertility Lab Sciences, Lone Tree,
CO; b Fertility Labs of Colorado, Lone Tree, CO; c Colorado Center for
Reproductive Medicine, Lone Tree, CO.
OBJECTIVE: Embryo biopsy has become common procedure for many
in vitro fertilization (IVF) laboratories. However, as one of the most invasive
techniques used, care must be taken to not compromise embryo
viability. Taking two blastomeres from cleavage stage embryos has been
shown to be detrimental compared to a single blastomere biopsy. Trophectoderm
(TE) biopsy has now become the approach of choice due to reduced
embryo damage as well as other advantages in analysis. Situations exist
where two TE biopsies may be warranted. However, it is unknown if a double
TE biopsy is detrimental to reproductive outcomes compared to a single
TE biopsy.
DESIGN: Retrospective data analysis.
MATERIALS AND METHODS: Per standard laboratory protocol for
patients undergoing simultaneous preimplantation genetic diagnosis
(PGD) for single gene defects coupled with comprehensive chromosome
screening (CCS), good quality blastocysts R3BB were subjected to two
TE biopsies (3-4 cells per biopsy). One biopsy was subjected to targeted
PGD for single gene defects, while the other biopsy was subjected to
WGA for CCS. Clinical outcomes were compared to age-matched patients
undergoing only CCS who had blastocysts subjected to one TE biopsy
(4-6 cells). All blastocysts were vitrified/warmed a single time prior
to transfer in a subsequent frozen cycle. This approach ensured similar
quality blastocysts were compared. Differences were determined using
Fisher’s Exact test, p

DESIGN: Prospective, randomized trial.
MATERIALS AND METHODS: From June 2014 to November 2014 data
was collected from 51 patients undergoing IVF treatment whose oocytes
were split and randomly allocated between sequential and 1-step mediums.
Fertilized oocytes were cultured up to day 6. A modified Gardner grading
system was used to score embryos. Good quality (GQ) blastocysts are those
with at least an ICM and TE of grade B. A total of 140 embryos (from 17 patients)
underwent a biopsy procedure on day 5/6 and aCGH testing. Statistical
analysis was performed using Fisher’s exact test, and the results were
considered significant if P

Table 1.
embryos
Y Chromosome
Day 3 biopsy
Trophectoderm
biopsy
Analysis
References:
1. Orzack et al,
2. Tarin et al, Reproductive
3. Maalouf et al,
P-240 Tuesday, October
TIME-LAPSE VIDEO
TED HATCHING
BLASTOCYST WARMING
BRYO’S ABILITY
C. Underberger, a
W. Chang, b C. Alexander,
Hills, CA; b Southern
OBJECTIVE: The
(ZP) hardening due
quence of vitrification.
ZP post-warming,
receptivity. We set
were able to fully escape
hatching (LAH) of
ing compared to embryos
DESIGN: Prospective
placed in an Embryoscope
MATERIALS AND
blastocysts vitrified
with a range of 23-46.
protocol on the same
ditions. Embryos were
ablated using LAH
All embryos were incubated
bator in GTL medium
the time it took to fully
embryo underwent
RESULTS: Assessment
hatched out. In Group
ing within 24 hours,
an average of 2.1 expansions.
complete hatching
expansions. Two of
at 37 and 48 hours with
plete hatching within
CONCLUSIONS:
tion regarding the
cantly higher percentage
the hatching process
that were not LAH.
completely escape
receptivity. The LAH
ergy to complete the
mized, thus ensuring
Conversely, an overwhelming
have LAH prior to embryo
process, even within
Overall, LAH of 33%
bryo’s escape and could
P-241 Tuesday, October
A TIME-LAPSE
FREEZING
EMBRYOS. M.
G. R. Navarrete, d Total Total
with without
Chromosome
1893 1966
1243 1291
_
www.pnas.org/cgi/doi/10.1073/pnas.1416546112,
Biology
F & S 101:
20, 2015
ANALYSIS
OF 33%
TO FULLY
D. L. Hill, J. Barritt.
California
human embryo
both to extended
This may
especially within
out to determine
the ZP
33% of the
that were
analysis
with
METHODS:
on Day 5
Warming
day, at the
randomized
on a Zilos laser
for
(Vitrolife).
hatch from
to escape the
was performed
1, 11 of 14
with an average
In
within 24 hours,
the Group 2
an average
24 hours was
Time lapse
hatching process
of warmed
when 33%
Importantly,
the ZP was within
embryos also
hatching process,
the energy
percentage
transfer
48 hours, which
of the ZP
significantly
20, 2015
EVALUATION
AND VITRIFICATION
Meintjes, a
B. Tilley, e embryos
Y Total embryo
gender ratio
0.95
0.96
Not significant
and Endocrinology
1321, 2014.
DEMONSTRATES
OF THE ZONA-PELLUCIDA
SIGNIFICANTLY
HATCH.
M. Surrey, ART
Reproductive
is known
culture to
affect the blastocysts
an optimal
if the percentage
post-warming
ZP was performed
not LAH.
of donated
and without LAH.
Overall,
and 6 were used.
of embryos was
same time, under
to either a group
at 400 ms (Group
48 hours in an
We recorded
the ZP and the
ZP.
to determine
embryos (78.6%)
of 17.0 hours
Group 2, none
after having
embryos (20%)
of 5 expansions.
significantly
video evidence
of blastocysts
embryos
of the ZP was
the average
the optimal
demonstrated
through
stores are not
of
are unsuccessful
is outside optimal
post-warming
increase implantation
OF CONVENTIONAL
S. Purcell,
K. L. Lee, f Euploid Euploid
XY XX gender
554 632
649 608
_ _
12:56,
LASER-ASSIS-
IMPROVES
K. Salmon, a D.
H. Danzer, b S.
Reproductive Center,
Center, Beverly Hills,
to undergo Zona-Pellucida
the blastocyst and
ability to
window of endometrial
of blastocysts
was different if laser
immediately after
vitrified/warmed
24 research good/fair
The mean age
performed per established
the same incubation
that had 33%
1) or no LAH
Embryoscope time
if the embryo escaped
number of expansions
if and when
achieved complete
elapsed to fully
of the 10 embryos
undergone an average
achieved complete
Group 1 vs. Group
different (p

sperm collected from cauda epididymis (C57BL6 inbred and B6D2F1
outbred) for 4-6h. Zygotes were cultured either in suboptimal Whitten’s
medium and 20% O2 (IVFWM) or in optimal KSOM medium with amino
acids and 5% O2 (IVFKAA) for 96h in 37 C. Control blastocysts were
flushed from the uterus of mated mice (FB mice). Blastocysts were transferred
to pseudo-pregnant recipients. Pups were reared to adulthood and peripheral
tissues were collected and serum corticosterone levels measured.
Total RNA and protein were isolated from fat, muscle and liver. mRNA
expression of GR and a selected group of GR-downstream target genes
were analyzed by qPCR; GR protein level by Western Blot. One-way AN-
OVA with post hoc correction was used as appropriate; p

RESULTS: The majority of PGS cases involve biopsy and cryopreservation
following an oocyte retrieval. In these patients we obtained a 68% implantation
rate and 62% ongoing pregnancy. The number of patients was
low and patients were younger in the re-frozen group, and these patients
had higher implantation (89%) and ongoing pregnancy rates (71%).
Table 1. Control and Re-frozen PGS FET results.
Group
Control
PGS
Re-frozen
PGS
N
Patients
Average
age
N
Embryos
Implantation
(%)
Ongoing
pregnancy (%)
78 36.1 98 68 62
7 34.7 9 89 71
CONCLUSIONS: Cryopreserved embryos that had not been previously biopsied
can be thawed, biopsied, and re-frozen effectively. This process does not
appear to have a detrimental effect on their subsequent post-thaw success. It is
not necessary transfer these embryos back immediately after thaw and biopsy,
which allows for greater flexibility to the patient and laboratory.
P-246 Tuesday, October 20, 2015
EVALUATION OF THE USEFULNESS OF REFROZEN-THAWED EM-
BRYO TRANSFER (R-FET). K. Takeuchi, Y. Homan, Y. Fukumoto,
Y. Kuroki, M. Tokudome, H. Setoyama, S. Awata, M. Takeuchi. Takeuchi
Ladies Clinic/Infertility Center, Aira-shi, Kagoshima, Japan.
OBJECTIVE: Multiple embryos are occasionally thawed for frozen
-thawed embryo transfer (FET). In the case that all the thawed embryos
are not used, the surplus embryos need to be refrozen. However, the number
of reports on R-FET is limited, and its usefulness is poorly understood. Due
to this, data on FET at our clinic was collected, and the clinical outcomes of
the embryo transfers that were frozen and thawed twice or three times, were
compared with those that were frozen and thawed once.
DESIGN: Retrospective study.
MATERIALS AND METHODS: The data for 590 frozen-thawed blastocyst
transfer cycles, performed from April 2012 to March 2014, was subjected
to assessment. To make a comparison, with regard to clinical outcomes, they
were classified into the following three groups: Embryos frozen once (A), embryos
frozen twice (B) and embryos frozen three times (C). There was no statistically
significant difference between these groups in terms of patients’ age,
the number of embryos transferred. Cryotip Safety Kit (Kitazato BioPharma
Co., Ltd.) was used for freezing/thawing of embryos.
RESULTS: The pregnancy rate was 45.2%(200/442) in A, 45.7%(64/140)
in B, and 37.5%(3/8) in C, while the abortion rate was 19.5%(39/200) in A,
18.8%(12/64) in B, and 0.0%(0/3) in C. There was no significant difference
between A and B. The pre-freeze embryo grade was compared with the postfreeze
embryo grade and the rate of decline in embryo grade was 19.2%(85/
442) in A, 21.4%(30/140) in B, and 12.5%(1/8) in C. There was no significant
intergroup difference. The pregnancy/abortion rates of the embryos that
showed elevation in grade were 45.7%(163/463) in A, 46.4%(51/110) in B
and 28.6%(2/7) in C, and 19.0%(13/163) in A, 17.6%(9/51) in B and
0.0%(0/2) in C respectively. The pregnancy/abortion rates of the embryos
that showed decline in grade were 43.5%(37/85) in A, 43.3%(13/30) in B
and 100.0%(1/1) in C, and 21.6%(8/37) in A, 23.1%(3/13) in B and
0.0%(0/1) in C respectively. In the assessed embryos that showed elevation
in grade and those that showed decline in grade, no significant difference
was seen between A and B in terms of the pregnancy/abortion rates. C was
cut from assessment because the total number of cycles was limited. However,
both the pregnancy and abortion rates obtained from C suggest the usefulness
of R-FET.
CONCLUSIONS: In this study, the pregnancy rate achieved by R-FETwas
comparable to that of pregnancy achieved by FET. Also no difference was
observed in terms of the abortion rate between the two treatments. These results
suggest the usefulness of R-FET.
P-247 Tuesday, October 20, 2015
UTILIZATION OF CRYOPRESERVED EMBRYOS FOR A SECOND
PREGNANCY. L. Kroener, a A. l. Akopians, a M. D. Pisarska, b
D. L. Hill, c J. Barritt, c E. T. Wang. b a Obstetrics and Gynecology, University
of California Los Angeles, Los Angeles, CA; b Cedars Sinai Medical Center,
Los Angeles, CA; c ART Reproductive Center, Beverly Hills, CA.
OBJECTIVE: As laboratory technology improves and elective single embryo
transfer rises, an increasing number of embryos are being cryopreserved.
However, there is little data on the utilization and fate of this
growing number of cryopreserved embryos. Our aim is to investigate the
use of cryopreserved embryos for a second pregnancy, as well as the overall
utilization and disposition of cryopreserved embryos on a per cycle basis.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: We reviewed 1,131 IVF cycles with at
least one embryo resulting in a clinical pregnancy and remaining cryopreserved
embryos between 1/2009 and 12/2012. Both autologous and donor oocyte cycles
were included. The primary outcome was use of embryos for a second
pregnancy over a mean follow-up time of 51.4 months. Disposition of cryopreserved
embryos was defined as continued cryopreservation, discarded, or
donated to research. Analysis was performed to assess the impact of age
(42) on utilization of cryopreserved embryos for
a second pregnancy, length of time between the first and second pregnancy
transfer, and disposition of cryopreserved embryos. Statistical analysis was performed
using the Chi-squared test and Kruskal Wallis test as appropriate.
RESULTS: A total of 3,994 embryos were cryopreserved from 1,131 IVF
cycles with at least one embryo transfer resulting in a clinical pregnancy. The
breakdown of cycles by age was 453 patients 42, with 46.8% and 94.3%
donor cycles in the 41-41 and >42 age groups respectively. Only 18.0% (204/
1,131) of patients attempted to use their remaining cryopreserved embryos for
a second pregnancy. There was no difference in the likelihood of returning for
a second pregnancy transfer across age groups (p¼0.136). There was an inverse
relationship between patient age and length of time between their first
and second pregnancy transfers (31.4 months for age 42, p¼0.001). There was
no impact of age on the disposition of embryos (p¼0.41). Of the 927 cycles
that patients have not come back to use their embryos for a second pregnancy,
12.9% chose to discard or donate their embryos (no difference across age
groups, p¼0.587) and 87.1% have continued cryopreservation.
CONCLUSIONS: Use of cryopreserved embryos for a second pregnancy
is lower than expected and is not influenced by age. Despite this low utilization
of embryos, the vast majority of patients continue to preserve rather than
discard or donate their embryos. This data provides evidence of an increasing
number of unused cryopreserved embryos and yields important insights for
counseling patients on use of cryopreserved embryo use for second pregnancy.
Studies with a longer follow-up time are needed to capture patients
coming back after more than 4 years.
P-248 Tuesday, October 20, 2015
DECREASED RATE OF EUPLOID EMBRYOS BY PREIMPLANTA-
TION GENETIC SCREENING IS ASSOCIATED WITH LOWER
LIVE BIRTH RATES. J. Lekovich, a P. Chung, a A. Lobel, a
N. Pereira, b J. Stewart, c Z. Rosenwaks. a a Weill Cornell Medical College,
New York, NY; b The Ronald O. Perelman and Claudia Cohen Center for
Reproductive Medicine, New York, NY; c Weill Cornell Medical Center,
New York, NY.
OBJECTIVE: To investigate whether the rate of euploid embryos by preimplantation
genetic screening (PGS) affects the outcome of pregnancy.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: In a one-year period, data of 102 pregnant
patients who underwent frozen embryo transfer (FET) of PGS-proven
euploid embryos were analyzed. They were divided into two groups based
on the cycle outcome: one with live births and the other whose cycle resulted
in a pregnancy loss (miscarriage or a biochemical pregnancy). Patients with a
history of recurrent miscarriages, thrombophilia, and uterine factor were not
included in the study. Main parameter of interest was to compare the rate (%)
of aneuploid embryos over total number of biopsied embryos (per patient)
between the two groups. Other confounding factors were also compared:
age, gravidity, parity, antimullerian hormone (AMH) levels, mid-cycle endometrial
thickness (ES), number of 2 pronuclear zygote (2PN) stage embryos,
the rate of progression to blastocyst by day 5, and the number of embryos
transferred.
RESULTS: Eighty one patients with live births were compared to 21 patients
who had a pregnancy loss (either a biochemical pregnancy or first
trimester miscarriage). The miscarriage group demonstrated significantly
lower AMH levels. All other confounding factors were comparable between
e190 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

Table 1
the two groups (Table 1). However, aneuploidy rate of all biopsied embryos
was significantly higher in the miscarriage group (0.62+0.28 vs. 0.49+0.28,
p¼0.03) (Table 1).
CONCLUSIONS: Despite the transfer of similar number of PGS-euploid
embryos and other confounding factors being controlled for, it appears that
the pregnancy loss is associated with decreased proportion of euploid embryos
among all embryos biopsied for PGS.
Supported by: Institutional.
P-249 Tuesday, October 20, 2015
Live birth
group
(81)
Miscarriage
group (21)
Age (years) 36.5 4.2 36 4.8 0.8
Gravidity 2 1.4 1.8 1.6 0.6
Parity 0.6 0.9 0.6 1 0.8
Mean endometrial stripe (mm) 9.7 þ 1.9 9.6 þ 2 0.7
AMH (ng/mL) 3.7 3.8 2.1 1.6 0.004
Mean # of 2PN 10.5 þ 6 9.6 þ 6.3 0.5
Rate of progression to 0.7 0.2 0.6 0.2 0.6
blastocyst stage by
day 5 (# day 5
blastocyst/# 2PN)
Mean # of embryos transferred 1.3 0.5 1.5 0.4 0.4
Aneuploidy rate
(#aneuploid embryos/total
# of biopsied day
5 embryos per patient)
0.49 0.25 0.62 0.28 0.03
EVALUATION OF EMBRYO IMPLANTATION POTENTIAL AFTER
VITRIFIED AND WARMED SINGLE BLASTOCYST
TRANSFER PER EMBRYO CATEGORIZATION USING THE
EEVAÔ TEST, AN AUTOMATED TIME-LAPSE QUANTITATIVE
ANALYSIS. S. De Gheselle, a B. Heindryckx, a P. De Sutter, b E. Van
den Abbeel. a a Department for Reproductive Medicine, University Hospital
Ghent, Ghent, Belgium; b Head Department for Reproductive Medicine, University
Hospital Ghent, Gent, Belgium.
OBJECTIVE: The Eeva Test provides a set of categories predicting embryo
developmental potential based on an automated analysis of early cell division
timings. The aim of this study was to evaluate the percentage of
embryos vitrified at the blastocyst stage and to compare the implantation
rate of the vitrified and warmed blastocysts in each Eeva Test category
(HIGH, MEDIUM, and LOW), in a group of supernumerary embryos after
single embryo transfer (SET) on day 3.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: This study included 76 patients who
underwent day 3 SET (Feb-Dec 2013). All embryos received Eeva Test
scores on day 3. Supernumerary embryos (N¼540) were cultured to the
blastocyst stage (day 5), and blastocysts exhibiting expansion status 1-2
and 3-5 with quality Inner cell mass (ICM) and Trophectoderm (TE)
grade A, B, C were vitrified (N¼209). Single vitrified and warmed blastocysts
were transferred and implantation potential was evaluated
(N¼95). The percentage of embryos vitrified and the implantation rate
of warmed blastocysts was calculated for each Eeva Test category and
compared.
RESULTS: Pregnancy rate for fresh day 3 SET was 31.6% . The percentage
of good and fair embryos on day 3 in each category was 53.4%
(78/146) for Eeva HIGH, 48.1% (39/81) for Eeva MEDIUM and 27.1%
(85/313) for Eeva LOW, respectively (p 0.05
re-expansion
Survival rate 58.1 63.2 33.3 > 0.05
Implantation rate 22.6 14 5.6 > 0.05
FERTILITY & STERILITY Ò
e191

P-251 Tuesday, October 20, 2015
CUMULATIVE PREGNANCY RATE BASED ON THE NUMBER OF
EMBRYO TRANSFERS IN ASSISTED REPRODUCTIVE
TECHNOLOGIES. Y. Mio, a K. Iwata, b K. Yumoto, b C. Mizoguchi, a
M. Sugishima, b M. Tsuneto, b Y. Iba. a a Reproductive Centre, Mio Fertility
Clinic, Yonago, Japan; b Mio Fertility Clinic, Yonago, Japan.
OBJECTIVE: The pregnancy rate (PR) of assisted reproductive technologies
(ART) was calculated as the number of pregnancies divided by the number
of embryo transfers (ET) cycles and/or patients treated within a specified
period. However, the population of these cycles and/or patients included
several unsuitable cases involving cancellation, failed oocyte pick-up
(OPU) or ET, and/or freeze-all cycles. Thus, it was difficult to evaluate the
actual success rate of ART programs in those couples having continued
ET. Considering that the goal of ART is to achieve successful pregnancy,
the most important issues on which to evaluate success are 1) the number
of ET required for successful pregnancy and 2) the cumulative pregnancy
rate (PR) with repeated ET. This study reviewed these data based on the number
of ET per couple.
DESIGN: Retrospective study based on the ART database in our Reproductive
Centre from January 2006 to December 2014.
MATERIALS AND METHODS: For the 2192 patients participating in
our ART programs, we reviewed the individual clinical outcomes of
ART, including the number of cycles required to achieve pregnancy, the cumulative
number of pregnancies and take-home babies, and the number of
ART cancelations. We then revised the data against each couple and their
age.
RESULTS: Of the 2192 study participants, 81.9% (n ¼ 1797) had ET and
73.8% (n ¼ 1327) achieved successful pregnancy, and of these, 79.1% (n ¼
1050) achieved successful pregnancy within three ET, 94.7% (n ¼ 1257)
within six ET, and 98.5% (n ¼ 1307) within nine ET. The number of patients
who experienced cancellation and no ETwere 317 and 395, respectively. The
revised PR [successful pregnancy (n ¼ 1327) divided by the revised population
(n ¼ 1480)] was 89.7%, and of these patients, 95.7% (201/210) were
aged under 30 years, 95.9% (523/547) 30-34 years, 88.1% (474/538) 35-39
years, and 71.9% (128/178) were aged 40-44 years. The number of takehome
babies in this period was 1171 (65.2%/ET and 88.2%/successful pregnancy).
CONCLUSIONS: The PR from individual ET cycles was limited and
declined dramatically with respect to advanced maternal age (32.6% in <
30 year-olds, 30.9% in 30-34 year-olds, 25.6% in 35-39 year-olds, and
12.1% in 40-44 year-olds). In contrast, the miscarriage rate increased markedly
depending on advanced maternal age (14.2% in < 30 year-olds, 17.8%
in 30-34 year-olds, 23.0% in 35-39 year-olds, and 38.8% in 40-44 year-olds).
This study of revised PR demonstrates clearly that ART provides a great opportunity
to achieve successful pregnancy by repeating the ET. To this end, it
is crucial to obtain a suitable number of cryopreserved embryos, for both
reducing the cost and the psychophysical burden of such techniques, and
increasing the expectation of successful pregnancy in participating ART patients.
P-252 Tuesday, October 20, 2015
DOES EGG VITRIFICATION DISPLACE THE FIRST POLAR
BODY - MEIOTIC SPINDLE AXIS? S. Phillips, a J. Graham, b
A. Coutinho, a M. Racicot, a C. Beauchamp. a a OVO Fertility, Montreal,
QC, Canada; b Shady Grove Fertility Centre, Rockville, MD.
OBJECTIVE: To assess whether the polar body-meiotic spindle axis is
affected by vitrification.
DESIGN: A prospective observational study carried out in one centre of a
donor egg bank programme.
MATERIALS AND METHODS: Vitrification, transport, storage and
warming are all carried out according to strict egg bank protocols and regularly
validated. We received 9 donor egg lots from 8 different clinics around
the US between November 2014 and January 2015. At the time of ICSI, all
oocytes were assessed for the presence and location of the meiotic spindle in
relation to the first polar body using a polscope. The degree of displacement
was noted. Embryo transfer was carried out on day 3 or day 5 according to the
number and quality of the embryos available. One embryo on day 3 or day 5
was transferred in all cases except one where two day 3 embryos were transferred.
Clinical pregnancy was confirmed by the presence of an intrauterine
fetal heart on ultrasound at 7 weeks.
RESULTS: A total of 65 MII eggs were warmed with a survival rate of
80%. The meiotic spindle was seen using a polscope in 84% of the eggs .
The spindle was within 45 of the first polar body in 34% of cases and was
more than 45 displaced in 50% of eggs. The fertilisation rate was 64%
with 7 cases reaching embryo transfer and 5 of these obtaining a clinical
pregnancy (71%).
CONCLUSIONS: In the literature, the spindle is visualised in approximately
80% of fresh oocytes (1) and this compares with the 80% visualisation
rate that we saw in cryopreserved-warmed oocytes. The proportion
of spindles from fresh oocytes which are within 45 of the first polar body
have been reported as approximately 70% (2,3). In our study it was noted
that only 34% of spindles were within 45 of the first polar body so half of
what would be expected from previous studies on fresh oocytes. It could
be suggested that the method of cumulus cell removal prior to ICSI can
have an effect on the displacement of the polar body, however in our
study oocytes came from 8 different centres all using their own technique
for denudation and displacement was seen from all centres. In addition,
the previous fresh oocyte studies all required cumulus cell removal prior
to polscope assessment. The process of oocyte vitrification causes the ooplasm
to shrink and re-expand within the zona pellucida which could
cause a mechanical displacement of the polar body as cryoprotectants
are introduced and then withdrawn. It has been shown that high doses
of FSH can affect the spindle (4) and it is therefore possible that strong
ovarian stimulation of egg donors attempting to maximize oocyte recovery
could have an impact. Unfortunately in these cases, spindle assessment
prior to vitrification was not possible. Further studies to eliminate
these variables are necessary to confirm that the process of vitrification
itself can impact on the placement of the meiotic spindle post warming
and therefore the use of a polscope when performing ICSI on these
eggs would be highly recommended.
References:
1. Rama Raju G. et al. Meiotic spindle and zona pellucida characteristics
as predictors of embryonic development: a preliminary study using Pol-
Scope imaging. RBMOnline. 2007. 14(2):166-74.
2. Rienzi L et al. Relationship between meiotic spindle location with regard
to the polar body position and oocyte developmental potential after
ICSI. Human Reproduction. 2003. 18(6):1289-93.
3. Moon JH et al. Visualization of the metaphase II meiotic spindle in
living human oocytes using the Polscope enables the prediction of embryonic
developmental competence after ICSI.Human Reproduction.
2003. 18(4):817-20.
4. Madaschi C et al. Zona pellucida birefringence score and meiotic spindle
visualization in relation to embryo development and ICSI outcomes.RBMOnline
2009. 18(5):681-6.
P-253 Tuesday, October 20, 2015
CRYOPRESERVED DONOR OOCYTES YIELD SIGNIFICANTLY
LOWER BIRTH RATES THAN FRESH OOCYTES. V. A. Kushnir, a
D. H. Barad, b D. F. Albertini, c S. K. Darmon, d N. Gleicher. b a Center for Human
Reproduction & Wake Forest Univer, New York, NY; b Center for Human
Reproduction & Foundation for Reproductive Medicine, New York,
NY; c Center for Human Reproduction & University of Kansas Medical Center,
New York, NY; d Center for Human Reproduction, New York, NY.
OBJECTIVE: To assess whether, as has been claimed, IVF live birth rates
(LBRs) in fresh (FDO) and thawed (TDO) donor oocyte cycles are similar.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: We reviewed aggregate national oocyte
donation data for 2013 in the annual Society for Assisted Reproductive
Technology (SART) outcome report for the U.S. IVF centers. LBRs per cycle
and per transfer in FDO and TDO were compared using the chi-square
test.
RESULTS: Of a total of 11,148 oocyte donation cycles, 2227 (20.0%)
involved use of TDO. Recipient LBRs, whether per cycle start (intent to treat)
or with reference point embryo transfer, were significantly higher for FDO
than TDO. Indeed, per cycle start there was a 6.4% absolute difference in
LBR, representing a 12.9% relative difference (P < 0.0001). This difference
further increased with reference point embryo transfer (absolute 9.0%; relative
16.0%; P < 0.0001). The cycle cancelation rate was higher for cycles utilizing
FDO than TDO (P < 0.0001) (Table).
e192 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

Donor oocytes
Fresh
Thawed
P-value
Number of cycles 8921 2227
Recipient starts resulting 4425/49.6 962/43.2

(53.8% vs. 22.5%; p¼ .027). In 19 cases, one or more contractions of the blastocyst
were observed but no relation with implantation was confirmed.
CONCLUSIONS: The analysis of warmed blastocysts by time lapse imaging
is providing new promising markers for blastocyst implantation potential
and represents a novel alternative to morphological evaluation, establishing
objective quantitative values linked with clinical outcome. However, further
studies are required to validate the predictive power of this technology.
P-257 Tuesday, October 20, 2015
LIPID PROFILE ANALYSIS FROM MICE EMBRYOS
VITRIFIED. A. A. de Melo, a J. Camillo, a T. D. Oleinki, a
F. B. Cordeiro, a E. G. Lo Turco, b T. G. Santos. c a Department of Surgery, Division
of Urology, Human Reproduction Section, S~ao Paulo Federal University,
S~ao Paulo, Brazil; b Head Embryologist, Sao Paulo, Brazil; c Student, Sao
Paulo, Brazil.
OBJECTIVE: This study aimed to identify potential lipid biomarkers
related to the embryo vitrification, since this procedure is extremely important
for assisted reproduction and may affect the development and the lipid
profile of embryos.
DESIGN: Prospective study.
MATERIALS AND METHODS: Mice females C57BL/6J were superovulated,
the embryos were collected after 60 hours in 8-cell stage and were
divided into two groups: the control group, in wich embryos were cultured
in 5% CO2 incubator until the blastocyst stage and evaluated based on their
development time, and the experimental group, composed of vitrified embryos,
that were thawed after 5 days, cultured in 5% CO2 until the blastocyst stage
and also evaluated based on their development time. The lipids were extracted
using the Bligh & Dyer method and were analyzed by Matrix-assisted laser
desorption/ionization (MALDI-TOF/MS). Statistical analysis was performed
using Principal Component Analysis (PCA) and partial least squares - discriminant
analysis (PLS-DA), and the 15 ions corresponding to the potential lipid
biomarkers were identified by the variable importance in projection (VIP).
RESULTS: There was a lower rate of vitrified blastocyst group of 75.60%
against 94.82% in the control group and variation in the rate at 24.40% of degenerated
embryos vitrified group and 5.18% in the control group and rate
variation in the degenerate embryos vitrified group of 24.40% and 5.18%
in the control group. The PCA demonstrated that the principal component
five better indicates the variance between the groups, presenting a prediction
model with 81% of accuracy. The experimental group presented ganglioside,
anthocyanins, phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol
as higher represented lipids and the experimental group presented flavonoids,
phosphatidylserine and anthocyanins hyper-represented.
CONCLUSIONS: There was a significant difference between the development
rate after vitrification and the lipid profile differs significantly between
embryos in the blastocyst stage and embryos subjected to cryopreservation.
The lipids found in this study are related to good embryo quality in the control
group and may present a broadly correlation with altered metabolism in
the experimental group, indicating that despite the vitrification is essential,
this is a procedure that affects the lipids embryo quality. This lipids are potential
biomarkers since they may help in the development of procedures that
may improve vitrification and reduce embryos damage.
Supported by: Capes.
type A (N¼567; 927 embryos), type B (N¼360; 575 embryos) and type C
(N¼121;196 embryos) were compared to fresh embryo transfers (fET) of
embryos type A (N¼667;1099 embryos), type B (N¼337; 568 embryos)
and type C (N¼84;135 embryos). Outcomes were compared by ANOVA
test or chi-square as appropriate. Logistic regression analysis including
age, own or donated cycles, fresh or vitrified oocytes and natural or HRT cycle
for endometrial preparation as potential confounders, was used to adjust
the Odds ratio (OR) for implantation.
RESULTS: Survival rate was not altered by embryo morphology in CT:
97.3% (96.3-98.3); 94.1% (92.2-97.1); and 95.9% (92.4-99.4) for type A,
B and C embryos respectively (NS). IR was higher for type A embryos in
both CT and fET and type B showed higher IR when compared to type C:
40.4% (36.9-43.9); 32.7% (28.9-36.5); 24.4% (18.1-28.3) and 41.1%
(38.5-44.3); 34.1% (30.2-38.0); 21.2% (14.4-28.1) for types A, B and C in
CT and fET respectively. IR was similar for each embryo category when
compared between CT vs fET (NS). Similarly CPR was comparable between
CTand fET groups (not shown). The OR for IR (0.932 (95% CI 0.789-1.090))
showed no impact of vitrification on implantation. Adjusted OR showed that
the only factor affecting IR was the embryo morphology (OR for IR (type A
with respect to type C)¼ 2.210 (1.669-2.926) and OR for IR (type B with
respect to type C)¼ 1.600 (1.194-2.143)).
CONCLUSIONS: Implantation potential is related to embryo quality
regardless whether they are fresh or vitrified, thus confirming the safety of
the vitrification procedure. This finding is of particular interest when evaluating
the prognosis of CTs especially when considering the ‘‘Freeze all’’ strategy.
P-259 Tuesday, October 20, 2015
THE EFFECT OF THE DAY OF CRYOPRESERVATION ON FET SUC-
CESS RATES IN PGS AND NON-PGS BLASTOCYSTS. G. Navarrete, a
S. Purcell, b B. Tilley, a S. J. Chantilis, a K. Lee, a M. Thomas, a R. Gada, a
M. Meintjes. c,a a Dallas-Fort Worth Fertility Associates, Dallas, TX; b Reproductive
Medicine and Surgery Center of Virgin, Charlottesville, VA; c Frisco
Institute for Reproductive Medicine, Frisco, TX.
OBJECTIVE: To determine if the day of blastocyst vitrification will have a
different impact on euploid blastocysts when compared with genetically unscreened
blastocysts.
DESIGN: A retrospective analysis.
MATERIALS AND METHODS: All biopsies and blastocyst transfers took
place over a 2-year period. Embryos were cultured in G-1/G-2 media, supplemented
with 10% SSS at 37.1 Cin6%CO 2 and 5% O 2 before vitrification on
day 5 (d5) and/or day 6 (d6). Vitrification and warming were performed with
Irvine’s vitrification/warming kit and a Rapid-I vitrification device. For PGS
patients, assisted hatching was done on day 3, followed by trophectoderm biopsy
on d5 and/or d6 and immediate vitrification. Standard 24-chromosome
aneuploidy screening was performed by genetic reference laboratories. Clinical
outcomes from mixed d5/d6 transfers were excluded from this study.
RESULTS: A total of 439 embryos were biopsied for the 179 patients evaluated
in this study. The proportion of euploid embryos was not different for
embryos biopsied on d5 (48%) compared with those biopsied on d6 (53%).
Similarly, an equivalent number (2.5%) of samples resulted in no DNA
amplification when comparing d5 and d6 biopsies. Degraded DNA was
observed in 5/203 d6 embryos but 0/236 d5 embryos.
P-258 Tuesday, October 20, 2015
IMPLANTATION POTENTIAL IS RELATED TO EMBRYO QUAL-
ITY REGARDLESS WHETHER THEY ARE FRESH OR
VITRIFIED. A. Cobo, a A. Coello, a M. Meseguer, b J. Remohi. c a IVI Valencia,
Valencia, Spain; b Clinical Embryology, Valencia, Spain; c Reproduction,
Valencia, Spain.
OBJECTIVE: Vitrification allows to the recovery of a high percentage of
fully intact embryos in which the pre-freeze morphological characteristics
are entirely preserved. The remaining question is if the ability to implant after
vitrification also remains unaltered. This study was aimed to evaluate the
impact of vitrification of day-3 embryos according to their morphological parameters
on survival implantation (IR) and clinical pregnancy (CPR).
DESIGN: Retrospective analysis.
MATERIALS AND METHODS: Embryo grading was addressed in accordance
with the Istanbul consensus on embryo assessment. All embryo transfers
(ET) in which the embryos belonged to the same morphological category
were selected from January 2012 to October 2013. ETs of mixed morphological
categories were excluded. Results after cryotransfers (CT) of embryos
Clinical Outcomes for Day of Cryopreservation With or Without PGS.
Treatment
Group
Number
of Patients
Implantation
Rate SEM
Ongoing Pregnancy
Rate SEM
Day-5 PGS 27 79.6 a 9.3 70.4 a 8.9
Day-6 PGS 20 67.5 a 8.9 65.0 a 9.4
Day-5 no PGS 73 62.3 A 6.1 63.0 A 5.7
Day-6 no PGS 59 47.4 B 5.4 50.8 A 5.9
CONCLUSIONS: Previous studies have consistently demonstrated a significant
increase (10-15%) in FET success when transferring non-PGS blastocysts
vitrified on d5 compared with blastocysts vitrified on d6. Similarly,
the implantation rate (IR) in this study was significantly higher for blastocysts
vitrified on d5 compared with blastocysts vitrified on d6. This difference
in IR between d5 and d6 frozen blastocysts was not observed for
confirmed euploid embryos. The equivalent aneuploidy rates for d5 and
e194 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

d6 biopsied blastocysts suggest that the difference in IR seen for non-PGS
blastocysts is not likely due to an increase in the aneuploidy rate of d6 embryos.
However, blastocysts vitrified without PGS is typically selected
based on a minimum morphology grade; whereas, blastocysts biopsied
for PGS are vitrified with much less emphasis on standard morphology.
The aneuploidy rate of d5 and d6 blastocysts that met minimum
morphology criteria (non-PGS blastocysts) may not necessarily be the
same as those of PGS blastocysts when less attention is paid to standard
morphology. Further studies are needed to confirm that there are also no
difference in aneuploidy rates of d5 and d6 blastocysts that meet a minimum
morphology score.
P-260 Tuesday, October 20, 2015
COMPARISON OF VITRIFICATION MEDIUM WITH TREHALOSE
AND HYDROXYPROPYLCELLULOSE TO THAT WITH SUCROSE
AND COMPLEX PROTEIN SUPPLEMENTATION. T. Schlenker, a
S. McCormick, a R. Smith, a W. B. Schoolcraft, b R. L. Krisher. c a Fertility
Labs of Colorado, Lone Tree, CO; b Colorado Center for Reproductive Medicine,
Lone Tree, CO; c National Foundation for Fertility Research, Lone
Tree, CO.
OBJECTIVE: Human oocytes have historically been difficult to cryopreserve.
Optimizing vitrification solutions is one method to improve outcomes.
Our objective was to compare the efficacy of two solutions for the vitrification
of donor eggs, one containing trehalose and hydroxypropylcellulose
(HPC) and the other with standard sucrose and complex protein supplementation.
DESIGN: Prospective randomized donor egg split.
MATERIALS AND METHODS: Eggs from each donor (n¼33) were
randomly assigned to vitrification using either Cryotech (CT; n¼244) or an
In House prepared (IH; n¼260) medium. IH vitrification solution consisted
of standard Tissue Culture Medium 199 (TCM199) with HEPES, bicarbonate,
serum protein substitute (SPS; Origio), sucrose, DMSO and ethylene glycol
(EG). Cryotech vitrification solution also contains DMSO and EG, but
substitutes trehalose for sucrose and HPC rather than SPS. Eggs were vitrified
using the CryoTop (Kitazato).
RESULTS: There was no difference in egg survival post-warm (CT,
95.5%; IH, 96.2%). Fertilization was significantly higher for eggs vitrified
and warmed in IH solutions (2PN/eggs survived, 85.2%) compared to those
in CT (74.2%). There were no differences between CT and IH in the percentage
of good quality blastocysts (GQ, R3BB) produced per 2PN (CT,
54.3%; IH, 51.3%) or per total eggs vitrified/warmed (CT, 41.8%; IH,
44.0%), or GQ blastocysts produced on D5 per 2PN (CT, 29.3%; IH,
29.0%). Pregnancy data was analyzed from those cases in which only embryos
produced from a single treatment were transferred (CT, n¼10, 1.3
embryos/ET; IH, n¼14, 1.4 embryos/ET). There were no significant differences
between % positive hCG (CT, 80%; IH, 64.3%) or % implantation
(CT, 84.6%; IH, 52.6%).
CONCLUSIONS: Both commercially available (Cryotech) and In House
prepared vitrification media are equally effective for egg vitrification, despite
differences in non-permeating cryoprotectants and protein content. Although
IH solutions resulted in higher fertilization success, the proportion of blastocysts
produced per fertilized zygote and per vitrified/warmed egg was equivalent.
In summary, trehalose with HPC or sucrose with complex protein can
both be used for egg vitrification, resulting in successful embryo development
and pregnancy.
P-261 Tuesday, October 20, 2015
INFLUENCE OF SEMINAL QUALITY IN DONOR EGG IVF PRO-
GRAM USING VITRIFIED OOCYTES. B. Barros, a
T. S. Domingues, b A. S. Belo, a R. Mazetto, b A. P. Aquino, b
E. L. Motta. c a Embryology, Huntington Medicina Reprodutiva, Sao Paulo,
Brazil; b Huntington Medicina Reprodutiva, Sao Paulo, Brazil; c Medical,
Huntington Medicina Reprodutiva, Sao Paulo, Brazil.
OBJECTIVE: To evaluate the influence of different seminal parameters to
fertilize donor banking oocytes and correlate to blastocyst development, implantation
and pregnancy rates.
DESIGN: Retrospective study.
MATERIALS AND METHODS: From July 2013 to April 2015, 320 cycles
of donor banking oocytes were fertilized by ICSI with ejaculated sperm and split
into three different groups: G1: Normozoopermia (n¼77); G2: Oligoteratozoospermia
(n¼34); G3: Teratozoospermia (n¼209). All oocytes were vitrified using
open system and warmed following standard protocols. After fertilization,
embryos were cultured as routine and blastocyst transfer placed on days 5 or
6. Endometrium preparation was performed with 4 mg of estradiol valerate
plus 800mg of micronized progesterone according to standard protocols.
RESULTS: A total of 2846 vitrified donor oocytes were used. The mean
numbers for sperm concentration were respectively (x106/mL): (107.4 vs 8.0
vs 64.8, p

Technology. Role of assisted hatching in in vitro fertilization: a guideline.
Fertil Steril. 2014 Aug;102(2):348-51. http://dx.doi.org/10.1016/
j.fertnstert.2014.05.034. Epub 2014 Jun 18.
3. Sunkara SK1, Siozos A, et al.The influence of delayed blastocyst formation
on the outcome of frozen-thawed blastocyst transfer: a systematic
review and meta-analysis. Hum Reprod. 2010 Aug;25(8):1906-15.
P-263 Tuesday, October 20, 2015
SINGLE EMBRYO TRANSFER OF FROZEN-THAWED EMBRYOS
IS ASSOCIATED WITH INCREASED MATERNAL
COMPLICATIONS. T. Shavit, a G. Oron, b T. Tulandi, c W. Son, b
H. Holzer, b W. Buckett. c a McGill University- MUHC Reproductive Center,
Montreal, QC, Canada; b Department of Obstetrics and Gynecology, McGill
University, Montreal, QC, Canada; c McGill University, Montreal, QC, Canada.
OBJECTIVE: Cryopreservation of embryos allows transfer of a single embryo
(SET) and storage of supernumerary embryos maximizing the cumulative
pregnancy rates. It has been reported that IVF conceived singletons are
prone to pregnancy complications including low birth weight (LBW), preterm
deliveries (PTD) and small for gestational age (SGA). The purpose of
our study was to compare the pregnancy outcome in singletons born after
fresh or frozen-thawed single blastocyst transfer (SBT).
DESIGN: A single center retrospective cohort study, a reproductive unit of
a tertiary university health center.
MATERIALS AND METHODS: We compared singleton live births resulting
from transfer of fresh or frozen-thawed single blastocyst embryo (SBT).
The primary outcomes were perinatal outcomes including SGA, LBW, very
LBW, PTD, early PTD, large for gestational age (LGA), hospitalization at
the neonatal intensive care unit, respiratory and gastrointestinal complications
and congenital malformations. Maternal complications included preeclampsia,
placenta previa, placental abruption, gestational diabetes mellitus (GDM) and
chorioamnionitis. Adjustment for confounding factors was done.
RESULTS: We studied 1886 fresh-SBTand 1200 FET-SBT cycles. SBTof
fresh embryo resulted in a clinical pregnancy rate of 52.2% and live birth rate
of 31.3% per embryo transfer (ET). These were significantly higher than
34.4% clinical pregnancy rate and 13.7% live birth rate per ET in the FET
group (p

OBJECTIVE: No significant differences in outcomes have been found using
various protocols of endometrial preparation for frozen embryo transfer
(FET), however existing data have not accounted for infertility diagnoses.
This study compares clinical outcomes of women with different diagnoses
in FET cycles using leuprolide acetate (L) suppression versus use of a
mid-cycle antagonist (Ant), both with estrogen/progesterone supplementation.
We hypothesized that prior L suppression versus mid-cycle Ant
blockade may benefit women with polycystic ovarian syndrome (PCOS) or
endometriosis.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: FET cycles performed at a single academic
center from January 2012 to December 2014 were reviewed; previous
FET cycles of identified women were also included. Patient demographics,
infertility diagnosis, in-vitro fertilization and FET cycle information were
collected. Characteristics and clinical outcomes of L versus Ant FET cycles
were compared. We controlled for patient age, use of donor oocytes, BMI,
use of ICSI, day of transfer and number of embryos transferred.
RESULTS: FET cycles (n¼872) were identified and detailed data was
available in 670 cycles of which L (n¼491) and Ant (n¼179) approaches
were prescribed. Further patient follow up and data collection continues to
date. Patients undergoing Ant cycles were older (406.7 vs. 374.9 years,
p

P-269 Tuesday, October 20, 2015
OOCYTE BIOLOGY
SIMILAR LEVELS OF PREMATURE SEPARATION OF SISTER
CHROMATIDS IN THE MOUSE AND HUMAN OOCYTE RE-
VEALED BY NEXT GENERATION SEQUENCING. N. R. Treff, a
R. L. Krisher, b X. Tao, c H. Garnsey, c C. Bohrer, c E. Silva, b J. Landis, c
R. T. Scott, a T. K. Woodruff, d F. E. Duncan. e a RMA, NJ, NJ; b National Foundation
for Fertility Research, LoneTree, CO; c FAEEC, Basking Ridge, NJ;
d Northwestern University, Chicago, IL; e University of Kansas Medical Center,
Kansas City, KS.
OBJECTIVE: Accumulating evidence supports the relevance of the mouse
model to human age-related oocyte aneuploidy. Still, many questions remain
including whether there is chromosome-specific susceptibility and a higher
incidence of premature separation of sister chromatids (PSSC) compared
to classical nondisjunction (ND) in the mouse as in the human. The present
study develops a method to address these and other ongoing questions associated
with the etiology of aneuploidy.
DESIGN: Observational.
MATERIALS AND METHODS: GenomePlex WGA4 (Sigma) and next
generation sequencing (NGS) based (Ion Proton) comprehensive chromosome
screening (CCS) was performed on known trisomy 16 single mouse
embryonic fibroblasts (positive controls), and paired 1st polar body (PB)
and remaining oocytes (to evaluate reciprocal aneuploidies) and whole
blastocysts from aged mice.
RESULTS: Trisomy 16 was clearly observed with NGS CCS in all positive
control (fibroblast) samples tested. 40 paired PB and oocyte samples from
mice aged 15-19 months displayed an aneuploidy incidence of 15% (chromosomes
2, 5, 7, 9, 11, 12, and 15), and PSSC in 5 of the 6 cases (83%). Reciprocal
errors were observed in the paired PBs and oocytes in all cases. 10 of 30
blastocysts (33%) from mice aged 13-14 months displayed aneuploidy
involving chromosomes 1, 2, 3, 5, 8, 10, 12, 13, 14, 17, and 19.
CONCLUSIONS: This study confirms the validity of a novel method for
aneuploidy screening in mouse polar bodies, oocytes, and embryos based
on detection of trisomy 16 in a positive control and reciprocal errors in sister
PBs and oocytes. It was also possible to distinguish PSSC from ND, which
indicated percentages similar to those observed in human oocytes. Increased
prevalence of aneuploidy in the blastocysts in this study may be related to
additional contribution of meiosis II errors or strain specific differences in
susceptibility. This model system and NGS CCS method of analysis represents
an important new tool to better understand the etiology of and factors
that may influence aneuploidy.
P-270 Tuesday, October 20, 2015
AN FSH-LOWERING ACTIVIN DISRUPTING THERAPY IN-
CREASES YIELD OF HEALTHY OOCYTES IN A MOUSE MODEL
OF FEMALE MIDLIFE REPRODUCTIVE
AGING. L. R. Bernstein, a,b,c,d A. Mackenzie, e C. L. Chaffin, f
I. J. Merchenthaler. g a Pregmama, LLC, Montgomery Village, MD; b Epidemiology
and Public Health, University of Maryland School of Medicine,
Baltimore, MD; c Gynecology and Obstetrics, Johns Hopkins School of Medicine,
Baltimore, MD; d Veterinary Integrative BioSciences, Texas A & M
College of Veterinary Medicine, College Station, TX; e University of Maryland,
Baltimore, Baltimore, MD; f University of Maryland School of Medicine,
Baltimore, MD; g Epidemiology, Baltimore, MD.
OBJECTIVE: Women with diminished ovarian reserve often exhibit poor
response to ovarian stimulation when given FSH stimulation. Alternative
treatments that stimulate oocyte production may reduce cycle cancellation
and improve pregnancy rates.
DESIGN: We developed SAMP8 mice as model with human-like reproductive
aging attributes, including diminished oocyte yield, elevated FSH,
increased rates of oocyte spindle misalignments, and diminished fertility
by midlife (age 7 months). ActRIIB:Fc is an activin decoy receptor that sequesters
activin and suppresses activin signaling. It was given to midlife
SAMP8 for 3 weeks, the duration of oocyte growth. A second test group
was treated for 3 weeks with high FSH activity in the form of PMSG.
MATERIALS AND METHODS: Freshly ovulated oocytes were collected
after hCG injection. Total yields of oocytes and of viable oocytes were scored.
RESULTS: ActRIIB:Fc lowered FSH in midlife SAMP8 to the levels of
young SAMP8. ActRIIB:Fc increased oocyte yield by 3.7/mouse, from
10.64 to 14.3/mouse (P¼0.0031), an increase of 34.7%. The yield was
greater than for young SAMP8. ActRIIB:Fc did not affect the yield of
non-viable oocytes. ActRIIB:Fc increased the yield of viable oocytes by
2.9/mouse, from 9.86 to 12.7 oocytes/mouse (P¼0.0051), an increase of
29.4%, a number greater than that of young mice.PMSG had no effect on
the total yield of oocytes in midlife SAMP8 (10.64 vs. 13.28/mouse;
P¼0.9829, NS), though it increased the total yield in young SAMP8 by 7.4
oocytes, from 12.1 to 19.5/mouse (P¼0.0063). This is analogous to DOR
AMA women, who are often more refractory to FSH stimulation than their
younger peers. PMSG caused a redistribution of oocytes from 9.86/10.64
(92.7%) viable oocytes in untreated mice, to just 4.24/13.28 viable oocytes
(31.9%; P 0.05 in bivariate analysis). Moreover,
there was no significant correlation between blood and oocyte telomere length
from the same patient (R2 < 0.01). No difference was detected in mean telomere
length from blood nor oocytes with pregnancy outcomes.
CONCLUSIONS: Telomere DNA content from single eggs was associated
with ovarian function independent of age, suggesting telomere DNA may be
a useful biomarker of reproductive age. This study was limited by the use of
spare, immature oocytes, and pregnancy outcomes could not be directly
e198 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

elated to telomere length from the transferred embryos. This is the first study
to compare telomere length in human oocytes with somatic tissues. Somatic
tissues from the same individual exhibit a consistent decline in telomere
length with age, but we did not find such an association between oocytes
and leukocyte telomere DNA. Like sperm, oocytes likely represent a unique
niche in telomere biology.
P-272 Tuesday, October 20, 2015
TELOMERE ATTRITION IN GERMINAL VESICLE ARRESTED
HUMAN OOCYTES. K. Kalmbach D. L. Keefe. New York University
Langone Medical Center, New York, NY.
OBJECTIVE: Telomere length is maintained in the ‘immortal’ germline
yet evidence from mouse and human studies suggests that telomere length
in oocytes is significantly shorter than in somatic tissues. We have proposed
that telomere attrition in oocytes contributes to infertility in women, but the
mechanisms underlying the effects of telomere length on reproduction in
women are poorly understood. Telomere shortening arrests mitotic cells so
we examined its effects during meiosis.
DESIGN: Prospective observational study.
MATERIALS AND METHODS: Human germinal vesicle oocytes (GVs)
were collected from patients at an academic fertility center and in-vitro
matured for up to 48 hours following oocyte retrieval. Samples were assessed
for polar body (PB) extrusion and germinal vesicle breakdown (GVBD) at 24
and 48 hours, then fixed for telomere length analysis. If no PB was observed
at 48 hours, the sample was processed and staged as meiosis I (M1) or GV by
absence or presence of GV nuclear structure respectively. Oocytes were
denuded, treated with pronase and washed to exclude granulosa cells. One
oocyte was delivered to each tube assuring no contamination with follicular
cells. Telomere length was measured by single cell telomere pre-PCR amplification
qPCR (SCT-pqPCR) and reported as telomere DNA (T) normalized
to reference DNA (R) in a T/R ratio. T/R measures were log-transformed for
statistical analyses to obtain a normal distribution.
RESULTS: Mean telomere length of GV arrested oocytes (n ¼ 35; 0.09
0.44) was significantly less than that of oocytes that had matured to metaphase
II (n ¼ 74; 0.50 0.54) oocytes (p < 0.001, paired t-test). Averaged
per patient, mean telomere length of GV (n ¼ 23; 0.00 0.43) and M2 (n
¼ 32; 0.50 0.43) also differed significantly (p < 0.001). Telomere length
of oocytes that had reached M1 by 48 hours was similar to that of M2 oocytes
(n ¼ 9; 0.28 0.68; p ¼0.38).
CONCLUSIONS: Arrested germinal vesicle oocytes exhibited less telomere
DNA content than MI or MII oocytes. Telomere length influences progression
through meiotic cell cycle, and this pilot study suggests oocyte
telomere length may be an important regulator of oocyte maturation. Future
studies will determine whether telomeres regulate mitotic and meiotic cell
cycle via common regulatory pathways.
P-273 Tuesday, October 20, 2015
OOCYTE SECRETION IS REGULATED BY SOMATIC CELLS DUR-
ING OOCYTE MATURATION AND MEDIATES CUMULUS CELL
FUNCTION. H. Cakmak, F. Franciosi, M. Cedars, M. Conti. UCSF
Dept. of Ob/Gyn, San Francisco, CA.
OBJECTIVE: De novo mRNA synthesis ceases during the final stages of
oocyte maturation and previously synthesized mRNAs are translated according
to a well-orchestrated program of recruitment to the polysomes. The aim
of this study is to investigate the regulation of oocyte mRNA translation and
protein secretion during maturation, using interleukin (IL)-7 as the prototypic
secreted factor, and to determine the role of this oocyte secreted factor
in cumulus cell (CC) function.
DESIGN: Prospective.
MATERIALS AND METHODS: Studies involving in vivo/in vitro maturation
and fertilization of oocytes from 21 day old C57BL/6 mice, and human
follicular fluid before and after hCG administration.
RESULTS: Microarray analysis of oocyte polysomal fraction showed that
191 of 7600 polysome-bound mRNAs encoded for secreted proteins. Specifically,
72 transcripts were constitutively present in the polysome fraction
throughout in vivo oocyte maturation, while 80 were decreased and 39 were
increased. IL-7 was selected as the prototype because its translation increased
the most during oocyte maturation. In qPCR analysis, the level of polysomebound
IL-7 mRNA enhanced in MI oocyte compared to GVoocyte, and further
increased with maturation to MII. Total IL-7 mRNA levels did not change during
oocyte maturation. Renilla luciferase reporter under the control of IL-7
3’UTR was injected into cumulus enclosed oocytes (CEOs). Translation of
the IL-7 reporter significantly increased as the oocytes progressed from GV
to MII, and was further stimulated by amphiregulin (AREG), a somatic cellderived
EGF-like growth factor that accumulates in the follicle after LH surge.
AREG-induced effect was not detected when the oocytes were denuded prior
to stimulation. IL-7 protein secretion increased with oocyte maturation in
CEOs and, to a less extent in denuded oocytes (DOs). AREG further enhanced
IL-7 secretion in CEOs, but not in DOs. IL-7 was not detectable in spent media
of CC only cultures. In humans, IL-7 levels were significantly higher in the
follicular fluid containing MII oocyte (n¼42) compared to those containing
GV oocyte (n¼8). IL-7 levels positively correlated with AREG concentration
in the follicular fluid. After fertilization, IL-7 secretion diminished in both
mouse and human embryos. In mouse, IL-7 receptor mRNA expression in
CCs increased during oocyte maturation. IL-7 increased the proliferation of
CCs, but did not affect CC expansion.
CONCLUSIONS: Oocyte secretion during maturation is highly dynamic
and regulated mRNA translation is the molecular mechanism underlying
this timed secretion. The oocyte secretion is sensitive to somatic cues and
modulates CC function supporting the concept that these regulated secretions
are the part of cross-talk between the oocyte and surrounding CCs, which is
crucial for the oocyte developmental competence.
Supported by: RO1-GM097165 and T32 HD007263-28.
P-274 Tuesday, October 20, 2015
IQGAP3 IS A CANDIDATE PROTEIN FOR CORRECT SPINDLE
POSITIONING DURING MEIOTIC MATURATION IN MAMMA-
LIAN OOCYTES. L. A. McGinnis J. P. Evans. Biochemistry and Molecular
Biology, Johns Hopkins Bloomberg School of Public Health, Baltimore,
MD.
OBJECTIVE: The progression of the mammalian oocyte through its two
meiotic divisions is accompanied by organization and orientation of the
meiotic spindle, as well as remodeling of the actin-rich cortex. These events
are crucial for oocyte quality because spindle positioning is key for equal
segregation of chromosomes and unequal distribution of cytoplasm to
daughter cells.
DESIGN: IQ motif-containing GTPase activating protein 3 (IQGAP3)
is a protein of interest because it is implicated in cytokinesis and contractile
ring organization in model organisms. The domain structure of IQ-
GAP3 includes an actin-binding domain at the C-terminus and a
microtubule-binding domain at the N-terminus, leading us to hypothesize
that IQGAP3 may serve as a scaffold between the microtubule-based
meiotic spindle and the actin-rich cortex. Mammals have three IQGAPs.
Transcriptome databases suggest that Iqgap3 mRNA is enriched in mouse
oocytes relative to other tissues, prompting us to examine IQGAP3 function
in oocytes.
MATERIALS AND METHODS: Mouse oocyte collection and in vitro
maturation: Prophase I oocytes are collected from ovaries of 6-8-week-old
CF1 female mice and cultured in medium containing dibutyryl-cAMP (dbcAMP)
to maintain meiotic arrest. Oocytes are then allowed to mature to
met I or to met II. Microinjection for RNAi-mediated knockdown: Prophase
I oocytes are microinjected with Iqgap3-targeting siRNA. Oocytes are
cultured in dbcAMP to maintain meiotic arrest for 48 hours to allow sufficient
time for RNA degradation and protein turnover. Knockdown is assessed
by RT-PCR, immunofluorescence or immunoblotting. Immunofluorescence:
The zona pellucida is removed through a brief exposure to acidic medium.
Oocytes and eggs are fixed in paraformaldehyde and stained using anti-IQ-
GAP3 and anti-tubulin antibodies.
RESULTS: IQGAP3 is localized to the cortex of prophase I oocytes, and
associated with the spindle in met II eggs. IQGAP3-deficient oocytes have
a range of phenotypes including failure to recruit the metaphase I spindle to
the cortex. This phenotype supports our hypothesis that IQGAP3 plays a
scaffolding role between the spindle and the cortex such that in an IQ-
GAP3-deficient oocyte, correct positioning of the spindle adjacent to the
cortex does not occur. Studies in HeLa cells show that IQGAP3 interacts
with anillin and this interaction is required for the localization of IQGAP3
to the contractile ring. Ongoing experiments are investigating anillin in IQ-
GAP3-deficient oocytes.
CONCLUSIONS: It is crucial that the meiotic spindle is positioned
correctly through the two rounds of cytokinesis to ensure equal segregation
of chromosomes and unequal segregation of cytoplasmic stores between
the egg and polar body. Our work shows that IQGAP3 plays an important
role in spindle positioning.
Supported by: NIH R03 HD074773 to JPE.
FERTILITY & STERILITY Ò
e199

P-275 Tuesday, October 20, 2015
NON-INVASIVE PREDICTION OF BLASTOCYST FORMATION BY
DAY THREE EMBRYO CULTURE MEDIUM MASS SPECTROM-
ETRY LIPID FINGERPRINTING. D. P. Braga, a,b,c A. S. Setti, a,b,d
E. C. Cabral, e M. N. Eberlin, e E. G. Lo Turco, c A. Iaconelli, Jr., f,b
E. Borges, Jr., f,b a Scientific Department, Fertility Medical Group, Sao Paulo,
Brazil; b Scientific Department, Instituto Sapientiae - Centro de Estudos e
Pesquisa em Reproduç~ao Assistida, Sao Paulo, Brazil; c Urology Deparment,
Universidade Federal de S~ao Paulo (UNIFESP), Sao Paulo, Brazil; d Faculdade
de Ciencias Medicas da Santa Casa de Sao Paulo, Sao Paulo, Brazil;
e Chemistry Institute - ThoMSon Mass Spectrometry Laboratory, Universidade
Estadual de Campinas, Campinas, Brazil;
f Clinical Department,
Fertility Medical Group, Sao Paulo, Brazil.
OBJECTIVE: To identify lipid markers of blastocyst formation by day
three culture medium mass spectrometry (MS) fingerprinting.
DESIGN: Case-control study.
MATERIALS AND METHODS: For this study 50 embryo culture media
samples were harvested on the day three, from patients undergoing day five
embryo transfers. Embryos were split into groups based on their degree of
expansion and hatching status on day five (Complete-Blastocysts, n¼25
and No-Blastocysts, n¼25) and its secretomes were analysed by MS. Mass
spectra fingerprinting was acquired using a Q-Tof spectrometer (LC-MS,
Agilent 6550 iFunnel Q-TOF) equipped with an automated injector. Data
were analysed using the principal component analysis (PCA) followed by
a partial least square discrimination analysis (PLS-DA), combined with variable
influence in the projection (VIP) scores. The statistical analysis was
performed using Metabo-Analyst 2.0 (http://www.metaboanalyst.ca).
RESULTS: Overall, 1657 ions were observed. When the univariate analysis
was performed, 165 ions were observed to be differentially expressed between
the groups, with a fold chance R 4x and p

DESIGN: Basic research comparative study.
MATERIALS AND METHODS: Cumulus cells clumps (CCs) were isolated
from MII oocytes collected from patients &lt 35 years (‘‘younger’’,
n¼10) and &gt 40 years old (‘‘older’’, n¼11) undergoing ICSI for male factor
infertility. CCs were individually processed for RNA extraction, library preparation
and sequenced on Illumina HiSeq 2000 platform. Gene enrichment
and gene ontology analysis were used to define upregulated gene networks
and interactions in the two cohorts. Significance of differentially expressed
genes was assigned when both p value and false discovery rate were

de Ciencias Medicas da Santa Casa de Sao Paulo, Sao Paulo, Brazil; f IVF
Laboratory, Fertility Medical Group, Sao Paulo, Brazil; g Biotechnology
institute, Universidade de Caxias do Sul, Caxias do Sul, Brazil.
OBJECTIVE: To investigate whether patients presenting with endometriosis
are more likely to have oocyte with dysmorphisms, embryos with poor
development and decreased assisted reproductive technologies (ART) cycles’
outcome.
DESIGN: Case-Control study.
MATERIALS AND METHODS: The influence of the presence of endometriosis
on oocyte and embryo quality and blastocyst formation chance was evaluated.
Moreover, the presence of endometriosis was correlated with cycles’
characteristics. To avoid any bias concerning the age of the female, in the first
analysis only patients % 36 years old were included and the cycles were split
into: endometriosis infertility cycles (n¼431 and 3172 oocytes/embryos) and
others (n¼2510 cycles and 24480 oocytes/embryos). In a second analysis,
endometriosis infertility cycles (n¼669 and 4993 oocytes/embryos) and tubal
factor infertility cycles (n¼380 and 5029 oocytes/embryos) were compared.
RESULTS: For the first analysis, the number of retrieved oocytes
(10.621.2 vs 14.621.1, p

Vitrified OVA Banking Success Rates.
2011 2012 2013 2014 OVERALL
NUMBER 5 19 34 38 96
OVA SURVIVAL RATE (%) 59.0 89.5 93.7 93.8 90.5
FERTILIZATION RATE (%) 73.9 73.6 75.8 79.5 76.8
CLINICAL PREGNANCY 20.0 63.2 52.9 63.2 57.3
RATE (%)
NO ET RATE (%) 40.0 0.0 11.8 7.9 9.4
Reference:
1. Apollini Lm, Wyatt Sl, Collazo I, Et al. Preparation Of Warming Medium
For Vitirfied Specimens - Effects On The Final Warming Temperature.
2014 Aab Conference And Crm Symposium; Abstracts:15.
P-285 Tuesday, October 20, 2015
MICROFILAMENTS PLAY CRITICAL ROLE ON MITOCHON-
DRIAL TRAFFIC IN PORCINE GV OOCYTE. T. Yamochi,
S. Hashimoto, A. Amo, H. Goto, M. Yamanaka, M. Inoue, Y. Nakaoka,
Y. Morimoto. IVF Namba Clinic, Osaka, Japan.
OBJECTIVE: Oocyte maturation requires a variety of ATP-dependent reactions,
such as germinal vesicle breakdown, spindle formation, and polar
body extrusion, which is required for fertilization. Mitochondria are accordingly
expected to be localized to subcellular sites of ATP utilization.
Although cytoskeleton-dependent traffic for mitochondria has been studied
extensively in somatic cells, the mechanism of mitochondrial dynamics in
mammalian oocytes remains obscure. This study describes dynamic aspects
of mitochondria in porcine oocytes at germinal vesicle stage.
DESIGN: Basic research study.
MATERIALS AND METHODS: Mitochondria in donor oocytes were
stained with MitoTracker-Orange (MTO). Donor oocytes were centrifuged
at 10,000 x g and 37 C for 15 min. Mitochondria-enriched ooplasm were micropunctured
and injected into either central or subcortical area of recipient
oocytes. Mitochondria-injected oocytes were cultured with or without colcemid,
cytochalasin B or cytochalasin D. The image of mitochondrial dynamics
in the recipient oocytes was captured every 15 min using a confocal microscopy
for 15 hours, and analyzed quantitatively with ImageJ. Relative area
and distribution of fluorescent mitochondria in recipient oocytes were calculated
on the basis of their initial values at 0 h of culture. Total of 227 mitochondria-injected
oocytes were observed.
RESULTS: Mitochondria injected centrally moved unidirectionally to
subcortical area and those injected subcortically moved along plasma membranes.
The area occupied with MTO fluorescence increased significantly
during 15 hours. The area occupied with MTO fluorescence at both sites of
injection was not affected by colcemid, but inhibited strongly by cytochalasin
B and cytochalasin D.
CONCLUSIONS: This study shows that central mitochondria move from
central to subcortical area and those in subcortical area move along plasma
membranes, and suggests that microfilaments play critical role in mitochondrial
traffic in porcine GV oocyte. The method established in this study may
permit studies of the pathophysiology of intracellular traffic of mitochondria
and other organelles in oocytes from patients with infertility.
P-286 Tuesday, October 20, 2015
OTHER: ART - CLINICAL
EXPANDING ESET USE TO ALLWOMEN

and the odds of having a blastocyst transfer, high quality embryos, or ongoing
pregnancy.
TABLE 1. Repeat length and age adjusted associations (n¼4590).
Variable
Adjusted Association Per
Single Increase in CGG
Repeat Length
P value
FSH (U/L) -0.26 (-0.46 - -0.01) 0.01
AMH (ng/ml) 0.02 (-0.03 - 0.06) 0.36
AFC 0.05 (-0.02 - 0.12) 0.16
Estradiol at trigger (pg/ml) 10.0 (3.1 - 17.0) 0.005
Total gonadotrophins (IU) -12.1 (-25.2 - 1.0) 0.07
Total Follicles 0.04 (0.02 - 0.07) 0.03
Oocytes retrieved 0.04 (-0.01 - 0.09) 0.07
Mature oocytes 0.04 (0.01 - 0.08) 0.04
# of vitrified embryos 0.02 (0.01 - 0.03) 0.03
# of good quality blastocysts 0.02 (0.01 - 0.04) 0.05
CONCLUSIONS: In a population undergoing autologous IVF, increasing
FMR1 repeat size was positively associated with ovarian reserve and
response to stimulation for repeats less than 200. However, FMR1 repeat
number was not associated with pregnancy. These findings suggest that previously
described diminished ovarian reserve with premutation may not
apply in all populations. Instead of a ‘‘dose response continuum’’ of toxic
FMR1 mRNA, there may instead be a threshold effect or influence from as
yet uncharacterized factors, such as favorable X-inactivation patterns within
the ovary.
Supported by: Work Supported in part by the NICHD intramural research
program.
P-288 Tuesday, October 20, 2015
COMPUTER-AUTOMATED TIME-LAPSE TEST RESULTS ARE
PREDICTIVE OF PREGNANCY FOLLOWING BLASTOCYST
TRANSFER. B. Behr, a A. N. Beltsos, b B. Yee, c C. Kingsland, d
C. A. Benadiva, e J. Liebermann. b a Stanford Fertility and Reproductive Medicine
Center, Palo Alto, CA; b Fertility Centers of Illinois, Chicago, IL; c Reproductive
Partners, Redondo Beach, CA;
d Hewitt Fertility Center,
Liverpool, United Kingdom; e University of Connecticut, Farmington, CT.
OBJECTIVE: The Eeva Test is an automated time-lapse enabled test that
generates predictions based on cell division timings. As blastocyst development
was used to build the model, Eeva Test has been shown to improve Day
3 embryo selection (1). However, there is evidence suggesting by incorporating
key cell division timings Eeva results reflect underlying molecular
health of embryos (2,3). The objective of the study is to assess whether
Eeva Test can be applied to blastocyst selection.
DESIGN: Retrospective multi-center study.
MATERIALS AND METHODS: Total of 342 patients underwent blastocyst
transfers from 7 centers consented to use Eeva Test in IVF. Patients were
separated into three groups, at least 1 Eeva High (H) transferred, at least 1
Medium (M) transferred and only Low (L) transferred, and clinical pregnancy
and implantation rates were compared. Spearman rank were used to
check correlations among the variables of interest (age, #eggs, #2PN,
morphology, Eeva results, #embryos transferred). Univariate logistic regression
was used to assess predictive power of the variables against pregnancy.
Multivariate logistic regression with backward selection was used to test the
predictive value of Eeva results with other key pregnancy predictors
included.
RESULTS: Patients with at least 1 Eeva H blastocyst transferred had
significantly higher pregnancy and embryo implantation rates than patients
with only L blastocysts transferred (54% vs 34%, p¼0.002 and 46% vs
29%, p¼0.002), while patient age and #embryos transferred was similar between
groups. Univariate logistic regression showed that age, #2PN and
Eeva results (H vs L) are significantly correlated with pregnancy
(p¼0.008, 0.02, 0.02), while Spearman correlations showed Eeva results
were not correlated with the other variables. Multivariate model revealed
that Eeva results remain as significant predictor of pregnancy after adjusting
for age and blastocyst morphology while the other variables were not
significant.
Logistic regression result.
Variable Odds ratio P-value 95% CI (odds ratio)
Eeva results (H vs. L) 2.15 0.007 (1.23,3.74)
Age 0.92 0.005 (0.86,0.97)
Blastocyst morphology 0.46 0.002 (0.29,0.75)
#embryos transferred 1.41 0.14 (0.88,2.42)
#eggs 0.98 0.69 (0.92,1.05)
#2PN 1.03 0.49 (0.94,1.12)
CONCLUSIONS: It is well known that age and blastocyst morphology are
predictors of pregnancy. The current study is the first to show early cell division
timing parameters have independent predictive value for blastocyst
transfer outcome. Not only does it indicate Eeva Test may be used to improve
blastocyst selection, but it also supports the hypothesis that normal embryo
development may follow programmed timings at early stage (2).
References:
1. Diamond et al, Using the Eeva TestÔ adjunctively to traditional day 3
morphology is informative for consistent embryo assessment within a
panel of embryologists with diverse experience. J Assist Reprod Genet.
2015;32(1):618.
2. Wong et al, Non-invasive imaging of human embryos before embryonic
genome activation predicts development to the blastocyst stage. Nature
Biotech. 2010; 28(10):1115-21.
3. Chavez et al, Dynamic blastomere behaviour reflects human embryo
ploidy by the four-cell stage.Nature Commun. 2012;3:1251.
Supported by: Progyny (formerly Auxogyn).
P-289 Tuesday, October 20, 2015
THE IMPACT OF COMPREHENSIVE CHROMOSOME
SCREENING ON EMBRYO SELECTION OF DAY 5
BLASTOCYSTS. M. Schweitz, a S. McReynolds, a M. McGarvey, a
J. M. Stevens, b W. B. Schoolcraft, c M. Katz-Jaffe. c a Fertility Genetics,
Lone Tree, CO; b Fertility Labs of Colorado, Lone Tree, CO; c Colorado Center
for Reproductive Medicine, Lone Tree, CO.
OBJECTIVE: Embryo selection is a critical component of successful
infertility treatment. With the development of robust blastocyst culture, biopsy
and vitrification techniques, comprehensive chromosome screening
(CCS) is being applied for embryo selection with significant increases in
live birth rates following a euploid blastocyst transfer. The aim of this study
was to examine the relationship between D5 blastocyst morphology, chromosome
constitution, and the impact on embryo selection.
DESIGN: Research study.
MATERIALS AND METHODS: A large consecutive cohort of 2,670 blastocyst
CCS cycles performed from 2010-2014 were included for analysis. Blastocysts
were graded using a standard morphological system on D5 of
development prior to trophectoderm biopsy for CCS using either qPCR
(RMA-NJ) or aCGH (Illumina). Chi-Square analysis for independence was performed
to determine the relationship between the highest grade D5 blastocyst in
an embryo cohort and its chromosome constitution, with significance at P

P-290 Tuesday, October 20, 2015
WITHDRAWN
While IVF is more cost effective for the general population, cost effectiveness
is most comparable in the less than 35 year old age group, in which the net monetary
benefit of the strategies is balanced. Although more expensive, the IVF
strategy is favored as cost effective due to decreased cycle cancellations and
the ability to freeze surplus embryos for an additional transfer.
CONCLUSIONS: Although IVF is a more cost effective ART strategy, the
discounted patient charge of the MET protocol makes it a reasonable alternative
to traditional IVF in younger women where clinical pregnancy rates are
highest with MET.
Table 1. Characteristics of patients undergoing MET and IVF cycles.
Characteristic
MET less
than
35 years
(N¼108)
IVF less
than
35 years
(N¼146)
MET 35-40
years
(N¼101)
IVF 35-40
years
(N¼115)
MET
greater
than 40
years
(N¼73)
IVF greater
than 40 years
(N¼54)
Age - mean (SD) 29.8 2.7 30.8 3.0 37.7 1.8 37.4 2.0 42.9 1.8 41.9 1.0
Oocytes retrieved 2.6 1.6 13.6 7.7 2.2 1.3 10.8 6.4 2.5 1.8 9.6 6.5
- mean (SD)
Embryos transferred 1.7 0.7 1.9 0.6 1.7 0.7 2.5 0.9 1.8 1.0 3.2 1.4
- mean (SD)
Clinical pregnancy 26 (24.1) 53 (36.3) 17 (16.8) 46 (40.0) 3 (4.1) 11 (20.4)
per cycle (%)
Live birth per 28 (25.9) 68 (46.6) 11 (10.9) 40 (34.8) 1 (1.4) 9 (16.7)
cycle (%)
# Multiples (5 / 0) 18 / 2) (0 / 0) (8 / 0) (0 / 0) (2 / 0)
(twins/triplets)
Cancelled - 15 (13.8) 0 (0) 18 (17.8) 0 (0) 12 (16.4) 0 (0)
no retrieval (%)
Cancelled -
no transfer (%)
15 (13.8) 4 (2.7) 18 (17.8) 3 (2.6) 10 (13.7) 1 (1.9)
P-292 Tuesday, October 20, 2015
P-291 Tuesday, October 20, 2015
IVF PROVES COST EFFECTIVE COMPARED TO MINIMAL STIM-
ULATION EMBRYO TRANSFER, BUT MINIMAL STIMULATION
IS A COST NEUTRAL ALTERNATIVE FOR WOMEN UNDER AGE
35. N. Resetkova, a K. C. Humm, b A. Penzias, c D. Sakkas, d
B. M. Lannon. e a Obstetrics and Gynecology, Beth Israel Deaconess Medical
Center, Boston, MA; b Obstetrics and Gynecology, George Washington University
School of Medicine, Washington, DC; c Boston IVF / Harvard Medical
School, Waltham, MA; d Boston IVF, Waltham, MA; e Boston IVF, South
Portland, ME.
OBJECTIVE: In vitro fertilization (IVF) using injectable gonadotropins is
an effective albeit expensive treatment for infertility. A minimal stimulation
embryo transfer (MET) protocol may offer a less costly alternative. Our
objective was to compare treatment outcomes and cost stratify by age for
MET and traditional IVF cycles. Decision analysis was performed to identify
which subset of patients may benefit most from MET.
DESIGN: Retrospective cohort study, Decision analysis.
MATERIALS AND METHODS: We collected data on demographics,
treatment protocols, and outcomes for women who underwent oocyte
retrieval from January 2011 through April 2013. Cost data for IVF and
MET cycles included stimulation medications and were expressed in terms
of typical patient charges expressed in 2015 US dollars. Decision analysis
was performed through the design of a decision tree using TreeAge Pro (Williamstown,
MA).
RESULTS: A total of 597 cycles were included in the study, composed of
282 MET and 315 IVF cycles. Patient ages and cycle characteristics are reported
in Table 1. Cancelled cycles were approximately six times more likely
in MET cycles (0.312) as compared to IVF cycles (0.057). Awillingness to pay
of $50,000 per live birth was applied to our model for the purpose of analyzing
cost effectiveness. The average patient charge for a cycle of METwas $4710 for
an expected likelihood of delivery of 0.12, while the expected total charges of
an IVF cycle was $16,824 for a 0.40 likelihood of delivery. A frozen embryo
transfer cycle was permitted if excess embryos were present from the prior cycle.
Looking at the entire cohort (age independent), one-way sensitivity analysis
demonstrated that IVF is cost effective compared to MET when it is
priced less than 2.86 times that of an MET cycle (including medications).
VALIDATION OF BIRTH DEFECTS DATA IN THE SART CORS TO
BIRTH DEFECTS REGISTRY DATA IN
MASSACHUSETTS. J. E. Stern, a D. Gopal, b M. Kotelchuck, c
B. Luke. d a Obstetrics and Gynecology, Geisel School of Medicine at Dartmouth,
Lebanon, NH; b BUSPH, Boston, MA; c MassGeneral Hospital for
Children, Boston, MA; d Michigan State University, East Lansing, MI.
OBJECTIVE: To validate data on birth defects in the Society for Assisted
Reproductive Technology Clinic Outcome Reporting System (SART CORS)
with birth defects data in Massachusetts Birth Defects Registry (MBDR) reported
for the same children.
DESIGN: Retrospective validation.
MATERIALS AND METHODS: Singleton live births and stillbirths reported
to the SART CORS from July 1, 2004-Dec 31, 2008 were linked to birth
and fetal death certificates in MA with an overall linkage rate of 89.7%, which
was performed using mothers’ names, dates of birth, fathers’ names, and dates
of delivery. The birth and fetal death certificates are linked to the MBDR within
the Pregnancy to Early Life Longitudinal database. Data from the Congenital
Abnormalities field in SART CORS were compared with data in the MBDR;
sensitivity and specificity were calculated using the MBDR as the reference.
In addition, death certificates for any neonatal death were linked to the birth certificates
and compared to the neonatal death field in SART CORS.
RESULTS: There were 6,503 linked singleton births (6,484 live births and
19 stillbirths) in the SART CORS dataset that reported on birth defects. Of
the defects reported, the sensitivity ranged from 21.4% to 61.5%; specificity
was 98.7% for all defects and >99% for each defect due to the low absolute
percent occurrence. Neonatal deaths were reported for 13 infants in both systems,
but 11infant deaths were reported in SART CORS that did not appear in
the vital record system.
SART
CORS
Mass
BDR
Reported
in Both Sensitivity (%)
Any birth defect 132 132 51 38.6
Cleft palate 6 4 2 50.0
Genetic 18 13 8 61.5
Cardiac 25 42 10 23.8
Limb 7 13 4 30.8
Other 54 103 22 21.4
Unknown 25 - - -
FERTILITY & STERILITY Ò
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CONCLUSIONS: The SART CORS field for birth defects contains information
that, for Massachusetts deliveries, does not agree with the gold standard.
Although under-reporting of some birth defects may be due to
clinicians primarily reporting only defects discovered within days of delivery,
the reason for over-reporting of other defects is unclear. These findings
indicate that the SART CORS data field for birth defects is unreliable for use
in studies of ART outcome.
Supported by: NIH grants: R01HD064595 and R01HD067270.
P-293 Tuesday, October 20, 2015
FOLLICULAR LYSOPHOSPHATIDIC ACID MAY BE IMPORTANT
FOR FOLLICULOGENESIS IN OBESE WOMEN UNDERGOING
CONTROLLED OVARIAN HYPERSTIMULATION. J. K. Riley a
E. Jungheim. b a Obstetrics and Gynecology, Washington University School
of Medicine, St. Louis, MO; b Obstetrics and Gynecology, Washington University,
St. Louis, MO.
OBJECTIVE: Studies suggest lysophosphatidic acid (LPA) may influence
ovarian folliculogenesis, but the clinical significance is largely unknown.
Thus we investigated associations between LPA species in ovarian follicular
fluid (FF) and important clinical parameters in women undergoing in vitro
fertilization (IVF).
DESIGN: Cross-sectional.
MATERIALS AND METHODS: FF was collected from women ages 18-
44 during oocyte retrieval for IVF. Women using gestational carriers or
whose cycles were cancelled were excluded. FF LPA species were quantified
by liquid chromatography-mass spectroscopy. Clinical parameters investigated
included patient age, body mass index (BMI), infertility diagnosis,
antral follicle count, length of controlled ovarian hyperstimulation (COH)
required to achieve mature follicle size, number of oocytes retrieved, percent
mature oocytes, oocyte fertilization rate, and percent good quality cleavage
stage embryos. Associations between LPA species and clinical parameters
were made using appropriate bivariate statistics and stratified analysis.
RESULTS: 197 women were included. A negative correlation was noted
between amounts of FF LPA (C14:0, C16:0, and C18:0 species) and duration
of COH (r¼-.2, r¼-.2, r¼-.2 respectively, p< .002 for all), whereas a positive
correlation was noted between FF LPA and BMI (r¼.22, r¼.3, r¼.3 respectively,
p< .002 for all). FF LPA species were not associated with any other variables.
Women were stratified by BMI into normal (BMI

of identified articles, according to pre-specified criteria, using EROS software.
As a second step, two randomly selected independent reviewers assessed
each included study, to finally include them in the analysis and to
do the data extraction. We analyzed separately in the abstract and in the results
section of the full text, if the authors mentioned the p-value and a confidence
interval for the main outcome and for secondary outcomes. For
binary outcomes, we evaluated if any relative measure (i.e. RR, OR or
RRR) or absolute measure (i.e. ARR or NNT) was used or mentioned. For
continuous measures, we evaluated if the mean difference and its confidence
interval was used. We analyzed if the MCID or any reference related to any
clinically relevant result was mentioned in the material and methods section.
We also evaluated the discussion and conclusions and described the interpretation
that the authors did from the results that they published.
RESULTS: In the abstracts of the evaluated RCTs, neither the p-value nor
the confidence interval of the central estimation was mentioned in 25% of the
studies. In the material and methods section, the minimal clinically important
difference appeared only in 50% of the articles. In the results, both the p-
value and the confidence intervals were used in 57% of the studies, while absolute
measures were used in 42%. Finally, the confidence intervals were interpreted
in the discussion in only 42% of the cases.
CONCLUSIONS: Our data showed that there is an underreporting of the
most objective statistics such as the absolute differences and the confidence
intervals. It is the responsibility of authors and editorial boards to emphasize
the implementation of adequate reporting guidelines by the use of CON-
SORT in these specific type of studies.
Reference:
1. Glujovsky D, Boggino C, Riestra B, Coscia A, Sueldo CE, Ciapponi A.
Quality of reporting in infertility journals. Fertil Steril. 2015
Jan;103(1):236-41.
P-297 Tuesday, October 20, 2015
PUBLIC FUNDING OF IVF WITHOUT AGE LIMITS: A
CAUTIONARY TALE FROM QUEBEC. S. Ouhilal, a H. Lachgar, a
F. Bissonnette, b B. Buckett, c I. Kadoch, d P. St-Michel, e N. Mahutte. a a Reproductive
Endocrinology and Infertility, Montreal Fertility Centre, Montreal,
QC, Canada; b Reproductive Endocrinology and Infertility, OVO, Montreal,
QC, Canada; c Reproductive Endocrinology and Infertility, McGill University,
Montreal, QC, Canada; d Reproductive Endocrinology and Infertility, Centre
Hospitalier Universitaire de Montreal, Montreal, QC, Canada; e Reproductive
Endocrinology and Infertility, Procrea Clinics, Montreal, QC, Canada.
OBJECTIVE: In August 2010 the Quebec Government began covering the
costs of in vitro fertilization. The goals of the program were to facilitate access
to IVF independent of socioeconomic status and to reduce the multiple
pregnancy rate below 10%. The Quebec program had no upper age limit
eligibility criteria. We sought to determine the outcome and costs of IVF
treatment in women over the age of 40 who used their own eggs.
DESIGN: Cohort study.
MATERIALS AND METHODS: We used the Quebec data from the
CARTR database coupled with Quebec IVF reimbursement rates.
RESULTS: Most clinics have an age cut off between age 42 and age 44.
We looked at data with live birth outcomes between August 2010 and
December 2012. During that time period Quebec had 3685 fresh IVF cycle
starts in women over age 40. There were 3385 egg retrievals performed
and 2681 embryos transferred. The mean number of embryos transferred
was 1.9. The live birth rate per transfer was 9.2%. There were 246 babies
born. The table below shows the outcomes and cost per birth when broken
down by age between age 40 and 44.
CONCLUSIONS: For women over the age of 40, live birth rates are low and
come at a substantial financial cost in a public program. Age eligibility criteria
should be considered by any government planning to introduce public funding.
Outcomes at 40 and older.
Age 40 41 42 43 44
Number of cycle starts 1049 1005 944 488 141
Number of egg retrievals 972 922 865 447 128
Number of embryo 770 722 692 356 99
transfers
Number of live births 105 69 51 20 0
Live birth rate per 10% 6.9% 5.4% 4.1% 0%
cycle start
Mean treatment cost $43,153 $62,290 $79,100 $103,994 $597,800
per birth
(excluding
medications)
P-298 Tuesday, October 20, 2015
VALIDATION OF INFERTILITY TREATMENT AND ASSISTED
REPRODUCTIVE TECHNOLOGY USE ON THE BIRTH CERTIFI-
CATE: A US STUDY IN EIGHT STATES. B. Luke, a M. B. Brown, b
L. G. Spector. c a Obstetrics, Gynecology, and Reproductive Biology, Michigan
State University, East Lansing, MI; b Biostatistics, University of Michigan,
Ann Arbor, MI; c Pediatrics, University of Minnesota, Minneapolis, MN.
OBJECTIVE: To evaluate the accuracy of infertility treatment and assisted
reproductive technology (ART) reported on the birth certificate.
DESIGN: Cohort linkage study.
MATERIALS AND METHODS: ART cycles from the Society for Assisted
Reproductive Technology Clinic Online Reporting System were linked to
certificates of live birth in Florida, Massachusetts, New York, and Pennsylvania
(2004-09), Texas (2005-09), California and Ohio (2006-09), and Colorado
(2007-09) (ART children). All other live births to the same woman
were also identified (ART siblings), as well as a 10:1 sample of deliveries
of non-ART children (controls). Three questions on the most recent version
of the birth certificate were evaluated: Q1) Pregnancy resulted from infertility
treatment; Q2) Fertility-enhancing drugs, artificial insemination (AI) or intrauterine
insemination (IUI); Q3) Assisted reproductive technology (e.g.,
in vitro fertilization (IVF), gamete intrafallopian transfer (GIFT)). Since
not all items were included by each State, we created a summary item:
Any infertility question checked ‘Yes’. Information on the birth certificate
was evaluated for each of the three groups of children, overall and by plurality
(singleton vs multiple birth).
RESULTS: The distribution (%) by study group, plurality, and checkbox
item on the birth certificate is shown below.
CONCLUSIONS: The sensitivity of Q3 was 28.2% and the specificity was
99.7%. Only 36.5% of births of ART children were identified by any checkbox
on the birth certificate; multiple pregnancies were more likely to be indicated
as the result of infertility treatment than singletons (43.4% vs 33.3%). If
this undercount is applied to the ART siblings, about 1/3 of the singleton and
most of the multiple births also resulted from some type of infertility treatment.
Q1: Pregnancy
resulted from
infertility
treatment
Q2: Fertilityenhancing
drugs, AI, or IUI
Q3: Assisted
reproductive
technology,
IVF, or GIFT
Summary: Any
infertility item
checked ‘Yes’
ART Children ART Siblings Controls
Overall 69,969 9,489 636,645
Singleton Births 47,737 8,890 623,030
Multiple Births 22,232 599 13,615
Overall 36.8 14.1 0.8
Singleton births, 33.8, 43.0 12.3, 38.4 0.6, 8.9
multiple births
Overall 11.5 6.2 0.4
Singleton births, 10.1, 14.4 5.2, 20.6 0.3, 5.3
multiple births
Overall 28.2 5.5 0.3
Singleton births, 26.0, 33.0 4.9, 15.0 0.2, 4.0
multiple births
Overall 36.5 12.8 0.7
Singleton births,
multiple births
P-299 Tuesday, October 20, 2015
33.3, 43.4 11.1, 37.1 0.5, 8.7
SEX RATIO AT BIRTH IN RELATION TO USE OF ASSISTED
REPRODUCTIVE TECHNOLOGIES (ART). D. Kumar, a
K. Hoeger, b E. S. Barrett, c D. Li, d T. Dye. d a University of Rochester, Henrietta,
NY; b University of Rochester Medical Center, Rochester, NY; c University
of Rochester, Rochester, NY;
d ObGyn, University of Rochester,
Rochester, NY.
OBJECTIVE: To examine secondary sex ratio, the ratio of males to females
at birth, in relation to use of Assisted Reproductive Therapy (ART)
to conceive.
DESIGN: Retrospective cohort study using birth registry data collected
from 22 counties in Central and Upstate New York.
FERTILITY & STERILITY Ò
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MATERIALS AND METHODS: From 2004-2013, data on 322,397
singleton births was collected as part of the Central and Upstate New York
Perinatal Data System. At birth women completed questionnaires related
to their pregnancies, including an item on the use of ART to conceive the index
pregnancy. Multivariable logistic regression models were used to
examine the association between ART use and secondary sex ratio, adjusting
for maternal (age, race, education, weight gain, employment, Medicaid status),
and paternal (age and ethnicity) characteristics.
RESULTS: In total, there were 322,397 singleton births during the study
period, of which 1978 (0.6%) were conceived by ART. The secondary sex ratio
of infants conceived using ARTwas 0.956, whereas the sex ratio of infants
conceived naturally was 1.054. In unadjusted models, the odds of having a
male infant was reduced among ART users compared to non-ART users
(OR: 0.907; 95% CI: 0.831, 0.991). The results of adjusted models were
similar (OR: 0.908; 95%, CI: 0.828, 0.999).
CONCLUSIONS: Our analyses demonstrate that after adjusting for covariates,
ART is associated with a lower secondary sex ratio, indicating relatively
fewer males at birth. The results of this large, population-based
study agree with some (but not all) previous work on this topic. A notable
limitation is that this data registry did not have information on the indications
for use of ART nor the particular procedures used (IVF versus ICSI). Future
research should consider these factors as well as the mechanisms underlying
the skewed sex ratio following ART.
Supported by: UL1 TR000042.
P-300 Tuesday, October 20, 2015
COMPARISON OF OOCYTES DERIVED FROM NON-DOMINANT
SMALL FOLLICLES COLLECTED 24 AND 48 HOURS AFTER
TRIGGERING THE LUTEINIZING HORMONE SURGE IN NATU-
RAL CYCLE IN-VITRO FERTILIZATION. O. Miyauchi, T. Ueno,
T. Okubo, T. Hayashi, M. Kuroda, K. Omi, Y. Watanabe, T. Segawa,
H. Osada, S. Teramoto. Shinbashi Yume Clinic, Tokyo, Japan.
OBJECTIVE: In natural cycle in-vitro fertilization (IVF), when the endogenous
luteinizing hormone (LH) surge has started, oocytes from the dominant
follicles (DF) are collected 24 hours after the administration of gonadotrophin-releasing
hormone-agonist (GnRHa) to enhance the LH surge. However,
the rate of mature oocytes from non-dominant small follicles (SF)
increases if they are collected 48 hours after the GnRHa administration. In
this study, the efficacy of the delayed collection of SF oocytes was investigated
by comparison with SF oocytes collected 24 hours after the GnRHa
administration.
DESIGN: Prospective cohort study from February 2011 to April 2013.
MATERIALS AND METHODS: A total of 340 patients (mean age 35.6
3.7 years) in whom the endogenous LH surge had started in the natural
cycle gave written informed consent to be included in the study. The study
was approved by Shinbashi Yume Clinic’s institutional review board. Both
DF and SF (diameter 3-10 mm) oocytes were collected 24 hours after
GnRHa administration in 185 patients (group A). DF and SF oocytes
were collected 24 hours and 48 hours after GnRHa administration, respectively,
in 155 patients (group B). There was no statistical difference in
average age between the two groups. Mature oocytes and in-vitro-matured
oocytes were cultured to blastocyst stage and vitrified. A thawed single
blastocyst was transferred in the subsequent natural cycle. The vitrified
blastocyst rate (VBR) and the live birth rate (LBR) were compared between
the DF and SF oocytes of the two groups. For statistical analysis, we used a
Chi square for an independence test. The difference was considered significant
at P < 0.05.
RESULTS: In group A, there was no difference in the VBR between the
DF and SF oocytes (DF: 45 blastocysts, 45 patients, 24.3% ¼ 45/185 vs
SF: 51 blastocysts, 42 patients, 22.7% ¼ 42/185). There was no difference
in the LBR (DF: 45 transfers, 19 live births, 10.3% ¼ 19/185 vs SF: 41 transfers,
12 live births, 6.5% ¼ 12/185). The cumulative LBR of the DF and SF
oocytes was 16.8% (31/188), showing no difference compared to the LBR of
the DF-only group (p¼0.189). In group B, there was no difference in the
VBR between the DF and SF oocytes (DF: 36 blastocysts, 36 patients,
23.2% ¼ 36/155 vs SF: 51 blastocysts, 36 patients, 23.2% ¼ 36/155). There
was no difference in the LBR (DF: 35 transfers, 16 live births, 10.3% ¼ 16/
155 vs SF: 40 transfers, 15 live births, 9.7% ¼ 15/155). The cumulative LBR
of the DF and SF oocytes was 20.0% (31/155), showing a significant difference
compared to the LBR of the DF-only group (p ¼ 0.018).
CONCLUSIONS: SF oocyte collection 48 hours after GnRHa administration
compared with 24 hours after in natural cycle IVF, when the endogenous
LH surge had started, was shown to be a more efficient infertility treatment.
P-301 Tuesday, October 20, 2015
THE LACK OF IMPACT OFAGE ON TREATMENT TERMINATION
IN INSURED IVF PATIENTS. D. Sakkas, a L. Dodge, b M. R. Hacker, b
A. D. Domar. a a Boston IVF, Waltham, MA; b Department of Obstetrics and
Gynecology, Beth Israel Deaconess Medical Center, Boston, MA.
OBJECTIVE: To determine if the age of the female patient influences
treatment termination among insured patients undergoing in vitro fertilization
(IVF).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All cycles among insured women who
were age 18-

donor (27.94.3 vs. 37.22.0; p

the factors that influenced the most the ICER. Sensitivity analyses (one-way
and probabilistic), performed to evaluate inaccuracies, corroborate these
findings and attest their robustness.
CONCLUSIONS: Whereas drugs used for COS have different prices,
there are still not enough medico-economic analyses. These results indicate
that HP-hMG is the dominant option for IVF/ICSI in France. They corroborate
data from previous published data from international studies and should
raise awareness of health authorities on cost-effectiveness ratio of different
gonadotropins for COS in the setting of their health policy on IVF/ICSI reimbursements.
Finally, results are consistent when using different sources
of effectiveness data (published sources and French data).
Supported by: This study was sponsored by Ferring SAS, Gentilly, France.
EA is Ferring SAS employee (Medical Advisor, Medical Affairs Dept). CA,
PB, RF, SH and GPB were consulted as experts of ART.
P-306 Tuesday, October 20, 2015
ASSISTED REPRODUCTIVE TECHNOLOGIES (ART) AFFECT
EARLY CHILDHOOD GROWTH IN LOWER-BIRTHWEIGHT
CHILDREN. A. Butz, a R. S. Weinerman, a S. Senapati, a C. Sapienza, b
C. Coutifaris, a M. A. Mainigi. a a Division of Reproductive Endocrinology
and Infertility, University of Pennsylvania, Philadelphia, PA; b Fels Institute
for Cancer Research & Molecular Biology, Temple University, Philadelphia,
PA.
OBJECTIVE: A pilot study to determine the effect of ART on childhood
growth and to determine whether ART-associated placental DNA methylation
differences impact childhood growth.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Using parental survey data, current
birthweight (BW) and body mass index (BMI) percentiles (%) were
computed using World Health Organization and Centers for Disease Control
standards, adjusting for age, sex, and gestational age at birth (GA), for children
(age 1-8) born from singleton pregnancies conceived via ART or natural
conception (controls). Birth information had been collected prospectively
during a previous study analyzing placental DNA methylation. Placental
DNA methylation of the ART subjects had been analyzed using a custom-designed
Illumina VeraCode array containing 96 cytosine-phosphate-guanine
(CpG) sites in 82 genes known or thought to play a role in fetal growth.
The ratio of current BMI% to BW% was computed as a measure of a child’s
growth trajectory and was modeled as a function of BW% in control and ART
children. This was also done for ART subjects with normal and abnormal
placental DNA methylation at specific CpG sites. Between-group differences
were assessed using Mann-Whitney and t-tests. Non-linear regression
modeling was assessed using the extra sum of squares F test.
RESULTS: ART (n¼45) and control (n¼54) children did not differ in sex,
current age, mean GA, BW% or current BMI%. The mean BMI:BW% ratio
did not differ between ART and control children. However, when BMI:BW%
ratio was modeled as a function of BW% using non-linear regression, there
were differences in the curves between ART and control children that were
most notable in children with BW

696), the primary outcome of this study. In a post-hoc descriptive summary of
vital pregnancy rates, 298 women with embryo transfer enrolled in the
follow-up FTET study demonstrated per-cycle vital pregnancy rates of
32.0% with CFA (58/181) versus 32.7% with rFSH (54/165). There were
346 cycles among the 298 women, translating to per subject rates of
37.5% for CFA (57/152) versus 36.3% for rFSH (53/146).
CONCLUSIONS: The cumulative vital pregnancy and live-birth rates (from
fresh cycles and FTET) were similar in older women treated with CFA and
rFSH. No new safety signals were detected in this follow-up FTET study.
Supported by: Merck & Co., Inc., Kenilworth, NJ, USA.
P-309 Tuesday, October 20, 2015
DOES ULTRASOUND MONITORING AND OVULATION TRIGGER
WITH HCG IMPROVE OUTCOMES OF INTRAUTERINE INSEMI-
NATIONS (IUI) PERFORMED IN NATURAL CYCLES (NC)? H. El
Hachem, a,b P. E. Bouet, a,b L. Lapensee, a,b F. Bissonnette, a,b J. Benoit, a,b
R. Antaki. a,b a Ovo Fertility Clinic, Montreal, QC, Canada; b Obstetrics and
Gynecology, University of Montreal, Montreal, QC, Canada.
OBJECTIVE: To compare two methods of timing IUI in NC: spontaneous
triggering of ovulation by detecting LH surge with urinary ovulation kits (u-
LH) and ultrasound monitoring of follicular growth followed by ovulation
trigger with hCG (US/hCG).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All women %40 years with no history of
infertility who underwent donor sperm IUI (DS-IUI) in NC from Janauray 2011
to June 2014 were included. The indications for DS-IUI were the absence of a
male partner (single women and same-sex couples) and azoospermia. All
women had ovarian reserve testing (FSH, antral follicle count, AMH) and at
least one patent tube. None of the patients received any treatment. Women in
the u-LH group self-monitored ovulation at home using urinary ovulation
kits starting cycle day 10, and IUI was performed the day following the LH
surge. Women in the US/hCG group had serial US starting cycle day 10, and
when a leading follicle R17 mm was noted, ovulation was triggered with recombinant
hCG (Choriogonadotropin a,250mg) and IUI performed 36 hours
later. The choice of protocol was left to the physician’s discretion. Primary outcomes
were live birth rate per started cycle (LBR) and cumulative LBR. Secondary
outcomes were clinical pregnancy rate per started cycle (CPR) (fetal
heartbeat at 7 weeks), cycle cancellation and miscarriage rates.
RESULTS: 538 DS-IUI in 232 women were included: 267 u-LH in 113
women and 271 US/hCG in 119 women. The two groups were comparable
for age, Body Mass Index, ovarian reserve and number/mobility of inseminated
sperm. There were no significant differences in the primary and secondary
outcomes (Results are shown in table 1).
CONCLUSIONS: In women with no history of infertility undergoing
donor sperm IUI in a natural cycle, US monitoring of follicular growth followed
by ovulation trigger with hCG does not improve CPR and LBR
when compared with u-LH monitoring. The latter has the advantage of being
non-invasive and easy to perform at home and significantly reduces costs by
limiting hospital visits. Therefore, u-LH monitoring might be the best option
available to these women.
Outcomes of DS-IUI.
u-LH group
(n¼267)
US/hCG group
(n¼271)
p value*
Cycle Cancellation 1.1% (3/267) 1.4% (4/271) p¼ 0.71
CPR 16.1% (43/267) 11.8% (32/271) p¼ 0.15
Miscarriage 22.7% (10/44) 21.8% (7/32) p¼ 0.93
LBR 12.4% (33/267) 9.2% (25/271) p¼ 0.24
Cumulative LBR 29.2% (33/113) 21% (25/119) p¼ 0.15
* X2 tests were used to assess for differences between groups.
P-310 Tuesday, October 20, 2015
SCREEN MORE OR SCREEN LESS? CARRIER SCREENING IN
GAMETE DONORS: IT’S TIME FOR A NEW
PARADIGM. S. Rodriguez, a N. Kumar, a S. Yarnall, a R. Shraga, a
S. S. Chuan, b C. Pascale, c F. Licciardi. d a Recombine, New York, NY;
b San Diego Fertility Center, San Diego, CA; c Mental Health, Livingston,
NJ; d New York University Langone Medical Center, New York, NY.
OBJECTIVE: Carrier screening in gamete donors varies. Protocols are
based on ASRM guidelines, positive screening rates, and recipient testing
protocols. We assessed screening practices of varying scope and analyzed
the number of donors identified as carriers. With this data, our goal is to propose
a carrier screening protocol for gamete donors.
DESIGN: Retrospective.
MATERIALS AND METHODS: Genotyping was performed for over
1500 mutations associated with over 200 autosomal recessive and X-linked
genetic diseases. Custom testing panels were developed for referring institutions;
this allows for results reporting of only those diseases within a specific
panel. The analysis includes data obtained from 621 gamete donors referred
by reproductive endocrinologists and donor banks/agencies. Documented
informed consent was obtained. Frequently ordered testing panels were identified.
Overall genotyping results were compared to reported results, as determined
by inclusion in the ordered testing panel.
RESULTS: Of 78 custom donor screening panels, 2 panels were used to
screen 86% of donors. A limited panel of 10 diseases and

was made 30 minutes after IVI (IVI +30). Each recording was accelerated 10-
fold and UC frequencies at the isthmic and fundal parts of the uterus were
rated by 3 independent observers who were blinded for study design. Overall
UC frequency was obtained by the average frequency between isthmic and
fundal contractions and average ratings of observers.
RESULTS: Median UC frequency (min-max) increased slightly but not
significantly over the 3 observation points (baseline, IUI +15, and IVI +30):
3.2 (2.7-4.1), 3.3 (2.5-4.9), and 3.4 (2.4-4.8) UC/minute, respectively. Further,
UC frequency was not influenced significantly by semen volume nor the magnitude
of ovarian response. UC frequency was negatively correlated (Spearman
test) with women’s ages at IUI +15 (r¼-0.42, P< 0.04) and IVI +30 (r¼-
0.51, P< 0.009) but not at baseline (r¼-0.20, NS). Yet, the % of increase of
UC frequency over the 3 observation points was not related to women’s ages.
Finally, pregnancy rates tended to be higher (30% vs. 15%) in patients showing
R3.5 UC/minute (n¼13) than those with

outcomes such as birth weight (g) 3163 (CI95%3035-3292) vs. 3074(CI95%
2913-3236), low birth weight (0.8-%1.2 ng/mL (group 3) and >1.2 ng/mL (group 4). Low birth weight
was defined as

MATERIALS AND METHODS: Included in the study were agonist
(n¼206) and antagonist (n¼43) down regulated cycles. All patients received
both HP-hMG (MenopurÒ) and HP-FSH (BravelleÒ) in an approximate 1:1
ratio (i.e an LH/FSH ratio of 0.5) from day one of stimulation. Only cycles
with blastocyst transfers were included.
RESULTS: Characteristics of the study population were as follows (mean
SD): age ¼ 33.2 years 4.2 (range 21-42 years), BMI ¼ 24.0 4.3, stimulation
days ¼ 9.7 0.7, total FSH dose (IU) ¼ 1541 521, total HP-hMG dose
(IU) ¼ 1359 582, oocytes ¼ 13.7 5.5, MII ¼ 10.2 4.2, 2PN ¼ 8.5 3.2,
peak E2 ¼ 2356.35 1888.73 pg/ml, and peak P4 ¼ 0.96 0.63. Cycles were
divided into peak (day of hCG administration) P4 %1.5 ng/ml compared to P4
>1.5 ng/ml. The incidence of PPR was 16.4%. The implantation rate, clinical
and ongoing pregnancy rates were comparable between groups. To analyze the
association among variables associated with increased P4 levels, linear regression
was performed. BMI, peak E2 levels on the day of hCG administration, and
number of retrieved oocytes were associated with PPR.
CONCLUSIONS: Despite a 16.4% incidence of PPR, elevated peak P4
levels were not associated with a negative effect on IVF outcomes following
a stimulation strategy employing HP-hMG and HP-FSH at a 0.5 LH/FSH ratio.
The use of HP-hMG may protect against a potential negative effect of
elevated P4 on implantation in fresh autologous blastocyst transfers.
Table 1.
%1.5 ng/ml
(n¼208)
>1.5 ng/ml
(n¼41)
Embryos transferred 1.9 0.7 1.8 0.5 0.254
Implantation rate 229/406 (57%) 45/75 (59%) 0.753
Clinical pregnancy/ 154/208 (74%) 28/41 (68%) 0.572
cycle started
LB/cycle started 144/208 (69%) 27/41 (66%) 0.809
Reference:
Warner et al, Fertil Steril 2014.
Supported by: VCRM.
P-318 Tuesday, October 20, 2015
WITHDRAWN
P-319 Tuesday, October 20, 2015
CHANGES IN SINGLETON LIVE BIRTH WEIGHTS IN A LARGE
IVF PRACTICE OVER AN 18 YEAR PERIOD. K. Maas, a,b
E. Galkina, c K. Thornton, a,b D. Sakkas. b a Beth Israel Deaconess Medical
Center, Boston, MA; b Boston IVF, Waltham, MA; c Biomedical Engineering
and Biotechnology, University of Massachusetts Lowell, Lowell, MA.
OBJECTIVE: The fetal origins hypothesis suggests that some diseases
originate in utero owing to adaptations made by the fetus to the environment
it encounters. This has allowed live birth weights (LBW) to be used as a surrogate
marker of these in utero environmental encounters. It has been shown
that babies born from IVF in a thaw cycle have higher LBW on average
compared with those born from a fresh cycle. It has also been hypothesized
that embryo culture media can impact LBW. Clinical IVF practices including
stimulation protocols, medication dosing, medication types, transfer day, and
cycle monitoring have evolved significantly since the inception of IVF. The
IVF laboratory including incubators, culture media, culture devices and the
introduction of micromanipulation techniques has also changed significantly
over time. The objective of this study is to investigate the association between
singleton LBW over an 18 year period in a large academic IVF clinic in the
US over time after autologous in vitro fertilization (IVF) in both fresh and
frozen cycles.
DESIGN: Retrospective cohort study of 7332 singleton live births from
patients who underwent autologous fresh or frozen IVF cycles at Boston
IVF between 1996 and 2013.
MATERIALS AND METHODS: 6265 fresh and 1067 frozen cycles were
analyzed. One way ANOVA and t-tests were performed to compare average
LBW in autologous fresh and frozen cycles as well as average LBW per cycle
type over time. Six month increments were compared over the study period.
P
RESULTS: A total of 7332 singleton deliveries were included from the
fresh and frozen cycles. The mean LBW SD in fresh cycle and frozen cycle
cohorts were 3282 620g (3267-3298g) and 3456 600g (3420-3492g)
respectively. ANOVA and t-tests demonstrated a statistically significant
mean difference (173g, p

OVERALL
HIGH FSH &
NORMAL AMH
NORMAL
FSH & LOW AMH
HIGH FSH
& LOW AMH
NORMAL FSH &
NORMAL AMH
P-VALUE
N(%) 33 (2) 647 (33) 166 (8) 1111 (57)
Total gonadotropins Predicted Mean SE 5761 288 a 5297 66 b 6075 129 b 3582 50 ab

D3eSET and 153 had elective single blastocyst transfer (eSBT). Embryology
profiles and clinical outcomes were assessed for both groups. Primary
outcome measures were implantation rate (IR), clinical pregnancy rate
(CPR), and spontaneous abortion rate (SAB). Categorical data are presented
as percentages and compared using the Fisher’s exact test. Continuous variables
are described as mean +/- SD and compared using the Students t-test,
assuming equal variances.
RESULTS: Patients undergoing D3eSET had significantly fewer total oocytes
retrieved, mature oocytes, fertilized oocytes, top quality embryos for
transfer and cryopreservation. However, patients in the D3eSET group
achieved equally high clinical pregnancy and implantation rates as patients
in the eSBT group.
D3eSET vs. eSBT Cycle Outcomes for Patients < 35 years of age.
D3eSET
N¼ 25
eSBT
N¼153
P value
Mean Age 31.12.1 31.02.3 NS
Mean No. Oocytes 10.04.8 16.66.3

OBJECTIVE: To assess trends in embryo transfer practices and outcomes
of assisted reproductive technology (ART) cycles performed in the United
States.
DESIGN: Retrospective population-based cohort study.
MATERIALS AND METHODS: We analyzed ART cycles initiated between
1996 and 2013 in the United States and reported to the National
ART Surveillance System (2013 data are preliminary). For each reporting
year, we calculated: (a) average number of embryos transferred and percentage
of transfers that involved one embryo, (b) percentage of singleton ART
live births, (c) percentage of ART-conceived infants born with normal birth
weight (R2,500 grams), and (d) average gestational age and percentage of
term (R37 weeks) births among all ART births. Linear regression was
used to test for the significance of linear and quadratic trends in the average
number of embryos transferred and average gestational age. Linear probability
models (with a binomial distribution and an identity link) were used to test
for the significance of linear and quadratic trends in the proportions of all
other outcomes. Generalized estimating equations were used to accommodate
clustering by clinic.
RESULTS: The average number of embryos transferred decreased from
3.9 to 1.9 and the proportion of single embryo transfers increased from
5.5% to 30.7% between 1996 and 2013. The all-time largest increase in
the proportion of single embryo transfers (23.8% increase) was observed between
2012 and 2013. The percentage of singleton live births among all ART
births increased from 62.6% to 75.4%. The percentage of normal birth weight
infants among ART-conceived infants increased from 58.0% to 71.5%. Similarly,
the average gestational age of ART births increased from 37.6 weeks to
37.8 weeks, and the percentage of term births increased from 68.0% to
73.8%. All trends were significant (P

in blastocyst development, aneuploidy, pregnancy or implantation rate between
patients with Low or High BMI.
CONCLUSIONS: The impact of maternal BMI on the kinetics of embryo
development, determined by time-lapse image analysis, is dependent on
maternal age. Maternal BMI does not influence the success of ART in patients
35 years or older, possibly due to the overriding negative impact of
maternal age on oocyte quality.
Table 1. Embryo morphokinetics analyzed by AGE, BMI and the interaction
AGE x BMI.
Age BMI t2-t4 t8 tM tSB tB tEB
Young Low 11.6 58.1 92.2 101.6 106.2 108.3
Young High 11.3 56.0 87.6 98.9 103.6 105.9
AMA Low 11.0 59.4 92.4 102.2 107.3 109.9
AMA High 10.4 58.2 93.6 104.7 110.0 113.0
Age 0.06 NS 0.01 0.01 0.005 0.01
pvalue BMI NS NS NS NS NS NS
AgexBMI NS NS 0.017 0.04 0.04 0.10
P-330 Tuesday, October 20, 2015
DOES PREVALENCE AND EFFECT OF ELEVATED PROGESTER-
ONE ON THE DAY OF OOCYTE MATURATION DIFFER BY
ETHNICITY? G. D. Royster, IV, a S. Zarek, a A. Christy, a
A. DeCherney, a K. Devine, b M. J. Hill. a a National Institutes of Health, Bethesda,
MD; b Shady Grove Fertility Center, Washington, DC.
OBJECTIVE: To compare the effect of elevated day-of-trigger progesterone
concentration on live birth rate in various ethnic groups.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Analysis included all fresh autologous
ART cycles from 2009-2013 in which progesterone concentration was
measured on the day of oocyte trigger. Generalized estimating equations
(GEE) were utilized with nesting for patients with multiple cycles and to account
for confounding variables. The primary outcome was clinical pregnancy.
RESULTS: 3289 fresh autologous ART cycles were analyzed. Caucasians
had a similar clinical pregnancy rate (46.2%) to Hispanics (44.7%) and
higher pregnancy rates than African American (41.0%) and Asians
(38.3%) (P

Table-1. CYCLE OUTCOME OF DIFFERENT GROUPS OF EMBRYO.
Groups (n) Pregnancies (n) PR(%) OR (95% CI) IR (sacs/total Et)(%) OR (95% CI)
Total cycles (744) 326 43.8 13.9 (292/2093)
1-Easy-E T (543) 248 45.6 1vs2
1.23
(0.9 to 1.8) (p>0.05)
14.7 (236/1604) 1 vs. 2
1.3
(0.9 to 1.8)(p>0.05)
2-All difficut-ET- (201) 78 39.8 11.4
3-Cx - Tr (83) 42 50.6 1
vs3
0.8
4-Bl-OS (39) 17 43.6 1
vs 4
1.1
(0.5 to 1.3)(p>0.05)
(0.5 to 2.1)(p>0.05)
5-Bl-TC (45) 11 24.4 1
vs.5
2.6
(1.3
to 5.2)(P0.05)
1 vs. 5
1.7
(0.9 to 3.2)(p>0.05)
1 vs. 6
1.9
(0.8 to 4.2)(p>0.05)
OBJECTIVE: Easy embryo transfer ( E-ET) in contrast to difficult (D- ET)
is thought to result in better cycle outcome. Although definition of E- ET is
agreed upon ,there is disagreement about what constitutes D- ET. We aimed
to compare the impact of individual elements of D-ET : cervical traction (Cx-
Tr), blood on outer sheath ( Bl-OS) , blood on transfer catheter (Bl-TC) and
the need for sounding (Snd) individually and in combination on clinical pregnancy
rate (CPR) and implantation rates (IR) of ICSI/IVF cycles.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: A cohort of 744 ICSI cycles were
included for women who fulfilled the following inclusion criteria :age <
38 years, fresh first ICSI trial, number of transferred embryos R 2 high quality
.The technique of ET used was the tactile method and the same type of
catheter (Labottect ). ET was considered easy if no resistance was encountered
during passing the preloaded TC and sheath through cervical canal
without Cx -Tr ,Bl-OS or Bl-TC nor Snd . It was considered difficult if
Cx-Tr or Snd were needed and /or , Bl-OS, Bl-TC were present . Cycle
outcome (CPR , IR) were compared for cycles with easy ET and those
with D- ET as a whole and individually with subgroups of Cx-Tr, Bl-
OS,Bl-TC & Snd, using Odds ratio (OR) and 95% CI. The differences
were considered significant if the P value was< 0.05.
RESULTS: As shown in the table CPR for E-ET (45.6%) and D- ET
(39.8%) are not statistically significantly different. Comparisons of E- ET
with cx-Tr, Bl-OS,Bl-TC, Snd components of D-ET showed significantly
lower CPR with Bl-TC and Snd transfer subgroups only. Although IR showed
no statistically significant differences between E- ET and overall and components
of D- ET, the Bl-TC, and Snd subgroups tended to have significantly
lower IR but owing to the small number of cycles in these subgroups the differences
did not reach level of statistical significance (type II error).
CONCLUSIONS: Cervical traction and /or blood on outer sheath do not
compromise cycle pregnancy rate or implantation rate. Only when blood
on transfer catheter and/ or sounding the uterus at time of ET is the CPR
significantly undermined and implantation rate tends to be lower.
P-334 Tuesday, October 20, 2015
SHOULD EMBRYO TRANSFER NUMBER BE UPGRADED IN
ETHNIC MINORITY GROUPS? C. Chatzicharalampous, a
J. Stelling, b,c J. Jenkins, c M. Saketos, c,b L. Sung, c,d M. A. Bray. c,e a Obstetrics
and Gynecology, The Brooklyn Hospital Center, Brooklyn, NY; b SUNY
Stony Brook School of Medicine, Stony Brook, NY; c Reproductive Specialists
of NY, Mineola, NY; d Obstetrics and Gynecology, Winthrop Univ Hosp,
Mineola, NY; e Obstetrics and Gynecology, The Brooklyn Hosp Center,
Brooklyn, NY.
OBJECTIVE: To determine if the number of transferred embryos influences
IVF clinical pregnancy rates (CPR) in different ethnic groups.
DESIGN: A retrospective study of 565 fresh embryo transfer cycles: 214
single and 351 double. We compared ethnicity, age, BMI, FSH, gravidity,
parity,infertility diagnoses, previous fresh cycles, oocytes retrieved, fertilization
rate, total and quality blastocysts, CPR and twin rate by number of embryos
transferred.
MATERIALS AND METHODS: Women undergoing IVF or ICSI from
2010 to 2013 who had an autologous, fresh, elective single (eSET) or elective
double (eDET) day 5 blastocyst embryo transfer were included in the study.
Clinical Pregnancy Rate by ethnic group and embryo transfer count
Single Embryo Transfer
Double Embryo Transfer
Caucasian % (N) Non-Caucasian % (N) Caucasian % (N) Non-Caucasian % (N)
Non- Pregnant Pregnant Non-Pregnant Pregnant Non-Pregnant Pregnant Non-Pregnant Pregnant
51 (88) 49 (85)* 73 (30) 27 (11)* 55 (144) 45 (118) 56 (50) 44 (39)
Twin rate: 3.5%
Twin rate: 0%
Twin rate: 35%
Twin rate: 26%
Ectopic rate: 2.3%
Ectopic rate: 0%
Ectopic rate: 0%
Ectopic rate: 10%**
* p ¼ 0.02
** p < 0.001
FERTILITY & STERILITY Ò
e219

All women were under 38 years of age and all had < 2 previous fresh IVF
cycles. We included four ethnic groups: Caucasian, African American, Asian
and Hispanic, which were dichotomized into two main groups: Caucasian
and non-Caucasian. Variables were analyzed using Chi-square, Fisher’s exact
test, Student’s t-test and logistic regression. p < 0.05 was considered statistically
significant.
RESULTS: Age, BMI, day 3 FSH, gravidity, parity, number of previous
fresh cycles, number of oocytes retrieved, fertilization rate and number of total
and quality blastocysts were similar in both ethnic groups. Peak estradiol
levels were higher in non-Caucasians (p¼0.02). Infertility diagnoses were
similar in both groups with the exception of more tubal factor in non-Caucasian
women (p¼0.001) and more cases of unexplained infertility in Caucasian
women (p¼0.02). In the eSET groups, Caucasian women had a
significantly higher CPR (49%) compared to their ethnic minority counterparts
(27%) (p ¼ 0.02). Differences in CPR between patient groups persisted
after accounting for differences in infertility diagnosis. For the eDET groups,
the CPR did not differ by ethnicity: Caucasian (45%) vs. non-Caucasian
(44%) (p ¼ N.S.). The multiple gestation rates in both groups were similar
and not statistically significant. Ectopic rate was markedly higher (10%) in
the DET ethnic minority group, likely due to the higher tubal factor infertility
diagnosis.
CONCLUSIONS: In spite of similar measured demographic variables, minority
women have a significantly lower CPR in eSET cycles. Further
research is needed to identify confounding factors and to determine if minority
women would benefit from an upgrade to eDET in selected cases.
P-335 Tuesday, October 20, 2015
USE OF ICSI IN IVF CYCLES IN WOMEN WITH TUBAL LIGA-
TION DOES NOT IMPROVE PREGNANCY OR LIVE BIRTH
RATES. F. Grimstad, a A. Nangia, b B. Luke, c J. E. Stern, d W. Mak. e a Obstetrics
and Gynecology, University of Kansas, Kansas City, KS; b University
of Kansas Medical Center; c Michigan State University, East Lansing, MI;
d Geisel School of Medicine at Dartmouth, Lebanon, NH; e Yale School of
Medicine, New Haven, CT.
OBJECTIVE: To compare the fertilization, pregnancy, and live birth rates
of in vitro Fertalization (IVF) cycles with and without intracytoplasmic
sperm injection (ICSI) among patients with tubal ligation only.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: Assisted reproductive technology
(ART) cycles from patients with tubal ligation only and no other male or
female factor diagnoses from the Society for Assisted Reproductive Technology
Clinic Outcome Reporting System (SART CORS) database (2004-
2012) were compared according to use of ICSI (some or all) versus conventional
IVF (ICSI-none). Adjusted odds ratios and 95% confidence intervals
(AOR, 95%CI) were calculated using logistic regression with tubal ligation
patients having no ICSI as the reference group, and adjusted for year of
treatment, maternal age, race and ethnicity, gravidity, ovarian hyperstimulation
syndrome, number of oocytes retrieved, day of transfer, and number
of embryos transferred. Outcome measures included fertilization rate (2PN/
total oocytes); clinical intrauterine gestation (CIG/cycle); and live births
(LB/cycle).
RESULTS: Of 1,099,978 fresh autologous cycles performed, 11,198 were
for tubal ligation only (1%); of these, 5,629 cycles (50%) used some or all
ICSI versus 5,569 (50%) conventional IVF. The mean fertilization rate was
49.0% and 57.2% for ICSI-none versus ICSI-some/all respectively. The clinical
intrauterine gestation rate and live birth rate are listed in the table. There
were no significant differences in birth weight or gestational age.
Outcomes of all/some ICSI compared to no ICSI.
N, Cycles
Clinical Intrauterine
Gestation
Live Birth
% AOR 95% CI % AOR 95% CI
No ICSI 5,629 45.0 1.00 Reference 37.5 1.00 Reference
All/Some 5,569 40.3 0.81 0.75, 0.88 32.2 0.79 0.73, 0.86
ICSI
CONCLUSIONS: Use of ICSI versus conventional IVF did not improve
outcomes for tubal ligation patients. These results suggest that the use of
ICSI is generally not warranted in this population.
P-336 Tuesday, October 20, 2015
IDENTIFICATION OF OVARIAN RESPONSE PREDICTORS FOR A
NEWLY AVAILABLE RECOMBINANT FSH
PREPARATION. C. M. Howles, a J. Jenkins, b G. Griesinger, c
D. W. Warne. d a Aries Consulting, Geneva, Switzerland; b R&D and Medical
Affairs, Finox Biotech A.G., Kirchberg, Switzerland;
c Sektion f€ur
Gyn€akologische Endokrinologie und Reproduktionsmedizin, Universit€atsklinikum
Schleswig-Holstein – Campus L€ubeck, L€ubeck, Germany; d Consultant
Biostatistician, Thonex, Switzerland.
OBJECTIVE: Predictors of ovarian response have been reported both for
GnRH agonist and antagonist treatment cycles using established FSH stimulation
agents. Various predictors have been identified in different analyses,
but typically the most reported are Age, FSH, AMH, BMI. With the availability
of newly registered recombinant FSH (r-FSH) preparations, it is important
to establish if the same predictors of ovarian response can be utilized with
similar predictive capacity in order to allow treating clinicians to effectively
and safely perform ovarian stimulation.
DESIGN: A post-hoc analysis was conducted on data obtained from a randomized
controlled Phase III registration trial comparing 2 r-FSH preparations
(clinicaltrials.gov; NCT01121666). As recently described (1), a low
response was defined less than 6 oocytes and a high response more than 18
oocytes retrieved. Prognostic models were constructed using data from all
treatment cycles (n¼482).
MATERIALS AND METHODS: Patients aged 20-38 years were treated
in a long luteal phase start GnRH agonist protocol with a fixed-dose daily
regimen, of 150 IU r-FSH s.c. The dose could be reduced after day 6 of
FSH stimulation only if there was a risk of OHSS. 15 centres in six European
countries participated. Recruited subjects underwent up to 2 treatment cycles
(total cycle 1 n ¼ 372; cycle 2 n ¼ 110). The trial was conducted between
July 2010 and April 2012. Logistic and linear regression were used to investigate
factors which influence low and high response, and number of oocytes
retrieved.
RESULTS: 1) 81 cycles were classified as a low response (16.8%) whilst
44 (9.1%) were classified as a high response. AMH was the best performing
predictor for both low and high response in both treatment groups (AMH low:
ROC AUC ¼ 0.751; AMH high: ROC AUC ¼ 0.700). 2) multivariate analysis
showed that the choice of gonadotrophin treatment had no impact on the
predictive model hence results were pooled irrespective of gonadotrophin
used to refine predictive value of other variables. 3) adding further biomarkers,
such as baseline FSH and AFC did not significantly increase the
AUC of prediction models.
CONCLUSIONS: The results demonstrated that for both the new and
reference r-FSH products, there were common predictors of ovarian response
(AMH, AFC, FSH). The new r-FSH product can be clinically used in the
same way as the reference product in ART practice. Additionally the findings
support the utility of AMH as the most relevant marker of ovarian response
and provides physicians with a reliable marker of ovarian response when using
these FSH preparations.
Reference:
1. Broekmans FJ, Verweij PJ, Eijkemans MJ, Mannaerts BM, Witjes H.
Prognostic models for high and low ovarian responses in controlled
ovarian stimulation using a GnRH antagonist protocol. Hum Reprod.
2014 Aug;29(8):1688-97.
Supported by: Finox AG, Technikumstrasse 1, CH-3401 Burgdorf,
Switzerland.
P-337 Tuesday, October 20, 2015
LIVE BIRTH CORRELATION OF EEVA TEST RESULTS IN PA-
TIENTS WITH DAY 5 TRANSFER: A PROSPECTIVE BLINDED
STUDY. M. VerMilyea, J. T. Anthony, M. A. Mainigi. University of Pennsylvania,
Philadelphia, PA.
OBJECTIVE: Time-lapse-based embryo selection algorithms have been
extensively assessed against blastocyst formation, aneuploidy and implantation.
However, the single most important outcome of assisted reproductive
e220 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

technology (ART) is live birth. The objective of this prospective blinded
study was to perform the first examination of the correlation between an automated
time-lapse algorithm (the Eeva Test) and live birth outcome.
DESIGN: Prospective blinded study.
MATERIALS AND METHODS: The study included 69 patients (Jan
2013-2014) who consented to have embryos imaged using the Eeva System,
a platform technology that automatically measures early cleavage timing parameters
and generates an Eeva Test result regarding developmental potential.
For this non-selection, blinded study, embryos were selected for fresh
transfer using only morphology evaluation on Day 5. Embryos of known
live birth were those from patients with #live births equal to #embryos transferred,
and those who did not have a live birth.
RESULTS: Of the 69 patients, 58 patients had embryos with known live
birth rate. A total of 80 Day5 embryos were transferred, resulting in 26 babies
born (overall known live birth rate 32%). When live birth rate was analyzed
based on Eeva Test results, we observed a 38% relative difference in live birth
rates between Eeva High vs. Eeva Low embryos, while egg age was similar
across the categories.
CONCLUSIONS: To our knowledge, this is the first prospectively designed
blinded study of time-lapse analysis results and their correlation to
live birth outcomes. Previous studies have shown correlation of Eeva Test results
to aneuploidy risk and Day 5 implantation, indicating that the Eeva Test
results may reflect embryo quality in a manner that is independent of age. The
study is currently ongoing and increased sample size will allow meaningful
evaluation of statistical significance.
Live Birth Rate
Age
Eeva High 36% (14/39) 33.1 4.8
Eeva Medium 36% (5/14) 34.4 3.7
Eeva Low 26% (7/27) 33.7 4.6
p-value NS NS
Supported by: Progyny (formerly Auxogyn).
P-338 Tuesday, October 20, 2015
PROGESTERONE (P4) ELEVATION ON DAY OF HUMAN CHORI-
ONIC GONADOTROPIN (HCG) TRIGGER PREDICTS LOWER
PREGNANCY RATE DURING IN VITRO FERTILIZATION (IVF)
CYCLES. S. Chang, K. Thornton, H. Lieman, S. K. Jindal, E. Buyuk.
Albert Einstein College of Medicine / Montefiore Medical Center, Bronx,
NY.
OBJECTIVE: The effect of late follicular P4 elevation on IVF success
has been previously evaluated, with conflicting results. Supraphysiologic
estradiol (E2) and P4 elevations may be detrimental to endometrial receptivity,
resulting in premature closure of the implantation window, and a
subsequent decrease in pregnancy rates. Though there is growing evidence
for the negative effect of elevated P4 on day of hCG trigger, fewer
studies have looked at P4 elevations leading up to day of trigger, which
may more accurately reflect changes in the endometrium than a single
value on day of trigger. We aimed to determine whether elevations in
P4 leading up to, and on day of hCG trigger were associated with
decreased pregnancy rates.
DESIGN: Historical cohort.
MATERIALS AND METHODS: The first IVF/intracytoplasmic sperm
injection (ICSI) cycles of 238 women from January 2009 to August 2014
were included for analysis. Demographic data, stimulation method, infertility
etiology, baseline follicle-stimulating hormone (FSH) and E2, antimullerian
hormone (AMH), number of eggs/embryos, embryo quality,
transfer day, progesterone levels, and clinical pregnancy data were
collected. Main outcome measure was clinical pregnancy rate. Student’s
t-test, Mann-Whitney U, chi-square and logistic regression analyses were
used. Data was expressed as mean SD or as % and p

questionnaires, including report of usual alcohol consumption, before the
first IVF cycle. Women were categorized as non-drinkers, social drinkers,
or daily drinkers. Poisson regression with robust variance adjusted for female
age and the number of embryos transferred and was used to quantify the relation
between alcohol and pregnancy and live birth outcomes of the first IVF
cycle.
RESULTS: In total, 1,961 women reported their typical alcohol consumption
prior to their first IVF cycle. There were 536 (27%) non-drinkers, 1,352
(69%) social drinkers, and 73 (4%) daily drinkers. Nearly half (44%) of social
drinkers reported consuming R5 drinks per week. Among non-drinkers,
34% achieved clinical pregnancy, and 29% had a live birth. These proportions
were greater than what we observed among social drinkers (32% and
26%, respectively), but were not statistically significant (both PR0.32).
Among daily drinkers, 30% achieved clinical pregnancy, but only 19% had
a live birth, which though not statistically significantly lower than that among
non-drinkers (P¼0.18), is clinically meaningful. Compared to non-drinkers,
the relative risk of live birth did not differ among social drinkers (RR¼0.97,
95% confidence interval [CI] ¼0.80-1.17) or daily drinkers (RR¼0.70,
CI¼0.40-1.21).
CONCLUSIONS: While alcohol consumption prior to IVF treatment was
not statistically associated with differences in the proportion of cycles that
achieved clinical pregnancy or live birth, the decreased proportion of daily
drinkers who achieved a live birth compared to non-drinkers appears clinically
meaningful. Ongoing analyses will examine intermediate outcomes
within the IVF treatment response, and future work should examine cumulative
pregnancy rates over additional cycles and patterns of alcohol consumption
during IVF treatment.
P-341 Tuesday, October 20, 2015
HIGH ESTRADIOL AND ICSI INCREASE THE RISK OF
PLACENTAL COMPLICATIONS IN PREGNANCY. G. D. Royster,
IV, a J. Csokmay, b B. Yauger, b A. DeCherney, a E. F. Wolff, a M. J. Hill. a
a National Institutes of Health, Bethesda, MD; b Walter Reed National Military
Medical Center, Bethesda, MD.
OBJECTIVE: Higher peak estradiol (pE2) during ART has been associated
with lower birth weight, but the role of ICSI is not known. Here we
assess the relationships among pE2 levels, ICSI, and adverse placental obstetric
outcomes.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: All fresh IVF cycles from 2010-2013
were analyzed. Inclusion criteria were singleton pregnancy. Exclusion
criteria were prior history of placental complication, diabetes or hypertension.
Hypertension disorders include gestational hypertension (GHTN),
mild and severe pre-eclampsia. Growth disorders were defined as small for
gestational age (SGA) or intrauterine growth restriction (IUGR). Composite
placental outcome was any growth or hypertensive disorder. Generalized
estimating equations (GEE) models were utilized to analyze associations
of estradiol on the day of trigger (pE2) and ICSI use with pregnancy complications
and control for confounding variables. Receiver operating characteristics
(ROC) curves and greater than efficiency curves were generated to
evaluate estradiol thresholds for predicting adverse outcomes.
RESULTS: 1595 consecutive ART cycles starts resulted in 466 singleton
live births with 393 patients meeting inclusion criteria. pE2 was positively
associated with SGA, GHTN, severe pre-eclampsia, and composite placental
outcomes. ROC analysis showed an area under the curve (AUC) of 0.65 and
0.66 for pE2 predicting HTN and growth disorders respectively. ICSI was
positively associated with SGA and composite placental complications. In
adjusted GEE models, only pE2 (OR 1.33, 95% CI 1.13-1.52) and ICSI
(OR 4.21, 95% CI 1.73-10.25) were associated with composite placental
complications, whereas male factor infertility and female age were not. Interaction
testing of ICSI versus IVF with pE2 and composite placental complications
showed P

499 IU/L were at 9.7-fold greater odds of failing single-dose
MTX than those with peak hCG 372 IU/L were at 4.7-fold greater odds of
requiring additional treatment than those with hCG

OBJECTIVE: To determine whether antim€ullerian hormone (AMH) predicts
good quality supernumerary blastocyst cryopreservation.
DESIGN: Retrospective study.
MATERIALS AND METHODS: First, fresh IVF cycles (n¼247) from
two fertility ceneters, grouped as follows: women < 35 years with AMH
< 1 ng/mL (n¼40) or AMH R 1 ng/mL (n¼77); women R 35 years with
AMH < 1 ng/mL (n¼62) or AMH R 1 ng/mL (n¼68). AMH level was
measured prior to IVF. Ovarian stimulation protocols based on patient age
and AMH level included short Gonadotropin Releasing Hormone (GnRH)
agonist, GnRH antagonist, or GnRH agonist microdose flare. Supernumerary
good quality blastocysts were cryopreserved on days 5 or 6 post-retrieval.
Primary outcomes measure was supernumerary good quality blastocyst cryopreservation
in relation to AMH levels. Logistic regression was used for statistical
analyses.
RESULTS: Among women < 35 years of age, there was a significant difference
in supernumerary good quality blastocyst cryopreservation between
groups of AMH < 1 ng/mL and AMH R 1 ng/mL (30.0 % vs 58.4 %) when
adjusted for age. Among women R 35 years of age, there was a significant
difference in supernumerary good quality blastocyst cryopreservation between
groups of AMH < 1 ng/mL and AMH R 1 ng/mL (16.1 % vs 42.6
%), when adjusted for age.
CONCLUSIONS: Low AMH levels are associated with a significantly
lower likelihood of blastocyst cryopreservation as compared to higher
AMH levels. This effect was seen among women < 35 years of age and those
R 35 years of age. Patient counseling should include realistic expectations
for the probability of good quality supernumerary blastocyst cryopreservation.
P-347 Tuesday, October 20, 2015
IS ADDING ESTRADIOLE (E2) TO PROGESTERONE (P) LUTEAL
SUPPORT IN HIGH RESPONDER LONG GNRH AGONIST ICSI CY-
CLES DETRIMENTAL TO OUTCOME? : RANDOMIZED
CONTROLLED TRIAL (RCT). M. Ghanem, a M. H. Bedairy, a
A. S. Helal, a A. Shaaban. b a Mansoura Faculty of Medicine, Mansoura,
Egypt; b Ob.Gyn, Al-Azhar Faculty of Medicine, Cairo, Egypt.
OBJECTIVE: Retrospective studies have shown that high response in
terms of excess egg production and high levels of sex steroids are detrimental
to cycle outcome. The primary aim of this RCT is to find if adding E2 to P
luteal support in high responders ICSI cycles impairs cycle pregnancy rate
(CPR) and implantation rate (IR) Secondary aim is to find predictors of
outcome.
DESIGN: RCT.
MATERIALS AND METHODS: Using computer generated random
numbers, group allocation was blindly done at time of ovum pick-up.Arm
I (102 cases) received E2 valerate 6 mg /day plus daily IM injection of 100
P from day of OPU to the time of pregnancy test , arm II (112 cases) received
only daily IM100 mg P. Inclusion criteria: Female age %39 ,fresh first ICSI
trial, basal FSH< 10 mIU/mL, low risk of OHSS, number of eggs retrieved
>15. E2,P assay on days of hCG, mid-luteal were done .Means were
compared by using T-test , ratios by Chi square tests, and predictive values
were measured by ROC curve. The differences were considered significant
if P was < 0.05.
RESULTS: Baseline features were comparable : female age, basal FSH,
number of eggs retrieved, hormone profile on day of hCG-day, number of
embryos transferred and proportions of blastocyst and cleavage embryoes
and ease of transfer (ET).The two arms had comparable CPR
(48.4,44.2& 44.1,42,4) and implantation rates(18.3,18.5) respectively
(p>0.05). Hormonal profiles showed no significant differences in E2 and P
levels on hCG and mid-luteal days. The only significant difference between
groups was significantly lower mean drop of E2 level in arm I from hCG-day
to midluteal level (p¼0.028). Study of predictors of CPR revealed that only
lower level hCG-day P {(AUC curve ¼0.357 ,(95% CI ¼0.270 to
0.445)(p¼0.003)} and lower mean E2 drop from hCG -day to midluteal
(AUC ¼ 0.390, 95% CI¼0.297 to 0.484) ,p¼0.024) predict pregnancy.
CONCLUSIONS: E2 luteal support in high responders does not impair
outcome , on the contrary pregnant outcome is more linked to lower drop
of E2 in luteal phase implying that E2 luteal support may be beneficial in
high responders contrary to what was expected. While lower P level on
hCG-day is significantly linked to pregnant outcome , no association was
found with the number of eggs retrieved, nor E2 level on hCG-day or midluteal
E2.
P-348 Tuesday, October 20, 2015
IN A YOUNG COHORT OF PATIENTS, NUMBER OF EMBRYOS
FROZEN INDICATES BETTER PROGNOSIS FOR FUTURE
FROZEN CYCLE. E. M. Murphy, S. Amrane, A. P. Melnick,
O. K. Davis, Z. Rosenwaks. The Ronald O. Perelman and Claudia Cohen
Center for Reproductive Medicine, Weill Cornell Medical College, New
York, NY.
OBJECTIVE: To compare in vitro fertilization (IVF) cycles of patients
who had a negative pregnancy test in both fresh and frozen cycles to those
who had a negative pregnancy test in the fresh cycle with a live birth in a subsequent
frozen cycle.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All patients undergoing a fresh IVF cycle
with a subsequent frozen IVF cycle between January 2001 and December 2013
were included. Patients who did not have an embryo transfer, had genetic
testing on the embryos, and/or were using donor embryos were excluded. Patients
meeting criteria were separated into two groups. Patients in group one
(n¼261) had a negative pregnancy test in the fresh and frozen cycle. Patients
in group two (n¼221) had a negative pregnancy test in the fresh and a live birth
in frozen cycle. Statistical analysis using STATA software included t-test and
Chi-square. P value

ate than those with non-eSET (79/115 (68.7%) vs 99/214 (46.3%), respectively,
P

cycles and stored at -20 C. Media samples were collected, shipped, stored
and analyzed between December 2014 and April 2015. Blastocyst grading
was carried out according to Gardner’s criteria before ET. The media samples
were shipped frozen to the analytical laboratory and subjected to matrix assisted
laser desorption ionization (MALDI), time of flight (ToF) mass spectrometry
(MS). MALDI ToF mass spectrometry to identify pattern
differences in aneuploid and non-aneuploid blastocysts. Data from spectra
were collected from the region 3,000 to 30,000 m/z, smoothed and normalized.
Qualitative characteristics of the data from spectral hotspots in 6000 m/
z to 9500 m/z regions were examined for difference.
RESULTS: After blastocyst grading 19 were assigned 5AA grades and
correlated well with positive pregnancy (16/19, 84.2%), 18 were given mixed
grades but resulted in a negative pregnancy, 3 had an abnormal PGS and were
not graded or transferred. Secretome patterns following MALDI ToF MS
analysis in the 6000 m/z to 9500 m/z regions show distinct pattern differences
between high grade embryos, embryos which resulted in a negative pregnancy,
and aneuploid embryos. In particular, aneuploid blastocysts give
rise to significant profile differences in the 8500-9000 m/z regions such
that they are completely distinct from either high grade embryos or embryos
which resulted in a negative pregnancy.
CONCLUSIONS: Non-invasive analysis of spent blastocyst culture media
by MALDI ToF MS can identify aneuploid blastocysts amongst other high
and low quality blastocysts.
P-353 Tuesday, October 20, 2015
PRESENCE OF DARK GRANULOSE CELLS FOLLOWING PRO-
LONGED ANTAGONIST ADMINISTRATION AND ITS IMPACT
ON EMBRYO QUALITY. S. Ghosh, a G. B.S., a R. Chattopadhyay, b
S. K. Goswami, b G. Bose, b M. Goswami, b B. Chakravarty. b a Assisted
Reproduction, Institute of Reproductive Medicine, Kolkata, India; b ART,
Institute of Reproductive Medicine, Kolkata, India.
OBJECTIVE: To evaluate the relation between duration of antagonists to
levels of reactive oxygen species in the follicular fluid containing dark granulosa
cells, and to examine its association with embryo quality.
DESIGN: Prospective study done between December 2013 to November
2014.
MATERIALS AND METHODS: A total of 796 women undergoing antagonist
cycle in vitro fertilization (IVF) for tubal factor were divided into two
groups. Women with polycystic ovary syndrome (PCOS), and endometriosis
were excluded from the study. All patients underwent standard controlled
ovarian stimulation with recombinant FSH from day 2 of the cycle and
were given antagonist as in flexible protocol. 254 women (Group A) received
antagonist (Inj. Orgalutran, 0.25mg s.c) for more than 4 days before ovulation
trigger. The rest 542 women (Group B) were administered antagonist
in same dosage for 1-3 days before hCG was given. In group A, 58 women
(22.83%) and in group B, 26 women (4.8%) showed dark granulosa cells
along with oocyte-corona-cumulus complex in the follicular aspirate. Such
follicular fluid was collected for estimation of reactive oxygen species
(ROS) and it was assessed by chemiluminescence method in these patients.
Levels of ROS and embryo quality were subsequently correlated. Statistical
comparisons were performed using student’s t test. ’P’ value < 0.05 was
considered statistically significant. Study was approved by Institutional Review
Board.
RESULTS: Women who had dark granulosa cells following antagonist for
more than 4 days showed significantly higher level of ROS and fragmentation
compared to women who had received antagonist for less than four days.
CONCLUSIONS: Increased duration of exposure to antagonist may have
detrimental effect on the follicular fluid milieu, as evidenced by high ROS
level, resulting in poor quality embryos.
Results.
Parameter Group A Group B P Value
Reactive oxygen
species (ROS) cpm/106
Significant fragmentation
(RGrade 3 embryo)
95.38 7.9 48.84 4.89 0.001
54.46 2.17 14.17 2.18 0.001
P-354 Tuesday, October 20, 2015
BLASTOCYST IMPLANTATION IS CORRELATED WITH OUT-
PUTS FROM AUTOMATED TIME-LAPSE ANALYSIS BY THE
EEVA TEST. J. Liebermann, a A. Bartolucci, b S. Troup, c C. Wagner
Coughlin, d B. Yee, e B. Behr. f a Fertility Centers of Illinois, Chicago, IL;
b CARS, Farmington, CT;
c Hewitt Fertility Center, Liverpool, United
Kingdom; d Aparent IVF Laboratory, Highland Park, IL; e Reproductive Partners,
Redondo Beach, CA; f Stanford Fertility and Reproductive Medicine
Center, Palo Alto, CA.
OBJECTIVE: The automated, time-lapse enabled Eeva Test provides test
scores that predict blastocyst formation (1) and correlate to implantation (2).
Specifically, positive correlation was demonstrated between Eeva Test results
and implantation/pregnancy following Day 3/Day 5 transfer (2). However,
the relationship between Eeva Test results and blastocyst implantation has
not yet been examined. Furthermore, there is no data to inform the implication
for patients in different age groups.
DESIGN: Retrospective multi-center study.
MATERIALS AND METHODS: The study included a total of 342 blastocyst
transfer patients from 7 centers (2012-2014) who consented to have
embryos imaged using the Eeva System, a platform technology that automatically
measures P2 (time between 2- and 3- cell) and P3 (time between 3- and
4- cell) and generates an Eeva Test score of High, Medium, or Low to indicate
the embryo developmental potential. Embryos with known implantation data
were included in this study. P-values were calculated using c2 or student’s t-
test (SAS 9.3).
RESULTS: The overall known implantation rate for the study was 38%
(148/386). Eeva High blastocysts had a relative 40% higher implantation
rate than Eeva Low blastocysts (p¼0.002), while the age was similar between
the two groups. Further analysis revealed that in older patients (ageR35),
Eeva High blastocysts had over twice the implantation rate than Eeva Low
blastocysts (p¼0.002).
Eeva
results
Implantation
rate
Age
Implantation
rate (Age

of this study were infertile couples underwent IVF. The measurement of
serum AMH and estradiol level was performed at the beginning of IVF cycles,
while follicular fluid AMH, were measured on the day of oocytes retrieval.
Oocytes quality was measured using Xia morphology criteria, including the
assessment of polar bodies, perivitelline space and cytoplasmic granulation.
Pearson correlation and linear regression statistical analyses were performed
to determine the predictive value of AMH for oocytes quality.
RESULTS: We obtained 102 IVF patients with antagonist protocol. Serum
AMH, follicular fluid AMH, serum estradiol, number of mature oocytes, and
oocytes morphological score were assessed. Serum AMH has better correlation
to oocytes morphological score compare to follicular fluid AMH and
serum estradiol (r¼0.804 vs r¼0.525 vs r¼0.278). On the other hand, age
has negative correlation to oocytes morphological score (r¼-0.389). Based
on multivariate analysis, we found that age and serum AMH level are the
best predictor for oocytes quality.
CONCLUSIONS: Serum AMH, but not the follicular AMH, can be used to
predict oocytes quality.
P-356 Tuesday, October 20, 2015
INVESTIGATION OF ZONA PELLUCIDATHICKNESS VARIATION
AND IMPLANTATION RATE - WITH AND WITHOUT ASSISTED
HATCHING. E. I. Lewis, a,b R. Farhadifar, c D. Needleman, c
S. A. Missmer, a,b,d L. V. Farland, a,b,d C. Racowsky. a,b a Center for Infertility
and Reproductive Surgery, Dept of OBGYN, Brigham and Women’s Hospital,
Boston, MA; b Harvard School of Medicine, Boston, MA; c School of Engineering
and Applied Sciences, Dept of Molecular and Cellular Biology,
Harvard University, Cambridge, MA; d Harvard School of Public Health,
Boston, MA.
OBJECTIVE: Recent literature has proposed that zona pellucida thickness
variation (ZPTV) in cleavage stage embryos is associated with implantation
rate (IR), but studies have been small and have only used a crude ZPTV
assessment. We utilized novel image processing software to assess ZPTV
to evaluate the relation between ZPTV and IR. In addition, we investigated
whether this relation was affected by assisted hatching (AH).
DESIGN: Pilot retrospective cohort study.
MATERIALS AND METHODS: 349 embryos (56% fertilized with ICSI)
with known implantation results (i.e. 0% or 100%) were assessed for ZPTV
from single and double day 3 embryo transfers performed between 3/2014-
12/2014. Embryo photographs were analyzed using an automated image processing
platform to segment the zona pellucida (ZP) with an active contour
technique. From each image, 100 data points were obtained of ZP thickness
(ZPT) equidistant around the perimeter of each embryo to calculate ZPTV
(¼ [maximum ZPT-minimum ZPT]/mean ZPT). Logistic regression adjusted
a priori for female age, embryo quality, ICSI, Day 3 FSH, male factor infertility,
IVF attempt number, and AH was used to calculate the odds ratio (OR) of IR by
tertile of ZPTVand 95% confidence intervals (CI) / 2-sided Wald p-values (p).
RESULTS: In the adjusted model, there was no significant association between
ZPTV and Sac-IR (sacs/embryos transferred). The highest tertile of
ZPTV (>30%) compared to the lowest tertile ZPTV (22%), was associated
with 57% lower odds of Fetus-IR (viable fetuses @ >7 wks/embryos transferred)
(p¼0.02). Minimum, maximum, and mean ZPT were not associated
with IR. Among embryos without AH, those in the highest tertile of ZPTV
had 88% lower odds of Fetus-IR compared to those in the lowest tertile of
ZPTV (p¼0.01). However, among embryos with AH, ZPTV was not associated
with Fetus-IR.
CONCLUSIONS: As high ZPTV was associated with lower Fetus-IR,
ZPTV measurement may be a useful adjunctive variable for selecting day
3 embryos for ET. In addition, AH may negate the inverse relation between
high ZPTVon Fetus-IR. Further investigation is warranted to establish the association
between ZPTV and IR and to evaluate the impact of AH.
Associations between ZPTV and IR with and without AH OR (95% CI).
ZPTV Tertile 1(T1) ZPTV Tertile 2(T2) ZPTV Tertile 3(T3)
Sac-IR 30 (26)1.00 (ref) 31 (27)1.06 (0.60-1.88)
p¼0.83
Fetus-IR 28 (24)1.00 (ref) 27 (23)0.90 (0.52-1.59)
p¼0.73
Fetus-IR: 17 (37)1.00 (ref) 16 (35)0.75 (0.33-1.70)
No AH
p¼0.49
Fetus-IR: AH 11 (16)1.00 (ref) 11 (16)1.07 (0.45-2.53)
p¼0.88
22-30% (n¼117) >30% (n¼115)
19 (17)0.56 (0.28-1.11)
p¼0.10
15 (13)0.43 (0.22-0.85)
p¼0.02
7 (15)0.22 (0.07-0.67)
p¼0.008
8 (12)0.76 (0.34-1.68)
p¼0.49
Supported by: Ferring REI Research Grant awarded by New England
Fertility Society.
P-357 Tuesday, October 20, 2015
RETROSPECTIVE CASE CONTROL STUDY TO EVALUATE THE
EFFECT OF ENDOMETRIOSIS AND RELATED CLINICAL CON-
DITIONS WITH ELEVATED TNFA LEVELS ON EMBRYO PRO-
DUCTION AND IMPLANTATION DURING CYCLES OF
ASSISTED REPRODUCTION. M. Paczkowski, a,b T. Wincek, a,b
J. F. Pliego, a,b T. J. Kuehl. a,b a Obstetrics and Gynecology, Baylor Scott &
White Healthcare, Temple, TX; b College of Medicine, Texas A&M Health
Science Center, Temple, TX.
OBJECTIVE: Conditions that lead to elevated peritoneal levels of tumor
necrosis factor alpha (TNFa) can result in ovarian dysfunction and impaired
implantation. This study aimed to test the hypothesis that patients with clinical
conditions associated with increased TNFa have reduced development
of in vitro fertilized embryos to the blastocyst stage and reduced implantation
and pregnancy.
DESIGN: Retrospective study.
MATERIALS AND METHODS: A retrospective case-controlled study
with 2 matched controls for each of 43 cases with endometriosis, Crohn’s disease,
or arthritis undergoing IVF/ICSI cycles for infertility was performed.
Controls matched affected patients for age within 3 years and were performed
close to the same time. Groups were compared for age, days of
FSH stimulation, total dose of FSH, number of ova retrieved, number of
mature ova, percent maturation, percent fertilized per mature ova, percent
developing to the blastocyst stage, number of embryos transferred, number
of embryos implanting, number of embryos cryopreserved, types of implantation
(singleton, twin or triplet), percent clinical pregnancies, and percent
maintaining pregnancy to 30 wks by March 2015. Comparisons used Student’s
t test, chi-square test and for pregnancy, Mantel-Haenszel matched
pairs comparison.
RESULTS: Affected patients and controls did not differ for any of the variables
related to treatment, ova maturation, fertilization or development (p >
0.19). Number of embryos implanting (p ¼ 0.06), type of implantation (p ¼
0.07), and patients with gestational sacs (p ¼ 0.08) approached significance
and tended to decrease in affected patients. Mantel-Haenszel odds ratio for
presence of gestational sacs for affected patients versus controls was 0.48
(95% CI of 0.22 to 1.05). However, as pregnancies progressed, more gestational
sacs in affected patients were lost so that 47% of affected patients
versus 58% in controls (p ¼ 0.21) maintained pregnancies to beyond 30 wks.
CONCLUSIONS: There was no difference in stimulation, oocyte recovery,
oocyte maturation, fertilization, and blastocyst development for affected
patients compared to control patients. When similar numbers of embryos
were transferred in the fresh cycle the pregnancy outcomes were similar between
affected patients and controls suggesting that diagnosis and treatment
for conditions with abnormal immune mediators is not a risk factor for
impaired outcome from ART cycles. While there is a tempting trend that patients
with endometriosis, Crohn’s disease, or arthritis could have reduced
implantation, this was not significantly different and the outcomes converged
as the pregnancies progressed.
Supported by: Baylor Scott & White Healthcare, Department of Obstetrics
and Gynecology.
P-358 Tuesday, October 20, 2015
PERINATAL OUTCOME USING TIME-LAPSE SYSTEM AND
REDUCED OXYGEN CULTURE IN IVF PATIENTS. N. Zaninovic,
Q. Zhan, R. Clarke, Z. Ye, J. Malmsten, Z. Rosenwaks. CRM, Weill Cornell
Medical College, New York, NY.
OBJECTIVE: To investigate and compare the perinatal outcomes of patients
using time-lapse system with reduced oxygen vs. standard incubator
with ambient oxygen.
DESIGN: A retrospective analysis of IVF patients with fresh ET resulting
in live births from 2012 to 5/2014 was performed. Frozen ET and PGD/S cycles
were excluded. The incidence of term (TB), pre-term (PTB,

(S; 20% O2, Thermo Forma, USA). In total, 1661 babies were born; 947 from
EmbryoScopeÒ and 714 from standard incubator. Statistical analyses were
performed using X2, ANOVA and logistic regression.
RESULTS: Live birth implantation rate of D3 transferred embryos was
significantly higher in EmbryoScopeÒ vs. standard incubator (48.9% vs.
44.6%, p

Twenty embryos did not survive the warming (96% survival rate). Of 508
transferred embryos, there were 291 pregnancies by serum hCG (57%).
The implantation rate was 250/502 (50%, with 6 cases lacking implantation
data). Blastocyst scoring was determined by Gardner’s criteria before trophectoderm
biopsy. For statistical analysis, the qualities of TRO or ICM
were semi-quantitatively assigned: A¼6, B¼4,C¼2, unclear quality¼1.
The quality of EXP was assigned as its number, from 1 to 6, i.e., 1¼very early
blastocyst to 6¼hatched blastocyst. TQS was the sum of all 3 attributes.
Receiver Operator Characteristic Curve (ROC) and Chi-square tests were
used for statistical analysis for correlation of attributes with pregnancy. Cochran-Mantel-Haenszel
Armitage Tests used to examine the linear trend of
TQS and pregnancy, also analyzed the cut line of TQS for significant better
pregnancy outcome. Due to space limitation, only the set of hCG analysis is
presented.
RESULTS: The significant level of each attributes is summarized in Table
1.TQS is the most sensitive predictor (p < 0.0001). There is a significant
linear trend of TQS with pregnancy (p < 0.0001). When TQS is more than
12, the pregnancy rate is significantly (p

maintain potency and block differentiation. Here we test viable first lineage
differentiation reporter ESCs for use in HTSs for IVF/ART stress and toxicological
stress.To validate the effectiveness of a first differentiated lineage
(Pdgfra-GFP) reporter in embryonic stem cells (ESC) by detecting stress
dose- and time-dependent responses, and detecting additional first lineage
and later lineage markers (Dab2, laminin, AFP, LRP2).
DESIGN: Experimental.
MATERIALS AND METHODS: We obtained published mouse (m)
ESCs transgenic with green fluorescent reporter (GFP) knocked into a
Pdgfra loci and regulated by endogenous factors binding the Pdgfra promoter.
These were tested for stress responses in the presence of LIF (a
potency-maintaining growth factor) or for normal differentiation by
removing LIF. Assays used to test Pdgfra-GFP ESCs included microplate
reader, FACS, cell counts, immunofluorescence, and immunoblots. Dose
and kinetic responses were tested in reporter ESCs for hyperosmotic sorbitol
as we have done previously as this is a widely-used positive control
for stress. Retinoic acid (RA) -a specific first lineage inducer was used as
a positive control.
RESULTS: Hyperosmotic stress in the presence of LIF, or LIF removal
(+/-RA), induces Pdgfra-GFP in a time- and dose-dependent manner as assayed
by immunofluorescence and microplate reader. These stress doses
decreased stem cell population expansion. Only Pdgfra-GFP positive cells
co-expressed first lineage marker DAB2 and stress induced only Pdgfra-
GFP+/DAB2+ cells. With LIF removal there were DAB2+/Pdgfra-GFP+ coexpressors
and also Pdgfra-GFP+/DAB2- cells. This suggests that stress
overrides potency to cause a limited, ‘‘prioritized’’ differentiation but LIF
removal enables a wider array of first lineage and later derivatives of first
lineage that are Pdgfra-GFP+.
CONCLUSIONS: The time and dose-dependence of stress induced differentiation
are a proof-of-concept that the Pdgfra-GFP reporter ESCs can be
used as an HTS for fluids derived from IVF-ART protocols (follicular fluid,
spent media, uterine fluids) and from toxicological substances or new Pharma
encountered by the embryo in vivo. The Pdgfra-GFP differentiation reporter
ESCs should complement the previously reported and patented Rex1-RFP
ESCs and reduce the frequency of false positive and false negative outcomes.
The more limited early lineages induced by hyperosmotic stress - compared
with LIF removal - supports the theory of stress induced prioritized differentiation.
P-365 Tuesday, October 20, 2015
THE COMMON OR CELL-SPECIFIC TRANSCRIPTION FACTORS
BETWEEN OOCYTES AND EMBRYONIC STEM CELLS IN
MICE. K. Kim, S. Lee, K. Park, J. Ko, K. Lee. CHA University,
Gyeonggi-do, Korea, Republic of.
OBJECTIVE: By Yamanaka and Daley group, the six genes (Oct4, Sox2,
c-Myc, Klf4, Nanog, and Lin28) have been shown to induce pluripotency in
somatic cells. Based on the fact that both oocytes and embryonic stem cells
(ESCs) have the potential to reprogram differentiated somatic cells into
pluripotent cells, we conducted the present study to compare gene expression
profiles, especially transcription factors, between oocytes and ESCs for
finding common or cell-specific genes as candidates of novel reprogramming
factors.
DESIGN: This study was conducted to compare gene expression profiles
between oocytes and ESCs for finding safer natural novel regulators of reprogramming.
MATERIALS AND METHODS: Previously, we identified a list of upand-down
regulated genes during oocyte maturation using AB 1700 Full
Genome Expression Mouse Array. Among these genes, 111 transcription factors
were selected and homemade SuperArray was made on which these transcription
factors were blotted. Total RNA was purified from the GV and MII
oocytes, mESCs, STO and NIH3T3 cells with ArrayGradeTM Total RNA
Isolation Kit, cRNA was synthesized and labeled with biotinylated-UTP
and TrueLabeling-AMPTM 2.0, and finally hybridized with SuperArray. After
selection of a list of common or cell-specific transcription factors, mRNA
expression was confirmed by RT-PCR. The common or specific factors expressed
in oocytes and/or stem cells were identified.
RESULTS: From the SuperArray data, we can select 7 oocyte-specific, 3
mESC-specific, and 6 genes common between oocytes and mESCs. Interestingly,
mESC-specific genes were not found. After confirmation by RT-PCR,
expression of Lhx8, Nobox, Isl2, Nr2e1 and H1foo mRNA was detected only
in oocyte. Three (Lhx8, Nobox and H1foo) were already well-known oocytespecific
genes that have an impact upon oocyte maturation and female
fertility, while two genes (Isl2 and Nr2e1) were firstly confirmed. Exciting
finding was the list of genes detected in both oocytes and mESCs, such as
Klf11, Spic, Spry1, Gbx2, Ell3, and Grhl1, but not in differentiated cells.
Among them, Gbx2 has already reported to have LIF/Stat3-mediated selfrenewal
capacity in stem cells, and its gain of function facilitates reprogramming
of differentiated cells.
CONCLUSIONS: Taken together, list of transcription factors of the present
study per se would be insightful in studying oocyte maturation as well
as reprogramming for stem cell research. Identification of potential key or
master regulators of reprogramming and their functional characteristics in
the oocytes and/or ESCs will not only provide novel way for studying reprogramming
and stem cell characteristics, but also greatly contribute to generating
induced pluripotent stem (iPS) cells that are more relevant and safer
for the future cell replacement therapy.
Supported by: This research was Supported by the Basic Science Research
Program through the National Research Foundation of Korea (NRF) funded
by the Ministry of Education (2009-0093821).
P-366 Tuesday, October 20, 2015
NURSING
ESTRADIOL AND PROGESTERONE SUPPLEMENTATION IN
UNITED STATES DONOR OOCYTE AND EMBRYO PROGRAMS:
A PRACTICE SURVEY OF FACTORS INFLUENCING ROUTE
AND TIMING OF ADMINISTRATION. K. R. Hammond,
N. A. Cataldo, M. P. Steinkampf. Alabama Fertility Specialists, Birmingham,
AL.
OBJECTIVE: Estradiol (E) and progesterone (P) supplements for recipients
of donor oocytes and embryos are available in various formulations.
The objective of this study was to survey practice patterns among donoroocyte
programs in regard to timing and route of hormone administration.
DESIGN: Cross-sectional survey.
MATERIALS AND METHODS: A one-page questionnaire was offered to
attendees at a 2015 national third-party reproduction symposium. Descriptive
statistics were compiled, and agreement between practices expressed
by Cohen’s kappa and Bowker’s test.
RESULTS: There were 48 unique responses from 22 states; 2 self-identified
as academic programs and 43 as private practices. The majority of
respondents (85%) were nurses. Programs annually performed 40 (median)
fresh donor egg cycles (range 1-400), 25 (0-150) cycles using cryopreserved
(cryo) embryos from donor oocytes, and 8 (0-24) cycles using
cryo donor oocytes.The modal preferred routes of administration were
oral for E and intramuscular (IM) for P, with little difference in preference
for route among fresh and cryo cycle types. In fresh cycles, oral E was
preferred by 54% over IM, vaginal, patch, and combinations for its ease
(92%), convenience (62%), effectiveness (65%), and cost (58%). In fresh
cycles, IM P was preferred by 66% over vaginal and combinations for its
effectiveness (97%), while programs preferring vaginal P (29%) cited its
effectiveness (71%) and ease (57%). There was strong concordance between
fresh and cryo ET cycles as to preferred E route (k¼0.96), but
only fair concordance between cycle types as to preferred P route
(k¼0.51). Changes in preference for IM in the last year were reported to
be uncommon for either E or P. Patients’ preferred E and P routes were
honored by 24% and 45% of respondents, respectively. Modal hormone
supplement discontinuation was at 10 weeks’ gestation for both E (52%)
and P (47%). Time of E discontinuation ranged from 6-13 weeks, while
P discontinuation ranged from 8-13 weeks. P was continued longer than
E by 11 programs, but shorter than E by none (p¼0.01).
CONCLUSIONS: Route of administration and timing of recipient E and P
supplementation in US third-party reproduction programs are moderately
consistent, with the modal prescription of oral E and IM P until 10 weeks’
gestation. The greatest divergence in practice was found between the IM
and vaginal route for P. While little difference was seen within programs
in E or P route between fresh and cryopreserved embryo/oocyte cycles, programs’
preferences appear entrenched.
e230 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

P-367 Tuesday, October 20, 2015
GENETIC COUNSELING
DEVELOPMENT, VALIDATION AND CLINICAL USE OF AN
EXPANDED PAN-ETHNIC PRECONCEPTION CARRIER GENETIC
SCREENING TEST IN ASSISTED REPRODUCTIVE TECHNOL-
OGY PATIENTS AND DONORS. J. Martin, a B. Rodriguez-Iglesias, a
A. San, b Y. Yuting, b J. Jimenez, a Q. Li, c H. Du, b C. Simon. d a Carrier Genetic
Test, Igenomix, Valencia, Spain; b Binhai Genomics Institute, Tianjin,
China; c Binhai Genomics Institute, Yantian, China; d FIVI Universidad de Valencia,
Igenomix-Valencia, Paterna, Spain.
OBJECTIVE: Development, preclinical validation and accuracy assessment
of NGS-based test for expanded pan-ethnic preconception carrier genetic
screening test for use in assisted reproductive technology patients
and donors.
DESIGN: Retrospective analysis of results obtained from 2,570 analysis
for 1,170 individuals from the gamete donor programs; 1,124 individuals corresponding
to the partner of the patient receiving the donated gamete; and
276 individuals from 138 couples seeking ART using their own gametes.
In addition we performed a short comparative study between our NGS-based
test and a commercially available array-based test when patients moved on to
our screening test.
MATERIALS AND METHODS: NGS of 549 recessive and X-linked
genes involved in severe childhood phenotypes reinforced with 5 complementary
tests covering high prevalent mutations not detected by NGS. Use
of a blinded matching system to achieve a very low risk match for patientdonor.
RESULTS: Preclinical validation included 48 DNA samples carrying
known mutations for 27 genes resulting in a sensitivity of 99%. In the clinical
dataset, 2,161 (84%) samples tested positive with an average carrier burden
of 2.3 per sample. Five percent of the couples using their own gametes were
found to have pathogenic variants conferring high risk for 6 different diseases.
These high-risk couples and patients received genetic counselling
and recommendations for preimplantation genetic diagnosis. For patients
receiving gamete donation, we applied a genetic testing and blinded matching
system to avoid high risk combinations regardless of their carrier burden.
For female donors, 1.94% were positive for X-linked conditions; they
received genetic counselling and were discarded. Interestingly, during our
unintended platform-based test comparative study we detected discordant results,
including an array-based test result positive for the common mutation
in the CF gene (F508del); a separate blinded test was performed always
confirmed NGS-based results.
CONCLUSIONS: We have developed a NGS-based expanded carrier genetic
screening test that, combined with our matching system and genetic
counselling, constitutes a powerful tool to avoid more than 600 Mendelian
diseases in the offspring of patients undergoing ART, achieving enhanced
precision than alternative array-based assays.
P-368 Tuesday, October 20, 2015
ASSESSING TRENDS IN EMBRYO GENDER AFTER PGS:
ARE THERE MALE OR FEMALE PROGENY-DOMINANT
COUPLES? J. Gingold, a,b M. C. Whitehouse, c J. A. Lee, c
T. A. VanWort, c M. Daneyko, c T. Mukherjee, c,a A. B. Copperman. c,a a Obstetrics,
Gynecology and Reproductive Science, Icahn School of Medicine at
Mount Sinai, New York, NY; b Obstetrics/Gynecology and Women’s Health
Institute, Cleveland Clinic Foundation, Cleveland, OH; c Reproductive Medicine
Associates of New York, New York, NY.
OBJECTIVE: Normal spermatogenesis produces equal frequencies of
male and female sperm(1). Couples using in vitro fertilization (IVF) and preimplantation
genetic screening (PGS) for gender selection have expressed
concern that their embryos may have a gender bias. The study aims to identity
whether clinical data support the existence of predominantly male or female
embryo-producing couples.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: IVF couples (n¼116) treated between
February, 2006 -November, 2014 who had R10 embryos (range 10-34)
screened by PGS were analyzed by a two-sided binomial test to calculate
the probability (p) of a comparably or more extreme embryo gender imbalance
due to chance. The male to female embryo ratio was assumed to be
1:1. P-values were adjusted for the false discovery rate (FDR) by the Benjamini-Hochberg
method with significance at p

P-370 Tuesday, October 20, 2015
DOES THE PARENTAL ORIGIN OF TRANSLOCATION IMPACT
RATES OF ABNORMAL EMBRYOS? C. K. Celestine, a
M. D. Werner, b J. M. Franasiak, b C. R. Juneau, b N. R. Treff, b
T. Molinaro, b R. T. Scott. b a JSUMC, Neptune, NJ; b RMA, NJ, NJ.
OBJECTIVE: Phenotypically normal patients with balanced translocations
often seek pre-implantation genetic diagnostic testing to prevent transmission
of an unbalanced translocation to offspring. This is a unique
subgroup of patients who are at increased risk for suboptimal outcomes
including clinical pregnancy loss which may be associated with chromosomally
abnormal embryos. The goal of the present analysis is to determine
whether the parental origin of the translocation impacts the degree of embryonic
aneuploidy.
DESIGN: Retrospective Cohort Study.
MATERIALS AND METHODS: Couples undergoing IVF with Preimplantation
Genetic Diagnosis at our center were identified where one partner
carried a balanced translocation. All embryos underwent trophectoderm biopsy
with single nucleotide polymorphism microarray-based screening for
detection of unbalanced translocations and 24-chromosome aneuploidy
screening. Possible outcomes were recorded and included: Balanced/
Euploid, Balanced/Aneuploid, Unbalanced/Euploid, Unbalanced/Aneuploid.
The term ‘balanced’ refers to an embryo that has either a balanced
translocation or no translocation as it was not possible to distinguish these
outcomes. Abnormal embryos comprised the groups of embryos which
were either aneuploid and/or unbalanced. Demographics and outcomes of
maternal and paternal translocation carriers were compared using chi squared
and Mann Whitney U tests. Logistic regression utilizing Generalized Estimating
Equations was used to control for between patient differences and
reduce confounding from maternal and paternal age differences.
RESULTS: 1501 embryos from 271 patients were included in this study
with 36.98% of tested embryos found to be normal or balanced.
734(48.9%) embryos were analyzed from couples with maternal translocation
carriers and 767 (51.1%) from paternal translocation carriers. There
was no significant difference in patient or partner ages or the number of
mean embryos between both groups. When controlling for patient identity,
patient age and partner age, maternal carriers of a chromosomal translocation
had an increased risk of having an unbalanced embryo (OR 1.36; 95%CI
1.04-1.77; p¼0.02). However, due to other aneuploidies, the probability of
being normal for both aneuploidy and translocation screening was equivalent
between maternal and paternal carriers (OR 0.87; 95%CI 0.69-1.15;
p¼0.38).
CONCLUSIONS: Significant difference exists between the number of unbalanced
embryos when translocation carrier origin is paternal compared to
maternal. However, no difference exists in the number of normal embryos.
.
Table 1: Cycle Characteristics.
Paternal (n¼138)
Mean SD
Maternal (n¼133)
Mean SD
p value
Patient age 36.074.71 35.974.46 0.77
Partner age 37.325.17 37.184.86 0.92
Biopsied Embryos 5.584.48 5.563.42 0.41
Embryos N (%) N (%)
Abnormal 475 (61.93) 471 (64.17) NS
Normal 292 (38.07) 263 (35.83) NS
CONTRACEPTION/FAMILY PLANNING
P-371 Wednesday, October 21, 2015
PHASE III STUDY OFA NEW, LOW-DOSE LEVONORGESTREL IN-
TRAUTERINE CONTRACEPTIVE SYSTEM (LNG-IUS19.5MG)
OVER 5 YEARS OF USE: EVALUATION OF BLEEDING AND
DYSMENORRHEA. T. Faustmann, a A. Nelson, b K. Rosen, c
T. Schmelter, a D. Apter. d a Bayer Pharma AG, Berlin, Germany; b Los Angeles
Biomedical Research Institute, Los Angeles, CA; c Bayer HealthCare
Pharmaceuticals, Whippany, NJ; d Sexual Health Clinic, Helsinki, Finland.
OBJECTIVE: To evaluate the bleeding profile and dysmenorrhea associated
with LNG-IUS19.5mg over 5 years of use.
DESIGN: Phase III study. Women used a new, intrauterine contraceptive
system (IUS; LNG-IUS19.5mg) for 3 years; thereafter they had the option
to continue using it in an extension study for up to 2 more years (5 years
in total).
MATERIALS AND METHODS: Nulliparous or parous women aged 18-
35 years with regular menstrual cycles (21-35 days) were recruited.
RESULTS: The full analysis set (n¼1452; mean age 27.1 years; 39.5%
nulliparous) included all women for whom at least one LNG-IUS19.5mg
placement attempt was made. Between the 1st and 2nd 90-day reference intervals
(RIs), the mean number of bleeding/spotting days decreased with
LNG-IUS19.5mg use (39.7 to 21.1 days); the number gradually declined
thereafter (9.3 days at the 20th 90-day RI). Between the 1st and 20th 90-
day RIs, the mean number and length of bleeding/spotting episodes
decreased from 3.6 to 2.2 and 10.14 to 4.15 days, respectively. Bleeding patterns
(World Health Organization criteria) are shown in the Table.
Table. World Health Organization criteria for menstrual bleeding patterns.
Amenorrhea*
(%)
Infrequent
bleeding y
(%)
Frequent
bleeding z
(%)
1st 90-day RI 0.2 10.4 24.6 60.2
2nd 90-day RI 5.1 21.3 10.2 16.8
4th 90-day RI 12.7 26.7 4.2 7.4
(end of Year 1)
12th 90-day RI 20.1 25.7 2.2 2.2
(end of Year 3)
20th 90-day RI
(end of Year 5)
22.6 26.4 2.3 1.5
Prolonged
bleeding x
(%)
* No bleeding/spotting throughout RI; y One or two bleeding/spotting episodes;
z >5 bleeding/spotting episodes; x Bleeding/spotting episodes lasting
>14 days.
Between the 1st and 20th 90-day RIs, the mean number of days with
dysmenorrhea (any severity) declined (14.7 to 3.3 days). The percentage of
women reporting R1 day of moderate/severe dysmenorrhea declined over
time with LNG-IUS19.5mg use (68.8% to 16.9% from the 1st to 20th 90-
day RI, respectively). Thirteen women (0.9%) discontinued due to dysmenorrhea
and 76 women (5.2%) due to bleeding/non bleeding problems.
CONCLUSIONS: Women using a next-generation,low-dose IUS (LNG-
IUS19.5mg) generally experienced shorter, less-frequent bleeding over
time. By the end of 5 years of use, almost a quarter of women were amenorrheic;
a further quarter had infrequent bleeding. In addition, dysmenorrheasubstantially
decreased over time.
Supported by: Study and abstract funded by Bayer HealthCare.
P-372 Wednesday, October 21, 2015
INTERPREGNANCY INTERVAL AND SUBSEQUENT PREG-
NANCY OUTCOMES AFTER DILATION AND
EVACUATION. M. K. Kuwahara. University of Hawaii - OGBYN, Honolulu,
HI.
OBJECTIVE: There is little published on the ideal interconceptual period
following a dilation and evacuation (D&E). The purpose of this study is to
compare outcomes for pregnancies conceived 6 months after D&E.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: The University of Hawaii Abortion
Database was used to identify women who underwent D&E between 14
and 26 weeks gestation and were readmitted with a subsequent pregnancy between
2008 to 2014. The first subsequent pregnancy after D&E was identified
through International Classification Diagnosis-9 (ICD-9) codes. Additional
demographic and clinical data were gathered through chart review. The primary
outcome was the rate of preterm deliveries less than 37 weeks gestational
age. Secondary outcomes included cervical insufficiency,
placentation abnormalities, intrauterine growth restriction, postpartum hemorrhage,
and mode of delivery. Chi-square test and student t-tests were used
to compare categorical and continuous variables, respectively. To demonstrate
a difference of 8.8% in preterm delivery with 80% power and a significance
of 0.05, a sample size of 246 patients in each study arm was required.
RESULTS: A total of 737 D&Es were performed during the study interval
and 214 subsequent pregnancies were identified. Outcomes were available
for 184 (86%) of these pregnancies, of which 90 (49%) resulted in live births
>20 weeks gestation. Among the live births, 32 (36%) were conceived 6
e232 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

months from the time of D&E. No differences in demographic or clinical variables
were found between the two groups, nor between those who delivered
preterm or term. The incidence of preterm delivery at less than 37 weeks
gestational age was 9.4% (n¼3) with an interpregnancy interval 6 months
(p¼0.168). No differences in preterm delivery below 34 weeks, postpartum
hemorrhage, placentation abnormalities, intrauterine growth restriction, cervical
insufficiency, or mode of delivery emerged between groups.
CONCLUSIONS: Adverse pregnancy outcomes were not higher in the
group of women who became pregnant less than 6 months after D&E
compared to those who waited longer than 6 months to become pregnant.
Larger studies are needed to confirm these findings and provide evidencebased
recommendations for women desiring pregnancy after D&E.
P-373 Wednesday, October 21, 2015
NO SCALPEL VASECTOMY: 20 YEAR OUTCOMES UTILIZING
COMBINED CAUTERY, CLIP AND FACIAL
INTERPOSITION. K. Chiles, M. Feliciano, M. Goldstein. Urology,
Weill Cornell Medical College, New York, NY.
OBJECTIVE: Vasectomy failure is common (1-10%) and a major cause of
malpractice suits. No-scalpel vasectomy (NSV) is a minimally invasive technique,
but failure rates are dependent on occlusion techniques. In this report,
a combination of 4 occlusion techniques were employed.
DESIGN: Retrospective chart review.
MATERIALS AND METHODS: All men who underwent NSV by a single
surgeon within the past 20 years were included. For NSV the vas was
delivered through a single midline puncture hole under local anesthesia,
excluding vasal vessels and nerves. The vas was hemi-transected in two places
½-1 cm apart using thermal cautery. Intraluminal cautery was performed
on both ends for a distance of 1cm until smoke was observed and the wire tip
then rotated for 10 seconds to ensure a 360o burn. A hemoclip was lightly
placed on the testicular end to prevent sperm leakage until cautery causes
a permanent seal. The abdominal end was completely transected and allowed
to retract into the vasal sheath. The sides of the sheath were grasped with a
hemostat and sealed over the abdominal end with a hemoclip, accomplishing
fascial interposition. The intervening vas segment was excised and the ends
were dabbed with Betadine before retraction into the scrotum. The contralateral
vas was accessed through the same puncture hole and occluded identically.
Post-vasectomy semen analysis (PVSA) is requested after 6 weeks
or 15 ejaculations, and a PVSA after the same interval.
RESULTS: 646 vasectomies were performed over 20 years. Mean patient
age in years was 41.55.8 and partner 38.73.7. Median # of children was 3.
377/646 men (58%) of men had at least one PVSA performed a median of 55
days after NSV. 177/377 (47%) required a second PVSA, of which 138/177
(78%) complied. No pregnancies were reported. One patient had redo NSV
for persistent non-motile sperm on PVSA. One abscess required I&D. Two
hematomas (2-3 cm) were managed conservatively. No chronic (>6 months)
post-vasectomy pain was reported.
CONCLUSIONS: Vasectomy failure can be virtually eliminated utilizing
a combination of 4 different occlusion techniques: 1) intraluminal cautery for
10 seconds; 2) testicular end clip occlusion; 3) facial interposition; and 4)
resection of a ½ - 1 cm segment. This approach can be employed using the
no-scalpel technique through a single midline puncture hole. Delivery of
vas cleanly from its sheath, excluding vasal vessels and nerves may minimize
the incidence of chronic post-vasectomy pain. This belt, suspenders, rope and
wire approach to vasal occlusion minimizes failure and complications.
P-374 Wednesday, October 21, 2015
UNDERSTANDING HOW WOMEN WITH CYSTIC FIBROSIS
MAKE DECISIONS ABOUT FAMILY PLANNING. S. A. Traxler, a
C. A. Schreiber, a V. Chavez, a D. Hadjiliadis, b C. Mollen. c a Obstetrics
and Gynecology, University of Pennsylvania, Philadelphia, PA; b Pulmonary,
Allergy, and Critical Care Division, University of Pennsylvania, Philadelphia,
PA; c Pediatrics/Division of Emergency Medicine, The Children’s Hospital
of Pennsylvania, Philadelphia, PA.
OBJECTIVE: Based on a few reports, contraceptive use in women with
cystic fibrosis (CF) appears to be well below that seen in the general population
of women in the United States. Given that more women with CF are
reaching reproductive age and pregnancy has health complications for this
population, our objective is to explore contraceptive decision-making and
investigate ways to meet contraceptive needs for women with CF.
DESIGN: Qualitative methods via one-on-one, semi-structured interviews.
MATERIALS AND METHODS: We conducted an in-depth interview
study. Participants were included if they were female between the ages of
18 and 45 with a diagnosis of CF. Purposive sampling was used to balance
the sample age range until thematic saturation was reached. Interviews
were conducted in person or on the phone by a trained interviewer. The interview
guide included the domains of pregnancy attitudes, contraception attitudes
and perceptions of fertility. Interviews were audio-taped and then
transcribed, coded independently by two study team members and analyzed
using a modified grounded theory approach.
RESULTS: Twenty-four women were interviewed. Participants reported
pregnancy intentions to be influenced by a sense of urgency to become pregnant
due to a shortened life span and reported that pregnancy intentions are
impacted by personal health status as well as ethical issues concerning impact
of CF consequences on a potential child. Participants reported misconceptions
about contraception such as interactions with other medications and pervasive
skepticism about long-acting reversible contraceptive (LARC) methods. Many
reported that beliefs around CF-related infertility led to non-use of highly
effective methods of contraception. While medical providers were described
as valuable participants in shared decision-making about family planning
and contraception, many reported that providers lack knowledge and miss critical
periods for communication, such as during transitions of care.
CONCLUSIONS: This sample of women with CF described thoughtful
considerations about pregnancy intentions and desire pregnancies to be
planned during times when health is optimal, but misconceptions about
fertility and contraception interfere with their ability to use highly effective
methods of contraception. Medical providers, valued for their expertise in
shared decision-making, can bridge this gap, facilitate fully informed decisions
about contraception, and determine the best recommendations for
women with CF around childbearing.
P-375 Wednesday, October 21, 2015
RELATIONSHIP BETWEEN COPPER IUD COMPLICATIONS AND
ULTRASOUND FINDINGS. S. Fadiloglu, a B. Dilbaz, b E. Fadiloglu, a
S. Dilbaz. a a Etlik Zubeyde Hanim Kadin Hastaliklari ve Dogum Hastanesi,
Ankara, Turkey; b Adiyaman University School of Medicine, Adiyaman,
Turkey.
OBJECTIVE: IUD is a widely used long acting contraceptive, however the
side-effects related to IUD lead to method discontinuation. The aim of this
study is to evalute the relationship between the most common side-effects
of IUD use such as dysmenorrhea, menorrhaghia, abdominal cramping and
the position of the IUD device and uterine dimensions evaluated by transvaginal
ultrasonography.
DESIGN: Prospective Trial.
MATERIALS AND METHODS: Two hundred and eighty four patients
who had IUD insertion at the Family Planning Clinic of a tertiary center
were evaluated at insertion, 6th and 12th week after the insertion. Demographic
characteristics, medical history, symptoms and findings of the gynaecological
examination were recorded. Trnasvaginal ultrasonographic
measurement of uterine dimensions, the distance between the tip of the
Cu-IUD and fundus, endometrium and myometrium were measured in order
to evaluate the displacement of the IUD and the relationship between
displacement and the symptoms and complaints were investigated.
RESULTS: No pregnancy occured during the study period and one patient
had the IUD removed due to menorrhgia. The patients who complained
of menorrhgia had statistically significantly shorter uterine length
in comparison to the ones who did not have this complaint (54.27 6.11
vs 60.25 10.52 mm p¼0.02). There was no difference in the uterine length
between the patients who had dysmenorrhea and who had not at 12 weeks
(59,60 10,25 vs 60,33 10,68mm, p ¼ 0.71 ). There was a significant
difference between the IUD tip-fundus, endometrium and myometrium
measurements of the patients who experienced abdominal cramping in
comparison to the ones who had not after 3 months confirming the downward
IUD movement. ( Endometrium; 0,29 0,72 vs 0,45 0,35 p ¼
0.02, Myometrium; 1,25 1,39 vs 2,38 2,26 p

P-376 Wednesday, October 21, 2015
BARRIERS AND MYTHS THAT LIMIT THE USE OF INTRAUTER-
INE CONTRACEPTION IN NULLIPAROUS WOMEN: A SURVEY
OF BRAZILIAN GYNECOLOGISTS. M. M. Carneiro, a J. Lira, b
A. L. Silva-filho. a a Obstetrics and Gynecology, Federal University of Minas
Gerais, Belo Horizonte, Brazil; b Department of Adolescent Gynecology, Instituto
Nacional de Perinatologia, Universidad Nacional Autonoma de
Mexico UNAM, Mexico City, Mexico.
OBJECTIVE: To understand the extent to which barriers and misperceptions
about intrauterine contraception (IUC) remain among Brazilian gynecolgysts,
particularly for nulliparous women.
DESIGN: Prospective web based survey.
MATERIALS AND METHODS: An online survey was developed to
assess Brazilian gynecologists’ knowledge and attitudes towards IUC. Data
collected included demographic and professional data, main barriers when
considering IUC for women in general, and or nulliparous women, attitudes
towards inclusion of IUC in contraceptive counselling, opinions on what
could increase IUC prescription for nulliparous women. A question
regarding the knowledge about the World Health Organization Medical
Eligibility Criteria (WHO MEC) was also included in the survey.
RESULTS: 101 respondents completed the survey. The insertion rate in
nulliparous women was 79.2%. Brazilian gynaecologists considered IUC
in counseling or provided it upon request more for parous than for nulliparous
women (p

DESIGN: A prospective three-group randomized controlled trial was performed
with three assessment points. Participants (n ¼265) were randomly
allocated to the intervention group (IG, fertility awareness video), control
group 1 (CG1, no intervention) and control group 2 (CG2, work-family conflict
video). All participants were assessed before the intervention (T0),
immediately after (T1) and one week after (T2).
MATERIALS AND METHODS: Data were collected Oct-Dec 2014 from
Portuguese University students. After excluding participants who were married,
had children or disclosed a fertility problem, the final sample had 254
participants (96 IG, 76 CG1, and 82 CG2). The questionnaire included sociodemographic
and fertility knowledge questions. The video gave information
about women’s fertility decline, infertility definition and risk factors, probability
of getting pregnant according to women’s age. Mixed Anovas for
repeated measures tested interaction, time and group effects on fertility
knowledge variables.
RESULTS: The large majority (83%) of participants (84% female, Mage ¼
19.7) expressed a desire to have children. There were no pretest differences
among groups concerning sociodemograhic characteristics. A significant interaction
was found between assessment time and group regarding knowledge
variables (infertility definition, women’s fertility decline, probability of pregnancy
depending on age, risk factors). Medium effect sizes of 0.33 were found
between IG and CG2 and 0.39 between IG and CG1. The IG had significantly
higher knowledge than both control groups in all variables between T0 and T1
and T0 and T2, but not between T1 and T2, indicating that knowledge remains
a week later. There were no significant differences over time within each one of
control groups, excepting for women’s most fertile age, women’s fertility
decline, and risk factors. However, these differences were smaller (higher
eta for IG) and in some cases indicated a decrease in knowledge.
CONCLUSIONS: Results Supported the efficacy of a low-cost intervention
in increasing fertility knowledge. Given the increasing childbearing
postponement and the reduced fertility knowledge of young adults who
desire to have children, this intervention can be of great use in western countries
to enable an informed and conscious decision-making process. Future
research should focus on wider longitudinal studies.
Supported by: Study Supported by FEDER/COMPETE and FCT (PTDC/
MHC-PSC/4195/2012 and SFRH/BPD/85789/2012).
MALE FACTOR
P-380 Wednesday, October 21, 2015
COMPARISON BETWEEN THE SEMEN PARAMETERS ACCORD-
ING TO 1999 WORLD HEALTH ORGANIZATION CRITERIA AND
2010 WHO CRITERIA IN MALES OF SUBFERTILE COUPLES IN
KOREA. E. Lee, a J. Shin, a,b C. Joo, c H. Kim, b,a J. Lee, d,b B. Jee, d,b
C. Suh, b,a S. Kim. a,b a Seoul National University Hospital, Seoul, Korea, Republic
of; b Seoul National University College of Medicine, Seoul, Korea, Republic
of; c Maria Fertility Hospital, Seoul, Korea, Republic of; d Seoul
National University Bundang Hospital, Seoul, Korea, Republic of.
OBJECTIVE: To assess the effects of the new 2010 WHO criteria for
semen analysis in comparison with the previous 1999 WHO criteria.
DESIGN: Retrospective study.
MATERIALS AND METHODS: We retrospectively reviewed the results
of semen analyses of men of 660 subfertile couples from April 2014 until
December 2014. The study was performed at the 6 fertility clinics in Korea.
All semen parameters were reviewed and compared according to 1999 WHO
and 2010 WHO criteria.
RESULTS: The mean values of semen analyses in 660 men of subfertile
couples were 3.6ml of volume, 102.3x10^6/mL of concentration, 54.9% of
motility and 4.5% of strict morphology. Two hundred and seventeen men
(32.8%) who were considered to be infertile by the 1999 WHO criteria
were reevaluated to be normal according to 2010 WHO reference values.
Of the men with semen analyses, 11.2%, 3.0%, 16.7% and 42.7% of men
were reclassified as normal for volume, concentration, motility and strict
morphology, respectively.
CONCLUSIONS: About one-third of men of subfertile couples who were
thought to be infertile by the old criteria now considered normal by the new
2010 WHO criteria. Many men who were considered infertile according to
the old criteria now should be reevaluated normal by the new criteria. Strict
morphology was the main cause of abnormal semen parameters.
References:
1. Baker K, Li J, Sabanegh E Jr, Edmund Sabanegh Jr. Analysis of
semen parameters in male referrals: impact of reference limits, stratification
by fertility categories, predictors of change, and comparison
of normal semen parameters in subfertile couples. Fertil Steril
2015;103(1):59-65.
2. Catanzariti F, Cantoro U, Lacetera V, Muzzonigro G, Polito M. Comparison
between WHO (World Health Organization) 2010 and WHO
1999 parameters for semen analysis - interpretation of 529 consecutive
samples. Arch Ital Urol Androl. 2013 Sep 26;85(3):125-9.
Supported by: This research was Supported by a fund(2014ER630500) by
research of Korea Centers for Disease Control and Prevention.
P-381 Wednesday, October 21, 2015
COMPARATIVE PROTEOMIC ANALYSIS INDICATES UNDEREX-
PRESSION OF MOLECULAR CHAPERONES IN SPERMATOZOA
OF INFERTILE MEN. R. Sharma, a A. Agarwal, a
D. Durairajanayagam, a,b L. Samanta, a,c M. Assidi, d S. Gupta, a
E. S. Sabanegh. e a Center for Reproductive Medicine, Cleveland Clinic,
Cleveland, OH; b Physiology, Universiti Teknologi MARA, Sungai Buloh,
Malaysia; c Redox Biology Laboratory, School of Life Science, Ravenshaw
University, Orissa, India; d Center of Excellence in Genomic Medicine,
King AbdulAziz University, Jeddah, Saudi Arabia; e Urology, Cleveland
Clinic, Cleveland, OH.
OBJECTIVE: Functional spermatozoa are produced by a highly regulated
process of sperm maturation. Human semen used in assisted reproductive
procedures is traditionally separated by density gradient to provide morphologically
and functionally viable spermatozoa. Often these spermatozoa are
unable to achieve fertilization. Chaperones are involved in processing of
spermatozoal proteins required for motility, zona penetration and stress
response. We therefore studied a comparative proteome profile of these proteins
to understand their role in sperm function.
DESIGN: Protein profiles from spermatozoa in fertile and infertile men
were compared.
MATERIALS AND METHODS: Proteins were extracted and separated
by 1-D gel. Bands were digested with trypsin and analyzed on a LTQ-Orbitrap
Elite hybrid mass spectrometer system. Protein identification was done
using Mascot (Matrix Science, London, UK; version 2.3.02), SEQUEST
(Thermo Fisher Scientific, San Jose, CA, USA; version 1.4.0.288) and X!
Tandem (TheGPM, thegpm.org; version CYCLONE (2010.12.01.1).
Mascot, Sequest and X! Tandem were set up to search the human reference
with database assuming trypsin as the digestion enzyme. Functional annotations
of proteins were obtained using bioinformatic tools and pathway databases.
RESULTS: Proteomic analysis revealed a total of 35 focus chaperone
proteins involved in post-translational modification and protein folding
that were underexpressed in the spermatozoa of infertile men. The pivotal
chaperones downregulated were HSPA4L, HSPA 1A/1B, mitochondrial
HSP90, subunits of the T-complex proteins 1 (Tcp1a, Tcp 1g, Tcp1d,
Tcp 1ε, Tcp 1h, Tcp 1z) as well as the sperm surface protein 17 responsible
for zona binding. The BAG family regulator 5 was upregulated in these patients,
indicating a negative regulation of oxidative stress and ubiquitination.
CONCLUSIONS: Our results suggest that there is a differential expression
of sperm chaperones between fertile and infertile males. These may be
responsible for defective expression of sperm surface proteins leading to
failed fertilization.
P-382 Wednesday, October 21, 2015
SEMEN PARAMETERS DO NOT HAVE A CLINICALLY SIGNIFI-
CANT EFFECT ON ICSI CYCLE OUTCOMES: AN ANALYSIS US-
ING 2,861 SHARED OOCYTE DONOR RECIPIENT CYCLES TO
CONTROL FOR EGG FACTORS. G. Patounakis, a K. Devine, b
B. W. Whitcomb, c A. DeCherney, a M. J. Levy. b a National Institutes of
Health, Bethesda, MD; b Shady Grove Fertility, Rockville, MD; c University
of Massachusetts, Amherst, MA.
OBJECTIVE: The true impact of male factor on ICSI cycle outcomes is
complicated by concurrent female variables. Our objective was to determine
the effect of semen parameters on ICSI cycle outcomes using a cohort of
shared oocyte donation cycles to completely control for oocyte factors.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Anonymous oocyte donation cycles
from January 2010 through December 2014 using non-donor sperm
FERTILITY & STERILITY Ò
e235

were included. Only the first cycle was included for recipients with multiple
cycles. Prewash volume, concentration, motility, and total motile
sperm count from the ICSI specimen and morphology from the baseline
specimen were analyzed. The change in morphology criteria to WHO
2010 in September 2010 was accounted for in the statistical model.
The primary outcome was percent top quality blastocyst (AA, AB,
BA) conversion on the last day of embryo evaluation per inseminated
oocyte. Secondary outcomes were fertilization, overall blastocyst conversion,
implantation, clinical pregnancy, and ongoing pregnancy. Generalized
estimating equations (GEE) were used to account for donor
oocyte sharing and repeated donor cycles. Outcomes were controlled
for donor age, oocytes retrieved, recipient female age, male age, recipient
endometrial thickness, embryo stage at transfer, embryo quality at
transfer, and number of embryos transferred.
RESULTS: There were 2861 recipients with inseminated oocytes in the
cohort of 1057 oocyte donors undergoing 1659 retrievals. The percent of
oocyte donors shared with 3, 2, and 1 recipients were 57%, 26%, and 17%,
respectively. Only WHO 2010 normal morphology showed a significant association
with the primary outcome. For every 1% increase in WHO 2010
normal morphology the top quality blastocyst conversion increased by
0.4% (p¼0.04). A 6% higher top quality blastocyst conversion was observed
when morphology was >2% relative to %2% (p¼0.001). No other effects of
normal morphology for both WHO 2010 criteria and pre-WHO 2010 were
statistically significant. Pre-wash motility was significantly associated with
secondary outcomes. For every 10% increase in pre-wash motility, fertilization
increased by 0.7% (p¼0.001), implantation increased by 1.6%
(p¼0.004), odds of clinical pregnancy increased by 1.07 (p¼0.004), and
odds of ongoing pregnancy increased by 1.06 (p¼0.01). Pre-wash motility
did not have a significant effect on top quality blastocyst conversion or overall
blastocyst conversion. All other semen parameters lacked statistically significant
associations.
CONCLUSIONS: Sperm motility and morphology have statistically
significant but clinically insignificant effects on ICSI outcomes. Couples
with male factor infertility can be reassured that currently measured
bulk semen parameters are not clinically useful outcome predictors of
ICSI.
Supported by: This work was Supported, in part, by the Program in Reproductive
and Adult Endocrinology, NICHD, NIH, Bethesda, MD.
P-383 Wednesday, October 21, 2015
ACHIEVING PREGNANCY IN MEN WITH ISOLATED
ASTHENOSPERMIA. C. Deibert, K. M. Zeeck, J. Sandlow. Medical
College of Wisconsin, Milwaukee, WI.
OBJECTIVE: Men with isolated asthenospermia present a clinical challenge
regarding treatment. To date no studies have evaluated the clinical outcomes
for couples with isolated asthenospermia.
DESIGN: This is a retrospective chart review study.
MATERIALS AND METHODS: This is an IRB-approved review of infertile
men who presented to our Reproductive Medicine Center with isolated
asthenospermia, defined as motility

DESIGN: A prospective study.
MATERIALS AND METHODS: Sperm from individuals diagnosed as
normozoospermia (n¼43) or asthenozoospermia (n¼32) were collected.
Liquid chromatography-tandem mass spectrometry were used to detect
levels of total 5-methylcytosine (5-mdC) and 5-hydroxymethylcytosine
(5-hmdC) in sperm DNA as well as N6-methyl-adenosine (m6A) and
5-methylcytosine (5-mC) in RNA. Levels of the above four indicators
were compared. Linear and logistic regression models were performed to
determine the factors affecting IVF outcomes, adjusting for confounders.
RESULTS: Demographic characteristics including fertilization rate (FR),
cleavage rate (CR), good quality embryo rate (GQER) and pregnancy rate
(PR) were similar between asthenozoospermia (AS) group and normozoospermia
(NM) group, except for that sperm motility and concentration
were decreased in AS group. The level of 5-mdC (representing global
DNA methylation status) were found to be significantly decreased in AS
group (3.430.65% versus 3.730.57% in NM group, P¼0.042). Linear
regression analysis revealed that 5-mdC level was positively correlated
with FR (b 8.11, 95% CI (1.02,15.20), P¼0.043) and GQER in AS group
(b 16.18, 95% CI (2.294,30.067), P¼0.024), but was not related to cleavage
rate (P¼0.084). In contrast, level of 5-hmdC (standing for DNA demethylation
status) was negatively correlated with FR in AS group (b -25.12, 95% CI
(-49.82,-0.42), P¼0.047)). The other two indicators were found to have no
influence on IVF outcomes in our study. Either of the four indicators had significant
influence on clinical pregnancy as logistic regression analysis revealed.
CONCLUSIONS: Global sperm DNA methylation level was significantly
decreased in asthenozoospermia samples. Besides, DNA methylation/demethylation
status posed a potential influence on fertilization and embryo development
in these patients. This indicates that even though current ART
techniques help to select highly motile sperm in asthenozoospermia patients,
epigenetic alterations may still exist in these sperm, which may affect the
fertilization process and embryo development in IVF.
P-386 Wednesday, October 21, 2015
ASSESSING A COMPREHENSIVE CHROMOSOMAL ANALYSIS
OF HUMAN SPERMATOZOA BY NEXT GENERATION
SEQUENCING. S. Cheung, Q. V. Neri, Z. Rosenwaks,
G. D. Palermo. Reproductive Medicine, Weill Cornell Medical College,
New York, NY.
OBJECTIVE: To carry out a complete molecular karyotype on sperm cells
by unraveling, extracting and amplifying the compacted male genome in
adequate quantity and quality in comparison to FISH.
DESIGN: Following extraction and amplification of good quality sperm
DNA copy number variations were compared between specimens to assess
the occurrence of spermatogenetic meiotic errors. Aneuploidy outcome
was compared to standard FISH assessment.
MATERIALS AND METHODS: DNA extraction was achieved by a commercial
kit. We processed 48 ejaculates where we extracted DNA and processed
for DNA amplification. In 12 specimens where only few
spermatozoa were processed, satisfactory DNA was obtained following
PCR-based random hexamer amplification. Fifteen of these specimens
were processed by NGS and the copy number variations were recorded
and compared using CASAVA and VarScan2 software programs. FISH on
chromosomes X, Y, 13, 15, 16, 17, 18, 21, and 22 was carried out on spermatozoa
from fertile men as well as men with history of recurrent pregnancy
loss. Following NGS, a patient sample was compared to that of a fertile anonymous
donor’s.
RESULTS: We extracted good quality DNA ranging from 500,000 to as
low as 250 spermatozoa.Assessment of 9 chromosome FISH on spermatozoa
obtained from fertile men (n¼7) the total aneuploidy rate was only 0.2%,
whereas men with recurrent ART failure (n¼36) had a total aneuploidy of
3.92.5%. When we ranked the data according to paternal age, we established
that the sperm aneuploidy rate progressively increased with advancing
age to reach 10.4% at the average age of 6811yrs particularly for chromosomes
15 and 17.Following NGS, our data presented a total percentage of
21.47% of aneuploidy rate. The most represented was gonosomal with Y
monosomy and disomy at 4.2%, followed by chromosome 15 at 3.3%, and
chromosome 14 at 1.6%. On the other extreme chromosomes 18 and 21
had the lowest disomy at 0.10 and 0.15, respectively.When compared to standard
FISH assessment, the NGS aneuploidy results depicted higher aneuploidy
than FISH results overall. Moreover, NGS copy number variations
for individual chromosomes indicated higher aneuploidy levels for all chromosomes
screened for by FISH, with the exception of chromosomes 13, 18,
and 21. The remaining chromosomes X, Y, 15, 16, 17, and 22, were actually
found to have more than twice the aneuploidy value when assessed by NGS.
CONCLUSIONS: Sperm aneuploidy assessment help in counseling male
factor infertile couples or those with recurrent pregnancy losses. FISH has
been the procedure used for this assessment, however, it has a limited number
of chromosomes evaluated and not to mention the inherent accuracy concerns
related to the procedure and the inability when assessing haploid specimen to
discern through nullisomy. NGS proved to be the procedure capable of assessing
aneuploidy. It can assess a wide range of cells from 500,000 to
10million, it assess all chromosomes including nullisomy. In addition, there
is the ability to assess the copy number variations.
Supported by: WCMC.
P-387 Wednesday, October 21, 2015
HIGH THROUGHPUT INTEGRATED PROTEOMIC ANALYSIS OF
SPERMATOZOAL PROTEINS IN PATHOPHYSIOLOGY OF VARI-
COCELE ASSOCIATED MALE INFERTILITY. A. Agarwal, a
D. Durairajanayagam, a,b R. Sharma, a L. Samanta, a,c R. F. Turki, d
E. Sabanegh. e a Center for Reproductive Medicine, Cleveland Clinic, Cleveland,
OH;
b Physiology, Universiti Teknologi MARA, Sungai Buloh,
Malaysia; c Redox Biology Laboratory, School of Life Sciences, Ravenshaw
University, Orissa, India; d Ob/Gyn Specialist, King AbdulAziz University,
Mobil, AL; e Urology, Cleveland Clinic, Cleveland, OH.
OBJECTIVE: Varicocele appears to affect later stages of spermatogenesis.
It causes scrotal hyperthermia, hypoxia, hormonal imbalances, and re-flow of
metabolites from renal and/or adrenal glands leading to oxidative stress. The
objective was to study the major differences in the distribution of spermatozoal
proteins in infertile men diagnosed with varicocele compared to fertile
men.
DESIGN: Prospective proteomic study.
MATERIALS AND METHODS: Proteins were extracted from infertile
men with unilateral and bilateral varicocele (n¼5) and men with proven
fertility (n¼5). 1-D gel electrophoresis followed by LC/MS-MS (LTQ-Orbitrap
Elite hybrid mass spectrometer) was used for protein identification.
Mascot (Matrix Science, London, UK), SEQUEST (Thermo Fisher Scientific,
San Jose, CA, USA) and X! Tandem (TheGPM, thegpm.org) were set up
to search the human reference with database assuming trypsin as the digestion
enzyme. Functional annotations of proteins were obtained using bioinformatics
tools and pathway databases.
RESULTS: Of the 99 proteins that were differentially expressed (DEP) in
the varicocele group, 9 were uniquely expressed in the fertile group
compared to 2 proteins that were unique to the varicocele groups. Over
87% of the DEP involved in major energy metabolism and key sperm functions
were underexpressed in varicocele group. Key protein functions
affected in the varicocele group were spermatogenesis, sperm motility
(ACRBP, SPA17, AKA7), and mitochondrial dysfunction (NDUFS1,
UQCRC2).
CONCLUSIONS: We have identified proteins that are underexpressed in
varicocele group. These proteins may be key players involved in the pathology
of varicocele and in the onset of infertility.
P-388 Wednesday, October 21, 2015
CLINICAL OUTCOME OF TREATMENTS FOR AZOO-
SPERMIA. A. Tanaka, a M. Nagayoshi, a I. Tanaka, a S. Ikuma, a
T. Miki, a T. Yamaguchi, a H. Kusunoki, b S. Watanabe. c a Saint Mother Hospital,
Kitakyusyu, Japan; b Faunal Diversity Sciences, Graduate School of
Agriculture, Kobe University, Kobe, Japan; c Anatomical Science, Hirosaki
University Graduate School of Medicine, Hirosaki, Japan.
OBJECTIVE: Azoospermia is found in approximately one out of one hundred
men and the ratio of obstructive azoospermia to non-obstructive one is
about 3:7. The sole treatment for azoospermia is the collection of sperms or
spermatids surgically. We analyzed the clinical outcome following each procedure
over a period of 14 (1999-2013) years. Microsurgical epididymal
sperm aspiration (MESA) for obstructive azoospermia and microscopic
testicular sperm extraction (Micro-TESE) for non-obstructive azoospermia
are generally recommended. However there has not been enough information
available for the clinical data following each procedure.
FERTILITY & STERILITY Ò
e237

DESIGN: Retrospective cohort study to treatments for azoospermia.
MATERIALS AND METHODS: MESA was performed to azoospermic
men who had normal FSH concentration (15mIU/ml).
RESULTS: Clinical outcome was summarized in Table. Clinical data
following MESAwas significantly higher than all other results following Micro-TESE
except miscarriage rates using spermatid. Total of 1171
(M:F¼563:608) babies were born (778 from MESA, 393 from Micro-
TESE) and rate of congenital defect and perinatal complication was 2.7%
(32/1171) (12 cases of heart diseases, 3 cases of annal atresia, 11 cases of
palatal cleft, 3 cases of still birth and 3 cases of 21 trisomy).
CONCLUSIONS: MESA showed a satisfactory clinical outcome. However,
Micro-TESE resulted in lower collection rate and clinical success rate
than reported in other parts of the world. This unexpected lower collection
rate might be related to the homogeneous racial background of the population
of our study. And main causes for lower successful rate might be high percentage
of sperm with low or no motility and heads with anomalies and
low survival after freezing / thawing procedures.
Clinical outcome for azoospermia following MESA or Micro-TESE over a 14
years period.
Pregnancy rates (%)
(per ET)
Miscarriage
rates (%)
Birth
rates (%)
MESA 35y < 36.0% (644/1787) 25.9% (167/644) 25.5% (456/1787)
35-39y 30.0% (393/1308) 14.2% (56/393) 24.6% (322/1308)
TOTAL 33.5% (1037/3095) 21.5% (223/1037) 25.1% (778/3095)
Micro-
TESE
(sperm)
Micro-
TESE
(spermatid)
35y < 10.3% (41/397) 24.4% (10/41) 8.8% (35/397)
35-39y 11.5% (46/401) 28.3% (13/46) 8.2% (33/401)
TOTAL 10.9% (87/798) 26.4% (23/87) 8.5% (68/798)
35y < 23.0% (526/2291) 59.1% (311/526) 10.3% (236/2291)
35-39y 17.5% (231/1320) 64.9% (150/231) 6.7% (89/1320)
TOTAL 20.9% (756/3611) 61.0% (461/756) 9.0% (325/3611)
P-389 Wednesday, October 21, 2015
COMPARISON OF SEMEN QUALITY AND FERTILIZATION PO-
TENTIAL BETWEEN CONSECUTIVE
EJACULATES. A. M. Ragheb. Urology, Faculty of Medicine, Beni Suef
University, Beni Suef, Egypt.
OBJECTIVE: There has been a growing body of evidence on the improvement
of semen quality in sequential semen samples. We aimed to verify this
finding and its implication in the assisted reproduction arena.
DESIGN: A retrospective review of records.
MATERIALS AND METHODS: The study included the records of 102
infertile males with idiopathic asthenozoospermia or oligoasthenozoospermia
attending the Egyptian International Infertility and IVF Center from
January 2014 till December 2014. All the patients had been previously asked
to provide two semen samples (1-3 hours apart) after an abstinence period of
3-7 days. The two consecutive semen samples were analyzed according to
2010 WHO criteria for semen analysis and their parameters were compared.
RESULTS: Average subject age was 34.3 years (22-53). Of 36 patients
who had zero sperm concentration in their first sample, 9 of them were
able to show mature sperm in their second sample (25%). Hence, these patients
were spared from performing surgical sperm retrieval whether for diagnostic
or therapeutic purposes. Mean seminal volume among the study group
was 2.9 1.6 mL in the 1st sample, with a mean concentration of 35.4*106
sperm /mL and mean motility of 4.97.8%. Although second sample mean
seminal volume decreased significantly compared to 1st (1.50.9 mL), yet
we detected a statistically significant incline in the mean sperm concentration
(3.6 6.3*106 sperm /mL) and mean motility (6.79.7%) of 2nd sample
sperm (p

(lutenizing hormone (LH), follicle stimulating hormone (FSH), testosterone),
and the period from SCI injury were also obtained and analyzed to
detect any associations with the presence of spermatogenesis.
RESULTS: In this series, SRR and PR were 89% and 62%, respectively.
Univariate analysis identified age, testicular volume, serum LH level and
serum FSH level as significant predictors of TESE outcome, of which only
serum FSH level appeared to be independently related to TESE outcome
on multivariate analysis (Table).
Univariate and multivariate analysis of several parameters as predictors of
micro-TESE.
Univariate Analysis Multivariate Analysis
Hazard Ratio P Value Hazard Ratio P Value
Age 0.89 0.098 - -
Testicular volume 1.56 0.015 0.065 0.10
LH 0.74 0.0019 7.329 1.00
FSH 0.56

2010 morphologic criteria. Beyond these thresholds, there was no additional
increase in post-wash TMSC. For every % increase in morphology
(below the threshold), the post-wash TMSC increased by 1.1 (p0.05
BFR, total 21 49.01 15.39 0.23 0.29 49 p>0.05
BFR, total X, Y 49.01 15.39 -0.06 0.29 49 p>0.05
BFR, high quality 13 31.77 11.72 -0.57 0.37 46 p0.05
blastocysts
BFR, high quality 18 31.59 12.13 -0.05 0.29 49 p>0.05
blastocysts
BFR, high quality 21 31.59 12.13 0.24 0.29 49 p>0.05
blastocysts
BFR, high quality
blastocysts
X, Y 31.59 12.13 0.08 0.29 49 p>0.05
P-397 Wednesday, October 21, 2015
LIVE BIRTH RATES AND MISCARRIAGE RATES WITH THE USE
OF CALCIUM IONOPHORE IN PATIENTS WITH PRIOR FAILED
OR POOR FERTILIZATION. E. Wood, a K. O. Pomeroy, a,b
J. Ricard, a N. Montalvo, c I. Collazo, a J. Eisermann. a a South Florida Institute
for Reproductive Medicine, Miami, FL; b The World Egg Bank, Phoenix, AZ;
c Clinica Montalvo, Santa Cruz, Bolivia, Plurinational State of.
OBJECTIVE: A critical step in fertilization is the activation of the oocyte
via a flux of calcium released from the cortical granules of the oocyte. Treatment
with calcium ionophore has been used to treat the ova of patients where
fertilization via ICSI fails. The aim of this study was to determine the efficacy
of calcium ionophore treatment for patients with poor or no fertilization after
ICSI. A literature review of 12 peer review publications and 4 abstracts since
1997 indicated that to date 64 healthy live births have occurred from the use
of calcium ionophore treatment.
DESIGN: This was a retrospective analysis of ICSI cycles where calcium
ionophore was used from 2004 to 2015 at SFIRM. Ionophore treatment was
used on 12 couples with 15 fresh ICSI cycles. (We also had 2 FET cycles using
ionophore-treated ova.) 10 of these couples had 12 prior cycles at our
clinic where 79 mature ova were retrieved and 17 fertilized normally
(21.5%).
MATERIALS AND METHODS: Ionophore treatment consisted of
exposing post-ICSI ova for 10 minutes to 10 uM concentration of
A23187 at 37 C. Ionophore was then rinsed out and ova were cultured using
our standard methods for IVF. Embryos were cultured for 5 or 6 days prior
to transfer.
RESULTS: Fertilization after ionophore treatment was 56.8%, compared
to 21.5% for the prior cycles. (Our normal fertilization rate for our patients is
about 70%.) All 15 cycles resulted in an embryo transfer with an average of
1.9 embryos transferred per patient. 11 patients (73.3%) had a clinical pregnancy
and 8 (53.3%) had a birth or ongoing pregnancy. (One patient that
delivered a healthy baby from the fresh transfer delivered a healthy baby
from a subsequent frozen embryo transfer). There were 3 miscarriages,
one with a normal karyotype, one with trisomy 20 and one with trisomy
21. The miscarriage rate was 27.3%. So far, 9 healthy babies have been delivered
from this technique - two sets of twins and 5 singletons. The trisomy 21
came from a patient 38 years old and the trisomy 20 from a 40 year old patient.
CONCLUSIONS: We found that this method performs favorably with
patients with no fertilization and even those with poor fertilization.
Although the procedure resulted in a high degeneration rate of ova (about
15%) and multicellular embryos with fragmentation, fertilization rates
improved with ionophore treatment and live birth/ongoing rates (53.3%)
are near normal for this age group of women in our practice (41.7%;
SART 2012). The clinical use of calcium ionophore is still considered
experimental. World literature has suggested no adverse outcome in the
offspring. Its use for poor fertilization and fertilization failure in ICSI patients
is promising. Calcium ionophore incubation may become a reasonable
and efficient option to assist couples with this rare condition so that
they may have a healthy child.
e240 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

P-398 Wednesday, October 21, 2015
TREATMENT WITH COMBINED ANTIOXIDANT FORMULATION
BEFORE ICSI IMPROVES PREGNANCY RATE IN COUPLES WITH
OBSTRUCTIVE AZOOSPERMIA. S. Gulati, R. Chattopadhyay,
B. Ghosh, S. Yasmin, S. Ghosh, G. Bose, P. Chakraborty, B. Chakravarty.
Assisted Reproduction, Institute of Reproductive Medicine, Kolkata, India.
OBJECTIVE: To evaluate the efficacy if any, of a combined antioxidant
formulation, in couples with obstructive azoospermia after failed intra cytoplasmic
injection (ICSI) cycle.
DESIGN: A single centre prospective cohort study of 210 patients with
azoospermia was performed from February 2012 to March 2015.
MATERIALS AND METHODS: Patients with documented azoospermia
were divided into non-obstructive (NOA) (group A; n ¼ 98) and obstructive
(OA) (group B; n ¼ 112) counterpart. Absence of sperm (n¼15) and unsatisfactory
endometrium (n ¼ 3) were the exclusion criteria in either arm/s.
Testicular sperm extraction (TESE) followed by ICSI confirmed pregnancy
in 25.3% (n ¼ 21) and 32.11% (n ¼ 32) in group A and B respectively. Patients
who failed TESE/ ICSI cycles were treated with a combined antioxidant
formulation comprising L-carnitine, CoQ10, zinc, folic acid, vitamin
B12 and selenium for 6 months. A placebo-controlled normozoospermic
group comprising 75 patients was maintained as controls. ROS-TAC score
was determined as biomarker of oxidative stress (OS) from homogenized
testicular tissue by standard methods before and after the treatment. Mitochondrial
membrane potential (Djm), sperm DNA fragmentation were evaluated
by flow-cytometry and comet assay respectively. Study was approved
by Institutional Review Board. Statistical comparisons were performed by
Stata10.0. The main outcome was clinical pregnancy rate.
RESULTS: 62 patients with NOA and 77 patients with OA were treated
with combined antioxidant therapy followed by TESE/ICSI. Improvement
in sperm morphology was observed in 8.06% (n ¼ 5) in group A compared
to 63.64% (n ¼ 49) cases in group B. ROS-TAC score decreased significantly
in group B only than controls (33.1 6.2 vs. 52.0 7.1; p < 0.04). Djm and
rate of apoptosis improved considerably after antioxidant regimen. Fertilization
rate was significantly lower (p< 0.04) in NOA group (56.45%) than the
OA group (74.32%). The clinical pregnancy rate was 27.42% (n ¼ 17) and
41.55% (n¼32) in NOA and OA cohorts respectively. This was statistically
significant adjusting for age with an odds ratio of 1.06 (95% CI 1.20-3.20,
p

CONCLUSIONS: TPMSC is an independent predictor for biochemical
pregnancy rate and LBR in non-donor sperm, but this association is not
observed in donor sperm; this lack of association in donor group may be
due to small sample size. Furthermore, our data suggest that male factor
infertility in cycles using non-donor sperm (despite high sperm counts)
continues to compromise pregnancy outcomes as compared to donor
sperm.
References:
1: Freour T, Jean M, Mirallie S, Langlois ML, Dubourdieu S, Barriere P. Predictive
value of CASA parameters in IUI with frozen donor sperm. Int J
Androl. 2009 Oct;32(5):498-504.
2: Marshburn PB, McIntire D, Carr BR, Byrd W. Spermatozoal characteristics
from fresh and frozen donor semen and their correlation with fertility
outcome after intrauterine insemination. Fertil Steril. 1992 Jul;58(1):179-
86.
3: Byrd W, Bradshaw K, Carr B, Edman C, Odom J, Ackerman G. A prospective
randomized study of pregnancy rates following intrauterine and intracervical
insemination using frozen donor sperm. Fertil Steril. 1990
Mar;53(3):521-7.
4: Tan O, Ha T, Carr BR, Nakonezny P, Doody KM, Doody KJ. Predictive
value of postwashed total progressively motile sperm count using
CASA estimates in 6871 non-donor intrauterine insemination cycles. J
Assist Reprod Genet. 2014 Sep;31(9):1147-53.
P-401 Wednesday, October 21, 2015
TESTICULAR SPERM EXTRACTION FOLLOWED BY INTRACY-
TOPLASMIC SPERM INJECTION CHOICE OF TREATMENT
FOR THE APPREHENSIVE COUPLES HAVING COLLECTION
PROBLEM. N. Prasad, a R. Chattopadhaya, b S. Ghosh, b
S. K. Goswami, b B. Ghosh, b P. Chakraborty, b B. Chakravarty. b a Reproductive
Medicine, Fellow in Reproductive Medicine, Kolkata, India; b Reproductive
Medicine, Institute of Reproductive Medicine, Kolkata, India.
OBJECTIVE: To compare the fertility potential of freshly retrieved testicular
sperm obtained from patients with azoospermia and collection failure
undergoing intracytoplasmic sperm injection (ICSI).
DESIGN: Prospective analysis of 130 testicular sperm extraction (TESE)
/ICSI cycles from January 2011 to March 2015.
MATERIALS AND METHODS: Eighty four patients with documented
azoospermia underwent micro-dissection TESE with concomitant ICSI at
the same time (Group A). Patients having previous history of failed collection
(n¼53) were also treated with TESE followed by ICSI (Group B). 7 cycles
were cancelled due to unsuccessful sperm retrieval in azoospermic couples
only. Comparison between the two groups in terms of fertilization, cleavage,
implantation, clinical pregnancy and miscarriage rates was done. The Ethical
Committee of Institute of Reproductive Medicine approved the study and all
the participants consented to enroll in this study . Statistical comparisons
were performed using chi-square and student’s T test as required. P value

at a private suburban Fertility Center by a single andrologist (GAA) were reviewed.
Only the first SA was evaluated per patient.
RESULTS: In total, 901 SA including 276 who worked in Business/
Finance/Sales (30.6%), 224 worked in Information Technology (24.9%),
77 were engineers (8.5%), 51 worked in government (5.7%), and 37 in Health
Care (4.1%). There were no differences in any semen parameter among
different occupations; specifically IT use was not associated with any negative
effects on seminal parameters.
CONCLUSIONS: Men working in IT have no differences in seminal parameters
compared to other white collar occupations. There were no differences
among the different occupations evaluated, a reassuring novel
finding. As far as we know, this is the largest study evaluating the effects
of various occupations, including those that require significant sitting such
as IT, on semen parameters.
P-404 Wednesday, October 21, 2015
IMPROVEMENT IN PREGNANCY RATES IN IMMUNOLOGI-
CALLY INFERTILE MEN UNDERGOING PREDNISOLONE AND
IVF PROGRAMS PRECEDED BY SPERM PENETRATION
ASSAY. A. M. Taiyeb, a M. T. Ridha-Albarzanchi, a A. Haji, a
S. N. Mahmood, b M. E. Kjelland, c N. S. Hawa, b S. A. Muhsen-Alansarri. a
a BARZ IVF Center for Infertility Treatment and Embro Research, Erbil, Iraq;
b Department of Obstetrics and Gynecology, Baghdad College of Medicine,
Baghdad, Iraq; c US Army Engineer Research and Development Center,
Vicksburg, MS.
OBJECTIVE: Anti-sperm antibodies (ASA) impair not only human sperm
motility but also capacitation, penetration of cervical mucus, and conception
rate. Many studies have confirmed the clinical therapeutic effect of corticosteroids
(CST) in treatment of men with ASA [1-3]; however, other studies
have not observed a therapeutic effect [4-5]. This controversy is extended
to include the usefulness of assisted reproductive technologies in patients
with ASA. For instance, some studies have shown that pregnancy rates
following IVF or ICSI were similar or not associated in men with or without
ASA [6-7], whereas other reports showed the superiority of ICSI over IVF [8]
and IUI over natural intercourse in men with ASA [9]. In order to address this
controversy, immunological infertile men, treated or not treated with CST,
were admitted to conventional IVF or ICSI as determined by the sperm penetration
assay (SPA) of hamster ovum. The latter assay can identify patients
with compromised fertilization related to sperm binding and fusion to oolema.
It is possible that some patients with ASA have compromised sperm
fertilization as an additional problem, which may interfere with the therapeutic
effect of CST in patients with ASA.
DESIGN: A prospective clinical comparative study.
MATERIALS AND METHODS: SPA was performed for immunologically
infertile men. Positive or negative SPA partners were then admitted
to conventional IVF or ICSI cycles, respectively. Each IVF or ICSI male
group was divided into treated and control groups. The treated male group
received a prednisolone program for three cycles.
RESULTS: Administration of prednisolone in men with good SPA results
improved sperm motility when compared to control men (P

Table 1.
Group 1 Group 2 P
Age 33.7 2.3 29.4 4.8 0.004
FSH 6.1 1.3 6.4 2.2 NS
LH 6.0 3.9 6.4 2.2 NS
E2 33.1 16.2 44.6 25.4 NS
PRL 19.5 10.4 28.2 11.2 NS
FOL 14.3 8.2 10.4 4.6 NS
INY 9.6 7.5 9.2 5.3 NS
FERT 5.0 3.9 4.6 3.0 NS
P-406 Wednesday, October 21, 2015
EJACULATED SPERM MAY NOT RESULT IN THE BEST CLIN-
ICAL OUTCOME FOR ICSI TREATMENT CYCLES. Y. Lin, a
L. Cai, b J. Dong, b J. Liu, c R. Chian. d a reproduction medicine, nanjing,
China; b The First Affiliated Hospital of Nanjing Medical U, Nanjing, China;
c the First Affiliated Hospital of Nanjing Medical U, Nanjing, China; d McGill
University, Montreal, QC, Canada.
OBJECTIVE: To evaluate the impact of different sperm sources on fertilization,
embryo quality, clinical pregnancy and implantation rates in intracytoplasmic
sperm injection (ICSI) treatment cycles.
DESIGN: A retrospective analysis was performed in 1,263 patients undergoing
ICSI treatment cycles in our center from January 2012 to March 2014.
MATERIALS AND METHODS: The patients were classified into 4
groups based on the sources of sperm obtained, namely fresh ejaculated
semen, (n¼950), fresh testicular sperm aspiration (TESA) (n¼12), fresh
percutaneous epididymal sperm aspiration (PESA) (n¼204), and frozen
PESA sperm (n¼97). Fertilization, high-quality embryo, implantation, clinical
pregnancy and live birth rates in the fresh embryo transferred cycle were
compared among the groups.
RESULTS: There were no differences in the fertilization rates among four
groups (74.4%, 75.6%, 75.1% and 74.7%, respectively). The patients with
fresh TESA sperm and frozen PESA sperm showed significantly higher
high-quality embryo rates (75.4 % and 73.9 %) than those of fresh ejaculated
semen (63.8%) and fresh PESA sperm (66.3%) groups (P

2. Ben-Ami I, Raziel A, Strassburger D, Komarovsky D, Ron-El R, Friedler
S. Intracytoplasmic sperm injection outcome of ejaculated versus
extracted testicular spermatozoa in cryptozoospermic men. Fertil.
Steril. 2013;99(7):1867-71.
3. Bendikson KA, Neri Q V, Takeuchi T, et al. The outcome of intracytoplasmic
sperm injection using occasional spermatozoa in the ejaculate
of men with spermatogenic failure. J. Urol. 2008;180(3):1060-4.
4. Hauser R, Bibi G, Yogev L, et al. Virtual Azoospermia and Cryptozoospermia–Fresh/Frozen
Testicular or Ejaculate Sperm for Better IVF
Outcome? J. Androl. 2011;32(5):484-490.
5. Weissman A, Horowitz E, Ravhon A, Nahum H, Golan A, Levran D.
Pregnancies and live births following ICSI with testicular spermatozoa
after repeated implantation failure using ejaculated spermatozoa. Reprod.
Biomed. Online 2008;17(5):605-609.
P-409 Wednesday, October 21, 2015
S. Alshahrani. Prince Sattam Uni-
ARE OBESE MEN SUBFERTILE?
versity, Alkharj, Saudi Arabia.
OBJECTIVE: Although it is well known that overweight and obesity can
affect female fertility, in men the negative effects on reproductive system
attributed to obesity are less evident and have been less often studied. The
aim of this study is to evaluate the effect of Body Mass Index (BMI) on
different semen parameters.
DESIGN: prospective study.
MATERIALS AND METHODS: After IRB approval, semen samples
from 439 male partners of couples presenting for evaluation of their infertility,
in the period from 2013-2014, were collected. Men were divided into
three BMI groups; normal , overweight and obese. Meticulous scrotal examination
plus scrotal ultrasound were performed. Hormonal profile, including
testosterone, FSH, LH and prolactin was done to exclude any possible causative
hormonal factor.
RESULTS: Mean BMI was 29.675.89. ANOVA testing revealed no significant
differences in semen parameters between the 3 different BMI groups.
Also, pair-wise multiple comparisons were found to be non-significant (P >
0.05). The distribution of patients with normal sperm concentration per BMI
group was as follows: normal weight, 46/75 (61.33%); overweight, 114/179
(63.69%); and obese, 116/185 (62.70%). BMI had a negative correlation with
semen volume, sperm concentration, sperm motility and sperm morphology.
However, this correlation reached a significant level only between BMI and
sperm concentration (r¼0.101; p¼0.035).
CONCLUSIONS: Although found to have no significant relation with
semen volume, sperm motility or sperm morphology; BMI was proved, in
our study, to have a significant effect on sperm concentration.
The effect of BMI on semen parameters.
Parameters Total patients Normal BMI Over weight Obese p-value
Number (%) 439 (100) 75 (17.08) 179 (40.77) 185 (42.14)
BMI 29.675.89 23.051.34 27.061.41 34.885.31 ˂0.001
Age (y) 36.856.73 35.646.56 36.836.82 37.376.69 0.173
Volume (ml) 3.071.49 3.121.42 3.181.47 2.941.53 0.297
Concentration (mil/ml) 41.4650.96 44.6457.25 41.3248.96 40.3050.40 0.824
Actively motile sperm (%) 44.5319.54 46.0018.50 44.1120.46 44.3519.10 0.770
Normal morphology (%) 3.212.95 3.202.92 3.243.02 3.152.91 0.957
P-410 Wednesday, October 21, 2015
INDEPENDENT PREDICTORS OF INTRAUTERINE INSEMINA-
TION (IUI) SUCCESS. R. D. Kastury a G. S. Taliadouros. b a Obstetrics
and Gynecology, Inspira Health Network, Vineland, NJ; b Reproductive
Endocrinology and Infertility, Inspira Health Network, Vineland, NJ.
OBJECTIVE: Various causes of infertility are treated with intrauterine
insemination (IUI). Many factors affect IUI outcomes. Previously published
studies have provided conflicting results regarding the independent predictors
for the success of this treatment (1). In this study, we aim to evaluate
these predictors in a well-defined patient population.
DESIGN: A retrospective study.
MATERIALS AND METHODS: Our analysis evaluated 509 couples, between
ages 24 and 44, that underwent infertility assessment and subsequent
intrauterine insemination. Fresh partner’s semen was processed through a
50%/90% discontinuous gradient and one IUI was performed at the time
of ovulation, confirmed by ultrasonography and laboratory values. Females
with tubal factor infertility and patients with severe male factor infertility
were excluded. All other major medical concerns were addressed prior to
initiating treatment. Anti-estrogen and gonadotropins were used to address
ovulatory aberrations. The primary outcome measure was the occurrence
of pregnancy. Univariate (EXPAND) and multivariate logistic regression analyses
were used. The strength of association between pregnancy and each
independent predictor is presented as an odds ratio (OR) with confidence interval
(CI). All statistical tests used a two-tailed alpha of 0.05.
RESULTS: Univariate analysis shows that motile sperm count in the unprocessed
semen (OR ¼ 0.0023, CI ¼ 0.0010 - 0.0037, p ¼ 0.0005) and total
motile sperm inseminated (OR ¼ 0.0064, CI ¼ 0.0029 - 0.0098, p ¼ 0.0003)
were significant factors associated with the occurrence of pregnancy. Sperm
morphology performed using Kruger’s strict criteria, did not have statistical
significance on the occurrence of pregnancy. The age of female was
extremely predictive of pregnancy occurrence (OR ¼ -0.047, CI ¼ -0.0792
to -0.0149, p ¼ 0.0041) and the type of ovarian stimulation, gonadotropins,
also proved to be statistically significant in predicting a positive outcome
(OR ¼ -0.3257, CI ¼ -0.6256 to -0.0248, p ¼ 0.0339). Multivariate analysis
demonstrated that female age and number of motile sperm in the ejaculate are
the strongest predictors of pregnancy, p ¼ 0.0009 & 0.0007 respectively.
Gonadotropin stimulation increased the chance of achieving pregnancy by
30%.
CONCLUSIONS: Our results indicate that the total number of motile
sperm in the ejaculate is a significant predictor for pregnancy occurrence.
As expected, the increased number of oocytes available during controlled
ovarian stimulation will have an additive effect on the results. It has been
widely accepted that female age has an impact on fecundity. These findings
corroborate previously reported data, which can be used to advise future couples
on the likelihood of achieving pregnancy and guide physicians with
appropriate treatment selection.
Reference:
1. Ombelet W, Dhont N, Thijssen A, Bosmans E, Kruger T. Semen quality
and prediction of IUI success in male subfertility: a systematic review.
Reprod Biomed Online. 2014;28:300-309.
P-411 Wednesday, October 21, 2015
IS IT NECESSARY TO DO INTRACYTOPLASMIC SPERM INJEC-
TION ON PATIENTS WITH ABNORMAL SPERM DNA FRAGMEN-
TATION VALUE IN ART? S. M. Kang, J. H. Kim, Y. J. Lee. I-Dream
Center, Mizmedi Women’s Hospital, Seoul, Korea, Republic of.
OBJECTIVE: The purpose of this study is to evaluate how effective intracytoplasmic
sperm injection (ICSI) could be in the pregnancy outcome of patients
with DNA-fragmented sperm by comparison to conventional in vitro
fertilization (IVF).
DESIGN: A retrospective clinical study.
MATERIALS AND METHODS: According to standard protocol, 137
(ICSI and IVF) cycles that were performed with at least 5 retrieved eggs,
normal count and motility of sperms (the World Health Organization criteria)
at each cycle were retrospectively analyzed. Sperms that were used in this
study were classified into three groups depending on the sperm DNA fragmentation
(SDF) value (by Halosperm kit) as follows: Abnormal,
SDF>30%; Sub-normal, 15%

OH; b Case Western Reserve University, Beachwood, OH; c USC Fertility
Center, Los Angeles, CA.
OBJECTIVE: To study the correlation between pre- and post-wash sperm
morphology indices and pregnancy outcomes in intrauterine insemination
(IUI) and in vitro fertilization (IVF).
DESIGN: Retrospective Cohort.
MATERIALS AND METHODS: A total of 305 IUI and 113 IVF fresh and
frozen cycles performed at a single center from January to October 2012 were
reviewed. Three morphology indices were completed on pre- and post-wash
semen samples. The three indices included multiple anomalies index (MAI),
teratozoospermia index (TZI), and sperm deformity index (SDI). Index data
were correlated with pregnancy outcomes for IUI and IVF cycles via a principle
component analysis and logistic regression. Student T-tests were used
for comparisons of MAI, TZI and SDI with pregnancy outcomes.
RESULTS: The analysis showed a high correlation between the MAI, TZI,
and SDI in the pre- and post-wash groups. Due to this similarity, the optimal
morphology index could not be determined. A principle component analysis
was utilized to determine the relationship between the group of indices and
pregnancy outcomes. The pre-wash indices were not significant in predicting
pregnancy (p ¼ 0.335), while the post-wash indices were significant
(p ¼ 0.0211). The post-wash indices were more predictive of pregnancy outcomes
in IUI cycles compared to IVF cycles (p ¼ < 0.0001). When MAI,
TZI, and SDI were evaluated independently in IUI cycles, there was a trend
toward statistical significance in pregnancy outcomes in the post-wash group
(p ¼ 0.056, 0.055, 0.051, respectively). However, in IVF fresh and frozen cycles
the three indices had no statistical significance in predicting pregnancy
outcomes (p ¼ 0.698, 0.704, 0.747, respectively).
CONCLUSIONS: Due to the high correlation of MAI, TZI, and SDI, no
optimal morphology index was identified in this study. Although the grouped
post-wash indices predicted pregnancy outcomes, the individual analyses
were not significant. As expected, the combined post-wash indices may
have higher utility in predicating pregnancy outcomes in IUI compared to
IVF cycles. Consistent with prior studies, sperm morphology indices may
have predictive potential in select groups; however, the data from this study
does not support generalized use.
SPERM PREPARATION
P-413 Wednesday, October 21, 2015
THE IMPACT OF SEMEN PROCESSING ON SPERM QUALITY
AND PREGNANCY RATES WITH INTRAUTERINE
INSEMINATIONS. J. J. Ruiter-Ligeti, a C. Agbo, b M. Dahan. a a McGill
University, Montreal, QC, Canada; b Stanford University, Stanford, CA.
OBJECTIVE: Semen processing is routinely performed prior to intrauterine
insemination (IUI). The expectation is that semen processing improves
semen quality parameters. However, no studies have evaluated the impact
of semen processing for IUI on those parameters. This study aims to evaluate
Table: Effect of processing on semen quality parameters and pregnancy rates.
Semen Quality
Parameter
Absolute
change
Mean
+/- SD
Pearson
correlation
coefficient
(r)
% of subjects
p-value of with parameter
correlation improvement
Concentration (M/ml) 66 74 0.043 0.04* 90%
Percent Motile (%) 24 22 -0.012 0.56 80%
Concentration Motile 27 58 0.038 0.08 70%
(M/ml)
Total Motile Sperm -55 81 -0.002 0.97

P-415 Wednesday, October 21, 2015
STATIC MAGNETIC FIELDS ON HUMAN SPERM. A SIDE EFFECT
OF MAGNETIC-ACTIVATED CELL SORTING? C. Avendano,
A. Mata, I. A. Anduaga Marchetti, C. Sanchez Sarmiento. Nascentis Medicina
Reproductiva, Cordoba, Argentina.
OBJECTIVE: Non-damaged sperm DNA is one of the prerequisites for
achieving successful fertilization and embryo development in assisted
reproductive technologies. Magnetic-activated cell sorting (MACS) using
paramagnetic annexin V-conjugated microbeads has been proposed as a
safe method to select non-apoptotic and viable sperm. The procedure involves
the magnetic labeling of sperm with damaged DNA and the passage
through a high power static magnetic field (SMF). Intact living
spermatozoa without DNA fragmentation pass through the column and
are collected for later use, while the fragmented cells are selectively retained.
Although there are some concerns about the possible selection
and use of sperm with paramegnetic beads, the potential impact of
SMF has never been mentioned. Therefore, the objective of this work
was to evaluate the possible effect of SMF on human sperm during in
vitro manipulation.
DESIGN: Prospective study.
MATERIALS AND METHODS: Motile sperm from healthy men (n¼5)
were selected by swim up. Each sperm suspension was separated in 4
fractions (F1, F2, F3 and C). F1, F2, F3 were exposed to a SMF (1.2
Tesla) during 1, 2 and 10 minutes, respectively. Fraction C was kept
as control without SMF exposure. Sperm parameters were evaluated
immediately after SMF exposure on each fraction and then incubated
in capacitating conditions for 3 hours at 37 C. Progressive sperm
motility (PG, %), curvilinear velocity (VCL, mm/s) and rectilinear velocity
(VSL, mm/s) were evaluated using videomicroscopy with computerized
image analysis. Spontaneous (sRA) and induced (iRA) acrosomal
reaction were evaluated using Coomassie brilliant blue staining. Sperm
DNA fragmentation was assessed by TUNEL assay.
RESULTS: PG was increased immediately after exposure to SMF in F1
compared with C (93.5 2.9 vs. 86.9 3.9, p

changes involved with impaired spermatogenesis were observed in
seminiferous tubule including the smaller size of tube, the scarce of
spermatocyte. [3] CASA detection of sperm showed that sperm count
and motility declined with age in Bnc1 -/- male mice. However Bnc1
+/ mouse showed a significant drop in sperm motility by 20 weeks
of age, suggesting a dosage effect of Bnc1 on testis development. [4]
Bnc1 knock down had no effects on proliferation of CRL-2196 and
C18-4 while promote the proliferation of CRL-2053 cell. Furthermore,
several spermatogonia stem cell markers such as Magea4,Ret,Plzf were
down-regulated while differentiation marker Kit were up-regulated in
C18-4 cell and CRL-2053 cell after Bnc1 knock down. Those data
suggested Bnc1 may play a role in maintaining undifferentiation of
spermatogonia cell.
CONCLUSIONS: The genetic mutation of human BNC1 results in male
infertility with the reduction of sperm motility and decreased sperm count
and that BNC1 is a functional molecule essential for mammalian sperm formation.
OVARIAN RESERVE
P-418 Wednesday, October 21, 2015
COMPARISON OF ANTRAL FOLLICLE COUNT ACROSS THE
MENSTRUAL CYCLE WITH ANTIMULLERIAN HORMONE AND
OVARIAN RESPONSE TO CONTROLLED HYPERSTIMULATION
RELATED TO OVARIAN RESERVE. N. Massin, G. Amand,
C. Villette, A. Dessapt, C. Pietin Vialle, H. Bry, M. Pasquier,
B. Haddad. Department of Gynecology, Obstetrics and Reproductive Medicine,
Intercommunal Hospital, University Paris 12, Creteil, France.
OBJECTIVE: To compare antral follicle count (AFC) at early follicular
(EF), late follicular (LF) and luteal (L) phases with AFC at day 1 of the
ovarian stimulation (S1), antimullerian hormone (AMH) and oocytes yield,
in women with low, normal or high ovarian reserve.
DESIGN: Monocentric retrospective analysis of consecutive patients undergoing
their first IVF cycle.
MATERIALS AND METHODS: A total of 131 patients aged from 20
to 41 were included in the analysis between 2014, jan and 2015, feb.
Ultrasound for basal AFC evaluation was performed in our center on
the day of patient’s visit, independent of her menstrual cycle phase.
The women were classified as EF-AFC from day 1 to 6 (N¼44), LF-
AFC from day 7 to 13 (N¼49) and L-AFC thereafter (N¼38). S1-
AFC was performed either with patient on agonist treatment or after
17b-estradiol treatment for antagonist protocol scheduling. AMH dosage
was performed from day 2 to 4. Comparison of AFCs, AMH and number
of oocytes retrieved was done with the Pearson correlation coefficient
in the whole population and according to ovarian reserve : low
for AMH

Statistical analysis included student’s t-test and chi-square test. P

Medicine, Feinberg SOM- Northwestern University, Chicago, IL; c NIEHS,
Research Triangle Park, NC.
OBJECTIVE: To determine what medical, lifestyle and reproductive factors
are associated with low AMH levels in young AAW.
DESIGN: Cross-sectional study.
MATERIALS AND METHODS: 1,654 AAW aged 23-35 years who
participated in the Study of Environment, Lifestyle & Fibroids (SELF)
were included. Medical history and lifestyle habit data were collected via
self-report. AMH levels were determined using an ultrasensitive ELISA
assay. AMH (ng/mL) was classified as ‘‘low’’ (2-7) or ‘‘high’’ (>7). Polytomous logistic regression was used
to identify factors that were associated with low AMH levels. The model
adjusted for age, body mass index (BMI) and use of hormonal contraception.
RESULTS: The mean age of the subjects was 28.73.5 years and the mean
AMH was 4.003.49 ng/mL. AMH levels were low, low normal, normal, and
high in 12.2%, 19.5%, 54.5% and 13.8% of women respectively. Hypertension,
present in 10.0% of women, was associated with a higher likelihood of
having a low AMH versus a normal AMH (OR 2.38, 95% CI 1.49-3.79).
AAW with diabetes were not significantly more likely to have reduced
AMH levels. Weight was also associated with AMH levels. As BMI
increased, AMH levels were significantly more likely to be low versus
normal (OR 1.03, 95% CI 1.01-1.04) or high (OR 1.04, 95% CI 1.02-
1.07). There was no significant association between AMH levels and various
smoking related factors. AAW who smoked, were cared for as a child by a
smoker, or had a mother who smoked while pregnant were not more likely
to have low AMH levels. Factors related to reproductive history were also
investigated to assess their relationship with AMH levels. Women with a history
of a tubal ligation were significantly more likely to have a low AMH than
a high AMH (OR 2.28, 95% CI 1.02-5.08). However, there was no significant
association between low AMH levels and a history of becoming pregnant,
having a live birth, or consulting with a physician for difficulty conceiving.
Previous infections due to gonorrhea, chlamydia or pelvic inflammatory disease
were also not significantly associated with low AMH levels.
CONCLUSIONS: The findings from this large study of young AAW suggest
that hypertension and obesity are two common conditions associated
with low AMH levels. Additional research is needed to determine whether
the impact of these factors on AMH is reversible. A history of tubal ligation
was also associated with lower AMH levels. Patients who present for IVF or
tubal reversal should be counseled that they may have a compromised
ovarian reserve due to their history of a tubal ligation.
Supported by: NIH R21 HD077479, NIH Women’s Reproductive Health
Research Scholar Award, Harold Amos Medical Faculty Development
Award, Robert Wood Johnson Foundation, Friends of Prentice, The Evergreen
Invitational, The Woman’s Board of Northwestern Memorial Hospital.
P-424 Wednesday, October 21, 2015
UTILITY OF ANTI-M€ULLERIAN HORMONE TO IDENTIFY
WOMEN WITH UNDIAGNOSED DIMINISHED OVARIAN
RESERVE. L. C. Grossman, a L. Zakarin Safier, a M. Kline, a C. Chan, b
R. Lobo, a M. V. Sauer, a N. C. Douglas. a a Columbia University Medical Center,
New York, NY; b Level, New York, NY.
OBJECTIVE: To determine the utility of anti-M€ullerian hormone (AMH)
to identify diminished ovarian reserve (DOR) in asymptomatic women
without personal or family risk factors.
DESIGN: Prospective cohort.
MATERIALS AND METHODS: Women ages 27-37 years (yrs) currently
deferring pregnancy were invited to take an AMH blood test and pre- and posttest
surveys. Women were recruited via alumni listservs, social media, and
flyers in gynecology offices. AMH level was determined for 97 women, median
age 31 (interquartile range (IQR) 29-34) yrs, using the Beckman Coulter
Gen II AMH assay with interassay variability of 0.3 ng/mL and lowest level of
detection of 0.17 ng/mL. For healthy 31 yr old women, the median AMH level
is 3.23 ng/mL, 10th percentile is 1.15 ng/mL, and 90th percentile is 6.59 ng/mL
(1). Thus we defined DOR as AMH

Omega-3 Fatty Acid Supplementation Lowers Serum FSH in Women.
Baseline
Following 1
month of omega-3
supplementation
p
Percent
change
LH mean serum level, IU/L 4.4(0.6) 4.1(0.4) 0.77 -4.9
FSH mean serum level, IU/L 4.8(0.3) 4.0(0.3) 0.06 -16.9
Peak serum FSH, IU/L 5.5(0.4) 4.6(0.4) 0.06 -16.2
LH response to GnRH,
790 (76) 795 (74) 0.96 0.58
area under the curve, IU/L
FSH response to GnRH, area 626 (42) 518 (38) 0.04 -18.8
under the curve, IU/L
Plasma
omega 6/omega 3
FA ratio
7.7 (0.8) 3.2 (0.4) 67.91) 53 32.8 7.4 17.0 11.1 5.7 41.2
Optimal(40.03-67.9) 141 32.9 6.0 14.0 13.3 6.4 31.2
Satisfactory(21.97-40.03) 362 34.2 6.7 12.0 12.0 4.4 39.5
Low(3.07-21.97) 1314 35.9 9.0 8.0 8.8 6.8 36.8
Very Low(0-3.07) 319 37.6 11.4 4.6 3.7 14.7 16.9
CPR/
Cycle
Started (%)
Reference:
1. La Marca A1, Sighinolfi G, Radi D et al. Anti-Mullerian hormone
(AMH) as a predictive marker in assisted reproductive technology (AR-
T).Hum Reprod Update. 2010 Mar-Apr;16(2):113-30.
P-427 Wednesday, October 21, 2015
ANTRAL FOLLICLE COUNT MEASURED AFTER PITUITARY
SUPPRESSION AS PREDICTOR OF OVARIAN RESPONSE AND
PREGNANCY IN CLINICAL PRACTICE: A REVIEW OF 2075 AS-
SISTED REPRODUCTION CYCLES. S. Peralta, a,b F. Fabregues, a,b
J. Penarrubia, c,b G. Casals, a,b M. Creus, a,b D. Manau, a,d I. Gonzalez-Foruria,
e,b A. Borras, a,b J. Balasch. a,b a Institut Clınic d’Obstetrıcia, Ginecologia
i Neonatologia, Hospital Clinic, Barcelona, Spain; b Institut d’Investigacions
Biomediques August Pi i Sunyer, IDIBAPS, Barcelona, Spain; c Institut
Clınic d’Obstetrıcia, Hospital Clinic, Barcelona, Spain; d IDIBAPS, Barcelona,
Spain; e Gynecologist, Barcelona, Spain.
OBJECTIVE: To investigate the usefulness of Antral Follicle Count
measured after pituitary suppression (AFCaps) in predicting ovarian
response and pregnancy in clinical practice.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All the IVF-ICSI cycles following a long
GnRH agonist protocol in which the AFCaps had been registered in our database
were included. A total of 2075 cycles between January 2011 and
December 2014 were analyzed. Cycles were divided into three subgroups according
to the following ovarian response categories: high (>15 oocytes),
normal (4-15 oocytes) and low (% 3 oocytes or cycle cancellation).
RESULTS: According to the predefined ovarian response categories, 170
cycles (8.9%) were classified as high, 1468 (70.7%) as normal and 437
(21.1%) as low. Table shows patient characteristic, AFCaps and basal FSH,
ovarian stimulation, ovum retrieval and cycle outcomes in high, normal
and poor respondes. AFC was significantly lower in poor responders and
significantly higher in high responders than normal responders. AFC was
significantly correlated with parameters of ovarian response (E2 on HCGr
day, number of oocytes retrieved, number of embryos obtained/) For the
prediction of high response, AFCaps had an area under the receiver-operating
characteristic curve (AUC) of 0.80. Both female age and basal FSH had lower
accuracy (AUC 0.66 and 0.63, respectively). For low response prediction,
Table: Baseline patient characteristics, stimulation and clinical outcomes by
subgroup.
VARIABLE
Group 1
High
responders
(n¼170)
Group 2
Normal
responders
(n¼1468)
Group 3
Poor
responders
(n¼437)
P value
Age (y) 34.393.54 35.963.40 37.723.03

again AFC had better accuracy (AUC 0.84) than age and basal FSH (AUC
0.67 and 0.62, respectively). When the likehood of pregnancy was analyzed
the AUC for AFCaps was 0.63, similar than AUC for age and basal FSH
(AUC 0.59 and 0.54, respectively).
CONCLUSIONS: AFCaps in a clinical setting is a good predictor of
ovarian response, better than age and basal FSH. However is not a good predictor
of pregnancy.
P-428 Wednesday, October 21, 2015
UTILITY OFAUTOLOGOUS FRESH IVF CYCLES IN EXTREMELY
POOR PROGNOSIS PATIENTS. M. G. Vega, a D. H. Barad, b
S. K. Darmon, c N. Gleicher, b V. A. Kushnir. d a Mount Sinai St. Lukes Roosevelt
Hospital & Center for Human Reproduction, New York, NY; b Center
for Human Reproduction & Foundation for Reproductive Medicine, New
York, NY; c Center for Human Reproduction, New York, NY; d Center for Human
Reproduction & Wake Forest University, New York, NY.
OBJECTIVE: To determine whether there still is utility of performing
autologous fresh in vitro fertilization (IVF) cycles in extremely poor prognosis
patients.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: We identified among 1,418 consecutive
fresh non-donor IVF cycles in our center’s anonymized electronic research
databank 768, which by Bologna Criteria qualified as poor responders
because they produced %3 oocytes, were > 40 years and/or had AMH

evaluated the oxidative DNA injury. Histopathologic examination for follicle
counting was observed following hematoxylin-eosin staining.
RESULTS: AMH levels were significantly decreased compared to the preoperative
and postoperative periods in I/R, I/R+NAC and I/R+Enoxaparine
groups (p¼0.004, p

controlled primordial follicle activation in vivo. Also, PPAR gamma agonist
induced PTEN expression and inhibited Akt phosphorylation in vitro. Ovary
specific PPAR gamma was predominantly expressed in ovarian tissue. The
aim of this study was to investigate whether PPAR gamma modulator might
induce the activation and growth of primordial follicles from 5day female
mouse after birth.
DESIGN: Animal study.
MATERIALS AND METHODS: Ovaries were collected from 5days old
B6D2F1 or ICR female mouse and cultured on six well plate and insert for
10 days in vitro. Ovaries were cultured in DMEM/F12 containing BSA,
ITS-X, and ascorbic acid, with or without PPAR gamma modulator for 2
days and transferred PPAR gamma modulator free medium. To evaluate
the expression of PPAR gamma, PTEN, and Akt1 on mouse ovary, we performed
real-time-PCR, immunohistochemistry, and western blot. Also, end
of culture, ovaries were fixed 10% neutral-buffered formalin and subjected
of HE staining, immunohistochemistry against Ki-67, AMH, PTEN, Akt1
and FOXO-3a, and apoptosis assay with TUNEL. Also, western blot was performed
to evaluate the changes of PTEN, Akt1.
RESULTS: The expression of PPAR gamma, PTEN, Akt1 and FOXO3a
were evaluated in ovaries from 5, 10, 15, 20 days and 8 weeks old mouse
by RT-PCR. PPAR gamma was expressed in ovaries from all the ages, it
increased in ovary in 20 days explosively. PTEN and Akt1 were increased
in ovary form 10 Days. In PPAR gamma antagonist treated ovaries, PTEN
was decreased and Akt1 was activated. Also nuclear exclusion of FOXO3a
was observed in oocytes of primordial follicles at 6hr treatment with PPAR
gamma antagonist. AMH expression was observed in PPAR gamma antagonist
treated ovaries, but was not in PPAR gamma agonist. After 12 days culture,
increases of ovarian sizes of PPAR gamma antagonist treated groups
were observed as compare to non-treated or PPAR gamma agonist treatment.
CONCLUSIONS: From these result, PPAR gamma may participate primordial
follicle activation and further development, possibly mediated in part of
PI3K-PTEN signaling pathway in vitro. Generation of activated primordial follicle
in cancer treatment patients, or other infertile women may allow fertility
preservation. Also Production of a large number of oocytes may facilitate
future derivation of embryonic stem cell for regenerative medicine.
Supported by: This work was Supported by a grant from the Korea Healthcare
Technology R&D Project, Ministry for Health, Welfare & Family affairs,
Republic of Korea (A120080).
P-434 Wednesday, October 21, 2015
THE COMPARISON OF THE EFFECTS OF SINGLE-DOSE METH-
OTREXATE AND SALPINGECTOMY ON OVARIAN RESERVE IN
TERMS OF THE ANTI-MULLERIAN HORMONE LEVEL
MEASUREMENT. G. Sahin Ersoy. Obstetrics and Gynecology, Dr. Lutfi
Kirdar Education and Research Hospital, Istanbul, Turkey.
OBJECTIVE: Ectopic pregnancy constitutes 1-2% of all pregnancies and
has 6% maternal mortality rate. Both surgical treatment and methotrexate
(MTX) administration for ectopic pregnancy is still controversial in terms
of ovarian reserve protection. Anti-mullerian hormone (AMH) is the best current
available measure of ovarian reserve for different clinical conditions.
(ref). To the best of our knowledge there is not any study dealing with the
long term effects of these two treatment methods on AMH levels. Therefore
we aimed to compare the effects of single-dose MTX and salpingectomy on
ovarian reserve in fertile women with ectopic pregnancy in the long term.
DESIGN: Cross-sectional study.
MATERIALS AND METHODS: A total of 181 patients have been
included into the study; 101 of them had received single-dose MTX or salpingectomy
treatment for ectopic pregnancy in the last 12 to 18 months. Patients
who have underwent salpingostomy, tubal milking, fimbriectomy and other
tuba-ovarian surgery were excluded from the study. Patients in the salpingectomy
group did not receive MTX. Their AMH, follicle stimulating hormone
(FSH), estrogen and antral follicle counts were evaluated.
RESULTS: The duration of time passed between application of treatment
and the measurement of AMH levels for MTX and salpingectomy groups
were 15.43 1.77 and 14.91 1.84, respectively (p¼0.144). The average
age was similar in both groups (p¼0.094). None of the three groups displayed
a significant difference in terms of FSH, E2 levels and antral follicle counts
(p¼0.393, p¼0.117, p¼0.528, respectively).There were no statistically significant
difference in AMH levels in all groups (2.96 0.85, 2.82 0.77
and 2.68 0.59 for control, MTX and salpingectomy groups respectively,
P¼0.147). (Table reference:aData are expressed as mean SD. AMH¼
Anti-mullerian hormone, FSH¼ Follicle stimulating hormone, E2¼ Estrogen,
AFC¼ Antral follicle count.
CONCLUSIONS: Neither single-dose MTX nor salpingectomy does have
any detrimental effects on ovarian reserve for the treatment of ectopic pregnancy
in terms of serum AMH levels and does alter antral follicle counts in
the long term.
Comparison of age, measures of ovarian reserve and smoking status.
Control
(n¼80)
Methotrexate
(n¼56)
Salpingectomy
(n¼45)
Age (years) a 27.15 3.49 27.18 2.97 28.33 3.01 0.094
AMH(ng/mL) a 2.96 0.85 2.82 0.77 2.68 0.59 0.147
FSH(mIU/mL) a 6.87 1.79 7.23 1.41 7.10 1.33 0.393
E2(pg/mL) a 44.61 13.94 45.41 13.95 40.11 7.63 0.117
AFCa 10.25 1.87 10.10 1.92 9.84 2.01 0.528
Smoking
(user/total)
17/80 11/56 10/45 0.949
P-435 Wednesday, October 21, 2015
IS A FREEZE-ALL CYCLE JUSTIFIED IN POOR
RESPONSE? M. Berkkanoglu, K. Coetzee, H. Bulut, K. Ozgur. Antalya
IVF, Antalya, Turkey.
OBJECTIVE: To investigate what the best treatment strategy is, if there
are 4 or less oocytes after an oocyte pick-up (OPU).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: The study group included 431 women
undergoing an antagonist protocol for intracytoplasmic sperm injection
and had 4 or less oocytes after OPU. Later, they were asked to have embryo
transfer (ET) on day 2 or day 5 or all freeze on day 5 and subsequent transfer
in a thawed cycle (FET).278 women (group A) wanted a cleavage stage day 2
embryo transfer and 153 women (group B) wanted their embryos cultured on
to day 5 to allow for blastocyst formation. 51 (group B1) of the day 5 group
had a fresh blastocyst transfer and 44 (group B2) had all their blastocysts
vitrified and had a vitrified-warmed blastocyst transfer. Blastocyst vitrification
was performed using the Cryotop method and technology.Student’s t
test and Chi-square test were used for statistical comparisons.
RESULTS: There was no significant difference in the mean age between
the groups (p¼0,288). 204 patients of group A and 95 of group B had ETs.
The cancellation ratio of ET after OPU was significantly lower in group A
than group B (26.1% vs 37.9%, p ¼ 0.02), but the pregnancy rates (PR)
per OPU were higher in group B than group A (31.3% vs 18.3%,
p¼0.003). PRs per ET were also higher in group B than group A (50.5%
vs 25%, p

P-436 Wednesday, October 21, 2015
DOES AGE MODIFY THE PREDICTIVE ABILITY OF
AMH? P. Kovacs, a B. Buzas, a F. Rarosi. b a Kaali Institute IVF Center, Budapest,
Hungary; b Dept. of Medical Physics, Bolyai Institue, Univ. of
Szeged, Szeged, Hungary.
OBJECTIVE: Anti Mullerian hormone (AMH) is one of the best
markers of ovarian reserve but it does not predict pregnancy well
either. Age is an important factor influencing fertility treatment
outcome. Our aim was to test the predictive ability of AMH in
different age categories.
DESIGN: Single IVF practice observational study.
MATERIALS AND METHODS: All IVf cycles performed between
Jan 1, 2014 and Dec 31, 2014 were considered for the analysis. Cycles
involving donor egg use and cycles that have not progressed to egg
retrieval were excluded. Cycles in which AMH result was not available
were excluded too. Three AMH categories were evaluated (low: under
1.1 ng/ml; normal: 1.11-3.5 ng/ml; high: over 3.5 ng/ml). IVF treatment
outcomes were compared in 3 age categories (under 35 yrs, 35-
40 yrs, over 40 yrs) based on AMH values. Egg yield, amount of gonadotropins
used per oocyte collected (gonadotropin IU/oocyte) and
ongoing pregnancy rate (OPR) were compared. The low AMH category
was further divided based on FSH/estradiol results (FSH>10IU/l and/
or E2>250 pmol/l and FSH

CONCLUSIONS: Compared to conventional IVF, ZACS permits DOR
patients to reap the benefits of trophectoderm biopsy aneuploidy screening
and offers logistical advantages over other embryo banking approaches. Patients
40 years
and/or had AMH

MATERIALS AND METHODS: The study was performed at the
Department of Obstetrics and Gynecology, Hacettepe University, between
November 2013 and March 2014. Total 2085 patients underwent
an ultrasound scan (Voluson 730, GE Healthcare, Istanbul, Turkey) by
one of the two physicians (C.P. or Z.D.) between days 1 and 12 of the
menstrual cycle. Inclusion criteria were (1) age 20 - 50, (2) regular
menstrual bleeding between 21 to 35 days, and (3) optimal visualization
of both ovaries. The exclusion criteria were (1) hirsutism or menstrual
irregularity, (2) any hormonal drug or oral contraceptive pill use
within the last 6 months, (3) history of endometrioma cystectomy or
detection of current endometrioma at the time of ultrasonography,
and (4) pregnancy. The fertility status was not a criterion while
deciding to include or exclude. The sum of both ovaries counts produced
the final AFC. The LMS method was preferred to produce the
smoothed centile curves of antral follicle count by age. For each set
of percentile curves, the initial smoothing methods were applied to
10th, 25th, 50th, 75th and 90th percentiles. For the comparison of
AFC across the age groups, statistical analyses were performed by
SPSS v21.0. Significance value was set at 0.05. Approval from institutional
review board was obtained.
RESULTS: Of the 2805 women that had been examined with ultrasonography
during the study period, 381 were appropriate for the final evaluation
according to inclusion and exclusion criterion (Table 1). The mean decrease
of AFC in each year was 0.41. Among the age groups, there were no statistical
significance between 20-24, 25-29 and 30-34, whereas decline in AFC
was obvious after 35 years.
CONCLUSIONS: The documentation of AFC nomogram might present
several benefits in theory. Initially, marking the current AFC of a given
woman across the nomogram might predict a woman’s total fertility potential
not only for assisted reproduction technologies (Polyzos, 2014) but also for
the chance of natural conception and pregnancy outcome. In conclusion,
comparison of our data and others suggest that AFC nomogram from
different reports and countries present similarity regarding the decline by
age. Those nomograms might be used to define age specific thresholds according
to the clinical conditions.
P-442 Wednesday, October 21, 2015
PATIENTS WITH LOW THEN NORMAL AMH MEASUREMENTS
DEMONSTRATE OVARIAN RESERVE COMPARABLE TO THOSE
WITH TWO NORMAL MEASUREMENTS. J. Gingold, a,b
j. M. knopman, c S. Talebian, d M. C. Whitehouse, e J. A. Lee, d
A. B. Copperman. d,a a Obstetrics, Gynecology and Reproductive Science,
Icahn School of Medicine at Mount Sinai, New York, NY; b Obstetrics/Gynecology
and Women’s Health Institute, Cleveland Clinic
Foundation, Cleveland, OH;
c Reproductive Medicine Associates of
New York, NY, NY;
d Reproductive Medicine Associates of New
York, New York, NY;
e Reproductive Medicine Associates of New
York, New York City, NY.
OBJECTIVE: There is a subset of patients who present initially with an
unexpectedly low AMH level. These situations precipitate concern about
short- and long-term female reproductive function. A later discordant confirmatory
test raises questions about whether the lower or the higher level is the
better predictor of ovarian reserve and responsiveness. This study sought to
evaluate whether oocyte yields in patients with discordant AMH levels were
more similar to those of patients with concordantly normal or abnormal
levels.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients who underwent two random
AMH measurements separated by > 30 days in the year prior to their
IVF start date from July 2007-March 2015 were included. Patients
with AMH >5 ng/mL were excluded. The first and second AMH
measurements were categorized as abnormal (

OBJECTIVE: This study was for aim, to identify circulating microRNAs
(miRNAs) in human follicular fluid (FF) in relation to
ovarian reserve disorders, providing new biomarkers of female infertility.
DESIGN: This prospective monocentric study included 131 patients
among which 91 women with normal ovarian reserve, 10 with ovarian
insufficiency (OI) and 30 affected by polycystic ovary syndrome
(PCOS). A pool of FF was retrieved for each patient during IVF/ICSI
procedure.
MATERIALS AND METHODS: At oocyte retrieval day, all FF were
collected and pooled for each patient. MicroRNAs were extracted from FF
pools and quantified by RT-qPCR, using TaqMan technology. The candidate
miRNAs, detected in FF samples were let-7b, miR-30a, miR-140 and miR-
191.
RESULTS: The miR-30a expression was significantly increased in FF
pools from women with PCOS compared to those with normal ovarian
reserve (p¼0.006). By contrast, miR-140 and let-7b levels were significantly
expressed at low levels in FF pools from PCOS patients than patients
with normal ovarian reserve (p¼0.01; p¼0.01, respectively). The
Receiving Operator Curve (ROC) analysis showed that the performance
of the combination of these three miRNAs in detecting PCOS reached
0.83 [0.73-0.92] with 83.8% specificity and 70% sensitivity
(p40g/m2,
or bone marrow transplant (n¼20) and others were classified as having
low dose treatment (n¼22). A matched set of randomly timed fingerstick
dried blood spot samples and venipuncture serum samples were
collected from each participant and sent to Ansh Labs for analysis using
the DBS AMH ELISA and picoAMH ELISA. These are three-step, sandwich-type
enzymatic microplate assays, the DBS was developed to measure
AMH levels in two 7.9 mm dried blood spot discs after extracting
the blood from the filter paper. Levels of serum and bloodspot AMH
were compared between groups using t-tests and multivariable linear
regression.
RESULTS: Compared to similar age controls (mean age 25.9), cancer
survivors had significantly lower serum and bloodspot AMH
values (bloodspot AMH 1101pg/ml vs. 2957pg/ml, p¼0.009) in unadjusted
and age adjusted models. High dose survivors had significantly
lower serum and bloodspot AMH values (bloodspot AMH 474pg/ml
vs. 2023pg/ml, p¼0.005) in unadjusted and age adjusted models.
Finally, late reproductive age controls (mean age 42.9) had significantly
lower serum and bloodspot AMH compared to mid-reproductive
age controls (mean age 25.9) (bloodspot AMH 759pg/ml vs.
2957pg/ml, p¼0.003).
CONCLUSIONS: Differences in ovarian reserve can be detected in
cancer survivors and late reproductive women compared to mid reproductive
controls using the Ansh Labs DBS AMH assay on bloodspot
samples, or picoAMH assay on serum samples. Blood spot collection
may be a useful, less invasive, low cost tool in evaluating ovarian
reserve and reproductive potential in populations at risk for ovarian
insufficiency.
Supported by: Ansh laboratories RO1-HD-062797-05.
P-446 Wednesday, October 21, 2015
CLINICAL USEFULNESS OF AMH APPLIED IN POOR OVARIAN
RESERVE PATIENTS IN AN ART PROGRAM. M. Chen, H. Guu,
Y. Chen, Y. Yih, M. Chou. Department of Obstetrics and Gynecology and
Women’ Health Health, Taichung Veterans General Hospital, Taichung,
Taiwan.
OBJECTIVE: The purpose of present study is to investigate the threshold
value of AMH to predict a poor ovarian response after COH and determine
appropriate protocol to maximize the successful rate of treatment in those
POR patients in our ART program.
DESIGN: A retrospective study with the ART database from Jan 2011 till
Oct 2013 was reviewed.
MATERIALS AND METHODS: We had a total 994 start cycles and
908 finished OPU cycles with total 36.8 % CPR and 28.5% OGPR/
LBR. 416 start cycles had AMH value within 6 months of COH. 161
(5 cancelled) long agonist cycles and 216 (27 cancelled) antagonist cycles
were eligible for analysis. POR was defined as cycles < 3 (mature)
oocytes retrieved or cancelled due to poor response after COH. ROC
curves for predicting POR by AMH value were created and threshold
levels were determined. Predicting factors for POR in these patients
were searched. The COH responses and the ART outcome of those patients
with AMH value around the threshold value were analyzed according
to the COH protocol and gonadotropin dosage administered.
Clinically useful opinions were derived from all these preliminary analyses.
Statistics were carried out by SPSS-PC ver.11.5 with p

CANCER
P-447 Wednesday, October 21, 2015
TUBAL LIGATION DOES NOT MODIFY OVARIAN CANCER RISK
IN BRCA MUTATION CARRIERS. J. Chan, a M. D. Sammel, b
T. M. Friebel, c C. Gracia, d T. R. Rebbeck. c a Reproductive Endocrinology
and Infertility, Hospital of the University of Pennsylvania, Philadelphia,
PA; b Univ. of Pennsylvania, Perelman School of Medicine, Rose valley,
PA; c Center for Clinical Epidemiology and Biostatistics, University of Pennsylvania,
Philadelphia, PA; d University of Pennsylvania.
OBJECTIVE: Risk-reducing salpingo-oophorectomy (RRSO) has been
shown to be protective against ovarian cancer (OvCa) in the BRCA mutation
carrier population and is recommended in women by age 40 or at completion
of childbearing(1). However, early RRSO is associated with premature
menopause. As tubal ligation (TL) has been demonstrated to reduce the
risk of OvCa in the non-BRCA population(2), we aimed to determine if
TL has a similar protective effect in BRCA carriers.
DESIGN: Cohort study
MATERIALS AND METHODS: Data from 5038 BRCA carriers were
collected between 1997 and 2011 from 23 centers. Demographics and reproductive
history/surgery were obtained. Occurrences of OvCa and vital status
were verified by medical records. Women who underwent TL were compared
with women who did not (non-TL). For failure-time analysis, observation
period started at date of birth and allocated all person-years of observation
(PYO) before TL to the non-TL group. PYO were thereafter allocated to
the TL group. Observation ended at age at OvCa diagnosis, RRSO, last contact,
death or study close, whichever came first. Log-rank test was used to
compare survival distributions. A Cox model with TL as a time-dependent
variable was used to obtain hazard ratios. Type of mutation, parity and oral
contraceptive pill (OCP) use were considered potential confounders. Ranksum
and Chi-square tests were used to compare demographic characteristics.
RESULTS: 392 underwent TL at a median age of 33. Groups were similar
with regards to BRCA mutation type (p¼0.5), Caucasian race (p¼0.7) and
Jewish heritage (p¼0.4). The TL group was older than the non-TL group
at the time of BRCA diagnosis (49 vs. 43, p

Histologic analysis of the tissue samples revealed no sign of microscopic
infiltration. Residual disease was detected by IgH monoclonality expression
in three of the mice from the DXM treated group and in three animals from
the CONTROL group. RCH-ACV cells were detected six months after injection
of amounts as little as 1000 cells, although only animals being injected
with 5x106 developed a clinical disease.
CONCLUSIONS: DXM incubation prior to re-transplantation of ovarian
tissue does not prevent the reintroduction of malignant disease. Although
the amount of cells introduced seem not to cause any relapse, caution in retrasplanting
ovarian tissue from patients with leukemia must be paid until
more secure system are developed.
Reference:
1. Kaspers GJ, Veerman AJ, Popp-Snijders C, et al. Comparison of the
antileukemic activity in vitro of dexamethasone and prednisolone in
childhood acute lymphoblastic leukemia. Med Pediatr Oncol.
1996;27(2):114-121.
P-450 Wednesday, October 21, 2015
DOES DIMINISHED OVARIAN RESERVE CONTRIBUTE TO
CANCER SURVIVORS HAVING FEWER CHILDREN THAN
DESIRED? A REPORT FROM THE FUCHSIA WOMEN’S
STUDY. P. P. Howards, a H. B. Chin, a A. Fothergill, a A. C. Mertens, b
J. B. Spencer. c a Epidemiology, Emory University, Atlanta, GA; b Pediatrics,
Emory University, Atlanta, GA; c Gynecology and Obstretrics, Emory University,
Atlanta, GA.
OBJECTIVE: To examine whether cancer survivors are failing to meet
their reproductive goals more than women never diagnosed with cancer,
and to compare antral follicle count (AFC) measurements between these 2
groups.
DESIGN: The FUCHSIA Women’s Study is a population-based cohort
study comparing reproductive outcomes among cancer survivors diagnosed
between 20 and 35 years to comparison women who were never diagnosed
with cancer, matched on age and place of residence.
MATERIALS AND METHODS: Cancer survivors were identified in the
Georgia Cancer Registry. Participants included 1,282 cancer survivors and
1,073 comparison women who were interviewed about their reproductive
history and desire for children. A subgroup of women also completed a clinic
visit where AFC was measured by ultrasound (n ¼ 374 cancer survivors, n ¼
376 comparison women).
RESULTS: Cancer survivors and comparison women were 22-45 years at
the time of the interview. During the interview, both groups of women reported
wanting an average of 2.5 children, but cancer survivors were less
likely to have met their reproductive goals compared with comparison
women (45% vs. 57%). There were also differences in cancer survivors’ ability
to meet their reproductive goals across cancer types. Women diagnosed
with a reproductive or blood cancer were least likely to achieve their desired
family size (42%). In a logistic model adjusting for age at interview, race, income,
and education, cancer survivors were significantly more likely to
report having fewer children than desired compared with comparison women
(OR ¼ 1.72, 95% CI: 1.45, 2.05). Cancer survivors also had a lower total
AFC (median ¼ 8, IQR: 3, 15) than comparison women (median ¼ 10,
IQR: 5, 18). Among women who had fewer children than desired, the median
AFC was 8, IQR: 4, 16 in cancer survivors and 11, IQR: 5, 19.5 in comparison
women. Women who never received chemotherapy or radiation were similar
to other cancer survivors with respect to meeting their reproductive goals
(44%), but had an AFC similar to the comparison women (median ¼ 10,
IQR: 5, 18).
CONCLUSIONS: Diminished ovarian reserve may contribute to cancer
survivors not having as many children as they want although other factors
may be important as well.
Supported by: NICHD 5R01HD066059 NICHD T32HD052460
P-451 Wednesday, October 21, 2015
SPERMATOZOA PROTEIN PROFILES IN CRYOBANKED SEMEN
SAMPLES FROM TESTICULAR CANCER PATIENTS BEFORE
TREATMENT. A. Agarwal, a E. Tvrda, a,b R. Sharma, a S. Gupta, a
G. Ahmad, a,c E. S. Sabanegh. d a Center For Reproductive Medicine, Cleveland
Clinic, Cleveland, OH; b Department of Animal Physiology, Faculty of
Biotechnology and Food Sciences, Nitra, Slovakia; c Physiology and Cell
Biology, University of Health Sciences, Lahore, Pakistan; d Urology, Cleveland
Clinic, Cleveland, OH.
OBJECTIVE: Patients diagnosed with cancer are encouraged to freeze
their sperm prior to the start of treatment. It is not clear how testicular cancer
impacts sperm quality at the proteomic level. Our goal was to identify differentially
expressed proteins in spermatozoa of patients with testicular cancer
who banked their specimens prior to start of their treatment and in spermatozoa
from infertile patients without cancer.
DESIGN: Prospective study examining proteomic profile of spermatozoa
from testicular cancer and non-cancer (infertile) patients.
MATERIALS AND METHODS: Semen samples were taken from 16
testicular cancer patients, prior to the start of their treatment and from 9
non-cancer patients (infertile men). Semen analysis was conducted before
the samples were banked. For proteomic analysis, cryoprotectant was
removed and the samples were pooled after normalizing for protein concentration.
Proteins were extracted and separated by 1-D gel. Bands were digested
and run on a LTQ-Orbitrap Elite hybrid mass spectrometer system.
Functional annotations of proteins were obtained using bioinformatics tools
and pathway databases.
RESULTS: The most prevalent testicular cancer diagnoses included
seminomas, mixed germ cell tumors, embryonal carcinomas and Sertoli
cell tumors. Significant differences in spermatozoa concentration were
observed between the testicular cancer and infertile patients (p

with cancer with and without embryo banking it was 31.0% and 36.7%,
respectively; for women without cancer, with and without embryo banking,
it was 31.0% and 31.3%, respectively. At cycle 2, live birth rates, and mean
birthweight and length of gestation did not differ significantly between
groups.
CONCLUSIONS: Women with cancer utilize embryo banking at higher
rates but are less likely to return for subsequent cycles and do so after longer
periods compared to women without cancer. Women with cancer who return
have comparable birth outcomes (live birth rates, birthweights and lengths of
gestation) to women without cancer. The use of embryo banking after cancer
may depend on several important factors, including cancer diagnosis, woman’s
age, and her partnership status.
P-453 Wednesday, October 21, 2015
TAMOXIFEN METABOLITES IN BREAST CANCER PATIENTS
UNDERGOING CONTROLLED OVARIAN HYPERSTIMULATION
(COH): ARE WE ACHIEVING THERAPEUTIC LEVELS?. L. Ross, a
N. J. Clarke, b F. Z. Stanczyk, a K. Chung. a a University of Southern California,
Los Angeles, CA; b Quest Diagnostics Nichols Institute, San Juan Capistrano,
CA.
OBJECTIVE: In breast cancer patients undergoing COH, tamoxifen is
used as an adjunct to gonadotropins to reduce the risk of stimulation of breast
cancer cells. The active metabolite, endoxifen, can be measured to assess
therapeutic activity. Our objective was to determine if the standard tamoxifen
dose used during COH in breast cancer patients results in therapeutic levels
of the active metabolite.
DESIGN: Retrospective analysis.
MATERIALS AND METHODS: Breast cancer patients with no previous
chemotherapy exposure who received tamoxifen during COH between
2008-2013 were identified. Tamoxifen 20 mg was initiated at
the start of COH and continued until oocyte retrieval. Samples were acquired
from serum stored from routine testing. Tamoxifen and its
metabolite levels were quantitatively measured throughout the COH cycle
using isotope dilution tandem mass spectrometry on a triple quadrupole
instrument. ANOVA and Fisher’s exact tests were used as
appropriate.
RESULTS: 9 women who met inclusion criteria were included in this
analysis. Mean age was 37.34.18. Mean BMI was 24.25.39. Mean number
of days on tamoxifen was 11.71.58. Tamoxifen and endoxifen serum
levels increased with increasing duration of treatment. Mean peak tamoxifen
level was 71.135.4 ng/mL and mean peak endoxifen level was
7.724.14 ng/mL. 7/9 women reached therapeutic endoxifen levels
(>5.97 ng/mL [1]). On average, therapeutic levels were reached by day 9
of stimulation (range day 7-10). Asian women were significantly less likely
than other ethnicities to reach therapeutic endoxifen levels (p¼0.03). While
there was no statistically significant difference in peak tamoxifen levels between
Asians compared to all other ethnicities (54.0vs76.0 ng/mL,
p¼0.47), there was a significant difference in peak endoxifen levels
(2.61vs9.19 ng/mL, p¼0.04).
CONCLUSIONS: Our data indicate that tamoxifen administered at 20mg
daily during COH achieved therapeutic endoxifen levels in the majority of
women, suggesting that this dose should provide adequate protection from
stimulation of breast cancer cells. Of concern, this dose did not reach therapeutic
levels in women of Asian ethnicity. It is possible that higher doses of
tamoxifen may be required in women of Asian ethnicity. Data on 8 more subjects
are currently being analyzed and will be included at the time of abstract
presentation.
Reference:
1. Madlensky L, Natarajan L, Simone T, Pu M, Mortimer J, Flatt S, Nikoloff
D, Hillman G, Fontecha M, Lawrence J, Parker B, Wu A, Pierce
J. Tamoxifen Metabolite Concentrations, CYP2D6 Genotype and
Breast Cancer Outcomes. Clin Pharmacol Ther 2011; 89(5): 718-725.
Supported by: Quest Diagnostics.
P-454 Wednesday, October 21, 2015
PATIENTS WITH LYMPHOMA DEMONSTRATE DIMINISHED
OVARIAN RESERVE BEFORE CANCER
TREATMENT. J. Lekovich, a A. Lobel, a J. Stewart, b N. Pereira, c
I. Kligman, a Z. Rosenwaks. a a Weill Cornell Medical College, New York,
NY; b Weill Cornell Medical Center, New York, NY; c The Ronald O. Perelman
and Claudia Cohen Center, New York, NY.
OBJECTIVE: Evidence shows that men with Hodgkin’s and non-Hodgkin’s
lymphoma have lower sperm quality before the start of chemotherapy.
We sought to investigate if women with lymphoma also demonstrate lower
ovarian reserve.
DESIGN: Retrospective cohort study
MATERIALS AND METHODS: The study group comprised 194 patients
with cancer who underwent fertility preservation before initiating
cancer treatment between 2010 and 2013. The control group consisted of
410 patients undergoing elective fertility preservation. Patients were
excluded if they had ovarian cancer, prior oophorectomy or were older
than 40. Cancer patients were compared to the control group. Then, a
sub-analysis within the cancer group compared breast cancer and lymphoma
patients. Primary outcomes included anti-mullerian hormone
(AMH) levels and number of eggs retrieved and cryopreserved. Secondary
outcomes included number of days of stimulation and the total amount of
gonadotropins used for stimulation. Mann-Whitney test was used for comparison
of continuous variables. Chi-Square and Fisher’s exact tests were
used for comparison of categorical variables as indicated. P

FERTILITY PRESERVATION
P-455 Wednesday, October 21, 2015
OVARIAN TRANSPLANTATION WITH SCAFFOLDS FOR DRUG
DELIVERY: AN IN VIVO TRANSGENIC MOUSE
MODEL. C. Chen, a,b S. Tan, a,c C. Tzeng. a,b a Center for Reproductive
Medicine and Sciences, Department of Obstetrics and Gynecology, Taipei
Medical University Hospital, Taipei, Taiwan; b Department of Obstetrics
and Gynecology, School of Medicine, College of Medicine, Taipei Medical
University, Taipei, Taiwan; c Graduate Institute of Clinical Medicine, College
of Medicine, Taipei Medical University, Taipei, Taiwan.
OBJECTIVE: Ovarian tissue cryopreservation and autotransplantation is a
promising option for fertility preservation of female cancer patients. However,
ischemia limits the life span of the ovarian grafts after grafting. Vascular
endothelial growth factor (VEGF) can promote angiogenesis, and sphingosine-1-phosphate
(S1P) can protect ovarian grafts from ischemic reperfusion
injury. This study aimed to investigate the efficacy of scaffolds for delivering
different drugs in promoting survival of ovarian grafts.
DESIGN: In vivo study with a transgenic mouse model.
MATERIALS AND METHODS: We use scaffolds served as vehicles for
drug delivery to promote the graft survival.Ovaries from 8-week-old FVB/N-
Tg(PolII-Luc)Ltc transgenic mice with or without scaffolds loaded with S1P
(2 mM, 5 mL) or VEGF (0.2 mg/ml, 5 mL) were transplanted into the peritoneum
of wild-type mice. The graft survival was tracked in vivo by bioluminescence
imaging (BLI) for 4 weeks, and histological examination was
performed at the end of the experiment.
RESULTS: Stronger signals of in vivo BLI were observed in the ovaries
with S1P- and VEGF-loaded scaffolds than those in the scaffolds without
drugs and those without scaffolds. Histological examination also showed
more follicles and surrounding vessels in the S1P group compared with other
groups. The above indicated better survival of the grafts.
CONCLUSIONS: We demonstrated that scaffolds loaded with drug can
promote ovarian graft survival.Scaffolds mimicking the structure and biological
function of native extracellular matrix are beneficial for tissue growth,
and applying tissue engineering technology may overcome some limitations
in regenerative medicine.
Supported by: This work was Supported by the grant MOST 103-2321-B-
038-008 from the Ministry of Science and Technology, Taiwan, R.O.C.
P-456 Wednesday, October 21, 2015
OBJECTIVE ASSESSMENT OF THE ONCOFERTILITY EDUCA-
TIONAL INFORMATION, AVAILABLE TO WOMEN, ON THE
WEBSITES OF NCI-DESIGNATED CANCER CENTERS IN THE
US: DO SOCIOECONOMIC DEMOGRAPHIC PROFILES BY STATE
MAKE A DIFFERENCE? C. de Haydu, a S. V. Eleswarapu, b
A. A. Dabaja, b C. M. Duke. a a Yale University School of Medicine, New Haven,
CT; b Henry Ford Health System, Detroit, MI.
OBJECTIVE: Current guidelines recommend that reproductive age
women, newly diagnosed with cancer, should be counseled on fertility preservation
(FP). Hospital websites increasingly serve as portals for reliable
web-based resources to supplement the knowledge of patients and families
regarding their diagnoses & treatments. This study aims to assess the quality
of hospital web-based resources which are available to women undergoing
cancer treatment at major cancer centers.
DESIGN: Prospective observational study.
MATERIALS AND METHODS: A validation rubric for FP/oncofertility
content quality standards using a scoring system for commonly accepted definitions
& terminology was developed. The publicly available websites of the National
Cancer Institute Designated Cancer Centers (NCICC) & the Cleveland
Clinic Foundation (CCF) were accessed by independent teams from two different
institutions between Nov. 1, 2014 & April 30, 2105 & queried in a systematic
fashion. Specific queries included: 1) Does the website discuss the effects of cancer
& cancer treatment on female fertility? 2) Are options for FP for all patients
discussed? 3) Is there a standalone page dedicated to educating patients on FP? 4)
Is parenting-related cancer survivorship addressed? 5) Is there a link to outside FP
information?. State & Region based demographic information on racial makeup,
household income & poverty status were obtained from the 2010 US Census Bureau’s
‘‘Geographic Level of Poverty & Health Estimates’’. Chi-square tests were
performed to assess for differences between FP website scores (individually &
within Regional groups); analysis was also performed to assess for any correlation
between socioeconomic/racial differences within States/Regions where NCICC
are located. Multivariate logistic regression analyses are ongoing.
RESULTS: 62 clinical NCICC were identified, including CFF. 92% of queried
sites were academic institutions. 84% of all websites mention the risk of cancer
treatment on a woman’s fertility potential but 44% do not discuss FP options
for women. 56% of websites have pages dedicated to discussion of non gender
specific FP options & 65% of the websites contain links to further resources. In
population based adjusted analyses, NCCIC in States where 50% of the population
identified as Non-Hispanic White (even after controlling for socioeconomic
status), p-value < 0.04. There were no differences observed when similar adjustments
& analyses were performed by for US census bureau geographic Regions.
CONCLUSIONS: Preliminary data suggest that NCICC websites are
inconsistent in the quality of oncofertility educational information for female
patients. Racial makeup of a State is associated differences in the quality of
patient centered oncofertility web-based resources for women. These findings
are concerning and suggest that more uniformed efforts aimed at attenuating
these racial gaps in patient education are needed.
P-457 Wednesday, October 21, 2015
THE EFFECT OF VITRIFICATION AND PROGRAMMED
FREEZING ON FROZEN-THAWED HUMAN OVARIAN CORTEX
TISSUE. X. Wang, a C. Fang, a C. Di, a H. Liu, b X. Liang. a a Reproductive
Medicine Research Center, The Sixth Affiliated Hospital of Sun Yat-sen University,
Guangzhou, China;
b Obstetrics and Gynecology Department,
Guangzhou Development District Hospital, Guangzhou, China.
OBJECTIVE: It has been long remained controversial whether vitrification,
compared with programmed freezing, is better for ovarian fertility preservation.
Therefore, this study aimed at comparison of the effects of these two techniques
on the frozen-thawed human ovarian tissues, to provide some
experimental evidences to select a better method to preserve women fertility.
DESIGN: Ovarian tissues came from cases of partial ovariectomy, with necessity
of diagnosis or treatment. And all the pathological reports told neither
tumor cell metastasis in the ovarian tissue nor existence of other nidi. In each
case, the cortex was cut into small pieces, followed by randomly divided into
Fresh (F) group, Vitrification (V) group and Programmed Freezing (PF)
group. Morphology changes, apoptosis in situ, follicle viability and secretion
function were compared with self-control method and in-vitro cultivation.
MATERIALS AND METHODS: Ovarian tissues came from 6 cases,
which met the above criteria. The morphology of tissues were examined
by HE staining. The cell apoptosis in situ was analyzed by TUNEL assay.
Neutral red staining and CaAM/EthD-1 staining were conducted respectively
after collagenase-1 digestion of frozen-thawed tissues, and the survival rate
of small follicles was analyzed. During in vitro culture of frozen-thawed
ovarian tissues, the secretion level of estrogen (E2) and progesterone (P4)
in media were examined and analyzed at Day 2, Day 4, Day 6 and Day 8.
RESULTS: The morphology of follicles and stroma cells in both V Group
and PF Group were similar as F Group. The apoptosis in situ showed no significance
between V Group and F Group, while apoptosis in PF Group is
significantly higher than other two groups. After frozen-thawed, no significant
differences were showed in assessment of viability of small follicles between
PF Group and V Group. Analysis of E2 and P4 level showed that secretion of
E2 had a tendency to change over time, and time effect varies with grouping. It
increased significantly higher in PF Group at Day 4 and Day 8, but similar at
Day 6, when compared with V Group. The secretion of P4 also had a tendency
to change over time, but no significant differences were found in the effect of
interaction of time and grouping, neither in the grouping effect.
CONCLUSIONS: Both of vitrification and programmed freezing can well
preserve the morphological characteristics of ovarian cortex tissue, but vitrification
may be better for protection of DNA integrity in cells. Both of vitrification
and programmed freezing can well preserve the viability of small
follicles in frozen-thawed tissues. Both of the frozen-thawed tissues after
vitrification and programmed freezing can recover secretion function. And
functional reconstruction of frozen-thawed ovarian tissues with these two
techniques still needs further researches.
Supported by: This study was Supported by the National Natural Science
Foundation of China (Grant No.81070495), and the Natural Science Foundation
of Guangdong Province (Grant No. S2013010013404).
P-458 Wednesday, October 21, 2015
MENSTRUATION IS AN UNRELIABLE SURROGATE IN THE
ASSESSMENT OF OVARIAN DAMAGE BY CHEMOTHERAPY: A
PROSPECTIVE LONGITUDINAL STUDY WITH AMH LEVELS
AS THE GOLD STANDARD. G. Bedoschi, a,b S. Goldfarb, c
J. Quistorff, c S. Goswami, d F. Moy, a M. Dickler, c K. Oktay. a,b a Obstetrics
and Gynecology, NYMC, Valhalla, NY; b Innovation Institute for Fertility
Preservation and IVF, Rye, NY; c Memorial Sloan Kettering Cancer Center,
New York, NY; d Yeshiva University, New York, NY.
e262 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

OBJECTIVE: The majority of studies assessing the impact of chemotherapy
(CT) on ovarian reserve continue to use menstruation as a surrogate. We aimed
to determine the reliability of menstrual status and pattern in assessing CTinduced
ovarian damage, using the serum AMH level as the gold standard.
DESIGN: Prospective longitudinal study.
MATERIALS AND METHODS: 81 women with breast cancer stage 1-3
were prospectively enrolled and followed up for 18 months (18mo). Sera
were frozen at baseline (BL) and 18mo post-CT, and were assayed for AMH
(ng/ml) at once. Women kept monthly menstrual calendars. Amenorrhea was
defined as no menses for >6 months, and regular periods as those with 21-35
day intervals. Results were analyzed with t-test or ANOVA for continuous variables
and chi square or Fisher’s exact test for discrete variables.
RESULTS: The median age at CT was 38 (range 27-44); 72.1% received
anthracycline-based, 13.2% received CMF, 13.2% received taxane-based and
1.5% received other CT regimen. Ten women did not complete menstrual calendars
and were excluded from the analysis. Seventeen (23.9%) patients
developed amenorrhea and 54 (76.1%) were menstruating at the end of
18mo follow up. The groups were similar in age, BL AMH , CT protocol,
and adjuvant tamoxifen use. Of those who were menstruating post-CT,
48.5% had RP and 51.5% had irregular periods (IP). Surprisingly, women
who developed amenorrhea post-CT had higher AMH levels than those
who retained menstruation post CT (0.770.34 vs. 0.230.1 ng/ml,
p¼0.049). However, women with RP showed a trend for higher AMH
post-CT than those with IP (0.310.15 vs. 0.050.02, p¼0.076). Of the
women who had detectable AMH levels at 18mo, a similar percentage had
amenorrhea vs. continued menstruation (41.18% vs. 58.82%, p¼0.393).
Moreover, their mean AMH levels were not significantly different.
CONCLUSIONS: Our data indicate that menstrual status has very little
value in assessing CT-induced ovarian damage. Studies that investigate the
impact of CT or interventions to preserve ovarian function should utilize
more reliable markers such as the serum AMH.
Supported by: NIH HD053112 (NICHD & NCI), Jodi Spiegel Fisher Cancer
Foundation and Susan G. Komen Foundation.
P-459 Wednesday, October 21, 2015
DEVELOPMENT OF A NEW LOCAL DRUG DELIVERY SYSTEM
FOR THE UTERUS USING BIO-NANOCAPSULE. K. Koizumi, a
H. Nakamura, a T. Matsuzaki, b S. Kuroda, c Y. Yasui, a K. Furuya, a
T. Miyake, a T. Takiuchi, a K. Kumasawa, a T. Kimura. a a Obstetrics and Gynecology,
Osaka University Graduate School of Medicine, Suita Osaka,
Japan; b Cardiovascular Medicine, Osaka University Graduate School of
Medicine, Suita Osaka, Japan; c The Institute of Scientific and Industrial
Research, Osaka University, Suita Osaka, Japan.
OBJECTIVE: Uterus is the applicable organ for local gene therapy
because it is not part of the peritoneum organ and it can be reached directly.
However, we still do not have any drug delivery system for uterus.Bio-nanocapsule
(BNC) containing hepatitis B virus surface antigen consists of
approximately 50-nm hollow particles displaying a human hepatocyte-recognizing
molecule (pre-S1 peptide). BNC has been used as an HB vaccine for
the last three decades. In this study, we optimized the BNC as a new local
drug delivery system for uterus.
DESIGN: Animal experiment.
MATERIALS AND METHODS: The N terminal of Pre-S1 peptide was
replaced with the TAT (trans-activating transcription factor) peptide.The
Cy7 labeled BNC was transferred into the murine uterine cavity. The distribution
of BNC was observed by in-vivo imaging system and also by immunohistochemistry.
The luciferase expression plasmid DNA was incorporated
into BNC using liposome. The luciferase expression plasmid DNAwas transferred
into uterine cavity using TAT-BNC-liposome complex. The efficiency
of gene transfection was analysed by luciferase assay.
RESULTS: BNCs were observed in the luminal and glandular epithelial
cells, but not in stroma and myometrium. The transfection efficiency of the
TAT-BNC-liposome complex was significantly higher than lipofection.
CONCLUSIONS: These results suggest that BNC could be an applicable
DDS for uterus. In this study, we replaced the N terminal of Pre-S1 peptide
with TAT peptide. However, it is replaceable with sugar chains and antibodies.
Recently there are some reports that some of special sugar chains
are expressed in the uterine endometrium during pregnancy and uterine cancer.
If we can find specific sugar chains or cell surface antibodies on uterine
endometrium for reproductive dysfunction and uterine cancer, this DDS system
can be more targetable.
Supported by: Grants-in-Aid for Scientific Research from the Ministry of
Education, Science and Culture of Japan (Tokyo, Japan).
P-460 Wednesday, October 21, 2015
SMALL ANTRAL FOLLICLE RESPONSIVENESS TO FSH, AS-
SESSED BY THE FOLLICULAR OUTPUT RATE (FORT), IS NOT
ALTERED IN CANCER PATIENTS, CANDIDATES FOR FERTILITY
PRESERVATION. S. Duros, a C. Sonigo, b J. Benard, a C. Sifer, c
M. Grynberg. d a Department of Reproductive Medicine, Hopital Jean Verdier,
APHP, BONDY, France; b Department of Reproductive Medicine, H^opital
Jean Verdier, APHP, BONDY, France; c Department of Cytogenetic and
Reproductive Biology, Hopital Jean Verdier, APHP, BONDY, France; d Department
of Reproductive Medicine, BONDY, France.
OBJECTIVE: To evaluate the small antral follicle responsiveness to exogenous
FSH, assessed by the Follicular Output RaTe (FORT), in cancer patients,
candidates for fertility preservation (FP) using oocyte vitrification
after controlled ovarian hyperstimulation (COH).
DESIGN: Prospective study.
MATERIALS AND METHODS: From July 2013 to December 2014, 71
cancer patients, aged 20-40 years, candidates for oocyte vitrification
following COH (FP group) were studied. Ovarian stimulation characteristics
and outcomes were compared with that of 91 infertilewomen (Control group),
included in an in vitro fertilization program in our centre during the same time
frame, matched for age, antral follicle count (AFC) and serum Anti-Mullerian
hormone (AMH) levels measured just before initiation of the stimulation (d0),
as well as FSH starting dose. All patients had 2 ovaries, no previous history of
chemotherapy and underwent COH using GnRH antagonist protocols. Antral
follicles were counted before FSH administration, and on the day of ovulation
triggering (dOT). FORT was determined by the ratio between the pre-ovulatory
follicle count (16-20 mm) on dOT 100/AFC on d0.
RESULTS: By design, mean age, AFC, AMH and FSH starting dose were
similar in FP and Control groups (31.5 3.6 vs. 32.2 4.9 years; 17.4 9.8
vs. 16.6 8.3 follicles, 2.9 2.4 vs. 3.0 1.7 ng/mL; 269 81 vs. 248 56
IU, respectively, NS). Characteristics and outcomes of the stimulation in both
groups are reported in the Table.
CONCLUSIONS: The present investigation shows that the cancer status
may not impact the responsiveness of small antral follicles to exogenous
FSH, assessed by the FORT, in candidates for oocytes vitrification. However,
alterations in the granulosa cell function in relation with the malignant disease
may account for the significantly lower levels of serum E 2 reached in the end
of the ovarian stimulation in these patients when compared to infertilewomen.
Characteristic and outcome of the stimulation in FP and Control groups.
FP group
(n¼71)
Control group
(n¼91)
Mean total dose of gonadotropin 29311095 26101188 NS
(IU)
Mean duration of stimulation 10.31.7 10.12.2 NS
(days)
E 2 on dOT (pg/mL) 1261 806 1904997 12 mm (pg/mL) 132.593.2 187.694.8 16 mm on
dOT x 100 / AFC on d0) (%)
3622 3720 NS
P-461 Wednesday, October 21, 2015
ANTI-M€ULLERIAN HORMONE PREVENTS CHEMOTHERAPY-
INDUCED FOLLICULAR BURNOUT. S. Tan, a,b C. Chen, a,c
C. Tzeng. a,c a Center for Reproductive Medicine, Department of Obstetrics
and Gynecology, Taipei Medical University Hospital, Taipei, Taiwan; b Graduate
Institute of Clinical Medicine, College of Medicine, Taipei Medical University,
Taipei, Taiwan; c Department of Obstetrics and Gynecology, School
of Medicine, College of Medicine, Taipei Medical University, Taipei,
Taiwan.
OBJECTIVE: Chemotherapeutic drugs may damage the reproductive system
and lead to infertility and premature ovarian failure. How to prevent
follicular loss is the key to preserve ovarian reserve. Anti-M€ullerian hormone
(AMH), which is produced by the granulosa cells of growing follicles, can
inhibit primordial follicle activation and follicle growth stimulated by follicle-stimulating
hormone. This study aimed to investigate the inhibitory effect
of recombinant AMH on chemotherapy-induced follicular burnout.
p
FERTILITY & STERILITY Ò
e263

DESIGN: In vivo mouse study.
MATERIALS AND METHODS: Eighteen 8-week old mice were randomly
divided into three groups: control, cyclophosphamide (Cy) group, and AMH + Cy
group. Mice in the AMH + Cy group were pretreated intraperitoneally with recombinant
AMH (2 mg) 1 hour before intraperitoneal injection of Cy (150 mg/
kg). Histological examination of ovaries was performed three and seven days later.
RESULTS: In the Cy-treated ovaries, the number of primordial follicles
was decreased, and the ratio of growing to primordial follicles was significantly
increased. Mice pretreated with recombinant AMH had similar ratio
of growing to primordial follicles to the controls, indicating that recombinant
AMH prevented the recruitment of follicles triggered by Cy treatment.
CONCLUSIONS: We demonstrated that pretreatment of recombinant AMH
inhibited the transition of the primordial follicles to growing follicles. Other tests
were under investigation to examine its application to fertility preservation.
P-462 Wednesday, October 21, 2015
VALUE OF ANTIMULLERIAN HORMONE AND ANTRAL FOLLI-
CLE COUNT IN PREDICTING FERTILITY PRESERVATION CY-
CLE OUTCOMES. V. Emirdar, a V. Turan, a F. Moy, b G. Bedoschi, a
K. H. Oktay. a,c a Department of Obstetrics&Gynecology, New York Medical
College, Valhalla, NY; b Biostatistics and Epidemiology, New York Medical
College, Valhalla, NY; c Innovation Institute for Fertility Preservation, New
York, NY.
OBJECTIVE: Appropriate strategy to predict the fertility preservation
(FP) cycle outcomes has not been determined. Our aim was to identify the
values of antimullerian hormone measurements (AMH) and antral follicle
counts (AFC) in the prediction of oocyte yield in response to controlled
ovarian stimulation with letrozole plus gonadotropins (LG).
DESIGN: Secondary analysis of prospectively collected database.
MATERIALS AND METHODS: One hundred and fifty two women with
breast cancer who underwent ovarian stimulation with LG protocol for
oocyte and/or embryo cryopreservation were included. Serum AMH and
AFC were determined on cycle day 2/3. The patients who produced %4 oocytes
or %2 embryos were considered low-responders (LoR).
RESULTS: The mean age of the patients was 34.94.5 years. AMH was
better correlated with oocyte yield and the number of embryos cryopreserved
than AFC (r¼0.625, p

significantly affected by the O2 levels (viability %: HV-PD, 68.4; LV-CD,
59.0; LV-PD; 51.6; HV-CD, 16.6). In a further experiment, follicle grading,
staging and viability in HV-PD and HV-CD was assessed at day 9. Results
clearly indicated the advantage of PD over CD in sustaining follicle quality
(grade 1, %: HV-PD, 40; HV-CD, 7. Viability, %: HV-PD, 89; HV-CD, 66),
and the latter also depended on the quality of the ovary.
CONCLUSIONS: Taken together, data indicate that an optimal level of O2
is required to support the quality and viability of follicles cultured in vitro.
The use of gas permeable dishes enabled us to optimally modulate O2 concentrations
in the tissue vicinity and establishing a healthier environment for
the cultured strips. The design of a bioreactor for modulation of O2 tension,
media supply and shear stress is under way to study the effects of fine tuning
the physicochemical parameters on follicle viability and growth.
P-465 Wednesday, October 21, 2015
SAFETY OF CONTROLLED OVARIAN STIMULATION WITH GO-
NADOTROPINS AND LETROZOLE IN WOMEN WITH BRCA
MUTATIONS. J. Kim, a,b V. Turan, a K. H. Oktay. a a Obstetrics & Gynecology,
NY Medical College, Valhalla, NY; b CHA University, Seoul, Korea, Republic
of.
OBJECTIVE: Safety of fertility preservation (FP) via controlled ovarian
stimulation with letrozole supplementation (COSTLES) prior to breast cancer
treatment has not been determined in BRCA mutation carriers. We sought
to determine whether women with BRCA-mutation-positive (BRCA+) cancer
were at higher risk for relapse after COSTLES.
DESIGN: Secondary analysis of a prospective cohort study.
MATERIALS AND METHODS: A total of 89 women diagnosed with
stage %3 breast cancer and underwent BRCA mutation screening test (26
BRCA+ and 63 BRCA-negative) and COSTLES for FP prior to chemotherapy
were enrolled. Follow-up information was collected either during return
visits, by phone interview, and in some cases by also contacting the patient’s
referring oncologist to confirm the information. Primary endpoint was cancer
recurrence defined as the detection of locoregional tumor (chest wall, regional
nodal disease), distant metastases, or contralateral invasive breast cancer.
RESULTS: There were 26 subjects with BRCA mutations (8 with BRCA1, 7
with BRCA2 and onewith both mutations). The BRCA + group was significantly
younger at cancer diagnosis (31.8 3.8 vs. 35.0 4.1 years, P ¼ 0.004) and FP
treatment (32.3 4.0 vs. 35.1 4.2 years, P ¼ 0.016) than the BRCA-negative
group. The BRCA+ women were more likely to have ER-negative breast cancer
than the BRCA-negative (68% vs. 32%, P ¼ 0.004). Consistent with the previous
reports of lower response in BRCA+ women, the peak E2 levels were lower in
BRCA mutation carriers compared to BRCA-negative (461.4306.0 vs.
604.3470.4 pg/ml, P ¼ 0.011), despite the younger age. The mean follow up
was 4.7 years in both groups. There was one recurrence among the BRCA+
women (BRCA1, 448insA) and four in the BRCA-negative. The two groups
had similar relapse-free survival (Kaplan-Meier method P ¼ 0.18).
CONCLUSIONS: This is the first data demonstrating the safety of ovarian
stimulation in BRCA+ women. COSTLES is unlikely to increase the risk of
breast cancer recurrence in women with BRCA mutations.
Supported by: NIH RO1 HD053112.
P-467 Wednesday, October 21, 2015
WITHDRAWN
P-466 Wednesday, October 21, 2015
WITHDRAWN
FERTILITY & STERILITY Ò
e265

P-468 Wednesday, October 21, 2015
FERTILITY PRESERVATION IN FEMALE PEDIATRIC PA-
TIENTS PRIOR TO GONADOTOXIC THERAPY: PREDICTIVE
FACTORS FOR RECEIVING FERTILITY PRESERVATION
TREATMENT. J. H. Selter, C. N. Cordeiro, F. S. Chuong, J. E. Garcia,
L. A. Kolp, M. S. Christianson. Johns Hopkins University School of Medicine,
Baltimore, MD.
OBJECTIVE: To identify predictive factors for receiving fertility preservation
treatment among female pediatric patients.
DESIGN: Retrospective analysis.
MATERIALS AND METHODS: We performed an analysis of female pediatric
patients seen for fertility preservation consultation at an academic
fertility center prior to gonadotoxic treatment from 2002-2014. Multivariate
logistic regression models were constructed to identify factors associated
with receiving fertility preservation treatment. Adjusted odds ratios (OR)
with 95%confidence intervals (CI) were calculated.
RESULTS: Over 12 years, 102 females ages 6-21 years old were seen for
fertility preservation consultation with a mean age of 16.7 years old; 14.7%
were prepubertal. Diagnoses included the following malignancies: hematologic
(61.8%, n¼63), neurologic (10.8%, n¼11), orthopedic tumors or sarcomas
(9.8%, n¼10), ovarian (8.8%, n¼9) and gastrointestinal (4.9%, n¼5). Remaining
patients had medical conditions requiring gonadotoxic treatment (3.9%,
n¼4). Of those seen for consultation, 25.5% (n¼26 pursued fertility preservation.
The most utilized treatment modality was ovarian tissue cryopreservation
(65.4%, n¼17) followed by oocyte cryopreservation (34.6%, n¼9). In
comparing patients who pursued ovarian tissue cryopreservation and oocyte
cryopreservation, there was no significant difference in age or pubertal status.
After adjustment, a history of prior chemotherapy was associated with
decreased odds of receiving fertility preservation treatment (OR¼0.03, 95%
CI 0.01-0.04). Age, race, pubertal status and type of cancer were not statistically
associated with odds of receiving fertility preservation treatment.
CONCLUSIONS: Ovarian tissue cryopreservation emerged as the most
utilized fertility preservation treatment modality in this patient population.
Future work is needed to investigate techniques to use cryopreserved ovarian
tissue for fertility for this group as they reach adulthood. The finding that
prior chemotherapy is negatively associated with fertility preservation in pediatric
patients undergoing gonadotoxic chemotherapy underscores the
importance of earlier referral and education regarding fertility preservation
in newly diagnosed pediatric cancer patients.
P-469 Wednesday, October 21, 2015
PATIENTS WITH SYSTEMIC CANCER HAVE LOWER OVARIAN
RESERVE AND REQUIRE HIGHER GONADOTROPIN
DOSES FOR FERTILITY PRESERVATION (FP). A. V. Dolinko, a,b
L. V. Farland, a,b,c S. A. Missmer, a,b,c C. Racowsky, a,b S. Srouji, a,b
E. Ginsburg. a,b a Center for Infertility and Reproductive Surgery, Dept of Ob-
Gyn, Brigham and Women’s Hospital, Boston, MA; b Harvard Medical
School, Boston, MA; c Harvard School of Public Health, Boston, MA.
OBJECTIVE: To evaluate ovarian stimulation (OS) outcomes in women
with systemic or local cancer
DESIGN: Retrospective cohort study of patients undergoing OS from June
2007 to October 2014
MATERIALS AND METHODS: The responses of cancer patients to their
1st OS cycle for FP were compared to patients without cancer undergoing
IVF/ICSI for male factor infertility. Multivariable linear, Poisson, and logistic
regressions were applied to calculate b-coefficients, relative rates, and
odds ratios, respectively, and 2-sided Wald p-values (p).
RESULTS: Of the 147 cancer patients, 105 had local cancer (stage I-III
solid malignancy) and 42 had systemic cancer (hematologic or stage IV solid
malignancy). Women with systemic cancer were significantly younger than
women with no cancer or local cancer. Adjusting for age and BMI, women
with systemic cancer had significantly lower baseline antral follicle count
(AFC) than women with no cancer or local cancer. Also adjusting for tamoxifen
or letrozole use, women with local cancer or systemic cancer required
higher total doses of FSH, and had lower serum E2 levels and total number
of follicles at hCG trigger than women without cancer. Women with systemic
cancer had 14.4 and 17.0 times greater odds of having a cycle cancellation as
compared to women with no or local cancer, respectively. 14 cancer patients
returned to use frozen embryos in 17 cycles; 7 used gestational carriers (GC)
in 9 cycles. Of these cycles, 7 resulted in live births, 2 as ongoing pregnancies,
and 1 SAB. To date, 19 of the 147 cancer patients have died; 1 of these
had a live birth via GC prior to death.
Characteristic
No Cancer
(n¼664)
Local Cancer
(n¼105)
Systemic Cancer
(n¼42)
Age at Stimulation (y) 34.6 (4.2) 33.6 (4.8) 27.1 (6.4)*^
BMI at Stimulation (kg/m2) 25.7 (4.2) 26.0 (6.3) 24.9 (5.9)
Baseline AFC 9.4 (7.2) 10.2 (7.4) 7.3 (7.5)*^
Total FSH dose (IU) 1839.2 (1294.7) 2813.6 (1785.6)* 3358.9 (2147.3)*^
Duration of Stimulation (days) 11.7 (2.0) 11.7 (2.3) 12.2 (2.2)
Serum E2 at hCG trigger 2229.0 (905.7) 1255.9 (973.4)* 1503.2 (812.6)*
(pg/mL)
Follicles at hCG trigger 12.9 (6.1) 12.2 (8.4)* 13.2 (5.9)*
Oocytes retrieved 15.7 (8.6) 16.8 (13.6) 20.6 (21.0)
Mature (MII) oocytes
76 (19) 76 (20) 78 (15)
retrieved (%)
2PN embryos (n) 8.9 (6.3) 8.8 (6.4) 12.3 (11.7)
Cycle cancelled [n, (%)] 14 (2.1%) 2 (1.9%) 9 (21.4%)*^
All values are Mean (SD), unless otherwise noted*Significant difference
compared to ‘‘No Cancer’’, all p%0.03^Significant difference compared to
‘‘Local Cancer’’, all p%0.04
CONCLUSIONS: These data suggest that women with cancer whose cycles
are not cancelled achieve similar oocyte and embryo yields compared to
women with infertile male partners, although they require higher FSH doses
to achieve those outcomes. Those with systemic cancer are at greater risk of
cycle cancellation.
P-470 Wednesday, October 21, 2015
VALIDITY OF SELF-REPORTED CANCER DIAGNOSIS AND GONA-
DOTOXIC TREATMENTS AMONG FEMALE YOUNG ADULT CAN-
CER SURVIVORS. A. N. Knight, a B. W. Whitcomb, b J. Gorman, c
I. Su. d a Reproductive Medicine, University of California San Diego, San Diego,
CA; b University of Massachusetts, Amherst, MA; c Reproductive Medicine,
Moore’s Cancer Center, La Jolla, CA; d UC San Diego, San Diego, CA.
OBJECTIVE: For reproductive endocrinologists, accurate cancer treatment
information is needed for fertility counseling of female young adult
cancer survivors. As primary medical records are hard to obtain, providers
rely on self-report by survivors. We assessed the validity of self-reported cancer
diagnosis and gonadotoxic treatments in this population.
DESIGN: Cross-sectional
MATERIALS AND METHODS: Female cancer survivors ages 18-45 were
recruited to a cohort study on reproductive health. Survivors reported cancer
diagnosis and treatments via an online questionnaire. Medical records were
abstracted for diagnosis and treatments. Self-reported cancer diagnosis and gonadotoxic
therapies (alkylating chemotherapy, stem cell transplant, pelvic radiation,
uterine and ovary surgery) were compared with medical record data.
Logistic regression models estimated odds ratios (OR) for characteristics associated
with accuracy of self-reporting for chemotherapy, radiation and surgery.
RESULTS: 101 survivors were included (mean age 28.2, SD 6.3; median 2.4
years since cancer diagnosis, IQR 1.2-4.4). Lymphoma (33%), breast cancer
(26%), and uterine or ovarian cancer (10%) were the most common cancer types.
98% of survivors accurately reported cancer type. Only 45% reported correct
stage. Sensitivities of self-reported gonadotoxic exposures ranged from 38-
77% (Table). Specificities and negative predictive values were higher. In adjusted
models, breast cancer diagnosis was associated with higher odds of accurate
chemotherapy reporting (OR 5.8, 95% CI 1.6, 21.2), while Hispanic ethnicity
was associated with lower odds (OR 0.14, 95% CI 0.02, 0.81). No demographic
characteristics were related to accuracy of radiation and surgery reporting.
CONCLUSIONS: Young adult female cancer survivors have poor recall of
gonadotoxic treatment exposures. Self report was accurate for type of cancer,
but not for stage, alkylating chemotherapy, pelvic radiation and surgery.
Healthcare providers who need this type of information will require primary
medical records or cancer treatment summaries.
Sensitivity, Specificity, Predictive values for self-reported gonadotoxic cancer
treatments.
Sensitivity
Alkylating
chemotherapy
Pelvic
Radiation
Stem cell
transplant
Uterine and/or
ovarian surgery
Sensitivity 25/39 (64) 8/13 (61) 5/13 (38) 10/13 (77)
Specificity 44/72 (71) 87/88 (99) 88/88 (100) 88/88 (100)
Positive Predictive Value 25/43 (58) 8/9 (89)
Negative Predictive Value 44/58 (76) 87/92 (92) 88/96 (92) 88/91 (97)
Supported by: Funding: UL1 RR024926 pilot, HD-058799.
e266 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

P-471 Wednesday, October 21, 2015
COMPARISON OF RANDOM START CONTROLLED OVARIAN
STIMULATION WITH STANDARD START IN LETROZOLE
GONADOTROPIN CYCLES FOR FERTILITY PRESERVATION IN
WOMEN WITH BREAST CANCER. G. Bedoschi, a V. Turan, a
V. Emirdar, a M. Sonmezer, b K. H. Oktay. a a New York Medical College,
Valhalla, NY; b Ankara University, Ankara, Turkey.
OBJECTIVE: When women are referred for fertility preservation prior to
chemotherapy, there may not be sufficient time to await menstruation to
initiate ovarian stimulation for embryo or oocyte freezing. In these patients,
random start controlled ovarian stimulation (RSCOH) may be an alternative.
Our aim was to compare the cycle characteristics and outcomes of RSCOH in
the late follicular or luteal phase of the menstrual cycle to the outcomes of
standard start (early follicular) controlled ovarian hyperstimulation
(SSCOH) cycles in women with breast cancer.
DESIGN: Secondary analysis of prospectively collected database in an academic
center
MATERIALS AND METHODS: One hundred and fifty women who underwent
SSCOH and 14 who underwent RSCOH for oocyte and/or embryo
cryopreservation before breast cancer chemotherapy were included. Ovarian
stimulation was performed with concurrent gonadotropin and letrozole-5 mg
treatment; an antagonist was added when there was a follicle>13-mm. Either
hCG or leuprolide acetate was used for trigger.
RESULTS: RSCOH was initiated either in the late follicular (50%) or luteal
phase (50%). The mean age was similar between the groups (RSCOH vs
SSCOH: 34.9 4.5 vs. 32.5 5.5 years, respectively; p ¼ 0.09). Total dose
of gonadotropins was significantly higher in the RSCOH group (4070.4
1968.1 vs. 2735.5 1219.0 IU, respectively; p < 0.041). Maturation and fertilization
rates were similar between the groups. The mean number of mature oocytes
was significantly higher in the RSCOH group (10.7 6.6 vs. 9.9 4.0;
p¼0.039). The mean number of oocytes retrieved (16.1 7.2 vs. 14.3 9.3,
respectively; p ¼ 0.073), and embryos cryopreserved (8.5 2.7 vs. 7.1 5.5,
respectively; p ¼ 0.053) trended higher in women undergoing RSCOH.
CONCLUSIONS: Our data indicate that the fertility preservation cycle
outcomes with RSCOH are not inferior to those with SSCOH. Further follow
up and larger prospective studies are required to determine the pregnancy
success rates of RSCOH in comparison to SSCOH cycles.
Cycle Characteristics and Outcomes.
Random start
(n¼14)
Standart start
(n¼150)
P value
Total letrozole dose (mg) 54.5 15.2 51.4 9.2 0.025
Total FSH dose (IU) 4070 1968 2735 1219 0.041
Ovarian stimulation length (days) 11.0 2.8 11.6 1.7 0.057
E2 level on trigger day (pg/mL) 678.3 434.1 595.9 410.0 0.575
No. of total oocytes 16.1 7.2 14.3 9.3 0.073
No. of mature oocytes 10.7 6.6 9.9 4.0 0.039
Maturity rate (%) 69.2 18.8 73.5 21.2 0.329
Fertilization rate (%) 80.1 30.5 72.3 27.6 0.909
No. of embryos frozen 8.5 2.7 7.1 5.5 0.053
P-472 Wednesday, October 21, 2015
FERTILITY PRESERVATION CHOICES & OUTCOMES OF
WOMEN WITH ADVANCED STAGE CANCERS. K. L. Palmerola, a
J. M. Choi, b M. V. Sauer. b a Columbia University Medical Center, New
York, NY; b Columbia University, New York, NY.
OBJECTIVE: To assess fertility preservation (FP) decisions and outcomes
from assisted reproductive technologies (ART) in newly diagnosed advanced
stage (stage III or IV) cancer patients.
DESIGN: Restrospective case control
MATERIALS AND METHODS: Patients (n¼26) presenting to a single
academic center for FP following a diagnosis of advanced stage cancer (stage
III or IV) between 2007 and 2015 were studied. Women identified as cancer
survivors were excluded. Demographic information, ART cycle data, and
ART outcomes were collected. FP decisions and outcomes were compared
with randomly selected, age-matched patients with stage I or II cancer undergoing
ART during the same period. Mann-Whitney rank sum tests were used
for analysis.
RESULTS: 26 patients with stage III or IV cancer presented for fertility
evaluation between September 2007 and March 2015. 9 (34.6%) patients
were cancer survivors and excluded from analysis. Of the 17 included patients,
9 (52.9%) were diagnosed with a stage III cancer (5 breast, 2 ovarian,
1 hematologic, 1 lung) and 8 (47.1%) with a stage IV cancer (3 hematologic,
3 gastrointestinal/colorectal, 2 breast). 5 (29.4%) patients pursued FP, collectively
undergoing 6 treatment cycles. The stage III/IV patients who pursued
FP demonstrated no statistically significant differences in baseline E2, FSH,
MIS levels, number of days of stimulation, total gonadotropins prescribed,
peak estradiol level, number of oocytes retrieved and fertilized, number of
embryos or oocytes cryopreserved or cycle cancellations compared to stage
I/II cancer patients. There were no complications from FP in these patients.
Advanced directive status was available for 3 of the 5 patients who pursued
FP. All 3 patients chose to discard unfertilized or cryopreserved eggs; 2 patients
opted to donate cryopreserved embryos to research; 1 patient opted to
donate cryopreserved embryos to other patients. Further information was
available for 8 of the 12 patients who did not pursue FP: 6 opted to defer
infertility treatment until after cancer treatment; 1 was recommended by their
oncologist not to delay cancer therapy; 1 had metastases to her ovaries, and
thus a concern that FP may worsen her disease status.
Comparison of Stage I/II vs. Stage III/IV Fertility Preservation Cycle
Outcomes.
Stage I/II
(medians)
Stage III/IV
(medians)
P-value
Age (years) 32 33 0.341
Baseline FSH (mIU/mL) 5.05 4.46 0.367
Baseline E2 (pg/mL) 40.6 46.2 0.176
AMH (ng/mL) 1.275 0.63 0.288
Days of Stimulation 10 11 0.078
Total Gonadotropins (IU) 4050 4500 0.123
Peak E2 (pg/mL) 1122 1429 0.397
Total # oocytes retrieved 13 12 0.484
MII 8 10 0.33
Fertilization Rate 71.4% 76.8% 0.409
Cancellations 0 0 -
CONCLUSIONS: Despite the potential poor prognosis associated with
advanced cancer, this rare cohort of patients may expect similar fertility preservation
outcomes to their earlier staged counterparts. Regardless of cancer
stage, all patients are candidates for fertility preservation, though prognosis
and advanced directives must be discussed before initiating care.
P-473 Wednesday, October 21, 2015
OUTCOMES OF UTILIZED CRYOPRESERVED AUTOLOGOUS
OOCYTES. J. P. Alvarez, a,b A. l. Akopians, a,b E. T. Wang, a
D. L. Hill, c J. Barritt, c M. Surrey, d,c H. Danzer, d,c M. D. Pisarska. a a Cedars
Sinai Medical Center, Los Angeles, CA; b UCLA, Los Angeles, CA; c ART
Reproductive Center, Beverly Hills, CA; d Southern California Reproductive
Center, Beverly Hills, CA.
OBJECTIVE: Oocyte cryopreservation is a rapidly developing technology
with reassuring outcomes from observational studies. However, there are
limited studies on utilization of cryopreserved oocytes. Thus, our objective
was to compare outcomes in women who returned to utilize their previously
cryopreserved oocytes to IVF cycles that utilized frozen embryos.
DESIGN: Retrospective cohort study from a large fertility center.
MATERIALS AND METHODS: Women who underwent autologous
oocyte cryopreservation for both medical and elective indications between
1/2010 and 12/2014 and returned to utilize their cryopreserved oocytes
were selected. Oocyte cryopreservation was performed either by slow freeze
or vitrification. Oocyte donation cycles were excluded. The control group
consisted of women who underwent IVF-freeze all cycles followed by a
frozen embryo transfer (FET) during the same time period. Clinical outcomes
of interest included number of mature oocytes, fertilization rate, blastocyst
progression (number of blastocysts per mature oocyte), and pregnancy
rate. Statistical analysis was performed using a student t test for continuous
variables and a fisher exact test for categorical variables.
RESULTS: Of 523 patients who underwent autologous oocyte cryopreservation,
29 (5.5%) returned to utilize their cryopreserved oocytes. The mean
age at oocyte cryopreservation was 37.3 4.5 years (range of 24-44), which
was comparable to the control group (P ¼0.24). The mean duration of cryopreservation
was 478.9 days with a range of 76-880 days. The oocyte cryopreservation
group had a similar number of mature oocytes (10.5 7.2 vs
13.1 8.1 P¼ 0.08) and fertilization rate (69% 24.3 vs 73.7% 20.14
FERTILITY & STERILITY Ò
e267

P¼ 0.31) compared to the control group. In addition, the blastocyst progression
was not significantly different between the oocyte cryopreservation and
the control group (0.2 0.19 vs 0.2 0.28 P¼ 0.30). The pregnancy rate per
embryo transfer was not significantly different from the IVF/FET group
(45.5% vs 52.3% P ¼ 0.65).
CONCLUSIONS: Utilization of cryopreserved autologous oocytes leads
to similar outcomes, including pregnancy rates compared to women undergoing
IVF with frozen embryo transfer. With greater utilization of cryopreserved
oocytes, larger studies are needed to confirm these findings.
P-474 Wednesday, October 21, 2015
WITHDRAWN
RESULTS: HLGCs rapidly underwent apoptosis after exposure to cyclophosphamide
and cisplatin. But they were resistant to 5-FU and paclitaxel as
evidenced by similar rates of apoptosis and comparable E2 and P productions
among control, 5-FU and paclitaxel groups. By contrast, paclitaxel inhibited
proliferation and induced apoptosis of COV434 and HGrC1 cells. This effect
was milder than cyclophosphamide and cisplatin. 5-FU had the slightest toxic
effects on the mitotic granulosa cells among the drugs tested.
CONCLUSIONS: The magnitude of cytotoxicity of chemotherapy drugs
varies depending upon their classes and the type of granulosa cells.
P-476 Wednesday, October 21, 2015
WITHDRAWN
P-475 Wednesday, October 21, 2015
HUMAN MITOTIC NON-LUTEINIZING AND NON-MITOTIC LU-
TEINIZED GRANULOSA CELLS HAVE DIFFERENT SENSITIVITY
TO CHEMOTHERAPY AGENTS. N. Akin, a G. Bildik, a Y. Guzel, b
B.-. Balaban, b B. Urman, c,b O. Oktem. c,b a Reproductive Biology, Koc University
Graduate School of Health Sciences, Istanbul, Turkey; b Women’s
Health Center Assisted Reproduction Unit, American Hospital, Istanbul,
Turkey; c Obstetrics and Gynecology, Koc University School of Medicine, Istanbul,
Turkey.
OBJECTIVE: Adult human ovary harbors different types of follicles and
granulosa cells. Of these, granulosa cells of preantral and early antral follicles
are proliferative but are not capable of undergoing luteinization whereas
the granulosa cells of corpus luteum are non-mitotic luteinized type. The aim
of the current study was to investigate if these two types of granulosa cells
differ in terms of their sensitivity to cytotoxic actions of chemotherapy drugs.
DESIGN: A translational research study
MATERIALS AND METHODS: Mitotic non-luteinizing cells (COV434
and HGrC1) and non-mitotic luteinized granulosa cells (HLGCs) retrieved
from IVF patients were used for the experiments. The cells were plated at
a density of 5000 cells/well and treated with 5-fluorouracil (50mg/mL), paclitaxel
(2mg/mL), cisplatin (100mg/mL) and cyclophosphamide (4-hydroperoxy
cyclophosphamide, active in vitro form of the drug, 100mM). The
drugs were used at the concentrations corresponding to their therapeutic
serum levels. Cytotoxic effect of these drugs were determined according to
their impact on the proliferation (real-time and quantitative cell proliferation
index xcelligence system; COV434 and HGrC1 cells only), viability/
apoptosis (YO-PRO-1/cleaved caspase-3 expression) and steroidogenic activity
(estradiol (E2) and progesterone (P) productions) of the granulosa cells.
The impact of different chemotherapy drugs on mitotic vs. non-mitotic human
granulosa cells.
Control Cyclophosphamide Cisplatin Paclitaxel 5-FU p-value
Mitotic non-luteinized granulosa cells
Proliferative
index
3.50.3 0.40.2 (a) 0.60.3
(a)
Apoptosis
(%)
4% 91% (a) 78%
(a)
Non-mitotic luteinized granulosa cells
Estradiol
(pg/ml)
1604116 38956 (a) 43767
(a)
Progesterone
(ng/ml)
Apoptosis
(%)
63092 27142 (a) 33838
(a)
2.50.4
(b)
56%
(b)
1458102
(b)
56956
(b)
3.10.2
(c)
12%
(c)
1412103
(b)
60098
(b)
a:

acceptability of one of the most effective, reversible forms of contraception,
the intrauterine device (IUD), in women with cancer is lacking, and
there are no studies on placement of an IUD at the time of oocyte retrieval
for fertility preservation. Thus, we surveyed women with cancer on their
satisfaction with their IUD and any noted side effects. The responses of
women with cancer were also compared to a smaller subset of women
without cancer.
DESIGN: A retrospective survey was performed of women age 18-
45 years who had an IUD placed at our institution between 2007
and 2015.
MATERIALS AND METHODS: Women who met inclusion criteria were
emailed a link to an internet-based survey. A 5-point Likert scale was used.
Standard descriptive statistics and univariate analyses were performed with
chi-square and T-tests as appropriate.
RESULTS: 46 women completed the survey. 29 had a history of cancer
with breast cancer being the most common cancer diagnosis (72.4%). For
the majority of cancer patients, the discussion about IUD placement was
initiated by a fertility specialist (44.8%) or by the patient herself
(27.6%). Women were overall satisfied (75.0%) with the counseling they
received, and 75.9% required no additional problem visits after IUD placement.
The majority of patients had a copper IUD (75.9%). After IUD placement,
the women described their menses as heavier (58.6%), more irregular
(51.7%) and associated with improved or unchanged menstrual cramps
(56.3%). 75% reported their sex life was improved or unchanged. At the
time of the survey, 84.4% still had their original IUD in place or had
kept the IUD in for over a year. 69.0% reported a positive attitude toward
the IUD, and 79.3% would recommend it to a friend. Thirteen women with
cancer had their IUDs placed at the time of oocyte retrieval. Compared to
those who did not have IUDs placed at the time of retrieval, these women
were more likely to report the procedure as not painful (85.7 vs. 34.4%
p¼0.001) and not stressful (84.6 vs. 43.8%, p¼0.024). Compared to
women without cancer, women with cancer were not more likely to report
stressful or painful IUD placement or changes in bleeding patterns or menstrual
symptoms. There were no differences in high levels of satisfaction
(58.1vs. 41.9%, p¼0.32).
CONCLUSIONS: Despite the increased menstrual symptoms compared to
baseline that cancer patients noted after IUD placement, satisfaction and IUD
retention remained high. This makes the IUD an option for women with cancer,
including women who are undergoing fertility preservation prior to initiating
chemotherapy.
CRYOPRESERVATION
P-478 Wednesday, October 21, 2015
DOES LASER ZONA BREACHING (LZB) OF WARMED BLASTO-
CYSTS IMPROVE HATCHING RATES? N. M. Sachdev, a
D. H. McCulloh, b C. McCaffrey, c J. Grifo. d a Obstetrics and Gynecology,
New York University Fertility Center, New York, NY; b New York University
Fertility Center, New York, NY; c NYU Fertility Center, New York, NY;
d NYU Langone Medical Center, New York, NY.
OBJECTIVE: To assess using time lapse microscopy (TLM) if post warming
LZB improves the ability of the blastocyst to completely exit the zona
pellucida (hatching).
DESIGN: Prospective Cohort study.
MATERIALS AND METHODS: Three groups were included in the
study; 1) non biopsied slow frozen blastocysts, 2) non biopsied vitrified
blastocysts and 3) biopsied aneuploid blastocysts, as determined by trophectoderm
biopsy (TE) and array comparative genomic hybridization
(aCGH) performed prior to vitrification. All blastocysts in group 3 had
an existing zona breach prior to cryopreservation. All blastocysts were
warmed using standard protocol. Each group was subdivided into a control
arm with no additional zona pellucida (ZP) manipulation, and a test arm
subjected to LZB following warming. This post warming LZB creates an
opening larger (approximately 1/3 of the zona) than that routinely created
for TE biopsy. Time lapse imaging was reviewed by a single observer. Primary
outcomes were number of successfully hatched blastocysts and
timing of hatching events.
RESULTS: A total of 144 blastocysts donated for research (IRB #H6902)
were included in the study. Survival rate post warming was 96%: An additional
7 embryos were zona free following warming and were excluded
from analysis. Hence, 131 blastocysts were observed using TLM. Group 1
consisted of 21 with LZB and 22 controls. Group 2 consisted of 21 with
LZB and 24 controls. Group 3 consisted of 22 with LZB and 21 controls.
A total of 55 (41.98%) warmed blastocysts completed hatching within 81
hours, while 76 (58%) did not complete hatching despite exhibiting viability.
There was no difference in patient ages among the blastocysts subjected to
LZB (34.58 4.89) and controls (35.95 4.24) p¼0.0895 or within groups.
Groups 1& 2 (non biopsied) showed a difference in the LZB groups versus
controls in the number of blastocysts that completed hatching. Group 3 (previously
biopsied) blastocysts showed no difference and were able to complete
hatching, likely due to the preexisting breach in the ZP created for
the TE biopsy. (Table 1).
Outcomes of Post Warm LZB vs Control Groups.
Group
Lazer Zona
Breaching
(LZB)
% Completed
Hatching (N)
Hours to
P Start of
(completed) Hatching P (start)
Hours to
Complete
Hatching
P
(end)
Group 1 + 66.7% (21) .00003 22 9 0.154 38 12 0.161
- 4.5% (22) 6 19
Group 2 + 76.2% (21) .0000009 18 5 0.140 26 9 0.732
- 4.2% (24) 27 33
Group 3 + 54.5% (22) 0.882 16 13 0.334 20 21 0.603
- 52.3% (21) 11 9 35 23
CONCLUSIONS: TLM revealed that non-biopsied embryos have difficulty
completing zona hatching, indicating that zona hardening may play a role. In
the absence of LZB, implantation may be impeded due to inability to escape
from the ZP. In contrast, previously biopsied blastocysts did not show a difference
in hatching rates whether subjected to LZB post warming or not.
Reference:
1. Vaccari & Conaghan, Fert & Ster Volume 103, Issue 2, Suppl, Pages
e7-e8.
P-479 Wednesday, October 21, 2015
ELECTIVE EMBRYO CRYOPRESERVATION AFTER PRO-
LONGED PREMATURE PROGESTERONE ELEVATION TO
IMPROVE IN VITRO FERTILIZATION (IVF) PREGNANCY
OUTCOMES. E. Anspach, B. Maslow, A. Bartolucci, C. Benadiva,
J. Nulsen, L. Engmann. University of Connecticut Health Center, Farmington,
CT.
OBJECTIVE: Studies demonstrate that prolonged premature elevations in
progesterone (P) levels during controlled ovarian stimulation have adverse
effects on IVF outcomes, potentially due to alterations in endometrial receptivity.
Cryopreservation with a subsequent frozen embryo transfer (FET) has
been proposed as a method of overcoming embryo-endometrium asynchrony.
The purpose of this study was to evaluate whether elective cryopreservation
(‘‘freeze all’’) improves pregnancy outcomes in patients with prolonged premature
P elevation during an IVF cycle.
DESIGN: Retrospective cohort
MATERIALS AND METHODS: 732 patients

Baseline characteristics and outcome.
Variable
Group 1
Fresh Transfer,
Elevated P
n¼233
Group2
FET
n¼43
Group 3
Fresh
Transfer,
Normal P
n¼456
P
Value
Age (SD) 34.0 3.6 32.9 4.0 33.6 3.8 0.16
AMH (SD) 3.4 3.9 3.9 5.6 3.6 3.9 0.62
Day of Trigger P (SD) 1.4 0.3 1.9 0.8 0.7 0.2

P-482 Wednesday, October 21, 2015
128 AUTOLOGOUS POST-VITRIFICATION OOCYTE THAW CY-
CLES: WARMING, EMBRYONIC DEVELOPMENT, AND PREG-
NANCY OUTCOMES. J. Doyle, K. S. Richter, J. Lim, J. Graham,
M. J. Tucker. Shady Grove Fertility Reproductive Science Center, Rockville,
MD.
OBJECTIVE: To evaluate a single treatment center’s experience with
autologous oocyte vitrification and warming, including embryonic development
and pregnancy outcomes.
DESIGN: Retrospective review
MATERIALS AND METHODS: Patient and cycle characteristics and
treatment outcomes were compared according to indication for autologous
oocyte vitrification. All MII oocytes that survived warming
were fertilized via ICSI. Outcomes were also compared to a reference
group of all fresh autologous ICSI cycles in 2013 (n¼2,963) using
generalized estimating equations analysis to control for repeat
cycles and adjust for age, blastocyst vs cleavage stage transfer, and
numbers of embryos transferred. Clinical pregnancy was defined as
ultrasound confirmation of an intrauterine gestational sac; implantation
rate was calculated as the number of gestational sacs per embryo
transferred.
RESULTS: Through January 2015, 128 oocyte warming cycles were
conducted following oocyte vitrification for the indications of elective
fertility preservation, unavailability of sperm at oocyte retrieval, or
desire for limited oocyte insemination, which compare as shown in the
table (within rows, values with different superscripts are significantly
different by post hoc Tukey-Kramer HSD, p

micronized progesterone 400 mg b.d. was started after 12 days when the
GnRh agonist was also stopped. Frozen-thawed embryos were transferred
on day+1 of their chronological age and when the endometrium reached
12 mm in thickness. Group B consisted of 100 patients (100 cycles)
who started daily estradiol valerate 6 mg administration from the second
day of the FET cycle and followed the same regimen but without prior
treatment with triptorelin
RESULTS: There was a significant increase in implantation rate in the
GnRh agonist group (group A) compared to the estrogen and progesterone
only group (group B) (44.1% vs 21.1%; P¼ 0.002*). The pregnancy rate
was also significantly higher in group A compared to group B (65.5% vs
42 %, P¼ 0.013*).
CONCLUSIONS: GnRh agonist administration during endometrial preparation
for FET increases the implantation rate and pregnancy rate.
CONCLUSIONS: The sperm Vitrification and conventional freezing have
negative impact on sperm parameters as well as sperm DNA oxidation, and
mitochondrial activity. Sperm Vitrification presented improved sperm
motility recovery, similar levels of DNA oxidation and a slightly increase
in mitochondrial activity when compared with sperm cryopreservation
with the conventional method. Suggesting Vas an optimal protocol for sperm
cryopreservation.
P-486 Wednesday, October 21, 2015
WITHDRAWN
Effect of GnRh agonist on implantation of FET.
GnRh group A Group B
P
Total number of cases 110 100
Total number of ET 322 285
Total number of implanted embryos 142 60
Implantation rate 44.1 % 21.1% 0.002*
Number of pregnant cases 72 42
Pregnancy rate 65.5% 42% 0.013*
* Statistically significant.
P-485 Wednesday, October 21, 2015
EFFECT OF HUMAN SPERM FREEZING-THAWING PROCESS ON
VITRIFICATION AND CONVENTIONAL FREEZING: EVALUA-
TION OF SURVIVAL, MOTILITY, DNA OXIDATION AND MITO-
CHONDRIAL ACTIVITY. D. Pabon, a M. Meseguer, a G. Sevillano, a
A. Cobo, a J. Romero, a N. Sota, b A. Mifsud, a J. de los Santos, a
J. Remohi, c M. de los Santos. a a Clinical Embryology-IVI, Valencia, Spain;
b Clinical Andrology-IVI, Valencia, Spain; c Reproduction-IVI, Valencia,
Spain.
OBJECTIVE: Sperm Vitrification is a method for cryopreservation,
without the use of conventional cryoprotectants, by plunging the sperm suspension
directly into liquid nitrogen. We developed and evaluated a new
method of vitrification, using a combination of 0,25 M trehalose-sucrose.
This study aimed to compare the vitrification (V) with conventional freezing
protocol (CF) on several sperm parameters: motility, vitality, mitocondryal
activity and DNA oxidation.
DESIGN: A prospective non-randomized study to evaluate spermatozoa
vitrification with respect to the conventional freezing protocol, using fresh
sperm as reference.
MATERIALS AND METHODS: The sperm vitrification solution was
0,25 M trehalose-sucrose and plunge directly in liquid nitrogen in microdroplets
of 5-10ml. Sperm mobility and viability rates were calculated
before and after cryopreservation. Mitochondrial function was evaluated
using JC-1 (fluorescent cationic dye, 5,5’,6,6’-tetrachloro-1-1’,3,3’-tetraethyl-benzamidazolocarbocyanin
iodide). Sperm DNA oxidation was
determined using a fluorescent assay (Oxy-DNA test) for the detection of
8-oxoguanine. The evaluation was carried out before and after cryopreservation
using flow cytometry analysis. Statistical analysis was performed using
ANOVA test and Chi square, p

TUNEL
PATIENTS
TUNEL
DONORS
ANNEXIN V
PATIENTS
ANNEXIN
V DONORS
BODIPY
PATIENTS
BODIPY
DONORS
8OH-dG
PATIENTS
FRESH-BASAL 20.112.8 14.74.2 8.93.2 6.72.8 12.44.8 9.63.1 65.313.6
POST THAWED 36.613.6 25.75.1 18.35.1 11.52.7 27.18.5 17.53.3 112.6 22.1
after thawed had higher values in compare to swim-up (fresh). Vitality
decreased after cryopreservation, contrary the control group samples presented
more tolerance to cryoinjury. It was determined that these parameters
significantly increase in men >45 y/o. All the comparisons performed on
these analysis were statistically significant (page 35 at time of treatment.
When successfully combined, these technologies afford postponement of
Total No. MI + MII 559/674 (83%) 238/292 (82%) 0.6
Oocytes Survived
/Thawed
Survived and
534 (79% ) 225 (77%) 0.4
Assessed as MII at
Thaw
2-Pronuclear (2PN) 373 (70%) 167 (74%) 0.3
Fertilization
No. Cycles Achieving 39/47 (83%) 15/22 (68%) 0.2
Blastocysts
Suitable for TEBX
No. Biopsied BL 145 (39%) 54 (32%) 0.2
No. Euploid BL 58 (40% ) 8 (15%) 0.0007
No. Aneuploid BL 86 (60%) 44 (85%) 0.004
No. TEBX Cycles 28/39 (72%) 7/15 (47%) 0.1
with At Least One
Euploid BL
No. TEBX Cycles 11/39 (28%) 8/15 (53%) 0.1
with All Aneuploid
BL
Embryo Implantation 14/22 (64%) 2/3 (67%) 1.0
Rate
Ongoing+Delivered
Rate/Transfer
12/21 (57%) 2/3 (67%) 1.0
FERTILITY & STERILITY Ò
e273

P-490 Wednesday, October 21, 2015
INCREASED INCIDENCE OF MOSAICISM AMONG BIOPSIED
TROPHECTODERM CELLS ANALYZED BY NEXT GENERATION
SEQUENCING. M. Smith, a D. Johnson, a D. L. Hill, a M. Surrey, b
S. Ghadir, b W. Chang, b H. Danzer, b C. Alexander, b S. Munne, c
J. Barritt. a a ART Reproductive Center, Beverly Hills, CA; b Southern California
Reproductive Center, Beverly Hills, CA; c Reprogenetics, Livingston, NJ.
OBJECTIVE: Preimplantation genetic screening (PGS) using Array CGH,
qPCR or SNP arrays provided very high levels of diagnostic accuracy, however,
very little ability to recognize low-level mosaicism. The newest Next
Generation Sequencing (NGS) technique provides the same level of diagnostic
accuracy and allows for an increased chance to detect chromosomal
mosaics, down to as low as 20%. We set out to determine the presence, quantity
and the types of genetic mosaicism in trophectoderm cells following embryo
biopsy in clinical PGS cases using NGS analysis.
DESIGN: Retrospective data analysis from a single, large, private fertility
clinic.
MATERIALS AND METHODS: Sixteen blastocyst biopsy PGS cycles
using NGS were analyzed. No ovum donor cycles were included. The
mean female age was 36. Trophectoderm biopsy was performed on 128 blastocysts
cultured 5, 6 and/or 7 days. All embryos were vitrified and the biopsied
cells were sent to a genetic testing laboratory (Reprogenetics, Los
Angeles, CA). Results were compiled, including the genetic diagnosis rate,
the quantity of mosaics, the types of mosaics, and the number of occurrences
for each chromosome involved in the mosaicism. Fisher’s Exact Test was
used for statistical analysis.
RESULTS: Genetic results were obtained in 124 of 128 (96.9%) blastocysts,
with 1 failure to amplify DNA and 3 chaotic profiles not allowing diagnoses.
Mosaicism was reported in 17 (13.7%) samples, as compared with
our previous experience with aCGH of 6728 blastocysts reporting only 10
(0.14%) mosaics (p

P-493 Wednesday, October 21, 2015
CLINICAL ERROR RATE OF ARRAY COMPARATIVE
GENOMIC HYBRIDIZATION (ACGH) IN EUPLOID
BLASTOCYSTS. A. W. Tiegs, B. Hodes-Wertz, D. H. McCulloh,
J. Grifo. NYU Langone Medical Center, New York, NY.
OBJECTIVE: Preimplantation genetic screening (PGS) and diagnosis
(PGD) with euploid embryo transfer is associated with improved implantation
and live birth rates as compared to routine in vitro fertilization. However,
misdiagnosis of the embryo is a potential risk. The purpose of this study was
to investigate the clinical misdiagnosis rate associated with transfer of trophectoderm-biopsied
blastocysts deemed to be euploid via array comparative
genomic hybridization (aCGH) at a large university-based fertility center.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All cycles utilizing PGS or PGD with
trophectoderm biopsy, aCGH and euploid embryo transfer at a large university-based
fertility center with known birth outcomes from 11/2010 through
7/2014 were included (n¼525). The number of embryos transferred, gestational
sacs, clinical pregnancies with a fetal heartbeat (FH), and cycle outcomes
(specifically live birth (LB) and spontaneous abortion (SAB)) were
recorded. Any products of conception (POCs) that were collected from dilation
and curettage of a SAB were reviewed.
RESULTS: There were 525 embryo transfers of 584 euploid embryos as designated
by aCGH. Implantation rate was 61%, clinical pregnancy rate with an FH
was 59%, and the first trimester SAB rate was 13% of pregnancies with a sac
(6% of pregnancies with an FH). There was one intrauterine fetal demise and
one periviable loss. Five clinical misdiagnoses were identified during this time.
Clinical aCGH errors.
Embryo aCGH
result Karyotype Outcome Error
46,XX 47,XX +13 SAB Biologic-Mosaic
46,XX 47,XX +7 SAB Test Error
46,XY 47,XY +21 SAB Biologic-Mosaic
46,XY 47,XYY LB Test Error
46,XX 46,XY LB Likely Contamination
Error
All aCGH specimens were analyzed again with aCGH and 2/5 were found
to be mosaic embryos. Error rate per embryo transfer cycle was 1.0%, 0.9%
per embryo transferred, 1.5% per pregnancy with a sac and 0.4% per euploid
embryo diagnosed by aCGH. The LB error rate was 0.7% (both sex chromosome
errors) and the SAB POCs error rate was 17.6% (3/17 POCs tested). Of
note, single gene disorders sequenced against using PGD were not mistakenly
selected for in any known cases.
CONCLUSIONS: Although aCGH has been shown to be a superior
method of comprehensive chromosome screening, several possible sources
of error still exist. These include contamination of the specimen and sampling
errors when mosaicism exists. Although the overall error rate is low
(60,000 BLASTO-
CYSTS TESTED VIA ARRAY CGH. E. Yeboah, a S. Munne, a
T. Escudero, a C. Wagner Coughlin, b M. Surrey, c S. Ghadir, c R. P. Marrs, d
J. Zhang, e G. Garzo. f a Reprogenetics, Livingston, NJ; b Aparent IVF Laboratory,
Highland Park, IL; c Southern California Reproductive Center, Beverly
Hills, CA; d California Fertility Partners, Los Angeles, CA; e New Hope
Fertility Center, New York, NY; f Reproductive Partners - UCSD, La Jolla, CA.
OBJECTIVE: To assess the results of blastocysts via aCGH by age and
type of abnormality
DESIGN: Retrospective Analysis
MATERIALS AND METHODS: 60,492 blastocyst biopsies were
analyzed from 7/24/2011 to 4/21/2015 from 12,275 cycles of preimplantation
genetic screening (PGS) using blastocyst biopsy and aCGH. Subsequently,
they were categorized by SART age groups to examine the percentage of embryos
that were euploid, monosomic, trisomic, had partial chromosome gains
or losses, double aneuploidies, orcomplex abnormal (more than two abnormalities),
using the PGS specific eIVF database (PracticeHwy, Dallas, TX).
RESULTS: The proportion of full monosomies to trisomies were 50:50,
however, for partial abnormalities it was skewed 69:31
CONCLUSIONS: As it is well known, chromosome abnormalities
increased with advanced maternal age, from 31% in egg donors to 85% in
women older than 42. The complexity of the abnormalities also increased
with advancing maternal age, with double and multiple abnormalities
increasing and single full aneuploidies and single partial aneuploidies
decreasing. Partial aneuploidies, that is, the presence or absence of a fragment
of a chromosome, had a ratio of partial monosomies to partial trisomies that
was significantly skewed, 69:31, while full aneuploidies had a 50:50 ratio of
monosomies to trisomies. In addition, the total rate of partial abnormalities
(alone or in combination with other abnormalities) does not change with
advancing maternal. These two very different tendencies indicate different origins
of partial and full aneuploidies, with full aneuploidies being mostly
meiotic in origin, while partials probably mitotic (non-age related) or paternal
in origin with the resulting chromosome fragments being lost more easily than
gained.
The results are shown in the below table:
EGD < 35 35-37 38-40 41-42 > 42
Embryos Analyzed 12832 15103 10302 11530 5744 3090
No Results 3% 4% 3% 3% 3% 4%
Euploid 69% 63% 55% 39% 23% 15%
Abnormal 31% 37% 45% 61% 77% 85%
1 Aneuploidy 52% 56% 54% 50% 39% 26%
2 Aneuploidies 17% 16% 22% 25% 29% 26%
>2 Abnormalities 16% 16% 16% 20% 30% 47%
Partial aneuploidy 15% 12% 8% 4% 2% 1%
(alone)
Partial total (as % of
abnormal)
15% 13.6% 16.3% 12% 15% 22%
FERTILITY & STERILITY Ò
e275

P-496 Wednesday, October 21, 2015
DOES BLASTOCYST MORPHOLOGY PREDICT PREGNANCY
OUTCOMES IN EUPLOID SINGLE EMBRYO
TRANSFERS? M. Shah, a J. Kort, b R. Lathi. c a Stanford University,
Palo Alto, CA; b Stanford University, Menlo Park, CA; c Stanford University,
Stanford, CA.
OBJECTIVE: Comprehensive chromosome screening (CCS) is a powerful
tool which allows for selection of euploid embryos to improve pregnancy
outcomes in IVF cycles. There is limited data suggesting a correlation between
blastocyst morphology of euploid embryos and pregnancy outcomes.
The objective of this study was to determine whether blastocyst morphology
could be used to predict implantation potential of a euploid embryo.
DESIGN: We performed a retrospective observational study performed
between January 2012 and December 2014 at the Stanford University
Fertility and Reproductive Health Center. The study included fresh and
frozen embryo transfer cycles of single euploid blastocysts.
MATERIALS AND METHODS: Trophectoderm biopsy was performed
for CCS analysis on day 5 blastocysts. Morphology, as defined by the Gardner
criteria, was assessed on the day of biopsy for frozen IVF cycles, and on
the day of transfer for fresh IVF cycles. The ongoing pregnancy rate (OPR)
was defined as presence of fetal cardiac activity beyond 20 weeks of gestation.
Chi squared analysis was used to test the relationship between blastocyst
morphology and OPR.
RESULTS: There were 133 single euploid embryo transfers included for
analysis from 125 unique patients. The mean patient age was 37.3 years
old and similar among patients with and without an ongoing pregnancy.
Fifty-five percent of these cycles employed a frozen-thaw embryo transfer,
while 45% were fresh embryo transfers. Forty-percent of euploid embryo
transfers resulted in an ongoing pregnancy or live birth. Chi-squared analyses
revealed that blastocysts with a greater expansion grade, and better inner cell
mass and trophectoderm quality grades were more likely to result in an
ongoing pregnancy or live birth, with the trend reaching significance for
the association with trophectoderm quality (Table I).
CONCLUSIONS: All components of the Gardner grading scale of blastocyst
assessment may help to select the embryo most likely to become a successful
pregnancy, even after assisted hatching and pre-implantation genetic
screening for euploid embryos.
Ongoing pregnancy/live birth rate (OPR) stratified by components of the
Gardner grading scale after.
Expansion grade 4 5 6 p-value
OPR 3% 22% 75% 0.06
Inner Cell Mass grade A B C
OPR 59% 41% 1% 0.09
Trophectoderm grade A B C
59% 35% 5% 0.04
P-497 Wednesday, October 21, 2015
SHOULD EGG DONORS BE SCREENED FOR X-LINKED
DISEASE? A. Hershlag, a O. A. Tusheva, a C. Mullin, a Y. Danovitch, b
T. Singer, a S. Munne, c N. Kumar. c a The Center for Human Reproduction,
North Shore LIJ Health System, Manhasset, NY; b Sackler School of Medicine,
New York, NY; c Recombine, New York, NY.
OBJECTIVE: Egg donors with negative family history of X-linked disorders
may still be carriers and thus put male offspring at risk. Carrier screening
for such disorders, with the exception of Fragile X, is currently limited. We
here present a case where an undiagnosed carrier state in the donor led to an
affected male child, review the literature, and propose a novel approach to
prevent transmission of X-linked disorders through egg donation.
DESIGN: A case report and literature review
MATERIALS AND METHODS: Case: An egg donation from a patient’s
sister led to the birth of a male child who experienced excessive bleeding at
circumcision and was diagnosed with Hemophilia A. The donor had a de
novo mutation. Second egg donor cycle using PGD for Hemophilia A was
performed and resulted in a birth of healthy unaffected girl.
RESULTS: Previously reported births of affected males have uncovered
unsuspected mutations for X-linked diseases in egg donors (hemophilia; adrenal
hypoplasia congenita; ocular albinism). While donors are screened for
the most common autosomal recessive disorders such as CF, SMA
(1:2500;1:6000 LB), they are not screened for the most common X-linkeddiseases,
such as Hemophilia A, Duchenne Muscular Dystrophy
(DMD)(1:5000;1:3300 male LB). Gender selection for females can reduce
the risk of symptoms but has ethical issues. Other approaches include molecular
testing (such as targeted mutation analysis or sequencing) in the donor to
determine carrier status and/or in the embryo to determine disease status. In
the case of Hemophilia A, with over 615 point mutations, 57 insertions, 270
deletions ranging from a single nucleotide to over 150 kb, and complex inversions,
testing techniques are cumbersome, technically challenging and
applicable only to a small number of families with a particular mutation.
Similarly, in the case of other X-linked disorders such as DMD, screening
is challenging due to deletions accounting for a significant proportion of disease,
large numbers of ‘‘private’’ mutations, and high de novo mutation rates.
Preimplantation genetic haplotyping (PGH), originally described in 2006,
combines a haplotyping approach and whole genome amplification, and
may be a robust, efficient and successful alternative method to detect X-
linked disorders in embryos.
CONCLUSIONS: It is incumbent upon us to do everything within the
reach of reproductive genetics to prevent X-linked diseases in male offspring
from egg donation, as well as from all ART in general. We need to incorporate
screening for X-linked disorders as part of regular carrier screening practice,
and overcome technical challenges to ensure we are effectively able to
test for the wide heterogeneity of mutations that cause X-linked disorders.
Preimplantation genetic haplotyping (PGH) may be a promising method
for X-linked disease screening in embryos.
P-498 Wednesday, October 21, 2015
CCS IMPROVES PREGNANCY OUTCOMES IN EGG DONOR FET
CYCLES. A. Coates, a C. Welch, b K. Ketterson, b S. Munne. b a Oregon
Reproductive Medicine, Portland, OR; b Reprogenetics, Livingston, NJ.
OBJECTIVE: Donor egg recipients are offered CCS as part of their IVF
cycle to optimize embryo selection. Aneuploidy rates of blastocyst embryos
from Egg donor cycles are around 25% and this alone may justify selection
by CCS. The purpose of this study is to determine if CCS can improve treatment
outcome in frozen /thawed egg donor cycles.
DESIGN: Retrospective data and sample analysis.
MATERIALS AND METHODS: Blastocyst embryos were biopsied on
day 5 or 6 and the cells were analyzed by array CGH (Illumina) or Next Generation
Sequencing (NGS, Life Technologies). All embryos were vitrified for
future use post biopsy. Egg donor cycles of frozen / thawed untested embryos
were used as a control group.
RESULTS: Results are shown in the table below:
CONCLUSIONS: 79% of frozen embryo transfer cycles resulted in
ongoing pregnancies after transfer of CCS selected embryos compared to
59% of transfers without selection. This significant difference justifies the extra
cost of the biopsy and testing procedure (compared to the cost of a subsequent
egg donor cycle) and helps reduce the risk of aneuploid
conceptions, which is still a significant concern after compensating for
advanced maternal age by using young donor eggs.
Frozen donor egg embryo transfers with and without CCS tested embryos.
CCS tested FET
Non tested
FET
P value
Average age(range) 25(21-27) 25(22-28)
# Frozen transfers 166 61
Total # embryos
246(1.5) 92(1.5)
transferred(ave/FET)
# positive tests (%/FET) 159*(96%) 52*(85%) *p¼0.02
# ongoing pregnancies 131*(79%) 36*(59%) *p¼0.006
(%/FET)
# embryos implanting (%
+ sac/embryo)
201*(82%) 54*(59%) *P¼0.0001
e276 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

P-499 Wednesday, October 21, 2015
TROPHECTODERM BLASTOCYST (TE) BIOPSY RESULTS IN
HIGHER PREGNANCY RATE, LOWER MULTIPLE RATE, AND
FEWER CYCLES TO PREGNANCY IN GESTATIONAL
CARRIERS. A. Nasseri, a K. Gleason, a C. Zollinger, a M. Post, a
D. Chen, a J. Grifo. b a The Valley Hospital Fertility Center, paramus, NJ;
b The New York University Fertility Center, New York, NY.
OBJECTIVE: To determine the usefulness of comprehensive chromosome
screeniong (CCS) by TE biopsy in gestational surrogacy
DESIGN: Retrospective case control study
MATERIALS AND METHODS: Data from all cycles involving nondonor
egg embryo transfers to gestational carriers (GC) at our center from
April 2008 to January 2015 were reviewed. FDA screening guidelines
were adhered to in all cases. Embryos of the intended parents (IP) were
created as per routine IVF protocol. There were a total of 88 embryo transfer
cycles. Twenty eight cycles involved the transfer of frozen eupolid blastocysts
after TE biopsy. Embryos were screened for all 24 chromosomes by
array CGH (group A). Of the remaining 60 cycles (group B), there were
35 fresh, and 25 frozen transfers. One fresh transfer was canceled due to embryo
arrest. Paired student t-test and the Chi-squared test of independence
were used for statistical analysis. P value

CONCLUSIONS: Aneuploidy screening was the most common indication
for PGD. Use of PGD was not associated with increased rates of pregnancy
and live birth for women 37 years, thereby suggesting
that more viable or euploid embryos were transferred after
screening.
P-502 Wednesday, October 21, 2015
WITHDRAWN
P-503 Wednesday, October 21, 2015
ELEVATED SERUM PROGESTERONE REDUCES THE LIKELI-
HOOD OF LIVE BIRTH AFTER FROZEN TRANSFER OF EUPLOID
EMBRYOS: TOO MUCH OF A GOOD THING? L. A. Kondapalli, a
L. Munkwitz, a R. Collins, a D. A. Minjarez, b W. B. Schoolcraft, a
M. Katz-Jaffe. a a Colorado Center for Reproductive Medicine, Lone Tree,
CO; b Colorado Center for Reproductive Medicine, Denver, CO.
OBJECTIVE: Although comprehensive chromosomal screening (CCS) allows
for selection of euploid embryos with high implantation and live birth
rates, IVF failures still occur. The objective of this study was to investigate
factors associated with negative pregnancy after frozen transfer.
DESIGN: Case control study
MATERIALS AND METHODS: Patients undergoing euploid frozen embryo
transfers (FET) between 2013-2014 were included (n¼441). All embryos
were biopsied at the blastocyst stage for CCS prior to vitrification.
Standard protocols were used for hormone supplementation prior to FET.
Patients were stratified by cycle outcome: successful live birth versus
non-pregnant (including biochemical pregnancy). Data on demographics,
ovarian reserve markers, IVF stimulation, and cycle outcomes were obtained
from medical records. Appropriate parametric and non- parametric
statistical analyses were performed using STATA version 13.1 (Stata
Corp; College Station, TX, USA) with P value

from a second biopsy for each embryo. Phase 2 involved clinical utilization
of combined SGD and CCS testing with follow-up.
RESULTS: Workup time approximated 1 month for each case. Phase 1
testing examined 152 embryos and demonstrated 99% concordance with
reference lab data with all discrepancies confirmed as an error with the reference
labs results. Phase 2 involved clinical application of these methods in 43
patients (304 embryos). A definitive result was reported for 99.7% (303/304)
embryos, with 0.3% (1/304) having an inconclusive result likely due to
recombination. All cases for which a sample was available in a newborn child
confirmed the PGD result. In patients receiving a transfer with follow-up,
clinical outcomes were excellent with an implantation rate of 75% (12/16),
with 92% of patients achieving pregnancy.
CONCLUSIONS: This methodology has demonstrated excellent concordance
with current methods of SGD and provides a unique opportunity to
avoid pitfalls of WGA when targeting additional loci in the genome in parallel
with CCS. Additional advantages of the method include the ability to
manage microdeletion and duplication cases and the potential for a 4 hour
turnaround time. In addition, it is well established that SNP markers are
denser than STRs thus generally reducing the distance of markers from mutations
and the potential impact of recombination.
P-506 Wednesday, October 21, 2015
THE CLINICAL OUTCOME OF PREIMPLANTATION GENETIC
DIAGNOSIS (PGD) CYCLES UNDER PRIVATELY FUNDED AND
PUBLICLY FUNDED PERIODS IN ONE CENTER. A. Ao, a
X. Zhang, b L. Zhang, b W. Buckett. b a Obstetrics and Gynecology, McGill
Univerisity, Montreal, QC, Canada; b McGill University, Montreal, QC,
Canada.
OBJECTIVE: To compare the overall clinical outcome of IVF-PGD cycles
during privately funded period and publicly funded period in one center.
DESIGN: A single center retrospective study.
MATERIALS AND METHODS: All couples that underwent IVF-PGD
cycles within specific study periods were divided under two groups: Privately
funded period (PRP) where the IVF and PGD treatment cycles were privately
funded and publicly funded period (PUP), where the treatment cycles were
funded by the provincial government healthcare system. Under Privately
funded period, 298 cycles (195 PGS and 42 translocation and 61 SGD)
and under publicly funded period, 221 cycles (59 PGS and 72 translocation
and 90 SGD) underwent IVF-PGD procedure. The outcomes were assessed
for pregnancy rate and live birth rate.
RESULTS: The average age of female patients for both private and
publicly funded periods were similar (35 vs 36 years old). The fertilization
rate (73% vs 72%), embryo biopsy rate (80% vs 80%) and genetic
diagnosis rate (89% vs 94%) were not significantly different between
these two groups. More embryos were transferred per cycle during
PRP than PUP (2.34 vs 1.12, p

RESULTS: The diagnostic rates were 98.7% for Karyomapping and
98.6% for CCS. The average numbers of embryos analyzed and eligible
for transfer per cycle were 5.3 and 2.5 for PGD alone, and 6.2 and 2.3 for
PGD+CCS. Batching of embryos prior to analysis was carried out for 11%
of PGD+CCS cases (2% for PGD alone).The CCS improved clinical
outcome. The implantation and clinical pregnancy rates were 70.5% (43/
61) and 64% (34/53) for PGD+CCS cycles, versus 54.5% (6/11) and
55.6% (5/9) for PGD alone. Most transfers involved a single thawed blastocyst,
except for ten patients (16%) who had two embryos transferred at once.
The average maternal ages were 32.3 and 34.2 years for PGD and PGD+CCS,
respectively. There are 13 live births and 36 ongoing pregnancies to date. Prenatal
and perinatal testing was performed in four cases confirming PGD in all
instances.
CONCLUSIONS: The high clinical pregnancy rates were similar to outcomes
achieved by the most successful in-vitro fertilization (IVF) programs.
As lengthy patient-specific test developments were unnecessary, greater
numbers of patients were treated with faster time to diagnosis, transfer and
pregnancy. SNP arrays thus allow PGD laboratories to deal with the rapidly
growing demand for embryo testing for inherited disorders. The application
of Karyomapping with simultaneous CCS speeds up the testing process and
further improves clinical outcomes.
P-509 Wednesday, October 21, 2015
BLASTOCYST MORPHOLOGY AND CHROMOSOME STATUS:
CORRELATION IN WOMEN < AGE 35. C. F. Boylan,
L. S. Morrison, S. M. Carney, G. Kovalevsky, A. B. Neithardt,
R. F. Feinberg. Reproductive Associates of Delaware, Newark, DE.
OBJECTIVE: To determine if day 5 and 6 blastocyst morphology correlate
with ploidy status in women < 35.
DESIGN: A retrospective study that compiled morphology and ploidy status
from day 5 and 6 blastocysts created from 131 women under age 35.
MATERIALS AND METHODS: All blastocysts were cultured in vitro to
day 5 or 6 of development. When stage appropriate, trophectoderm biopsy
was performed. Approximately 2-10 cells were removed by laser excision
and sent to a PGS/PGD laboratory. The inner cell mass (ICM) and trophectoderm
cells of each embryo were graded prior to biopsy according to Gardner’s
grading scale. Each embryologist in the laboratory was extensively
trained on the scale to lessen subjectivity. Blastocysts were grouped into
‘‘AA’’, ‘‘BB’’ or ‘‘CC’’ according to their overall quality. They were then
grouped exclusively by their ICM and trophectoderm. Chi Square analysis
was performed to test significance between the grade and ploidy status.
RESULTS:
Overall Grade Euploid Aneuploid
‘‘AA’’ Blastocysts 81/99 (81.8%) 1 18/99 (18.2%)
‘‘BB’’ Blastocysts 306/454 (67.4%) 1 148/454 (32.6%)
‘‘CC’’ Blastocysts 34/79 (43.0%) 1 45/79 (57.0%)
ICM Grade
‘‘A’’ ICM 123/158 (77.8%) 2 35/158 (22.2%)
‘‘B’’ ICM 382/582 (65.6%) 2 200/582 (34.4%)
‘‘C’’ ICM 52/111 (46.8%) 2 59/111 (53.2%)
Trophectoderm Grade
‘‘A’’ Trophectoderm 94/115 (81.7%) 3 21/115 (18.3%)
‘‘B’’ Trophectoderm 365/543 (65.6%) 3 178/543 (34.4%)
‘‘C’’ Trophectoderm 98/193 (50.8%) 3 95/193 (49.2%)
1,2,3 p-value¼

well characterized. The present study sought to develop a working model of
mosaicism through the use of aneuploid cell lines and to evaluate detection
limits of an NGS based CCS methodology.
DESIGN: Observational
MATERIALS AND METHODS: Cell lines with single chromosomal aneuploidies
(+15 or +18) were mixed with a total of 6 cells in various ratios
(16%, 33%, 50%, 66%, 83% and 100%) with euploid cells in order to model
a range of mosaicism levels that might be observed in a typical trophectoderm
biopsy. NGS based CCS was performed using a previously validated
custom amplification protocol and an Ion Proton. Statistical outlier analysis
of chromosome specific copy number assignments, with values above the 3
interquartile range (IQR) being identified as outliers, was used to identify
mosaic chromosomes.
RESULTS: Examination of mosaicism with artificial 6-cell mixtures revealed
that NGS could detect mosaicism at 16%, was relatively reliable at
50%, and showed high specificity throughout each mixture (Table 1).
CONCLUSIONS: This is study represents the most extensive evaluation
of CCS sensitivity to mosaicism reported to date. Most importantly, this
study defined the limits of detection of aneuploidy mosaicism within a 6-
cell sample, and can be used to set evidence-based expectations when counseling
patients. Future work will involve evaluating whether detection
criteria established in this model may be predictive of actual clinical outcomes
when applied to trophectoderm biopsies.
TABLE 1. Results of NGS CCS detection of mosaicism in a model system.
Mosaicism Spike-in 16% 33% 50% 66% 83% 100%
(n) 12 12 12 12 12 12
Sensitivity 8% 33% 67% 75% 75% 100%
Specificity 100% 91% 91% 100% 91% 100%
P-512 Wednesday, October 21, 2015
PUSHING THE LIMITS OF PREIMPLANTATION GENETIC DIAG-
NOSIS (PGD): A KARYOMAPPING AND NEXT GENERATION
SEQUENCING (NGS) STRESS TEST. M. R. Lindeman,
E. Czuprenski, J. Kitchen, T. K. McWilliams, M. Wyatt, T. T. Gordon,
M. Hughes. Genesis Genetics, Plymouth, MI.
OBJECTIVE: We sought to examine the limits of embryo genetic analysis
by performing PGD and aneuploidy screening (PGS) in 1) a consanguineous
mating for, 2) two disease-causing point mutations and 3) HLA-genotyping
to match 3 affected children while 4) detecting genetic recombination and
potential maternal cell contamination, and 5) 24-chromosome aneuploidy
screening with NGS.
DESIGN: Karyomapping utilizes 300,000 Single Nucleotide Polymorphisms
(SNPs) across the human genome and allows high-resolution analysis
for nearly every gene. We used karyomapping to rapidly design a PGD test
targeting the HBB gene and the highly recombinant HLA locus to identify
embryos with one or both healthy HBB alleles and an HLA genotype matching
the children in need of transplantation.
MATERIALS AND METHODS: Cheek-swab kits were used to collect
DNA samples from the parents and affected children. Mutation status of
each individual was confirmed by direct sequencing. DNA samples were
analyzed using the Infinium HumanKaryomap-12 DNA Analysis Kit from Illumina.
BluFuse Multi software was used to compare the SNP profiles of
each family member to identify informative SNPs, regions of consanguinity,
and determine HLA phase for each affected child. NGS was used for 24-chromosome
aneuploidy screening.
RESULTS: We identified sufficient informative SNPs surrounding the HBB
gene and HLA region to track the inheritance of familial haplotypes. As expected,
the same HBB haplotypes were observed in each affected child. Two children
were found to have the same set of HLA haplotypes, while the third inherited
the opposite paternal HLA haplotype. This is consistent with the HLA-typing reported
by an external laboratory. Recombination and consanguineous regions
were observed but did not impact our ability to determine genetic phase.
CONCLUSIONS: We successfully used karyomapping to build a complex
PGD test, which would have been highly challenging using previous methods.
Moreover, we determined the HLA phases of all three children simultaneously
and confirmed that recombination and consanguinity would not interfere
with diagnosis or HLA matching. Our success addresses concerns about
using karyomapping for HLA matching and diagnosing embryos of consanguineous
families. Karyomapping combined with PGS by Next-Generation
Sequencing represents a great advance to the field, 25 years after its inception.
P-513 Wednesday, October 21, 2015
REPRODUCTIVE OUTCOME FOLLOWING PRE-IMPLANTATION
GENETIC DIAGNOSIS (PGD) - AN ANALYSIS OF UK NATIONAL
DATABASE OVER TWO DECADES. A. N. Sharpe a M. Choudhary. b
a Newcastle Fertility Centre at Life, Newcastle-upon-Tyne, United Kingdom;
b Consultant Gynaecologist & Sub Specialist in Reproduction, Newcastle
upon Tyne, United Kingdom.
OBJECTIVE: In 2013, as a drive to ensure an equal and consistent policy,
the National Health Service Commissioning board centralised the funding in
England for up to 3 PGD cycles for couples who have or are carriers of a specific
genetic disorder and wish to avoid having an affected child. This study
aims to determine the UK incidence of PGD over the last 20 years and associated
clinical outcome.
DESIGN: Retrospective national database cohort.
MATERIALS AND METHODS: Data from 1991 to 2012 was analysed
from the Human Fertilisation Embryology Authority (HFEA) database to
determine the incidence and clinical outcomes of couples who had PGD. Binary
logistic regression was used to compare PGD cycles versus non-PGD
cycles for age matched women over the years.
RESULTS: Of a total 1,071,040 cycles of assisted reproduction techniques
(ART) from 1991 to 2012, 2974 had PGD (0.28%). The incidence has risen
127 fold from 1/15953 cycles in 1991 to 530/66553 cycles in 2012.There was
2443 PGD cycles from 1999 to 2011, where 585 resulted in live birth (23.9%)
compared to 137588 live births from 659622 non-PGD cycles (20.9%,
p¼0.15). The live birth rate increased over time and was greater in women
aged 18-34 years for both PGD and non-PGD cycles (p

P-514 Wednesday, October 21, 2015
LASER-ASSISTED HATCHING IN EARLY BLAST IS OPTIMAL
FOR TROPHECTODERM BIOPSY OF MOUSE
EMBRYOS. Y. Chiang, D. Russell, M. Rosario, S. Wang. Obetetric
and Gynecology, University of Colorado Denver, Aurora, CO.
OBJECTIVE: To determine which strategy of laser-assisted hatching is
optimal for trophectoderm biopsy in mouse embryos
DESIGN: Prospective in vitro study
MATERIALS AND METHODS: Totally 280 mouse embryos were randomized
to laser-assisted hatching in different cleavage stages, including
89 embryos without laser-assisted hatching, 76 embryos with laser-assisted
hatching in 8 cell stage, 46 embryos with laser-assisted hatching in morula
stage and 69 embryos with laser-assisted hatching in early blast stage. The
morphology of embryo in trophectoderm biopsy, biopsy time and atretic
rate in 24 and 48 hours after biopsy were analyzed.
RESULTS: Significant higher percentages of hatching in trophectoderm
was noted in embryos with laser-assisted hatching in early blast stage than
embryos without laser-assisted hatching, embryos with laser-assisted hatching
in 8 cell and morula stages (94.2% versus 18.0%, 43.4%, 32.6%, Chisquare
test, p

CONCLUSIONS: Our data suggest that mosaicism may be present in the trophectoderm,
at a rate of approximately 20.0%. More importantly, although mosaicism
is present, this phenomena only resulted in a clinically significant rate of
approximately 2.0% (euploid vs. aneuploid). Therefore, mosaicism may be present
in the human blastocyst; however at a low rate of clinical significance.
P-517 Wednesday, October 21, 2015
ANEUPLOIDY PARENT OF ORIGIN IN BLASTOCYST BIOPSIES
USING KARYOMAPPING TECHNOLOGY. K. McWilliams,
T. K. McWilliams, M. Wyatt, M. Hughes. Genesis Genetics, Plymouth, MI.
OBJECTIVE: Characterization of parental derivation of whole chromosome
aneuploidies by the single nucleotide polymorphism (SNP)-based karyomapping
platform.
DESIGN: Retrospective, multi-faceted analysis of blastocyst biopsies submitted
for single-gene preimplantation genetic diagnosis (PGD) and chromosome
screening (PGS) analysis.
MATERIALS AND METHODS: 90 blastocyst biopsies from 54 families
were tested by PGD and PGS by comparative genomic hybridization microarray
(aCGH) or Next Generation Sequencing. Maternal ages ranged from
27-39 (mean of 32). Paternal ages ranged from 27-54 (mean of 36). Aneuploidies
detected by PGS were analyzed by Illumina’s karyomapping platform,
which contains 300,000 genome-wide SNPs on a BeadChip array.
Embryos with segmental duplications/deletions or greater than 2 whole chromosome
aneuploidies were excluded from analysis. BlueFuse Multi software
data analysis revealed familial haploblock inheritance. Monosomies showed
lack of inheritance from one parent; whereas trisomies demonstrated inheritance
of 3 parental haploblocks. Data was collected on the basis of aneuploidy
type, maternal vs. paternal inheritance, and parental age.
RESULTS: To date, 49.6% of embryo biopsies studied by karyomapping
and PGS combined testing have been aneuploid. Ninety-eight aneuploid
events were studied by karyomapping. The majority (85.7%) of all detectable
whole chromosome aneuplodies were maternally-derived (p¼35 years) and embryo euploidy, and suggest
that variation in maternal genes may explain differences in aneuploidy incidence.
24-chromosome SNP-based PGS helps infertility patients achieve
high rates of implantation, and viable, healthy pregnancies.
Supported by: This work was partly Supported by National Institutes of
Health grants R01 GM100366, R01 GM097415, and R01 GM089926.
P-520 Wednesday, October 21, 2015
DESCRIPTION OF EUPLOID EMBRYO IMPLANTATION
OUTCOME BY MORPHOKINETIC INVESTIGATION. A. Tejera, a
M. Stoppa, b M. Meseguer, a A. Capalbo, b M. Florensa, c F. Ubaldi, b
Y. Galiana, a L. Rienzi. d a IVI Valencia, Valencia, Spain; b Genera, Rome,
Italy; c IVF Lab Director, Barcelona, Spain; d Genera Centres for Reproductive
Medicine, Rome, Italy.
FERTILITY & STERILITY Ò
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OBJECTIVE: To define euploid implanted morphokinetic parameters and
propose an algorithm for embryo selection based on this analysis.
DESIGN: Retrospective observational study.
MATERIALS AND METHODS: Embryo development from 265 patients
undergoing pre-implantation genetic screening, euploid transferred embryos
(n¼342) were analyzed with time-lapse imaging (Embryoscope, Vitrolife,
Denmark). Chromosomal analysis was performed through array-comparative
genome hybridization and qPCR. Timing of each cellular event was annotated
along embryo development (tn) in hours post ICSI, also we
calculated cell cycle duration (ccn¼tn+1-tn) and synchrony between blastomeres
(sn¼tn+1-tn). In addition, timing of compaction (tM), blastocyst formation
(tB) and expansion (tEB) were recorded. The data to define
morphokinetic behaviour was obtained from a total of transferred embryos,
and embryos with known implantation data (KID). Only KID embryos
with either full implantation (number of sacs matches the number of transferred
embryos n¼130) or no implantation (no biochemical pregnancy
achieved n¼169) were included in this analysis.
RESULTS: A logistic regression analysis identified tM (

In this study we found out that patients older than 36 years old with AMH
level less than 1 ng/ml, have significantly lower number of blastocyst suitable
for biopsy, compared to the patients with AMH level great than 1 ng/ml
(0.270.20 vs. 2.470.32). Patient in the lower AMH group also has significantly
higher BMI compared to the group with higher AMH group
(28.311.44 vs. 24.850.63).
P-523 Wednesday, October 21, 2015
USING 24-CHROMOSOME ANEUPLOIDY TEST AND FROZEN
BLASTOCYST TRANSFER CYCLES SIGNIFICANTLY IMPROVES
PREGNANCY RATE AND DECREASES MISCARRIAGE RATE IN
IVF PROGRAMS. K. Kirienko, a A. Strashnova, a E. Uvarova, a
N. Voronich, a Z. Zlatopolsky, b O. Verlinsky, b T. Pakhalchuk, b
V. Apryshko, a S. Yakovenko. a a Altravita IVF Clinic, Moscow, Russian
Federation; b Reproductive Genetic Institute, Chicago, IL.
OBJECTIVE: The purpose of this study was to compare clinical outcomes
(pregnancy and miscarriage rates) between frozen blastocyst transfer (FBT)
cycles without preimplantation genetic screening (PGS), FBT cycles with
24-chromosome aneuploidy test and FBT cycles with 9-chromosome
screening for infertile patients with normal karyotype aged 25-45.
DESIGN: Retrospective comparative study.
MATERIALS AND METHODS: Embryos were cultured to the blastocyst
stage and vitrified shortly (Cryotech, Japan) after trophectoderm (TE)
mechanical biopsy. Mechanically biopsied cells were analyzed by using 24-Chromosome
Microarray - aCGH or by QF-PCR with the focus on 9-chromosomes
(13,14,15,16,18,21,22,X,Y). A total of 503 couples were involved in the study.
The experimental group included 203 couples (average female age is 37), 1022
blastocyst stage (BS) embryos were obtained from this group. Of these embryos,
489 were analyzed by aCGH and 533 were analyzed by QS-PCR. The control
group included 300 couples (average female age is 33) undergoing IVF program
without PGS. Ongoing pregnancies after single embryo transfer (SET) were
confirmed by one fetal sac and heartbeat. Data was analyzed using Student’s t-test.
RESULTS: Results are shown in the table. In IVF with 24-chromosome
aneuploidy test group the risk of aneuploidies was strongly age-dependent
(p39
SET cancellation in %39
connection with >39
abscence of
PGS-normal
embryos, %
Clinical pregnancy %39
rate/SET, % >39
Miscarriage rate/ %39
pregnancy, %
The average cost of
the program,
USD (% of IVF)
>39
%39
>39
IVF
-
-
-
-
35.0
20.0
34.8
57.5
6521
6911
IVF+PGS 9-
chromosome
75.6
72.5
6.4
10.4
35.5
16.0
22.0
43.0
7911 (121%)
7184 (104%)
IVF+24-
chromosome
aneuploidy test
50.5
24.5
11.8
45.0
58.7
26.0
11.4
16.7
8147 (125%)
8562 (124%)
OBJECTIVE: To identify if pregnancy outcomes differ in patients that
make multiple euploid embryos from a single in vitro fertilization (IVF) cycle
in comparison to patients who make only one.
DESIGN: Retrospective Cohort Study
MATERIALS AND METHODS: Global retrospective IRB was obtained.
All patients who underwent IVF with preimplantation genetic screening
(PGS) with array comparative genomic hybridization (aCGH), with at least
one euploid embryo were included. The first single euploid frozen embryo
transfer cycle following the IVF cycle was included. All causes of infertility,
all indications for PGS, autologous and donor cycles were included. Patients
who elected to undergo two or more euploid embryo transfer cycles were
excluded. The primary outcome of the study included pregnancy (presence
of a gestational sac) and clinical pregnancy (presence of fetal heart beat
through cycle day 63 or a live birth). Statistical analysis was done using AN-
OVA, t-test and a linear regression model.
RESULTS: A total of 327 single euploid embryo transfers were included.
The average age of patients included in the study was 37.37 4.90. The overall
pregnancy rate was 68.50%(224/327), of which 61.47% (201/327) led to
ongoing clinical pregnancies or live births. The most common number of
euploid embryos per cohort was one (n¼108) with the highest number being
FERTILITY & STERILITY Ò
e285

20 (n¼1). There was no difference in pregnancy rates when embryos from
cycles with multiple euploid embryos were compared to cycles with only
one euploid embryo (RR 0.9965, CI 0.8872-1.119, p 0.9457). Linear regression
showed no difference was found in pregnancy rates when the percentage
of euploid embryos per cohort was compared as well (R ¼0.0905). There was
a significant difference in pregnancy rates in embryos of higher-grade inner
cell mass, however when controlled for embryo grades no difference was
noted in pregnancy rates in lower numbered euploid embryo cohorts.
Pregnancy Outcomes by Number of Euploid Embryos per IVF cycle.
Number
of Euploid
Tota
Patients
(n ¼ 327)
Average
Number of
Eggs
Retrieved
Total
Pregnant
(presence of
gestational
sac)
Percent
Total
Clinical
Pregnancies
(Fetal heart
beat or live
birth)
Percent
1 108 12.32 +/- 5.75 70 65 65 60
2 84 15.60 +/- 7.23 62 74 55 65
3 42 15.79 +/- 6.91 29 69 25 60
4 30 19.30 +/- 7.63 22 73 21 70
5 16 22.56 +/- 9.92 10 63 10 63
6 13 22.38 +/- 8.91 9 69 7 54
7 13 27.50 +/- 5.09 6 46 5 38
8 or more 21 30.0 +/- 10.78 16 76 13 62
p value p 0.552 p 0.552 p 0.676
CONCLUSIONS: The number or percentage of euploid embryos per IVF
cycle, is not associated with the pregnancy outcome of STEET.
P-526 Wednesday, October 21, 2015
FIRST VALIDATED METHOD FOR DISTINGUISHING NORMAL
FROM BALANCED TRANSLOCATION CARRIER
EMBRYOS. N. R. Treff, a K. Thompson, a M. Rafizadeh, a X. Tao, b
H. Garnsey, b C. V. Reda, a T. Metzgar, a E. Forman, a R. T. Scott. a a RMA,
NJ, NJ; b FAEEC, Basking Ridge, NJ.
OBJECTIVE: Carriers of balanced translocations often wish to prevent inheritance
in their offspring in order for their children to eventually avoid the
same reproductive challenges. Although identifying unbalanced embryos has
been possible for many years, current PGS methods are unable to reliably
distinguish a truly normal embryo from one that carries a balanced translocation.
The present study was conducted to validate a method that provides
the opportunity to make this distinction for the first time.
DESIGN: Prospective blinded.
MATERIALS AND METHODS: A series of translocation carrier couples
that underwent IVF with SNP array based PGD were included. Embryos that
were transferred in these cases could have been balanced or normal. SNP
array analysis was performed on parental DNA and unbalanced embryos
from each case to define informative SNPs whose alleles were linked with
the derivative chromosomes (phasing). The transferred sibling embryo array
data were then evaluated at the informative SNP loci to predict whether they
were truly normal (did not inherit the derivative chromosomes) or translocation
carriers. IRB approval was obtained to perform conventional karyotyping
on the newborns in order to establish the true genetic status of the original
transferred embryo (balanced or normal). Embryonic SNP array predictions
were then compared to the newborn karyotypes obtained.
RESULTS: Phasing SNPs using unbalanced embryos allowed accurate
prediction of whether transferred embryos were balanced translocation carriers
or truly normal in all 10 cases completed to date (100% concordance
with conventional karyotyping of newborns).
CONCLUSIONS: This study demonstrates the validity of the first method
capable of distinguishing normal from balanced translocation carrier embryos.
The only prerequisite is the availability of parental DNA and unbalanced
IVF embryos, making the method applicable to the majority of
carrier couples. In addition, the SNP array platform allows simultaneous
evaluation of comprehensive chromosome screening for aneuploidy, in parallel,
from the same biopsy. Future work will involve prospective predictions
to select normal embryos with subsequent karyotyping of the resulting newborns.
P-527 Wednesday, October 21, 2015
EMBRYOS FROM FEMALE CARRIERS OF BALANCED TRANS-
LOCATIONS DO NOT HAVE A HIGHER RISK OF WHOLE-CHRO-
MOSOME ANEUPLOIDY THAN THEIR AGE-RELATED
RISK. E. J. Forman, a N. R. Treff, b J. M. Franasiak, c R. T. Scott. b
a RMA, NJ, NJ; b RMANJ, Rutgers-RWJ, Basking Ridge, NJ; c RMANJ, NJ,
NJ.
OBJECTIVE: Given the altered chromosomal configuration of tetrads in
the synaptonemal complexes of translocation carriers during meiosis, there
has been concern that these couples may harbor increased risk for aneuploidy
in chromosomes not directly affected by the translocation. The purpose of
this study is to determine whether this interchromosomal effect (ICE) in
translocation carriers increases the risk of aneuploidy beyond their agerelated
risk when compared to a large data set of embryos tested with
comprehensive chromosome screening (CCS) at the blastocyst stage.
DESIGN: Case control.
MATERIALS AND METHODS: The baseline aneuploidy rate per chromosome
was calculated for all patients undergoing IVF with CCS and stratified
into

Frequency distribution analysis was performed. Log-regression analysis was
performed for the pre-treatment variables.
RESULTS: 105 patients met inclusion criteria. Treatment indications
included isolated HG in 7 patients and combined infertility and HG in 98 patients.
22 patients (21.0%) had inadequate androgenization after treatment
with CC, of which 13 patients paradoxically experienced a decrease in androgenization.
Pre- and post-treatment characteristics and univariate analyses
are reported in Table 1. The logistic regression model was statistically significant
22 patients (21.0%) had inadequate androgenization after treatment
with CC, of which 13 patients paradoxically experienced a decrease in androgenization.
Pre-treatment characteristics and univariate analyses are reported
in Table 1. The logistic regression model was statistically significant (c2(9)¼
45.12, p¼

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P-531 Wednesday, October 21, 2015
ROLE OF DIETARY ANTIOXIDANT SUPPLEMENTATION IN
TREATMENT OF IDIOPATHIC MALE INFERTILITY: PROMISING
EVIDENCE FROM A SUB-CONTINENTAL STUDY. S. K. Goswami, a
S. Yasmin, b P. Chakraborty, c R. Chattopadhyay, b S. Ghosh, b M. Goswami, b
B. Ghosh, d B. Chakravarty. b a Reproductive Medicine, Consultant, Kolkata,
India; b Assisted Reproduction, Institute of Reproductive Medicine, Kolkata,
India; c Infertility, Institute of Reproductive Medicine, Kolkata, India; d Andrology,
Institute of Reproductive Medicine, Kolkata, India.
OBJECTIVE: To evaluate the ameliorating potential of a diet rich in antioxidants
in couples with idiopathic male infertility with high reactive oxygen
species in comparison to a standard combined antioxidant formulation.
DESIGN: Prospective observational study from March 2013 to April
2015.
MATERIALS AND METHODS: 175 patients with idiopathic male infertility
(IMI) with high reactive oxygen species (ROS) were recruited, of which
80 patients were advised to take a diet rich in antioxidants along with few
changes in lifestyle (group A) and remaining 95 were prescribed a combined
oral antioxidant therapy (group B). A placebo-controlled group comprising
75 patients was maintained in parallel. Semen parameters, antioxidant concentrations
(CoQ-10, L-carnitine, zinc) with biomarkers of oxidative stress
including plasma total antioxidant capacity (TAC), total glutathione (GSH)
were analyzed before and after the treatment. Mitochondrial membrane potential
(Djm), ROS and sperm DNA fragmentation were evaluated by flowcytometry
and TUNEL assay respectively. MnSOD and PRDX5 expressions
were determined by real time PCR to evaluate the increased antioxidant effect.
Study was approved by Institutional Review Board. Statistical comparisons
were performed using Stata10.0.
RESULTS: Increased concentrations of plasma TAC and total GSH with a
significant increment in sperm motility (p

chemotherapy agents and the interval from chemotherapy were also
obtained and analyzed to detect any associations with the success in sperm
retrieval.
RESULTS: In a cohort of 66 NOA patients after chemotherapy, sperm
were successfully retrieved in 31 patients, with clinical pregnancy occurring
in 23 couples. Therefore, per-patient SRR was 47% and per-patient pregnancy
rate was 35%. Per-patient live birth rate was 27% (18/66). Our patients
had received chemotherapy for testicular cancer (21 patients), Hodgkin lymphoma
(9 patients), non-Hodgkin lymphoma (7 patients), acute lymphoblastic
leukemia (9 patients), acute myeloblastic leukemia (7 patients),
rhabdomyosarcoma (7 patients), bladder cancer (3 patients), osteosarcoma
(2 patients), and anaplastic anemia (1 patient). Table shows detailed information
of each cancer type. A multivariate analysis showed no significant predictors
of micro-TESE outcome.
CONCLUSIONS: Micro-TESE is the only method for fertility revival in
chemotherapy induced permanent NOA patients. However, our results suggest
micro-TESE and following intracytoplasmic sperm injection could
rescue only 27% of NOA patients after chemotherapy. No predictive values
of sperm retrieval were determined in this analysis. Sperm banking should be
offered before any chemotherapy even if the possibility of permanent azoospermia
is thought to be low.
P-533 Wednesday, October 21, 2015
SUPPLEMENTATION OF CRYOBUFFER WITH CATALASE AND
N-ACETYL CYSTEINE IMPROVES HUMAN SPERM POST-THAW
MOTILITY, VIABILITY AND DNA INTEGRITY. Y. Kobori,
M. Kathrins, C. Niederberger, G. S. Prins. University of Illinois at Chicago,
Chicago, IL.
OBJECTIVE: Cryopreservation of human sperm is a routine procedure
in assisted reproductive technology. The cryopreservation process can
lead to structural and functional alterations in spermatozoa, impairing
fertilization potential. Reactive oxygen species has been suggested as a
major contributing factor for cryodamage to spermatozoa. Accordingly,
antioxidant supplementation has been used to yield significantly
improved quality of frozen sperm post-thaw. We sought to investigate
improved outcomes with a combination of antioxidants Catalase and
N-Acetyl Cysteine (NAC), which function as intracellular and extracellular
antioxidants, respectively.
DESIGN: Laboratory investigation
MATERIALS AND METHODS: SpermFreeze (Vitrolife) with 15% glycerol
was used as cryobuffer supplemented with 0.0mM (control), 2.5mM,
5mM, and 10mM NAC with/without 200U/ml Catalase. Semen samples
were collected from normospermic men (n¼20) and aliquots frozen in
each dose of antioxidant tested. Post-thaw semen analysis by CASAwas performed
at 30 min and 2 hr. In addition, sperm viability (Eosin-Y staining) and
sperm DNA fragmentation (TUNEL assay) were quantitated at 2 hr postthaw.
RESULTS: A significant increase in total motility, progressive motility
and sperm viability was observed in all samples frozen with antioxidants
as compared to control. The cryobuffer containing 5mM NAC achieved the
highest recovered motility post-thaw among NAC doses. At 30 minutes
post-thaw, recovered motility (post thaw motility /pre-freeze motility
100%) in control, 200U/ml Catalase, 5 mM NAC and combined Catalase/
NAC supplemented buffer of sperm was 40.3%, 44.8%, 46.2%, and 47.8%,
respectively. Similar increased post-thaw progressive motility, viability
was observed with best results observed using the NAC/Catalase combination.
Preliminary results with DNA integrity analysis for double-stranded
breaks using TUNEL show similar trends and complete analysis will be presented.
CONCLUSIONS: Combined Catalase and N-Acetyl Cysteine supplementation
during cryopreservation results in improved post-thaw recovery of total
motility, progressive motility, percentage of sperm vitality and improved
DNA integrity. The results indicate that the combination of intracellular and
extracellular antioxidants results in the most pronounced effect in improving
post-thaw quality of human spermatozoa.
P-534 Wednesday, October 21, 2015
IDENTIFICATION OF SERTOLI CELL MARKERS IN MEN WITH
NON-OBSTRUCTIVE AZOOSPERMIA. A. Agarwal, a R. Sharma, a
Z. Cui, a,b E. S. Sabanegh. c a Center for Reproductive Medicine, Cleveland
Clinic, Cleveland, OH; b Institute of Toxicology, Third Military Medical University,
Chongqing, China; c Urology, Cleveland Clinic, Cleveland, OH.
OBJECTIVE: Dysfunctional spermatogenesis is presumed to be the main
cause of non-obstructive azoospermia (NOA). At present, serum follicle
stimulating hormone (FSH) levels and testicular volume are used to diagnose
NOA, however their sensitivity and specificity are limited as predictors of
testicular sperm extraction outcomes. The development of a reliable, noninvasive
biomarker that can predict NOA, especially in patients with normal
FSH, is the ultimate aim. We utilized proteomic analysis to identify proteins
that are differentially expressed in the seminal plasma of NOA patients with
normal FSH and NOA patients with increased FSH to help our understanding
of the etiology and mechanism of FSH in normal NOA. This might provide
new markers that can help in the diagnosis of NOA patients and better predict
the outcome of testicular biopsy.
DESIGN: Proteomic analysis of seminal plasma proteins from NOA men
with normal and elevated FSH levels in comparison with fertile men.
MATERIALS AND METHODS: Seminal plasma was obtained from 10
NOA patients with a high FSH level, and 4 NOA patients with a normal
FSH level and 7 fertile men. Short and long gels were run. Bands were cut,
and the gels (1mm 3 pieces) were trypsinized and run on a LTQ-Orbitrap
Elite hybrid mass spectrometer system. Functional annotations of proteins
were obtained using bioinformatic tools and pathway databases. Western
Blotting was performed to verify the expression levels of 3 proteins of interest.
RESULTS: We identified 493 proteins in the seminal plasma of fertile controls,
448 in NOAwith normal FSH and 436 in NOAwith high FSH levels. Of
these, 68 were differentially expressed proteins (DEPs) in NOA and normal
FSH and 15 between NOA and high FSH. DEPs that were overexpressed in
NOA and normal FSH levels were involved in glycosylation, cell-adhesion,
cell proliferation regulation, and secretion. DEPs that were underexpressed
were involved in stress response, glycosylation, secretion, spermatogenesis
and sterol transport. DEPs that were overexpressed in NOA with normal
and elevated FSH levels were involved in the processes of glycosylation,
secretion, defense response, cellular homeostasis, response to hormone stimulus,
and response to hypoxia. Proteins that were underexpressed were
involved in the processes of secretion, cell adhesion, cell skeleton, ECM-receptor
interaction, and oxidoreductase activity. We identified 4 proteins that
were related to Sertoli function such as secretion, Sertoli-germ cell junction
and hormone receptor function. These were lactotransferrin, desmoglein,
desmoplakin, and junction plakoglobin.
CONCLUSIONS: We have identified proteins that may serve as markers
of non-obstructive azoospermia. They participate in important Sertoli cell -
related functions and may play an important role in identifying patients
with NOA who are likely to have spermatozoa.
P-535 Wednesday, October 21, 2015
DIAGNOSTIC ACCURACY OF PHYSICAL EXAMINATION
COMPARED WITH COLOR DOPPLER ULTRASOUND IN THE
DETERMINATION OF TESTIS SIZE AND VARICOCELE
DIAGNOSIS. B. C. Tiseo, a M. Cocuzza, a V. Srougi, a G. J. Wood, a
S. Esteves, b M. Srougi. a a Division of Urology, Hospital das Clinicas, University
of Sao Paulo Medical School, Sao Paulo, Brazil; b ANDROFERT, Andrology
and Human Reproduction Clinic, Campinas, Brazil.
OBJECTIVE: To evaluate the accuracy of physical examination on the
diagnosis and grading of varicocele and on testicular volume measurement
comparing to ultrasonographic assessment. Varicocele treatment should be
offered to patient with palpable varicocele, provided it is associated with
infertility. Also, varicocele should be repaired in patients with testicular
growth impairment. Therefore, physical examination is the most important
parameter to deselect men with subclinical varicocele or testicular size alteration
to undergo treatment, as shown by the equivocal beneficial effect of
such interventions in such cases.
DESIGN: Prospective cohort.
MATERIALS AND METHODS: We consecutively evaluated 78 patients
attending at the University-based Urology Unit. All patients were examined
for presence of varicocele and testis size estimation by a group of two
non-experienced urologists and two experienced infertility specialists.
Anthropometric data was acquired for each patient. All of them were submitted
to Color Doppler Ultrasonography (CDU) to evaluate the presence of
varicocele and measure testicular volume.
RESULTS: There were 74 patients who completed the protocol with CDU
totalizing 147 testicular units evaluated. Mean testicular volume measured at
CDU was 13.6 5.6. Varicocele was diagnosed in 61.2% of the units by CDU.
Non-experienced urologists diagnosed varicocele in 47.6% and 48.9% and
false positive rate was 25.7% and 31.9%, while experienced urologists had
47.6% and 35.3% varicocele prevalence and false positive rate of 24.3%
FERTILITY & STERILITY Ò
e289

and 11.5%. The ROC curve analysis of physical examination for varicocele
presence revealed area under the curve of 0.570 and 0.631 within non-experienced
urologists and 0.645 and 0.703 within specialists.Regarding to volume
estimation, all examiners did not have correctly assessed testicular
volume with its difference form CDU measurement ranging from 1.30 to
2.41cc (p

P-539 Wednesday, October 21, 2015
OPTIMAL TIME TO BANK SPERM FROM PATIENTS WITH
TESTICULAR TUMORS. M. Ribeiro de Andrade, P. Ferigolo,
M. P. Antoniassi, M. Camargo, P. Intasqui, D. S. Tibaldi, R. Bertolla,
D. M. Spaine. Departament of Surgery, Division of Urology, Sao Paulo Federal
University, Sao Paulo, Brazil.
OBJECTIVE: While testicular tumors present high survival rates, post-orchiectomy
adjuvant treatment often utilizes gonadotoxic agents, which renders
sperm banking of great importance in these patients. We wished to verify
if sperm banking should be performed before or after orchiectomy (before
adjuvant therapy) in patients with testicular tumors.
DESIGN: Prospective Study.
MATERIALS AND METHODS: This prospective study was carried out
including 17 patients with testicular germ cell tumors, who provided one
semen sample before they were submitted to orchiectomy of the affected
testis, and one other semen sample 30 days after the surgery. An aliquot of
each sample was used to seminal analysis and other to determinate sperm
function. Sperm DNA fragmentation was assessed by an alkaline Comet
assay, in which sperm were classified as high DNA integrity (class I) to
high DNA fragmentation (class IV). Acrosome integrity was determined
by a fluorescent probe (PNA-FITC) and assessed by fluorescent microscopy.
Mitochondrial activity was determined by a colorimetric stain (3,3’-diaminobenzidine
- DAB) and classified as DAB I (all mitochondria active) to DAB
IV (all mitochondria inactive). Seminal plasma was also tested for oxidative
stress with a TBARS assay. Pre- and Post-orchiectomy samples were
compared using a paired Wilcoxon test (p

was a trend towards a higher BMI with worsening semen concentration.
When controlled for the multiple factors listed above, we did not find any significant
differences in birth weight or BMI at time of first driver’s license for
children of men with semen analyses relative to fertile controls.
CONCLUSIONS: Although obesity is associated with poor semen quality,
our study demonstrates that the BMI of offspring is not associated with
abnormal semen parameters of the father.
P-542 Wednesday, October 21, 2015
PATERNAL AGE AND VASECTOMY REVERSAL
SUCCESS. B. E. Kahn, a D. L. Davenport, b C. G. Schrepferman. c a Urology,
University of Kentucky, Lexington, KY; b University of Kentucky, Lexington,
KY; c University of Louisville, Louisville, KY.
OBJECTIVE: To evaluate the effect of paternal age on patency rates
following vasectomy reversals.
DESIGN: Retrospective review of all vasectomy reversals performed by a
single surgeon from 2005 to 2014. For inclusion, patients supplied at least
one post-operative semen analysis or a reported a pregnancy. Vasectomy reversals
performed for pain were excluded (n¼3). Patency was defined as the
presence of > 500,000 motile sperm on a post-operative semen analysis or
report of pregnancy.
MATERIALS AND METHODS: Patency rates were assessed by vasectomy
reversal procedure performed, which included microsurgical bilateral
vasovasostomies (VV/VV), unilateral vasovasostomy with contralateral vasoepididymostomy
(VV/VE), bilateral vasoepididymostomy (VE/VE), unilateral
VV, and unilateral VE. The patency rate was stratified by decade of
paternal age (20-29, 30-39, 40-49, 50-59). Statistical analysis was performed
using Fisher’s Exact test, Chi-square test, and ANOVA linear trend tests.
P-values

x10⁶ sperm and 55.028,57x10⁶/ mL, respectively. The lowest and highest
catalase concentration detected was 11.39 (catalase units /total protein)
and 101.84 (catalase units/total protein), respectively. There was no significant
correlation between ROS levels and catalase levels in spermatozoa.
Methylation status of CpG island I and CpG island II of CAT gene was evaluated.
CpG island I hypermethylation was found in all samples, however
CpG island II of CAT gene was not methylated in the 20 samples studied.
CONCLUSIONS: Our results demonstrate hypermethylation of CpG island
I and unmethylation of CpG island II of CAT gene in spermatozoa. In
addition, no significant association was seen between ROS level and methylation
status of CAT gene.
Supported by: Cleveland Clinic.
P-546 Wednesday, October 21, 2015
COMPROMISED TOTAL ANTIOXIDANT CAPACITY SUPPORTED
BY HIGH DFI TO GUIDE SPERM SOURCING. T. Paniza, a L. Park, a
L. Reisman, a Q. V. Neri, a T. Cozzubbo, a M. Goldstein, b Z. Rosenwaks, a
G. D. Palermo. a a Reproductive Medicine, Weill Cornell Medical College,
New York, NY; b Urology, Weill Cornell Medical College, New York Presbyterian,
New York, NY.
OBJECTIVE: To assess the impact of seminal antioxidant capacity on
sperm DNA integrity. To measure embryo developmental competence of
spermatozoa retrieved from a healthier micro-environment through microsurgery.
DESIGN: Men with extremely high DFI in their ejaculates (n¼45) simultaneously
received a TAC analysis on seminal fluid. Following counseling,
men underwent surgical sampling and DNA integrity assessment on spermatozoa
retrieved from vassal fluid, epididymis, and testis. TAC, sperm parameters,
DFI and clinical outcome were recorded and compared for each
individual sperm source.
MATERIALS AND METHODS: Ejaculates were processed in standard
fashion, assessed for TAC, DFI by TUNEL and/or SCSA. TAC was determined
by a colorimetric assay on an automated microplate reader. Surgical
specimens were isolated from different sites of the male genital tract and
DFI measured exclusively by TUNEL then cryopreserved for later use.
RESULTS: In 51 ejaculates the average DFI was 32.0% for TUNEL
and 40.6% with SCSA with a good concordancy between the two assays.
While proximal surgical sampling improved DFI, it inversely yielded
lower concentration of spermatozoa and with somewhat decreased
motility. The antioxidant buffering capacity of the ejaculates inversely
correlated with the DFI (P

formulations. 42 (48.3%) used cutaneous testosterone gels, while 23 (26.4%)
used testosterone injections. 49 (56.3%) respondents found their patients to
be satisfied ‘‘most times’’ and 30 (34.5%) said their patients are ‘‘almost always’’
satisfied with Testopel therapy. When Testopel therapy was discontinued,
cost was cited as the most common reason.
CONCLUSIONS: Testopel implantation appears to be a highly successful
therapy for the treatment of hypogonadism. This study categorizes common
practices that have yet to be standardized. Based on this specialty society
questionnaire, management of hypogonadism may require 10 or more pellets
in the majority of cases.
P-549 Wednesday, October 21, 2015
PREDICTORS FOR SPERM RETRIEVAL IN MICRODISSECTION
SPERM EXTRACTION FOR NON-OBSTRUCTIVE
AZOOSPERMIA. T. Ishikawa, K. Yamaguchi, Y. Takaya,
R. Nishiyama, K. Kitaya, H. Matsubayashi. Reproduction Clinic Osaka,
Osaka, Japan.
OBJECTIVE: Recently, the most popular treatment in patients with nonobstructive
azoospermia (NOA) has been micro TESE with subsequent assisted
fertilization by intracytoplasmic sperm injection (ICSI). With the spread of ICSI,
the presence of a minimum number of spermatozoa is required for fertilization.
Micro TESE and ICSI cycles expose the couple to an emotional and financial
burden, so it would be beneficial to predict the success of sperm retrieval using
noninvasive parameters before attempted procedure. The aim of this study is to
assess the predictors of sperm retrieval by micro TESE in NOA patients.
DESIGN: A retrospective study.
MATERIALS AND METHODS: A total of 1323 micro TESE attempts
were done in 1275 men with confirmed cryptozoospermia and NOA to
recover spermatozoa for ICSI between January 2006 and March 2015 by a
single surgeon. Micro TESE was used in which seminiferous tubule are
directly examined throughout the testicle using an operating microscope
and selectively biopsied for all of the NOA patients. We analyzed sperm
retrieval rate (SRR) of the patients with NOA. Serum follicle stimulating hormone
(FSH), luteinizing hormone (LH), testosterone (T), testicular volume,
age at micro TESE, chromosomal analysis, AZF microdeletions analysis, and
past history were examined as predictive factors for sperm recovery. Chromosomal
analysis was performed on all patients on cultured lymphocytes
from peripheral blood.
RESULTS: Testicular sperm were successfully retrieved by micro-TESE
in 560 of 1323 (42.5%). No correlation was found between serum FSH,
LH, and T level with the success of sperm retrieval. Testicular volume and
patient age also did not affect the SRR for micro-TESE. Good candidates
of sperm retrieval by micro TESE were cryptozoospermia (114/120:
95.0%), AZFc microdeletion (22/28:78.6%), associated with cryptorchidism
(50/69: 72.5%), and non-mosaic Klinefelter syndrome (77/144: 53.5%).
Worse candidates of sperm retrieval were AZFa or b microdeletions (0/
10:0%), 46XY male with NOA without past history (180/643: 28.0%), and
after chemotherapy (25/70: 35.7%).
CONCLUSIONS: A prognostic parameter for successful sperm retrieval
in TESE seems to be decisive for male fertility. FSH is not able to resolve
spermatogenesis on an individual tubule level, and, therefore, they should
not be used as predictors of sperm recovery. We conclude that at the present
time there are no absolute predictors of sperm yield for micro TESE. However,
we could predict good candidates for micro TESE by past history
and genetic analysis.
P-550 Wednesday, October 21, 2015
THE RELATIONSHIP BETWEEN A MAN’S SOMATIC HEALTH
AND ART OUTCOMES. M. Eisenberg, a S. Li, b B. Behr, c
S. Nakajima, a V. L. Baker. d a Urology, Stanford University, Stanford, CA;
b Stanford University, Palo Alto, CA; c Stanford Fertility and Reproductive
Medicine Cente, Pali alto, CA; d Stanford University, Stanford, CA.
OBJECTIVE: As medical comorbidity and medication use increases,
semen quality declines. However, less is known about how a man’s somatic
health may impact the outcomes of assisted reproductive techniques.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: After IRB approval, we identified couples
undergoing assisted reproductive technology (ART) cycles at our center
from 2004 until 2014. We only fresh IVF cycles utilizing fresh ejaculated
sperm from the male partner. We recorded patient and partner demographic
characteristics. The cohort was linked to administrative data to obtain information
on the male partners’ comorbidities identified using ICD-9-CM codes
and limited to men evaluated within the health care system. Cycle outcomes
were queried from our clinical database. We calculated fertilization rate, clinical
pregnancy rate, miscarriage rate, implantation rate, live birth rate, and
singleton birth weight. Regression models were adjusted for male and female
covariates.
RESULTS: In all, we identified 772 men who had outpatient data available.
Those men underwent 1,503 fresh ART cycles - 702 were IVF only
and 801 utilized ICSI. Overall, the mean age of the man was 39.6 and 37.7
for his female partner. 67 % of men had at least one medical diagnosis.
96% of men had a CCI of 0. After stratifying by organ system, differences
were noted for ART outcomes based on any male diagnosis. Men with neurologic
diseases had a lower live birth rate (15% vs 23%, p¼0.02) while men
with endocrine diseases had a higher implantation rate (73% vs 62%,
p¼0.02). The associations were similar for unadjusted and adjusted models.
When examining singleton birth weights after all forms of ART, men with
diseases of the nervous system (3270g vs 2990g, p

function tests. Fourteen of 32 subjects (43.8%) achieved pregnancy,
including 10/13 (76.9%) conceiving through in vitro fertilization, 2/3
(66.7%) through intrauterine insemination and 2/16 (12.5%) through natural
intercourse.
CONCLUSIONS: Significant improvement in testosterone levels and
sperm concentration is seen with the use of anastrozole in the treatment
of the overweight or obese, subfertile male. Anastrozole is well tolerated
with minimal side effects in overweight or obese male subjects.
Concomitant anastrozole therapy in conjunction with assisted reproductive
techniques appears to be associated with successful conception for
infertile couples.
P-552 Wednesday, October 21, 2015
EFFECT OF TIME ON OXIDATION-REDUCTION POTENTIAL
IN SEMEN AND SEMINAL PLASMA. R. Sharma, a S. S. Du Plessis,
a,b A. Agarwal, a A. Harlev, a,c L. Samanta, a,d G. Ahmad, a,e S. Gupta, a
E. S. Sabanegh. f a Center for Reproductive Medicine, Cleveland Clinic,
Cleveland, OH; b Medical Physiology, Stellenbosch University, Tygerberg,
South Africa; c Soroka Medical Center, Ben-Gurion University, Beer Sheva,
Israel; d Redox Biology Laboratory, School of Life Science, Ravenshaw
University, Orissa, India; e Physiology and Cell Biology, University of
Health Sciences, Lahore, Pakistan; f Urology, Cleveland Clinic, Cleveland,
OH.
OBJECTIVE: Oxidation-reduction potential (ORP) is a novel measure of
oxidative stress or redox imbalance in biological fluids. Reactive oxygen species
(ROS) are highly reactive and have a very short half-life. ROS levels in
the seminal ejaculate should be measured within an hour after collection to
prevent a reduction in ROS levels over time. The traditional methods of
measuring seminal ROS are time sensitive and time consuming, making it
difficult to use them for diagnostic purposes. It would be highly advantageous
to employ a method that is independent of semen age and provides results in
real time. The objective was to assess the effect of time on static ORP
(sORP), which provides a snapshot of current redox balance, and capacity
ORP (cORP) which is indicative of the amount of antioxidant reserves available.
DESIGN: Prospective study measuring ORP in semen and seminal plasma
samples at time 0 and 120 minutes.
MATERIALS AND METHODS: The sORP and cORP of both semen
(n¼18) and seminal plasma (n¼15) samples from normal control subjects
were measured after liquefaction (time 0) and after 120 minutes
of incubation at room temperature (RedoxSYSÒ, Aytu BioScience).
Values are mean SEM. Spearman correlation was used for statistical
analysis.
RESULTS: A significant correlation was seen between sORP at time 0 and
120 minutes in semen and seminal plasma. Similar correlations were found
for cORP values at both time intervals.
CONCLUSIONS: ORP values are not affected by the age of semen or seminal
plasma for up to 120 minutes, making it easier to employ this new technology
for diagnostic use.
Effect of time on semen and seminal plasma ORP levels.
Semen
sORP
(mV/10 6
sperm)
Semen
cORP
(mC/10 6
sperm)
Seminal
Plasma
sORP
(mV/10 6
sperm)
Seminal
Plasma
cORP
(mC/10 6
sperm)
Time 0’ 2.790.66 0.200.08 2.670.71 0.310.16
Time 120’ 2.840.67 0.240.11 2.790.79 0.260.11
Correlation (r) 0.97 0.88 0.97 0.98
Median absolute
value of
difference
0.175 0.01 0.13 0.01
P-553 Wednesday, October 21, 2015
THE EFFECT OF VARICOCELE ON EMBRYO DEVELOPMENT
AND IVF OUTCOMES. A. S. Polackwich, a E. S. Sabanegh, b
N. Desai. c a Urology, Cleveland Clinic, Cleveland Heights, OH; b Cleveland
Clinic, Cleveland, OH; c Cleveland Clinic, Beachwood, OH.
OBJECTIVE: The treatment of male factor infertility secondary to varicocele
is controversial in the era of Intracytoplasmic Sperm Injection. Utilizing
this technique, all but the most severe male factor can be overcome. In some
patients, treatment of varicoceles can help a couple avoid IVF, but for those
who require it, the impact of a varicocele on outcomes is not well defined. We
evaluated the influence of a clinically relevant varicocele on IVF.
DESIGN: Retrospective chart review of all couples treated by both Cleveland
Clinic Andrology and Reproductive Endocrinology
MATERIALS AND METHODS: After IRB approval, we cross-referenced
our database of patients who have undergone IVF with all patients seen in our
male infertility clinic. We then extracted data on IVF outcomes, demographics,
seminal parameters, physical exam and surgical history. All patients
who underwent a varicocele ligation prior to IVF were excluded. All
grading of varicoceles was confirmed by one of two specialists in male infertility
(AT and ES). Ultrasound findings were not included. We evaluated the
effect of the presence of a varicocele on IVF outcome parameters. Outcomes
evaluated were: fertilization rate, number of embryos transferred, clinical
pregnancy rate, implantation rate, number of embryos frozen, and blast formation
rate.
RESULTS: There were a total of 194 cycles performed. Of those, 11 had a
grade 3 varicocele, 54 had a Grade 2, 40 had a grade 1 and 89 had no varicocele.
There were no statistically significant differences in seminal parameters.
Comparing those with a varicocele to those without, patient ages were
37.3 +/- 5.4 vs 37.4 +/- 6 yrs. Partner ages were 34.5 +/- 4.1 vs 34.1 +/- 4
yrs. Results are listed in the table below. There were a higher number of embryos
transferred in those with a varicocele, but less embryos frozen.
CONCLUSIONS: Clinically significant varicoceles may affect IVF cycle
outcomes. For those patients who require IVF, varicocele may alter the outcomes,
but significant differences were not seen, though larger studies need
to be conducted to evaluate for more subtle effects.
IVF Cycle outcomes by varicocele presence and grade
Fertilization
Rate (%)
Embryos
Transferred
Clinical
Pregnancy
rate (%)
Implantation
rate (%)
Number
of
Embryos
frozen
Blastocyst
Formation
rate (%)
No Varicocele
76.7 1.8 +/- 0.77 53.9 44.7 2.6 +/- 2.8 49.4
(89)
Any Varicocele
75.1 2.1 +/- 0.62 43.2 30.1 1.8 +/- 2.54 39.4
(105)
Grade 1 (40) 76.1 2.3 43.6 27.4 1.7 37.7
Grade 2 (54) 72.9 2.1 44.4 32.7 1.7 42.6
Grade 3 (11) 80.8 1.9 36.4 27.2 2.9 32.7
P value (Any
v None)
0.69 0.006 0.16 0.11 0.06 0.06
FERTILITY & STERILITY Ò
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SPERM BIOLOGY
P-554 Wednesday, October 21, 2015
WITHDRAWN
RESULTS: A total of 1202 proteins were identified in the F1 fraction while
1140, 1025 and 890 proteins were recovered from the three other fractions,
F2, F3 and F4 respectively. With respect to the differentially expressed proteins,
F1 exhibited the highest number (522), followed by F2 (362) and
lowest was observed in F3 (188) as compared to F4. Ingenuity Pathway analysis
revealed 162 pathways showing a decreasing trend from F1 through F4.
The principal pathways were spermatogenesis, protein metabolism, cell cycle,
integration of energy metabolism, regulation of apoptosis, cell redox homeostasis
and translational elongation. The proteins detected in our data set
belonging to the top 15 pathways shared many proteins that were downregulated,
suggesting failure of signaling cascade in the process of maturation in
infertile patients.
CONCLUSIONS: We conclude that a defective signaling cascade is
responsible for the faulty sperm function in infertile patients. Particularly,
a decline in mitochondrial function and oxidative phosphorylation in the
F4 fraction implies an energy deprived hypoxic state. Dysregulated protein
turnover and protein folding may lead to accumulation of defective proteins
and can potentially impair of sperm function.
P-556 Wednesday, October 21, 2015
A COMPREHENSIVE ASSESSMENT OF HISTONE MODIFICA-
TIONS IN HUMAN SPERM. S. B. Schon, a L. J. Luense, b X. Wang, b
G. Donahue, b B. A. Garcia, b M. S. Bartolomei, b S. L. Berger. b a Reproductive
Endocrinology & Infertility, University of Pennsylvania, Philadelphia,
PA; b University of Pennsylvania, Philadelphia, PA.
P-555 Wednesday, October 21, 2015
MORPHOLOGICALLY DISTINCT PHENOTYPES OF SPERMATO-
ZOA IN INFERTILE MEN REVEAL DOWN REGULATION OF
MULTIPLE SIGNALING PATHWAYS. A. Agarwal, a Z. Cui, a,b
R. Sharma, a L. Samanta, a,c R. Turki, d M. Abu-Elmagd. e a Center For Reproductive
Medicine, Cleveland Clinic, Cleveland, OH; b Institute of Toxicology,
Third Military Medical University, Chongqing, China; c Redox Biology Laboratory,
School of Life Science, Ravenshaw University, Orissa, India; d Ob/
Gyn, King AbdulAziz University, Mobil, AL; e CEGMR, Center of Excellence
in Genomic Medicine Research, Jeddah, Saudi Arabia.
OBJECTIVE: Seven morphologically distinct spermatozoal phenotypes
can be detected in human semen under electron microscopy: sperm with
dysplasia of the fibrous sheath, non-specific flagellar defects, immotile cilia
syndrome, acrosomal hypoplasia, defective chromatin condensation and
compaction, pin head and sperm without heads. These conditions cannot
be identified by routine semen analysis because they are but secondary manifestations
of underlying pathology. We hypothesize that proteomics would
enable us to better understand the mechanism(s) underlying these pathologies.
DESIGN: This proteomic study analyzed proteins in spermatozoa from
infertile men fractionated on three layers of density gradient (80, 60 and
40%). Fraction 1 (F1) refers to the least mature stage having the lowest density
whereas fraction 4 (F4) contains the most dense and morphologically
mature motile spermatozoa. Fraction 2 (F2) and fraction 3 (F3) are the intermediate
stages.
MATERIALS AND METHODS: 1-D gel electrophoresis followed by LC/
MS-MS (LTQ-Orbitrap Elite hybrid mass spectrometer) was used for protein
identification. Mascot (Matrix Science, London, UK), SEQUEST (Thermo
Fisher Scientific, San Jose, CA, USA) and X! Tandem (TheGPM, thegpm.org)
were set up to search the human reference with database assuming
trypsin as the digestion enzyme. Functional annotations of proteins were obtained
using bioinformatic tools and pathway databases.
OBJECTIVE: The unique process of spermiogenesis involves a dramatic
reorganization of paternal chromatin, whereby 90% of histones
are evicted and replaced with protamines. Proper exchange is critical
for nuclear compaction and abnormalities in this process have been associated
with male infertility [1,2]. Recent evidence suggests that the 10%
of histones retained in mature sperm are located at specific genomic loci.
Furthermore, selective histone modifications (i.e. acetylation, methylation)
are enriched at genes encoding master regulators of embryo development,
suggesting a role for paternal transmission of epigenetic information
[3,4]. To date, our understanding of histone modifications in human
sperm has largely been limited to descriptive data of specific modifications.
Our objective was to perform a comprehensive evaluation of histone
modifications in normal sperm, to quantify their abundance, and to
assess variation in modifications between individuals and in fresh vs.
frozen sperm.
DESIGN: Descriptive study of human sperm with normal semen analysis
parameters utilizing acid extraction of histones and bottom up mass spectrometry
to identify and quantitate histone modifications.
MATERIALS AND METHODS: Normal semen samples were obtained
from men undergoing routine semen analysis (n¼8). 4 samples were
equally divided with one portion cryopreserved at our Andrology Laboratory
and one half processed immediately. Following acid extraction, histones
were digested into peptides for bottom up LC-tandem mass spectrometry
by treatment with propionic acid and trypsin. The identities and
relative abundance of peptides and associated modifications were analyzed
based on retention time, MS1 and MS2 of ion peaks. Kruskal-Walis and
ANOVA tests were used as appropriate for assessment of inter-individual
variability in modification abundance and to compare fresh vs. frozen
samples.
RESULTS: We identified modifications on all four canonical histones
(H2A, H2B, H3, H4), the linker histone H1 and histone variants. We describe
a total of 102 modifications identified for the first time in human sperm. Variability
in modifications on H3 was noted between individuals, while the
abundance of modifications on H4 was consistent. No differences were
observed between fresh vs. frozen sperm.
CONCLUSIONS: Our study is the first to provide a comprehensive and
quantitative analysis of histone modifications in human sperm. We demonstrate
significant variation between individuals in specific modifications on
H3 and relative conservation in the abundance of H4. The conservation of
specific modifications further supports the hypothesis that the paternal epigenome
plays a role in early developmental programming and we further postulate
that aberrations in histone modifications may play a role in decreased
fertility. Future analysis will focus on modification profiles of abnormal
sperm.
References:
1. Carrell, D.T. and L. Liu, Altered protamine 2 expression is uncommon
in donors of known fertility, but common among men with poor fertilizing
capacity, and may reflect other abnormalities of spermiogenesis. J
Androl, 2001. 22(4): p. 604-10.
e296 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

2. Aoki, V.W., et al., Sperm protamine 1/protamine 2 ratios are related to
in vitro fertilization pregnancy rates and predictive of fertilization ability.
Fertil Steril, 2006. 86(5): p. 1408-15.
3. Hammoud, S.S., et al., Distinctive chromatin in human sperm packages
genes for embryo development. Nature, 2009. 460(7254): p. 473-8.
4. Arpanahi, A., et al., Endonuclease-sensitive regions of human spermatozoal
chromatin are highly enriched in promoter and CTCF binding
sequences. Genome Res, 2009. 19(8): p. 1338-49.
Supported by: T32HD040135, U54HD068157.
P-557 Wednesday, October 21, 2015
SIGNIFICANT CORRELATION BETWEEN SPERM DNA DAMAGE
AND ACHIEVING A PREGNANCY IN 1847 NON-IVF FERTILITY
TREATMENT CYCLES. J. Brachtchenko, S. Moskovstev,
S. Swanson, E. Shlush, L. Kan-Ool, P. Sharma, A. Y. Baratz, K. Glass,
C. L. Librach. CReATe Fertility Centre, Toronto, ON, Canada.
OBJECTIVE: Timed intercourse (TI) or intrauterine insemination (IUI),
with or without ovulation induction (OI), are often attempted before in-vitro
fertilization (IVF) when the tubes are patent. Despite much research, controversy
remains regarding the effectiveness of these treatments as well as the
effect of sperm quality on the outcomes of TI and IUI. The objective of
this study was to identify semen parameters that might predict success of
these first-line fertility treatments.
DESIGN: A retrospective chart review.
MATERIALS AND METHODS: A retrospective chart review of patients
who underwent TI or IUI with or without OI at the CReATe Fertility Centre in
Toronto, Canada, was performed. Only cases where the male partner
completed an extensive laboratory evaluation of semen (defined as standard
semen analysis, computer-aided sperm analyses (CASA; with calculation of
mucus-penetrating kinetics [MPK] and sperm DNA damage (DNA fragmentation
index [DFI]) prior to the treatment were included. The patient clinical
data and cycle information was collected for both partners and analyzed by
SSSP statistical software.
RESULTS: In total, data was collected from 362 couples who underwent
1847 treatment cycles; 354 TI (19%) and 1493 IUI (79%); while
532 (28%) were ‘natural’ cycles and the rest were underwent OI. The
likelihood of pregnancy was not statistically different between OI and
‘natural’ cycles. In total, 132 couples were able to achieve a pregnancy
(36.5%) with 96 couples having a successful birth (26.5%) and 37
(10.2%) experiencing miscarriage(s) only. 230 (63.3%) did not achieve
a pregnancy using TI or IUI. The multiple pregnancy rate was 6.3%
(6/96). The pregnancy and miscarriage rate was stable from 1 to 6 cycles
performed, however, once the number of cycles surpassed 7, the likelihood
of achieving a pregnancy was greatly diminished (P < 0.05),
with no patients who attempted 8-10 cycles achieving a pregnancy.
DFI ranged from 0.8 to 77.6% (18.3% 12) and was significantly correlated
to patient’s age, sperm concentration, motility, MPK and total
motile sperm concentration (TMSC) inseminated during IUI (p 30%
DFI) was significantly less likely to eventually achieve a pregnancy
following TI and IUI when compared to the Low (

organ development, development and function of the reproductive system
were uniquely expressed in fraction F4 of the fertile donors.
CONCLUSIONS: The study suggests that altered protein profiles in infertile
men may be responsible for their sperm dysfunction. Further work is in
progress to validate the expression profile of these proteins.
Proteomic profile of different fractions of spermatozoa from infertile men.
Proteins identified
No. of
Proteins
No. of
Proteins
No. of
Proteins
No. of
Proteins
F1 F2 F3 F4
Total 1366 1285 1325 1100
No. of DEP 119 167 373 262
Overexpressed 38 79 135 106
Underexpressed 37 46 79 82
Unique to
21 18 91 44
fertile group
Unique to
infertile group
23 24 68 30
P-560 Wednesday, October 21, 2015
PATERNALTRANSCRIPTOME ANALYSIS BY RNA-SEQUENCING
AS A MEASURE OF EMBRYONIC DEVELOPMENTAL
POTENTIAL. T. Cozzubbo, Q. V. Neri, Z. Rosenwaks,
G. D. Palermo. Reproductive Medicine, Weill Cornell Medical College,
New York, NY.
OBJECTIVE: To classify and predict the value of the contribution by the
paternal gamete to the embryonic epigenome. By identifying vital spermatozoa
RNA transcripts, we can predict their influence upon delivery to the
oocyte and how they link to embryonic development.
DESIGN: Beyond transporting the genetic material to the oocyte, spermatozoa
deliver coding and non-coding RNA that ordain early embryonic development.
We attempted to profile the paternal transcriptome and categorize
key regulating factors involved in syngamy by controlling vital checkpoints
during embryogenesis. Expression of specific genes were related to fertile
and infertile individuals.
MATERIALS AND METHODS: Men undergoing infertility treatment
were screened and semen parameters were assessed according to WHO
2010. RNA was isolated from 25x106 human spermatozoa using a spin column
commercial kit. The nucleic acid quality, RNA integrity number (RIN),
and spermatozoal RNA concentration were assessed. The RNA samples were
then made into paired-end libraries. Pilot paired-end 36bp RNA-Sequencing
(RNA-Seq) using an Illumina platform (NextSeq 500) was carried out and
then expanded to 60M reads.
RESULTS: In 19 consenting infertile men with a mean age of
39.65.1yrs, an average sperm concentration of 47.916.0x106/mL,
motility of 47.15.5%, and normal morphology of 2.91.6%. The RNA
isolated from the samples yielded an average concentration of 14.055.9
ng/mL and a RIN of 5.91.7. BOK, a regulating factor of apoptosis during
the cell cycle, had an excess of transcripts in the sperm of infertile patients,
corresponding to a higher (14.3%) embryonic fragmentation rate. RANBP2
contributes to the reorganization of the paternal genome post-fertilization
but was found to be comparably expressed in both fertile and infertile
men; coinciding with acceptable embryo cleavage (85.7%) in the infertile
cohort. PLK4 and BUB1, which dictate chromosome stability & mitotic
segregation by regulating centriole development, displayed a decreased
expression in the infertile group and indeed this group yielded 66.7%
genetically abnormal embryos as determined by preimplatation genetic
diagnosis.
CONCLUSIONS: Sperm RNA-Seq is an ideal technique capable of reaching
beyond microscopic evaluation, morphometric assessment, and traditional
diagnostic assays to help predict the contribution of the male gamete
to early pre-implantation development. This novel technique, by assessing
coding and non-coding RNA, may further classify the transcripts that play
an influential role in pre and post fertilization and informs on embryo developmental
competence of a specific paternal epigenome.
Supported by: WCMC.
OOCYTE MATURATION
P-561 Wednesday, October 21, 2015
THE FGF 23 LEVELS IN HUMAN FOLLICULAR FLUIDS IN THE
FOLLICULAR PHASE OF PREIMPLANTATION ARE ASSOCI-
ATED WITH THE AGING OR QUALITY OF HUMAN
OOCYTES. F. Saji, S. Taguchi, M. Funabiki, N. Amano, T. Takano,
L. K. Young, T. Hayashi, Y. Tada, Y. Iwaki, M. Karita, Y. Nakamura.
Oak Clinic, Osaka, Japan.
OBJECTIVE: Though approaches to evaluate the aging of human oocytes
have been investigated, a simple biomarker to evaluate the aging or quality of
human oocytes is required. Therefore, the present study aimed to find such a
biomarker.
DESIGN: An experimental prospective cohort study.
MATERIALS AND METHODS: The associations between the fibroblast
growth factor (FGF) 23 levels in human follicular fluids in the follicular
phase of preimplantation and outcome data were investigated from June
2013 to January 2015 in 60 patients (median age 37.3 years) who provided
informed consent at an IVF clinic. The primary outcomes in the present study
were the maturation rates in human oocytes and fertilization rates. The FGF
23 levels in the human follicular fluids obtained during the follicular phase of
preimplantation were evaluated using an FGF-23 ELISA Kit. Statistical analyses
were investigated using ANOVA, Welch’s t test and multivariate analysis.
RESULTS: The FGF 23 levels in human follicular fluids in the follicular
phase of preimplantation were 20.78 6.84 pg/ml (mean SD). Furthermore,
the FGF 23 levels in human follicular fluids in the follicular phase
of preimplantation were elevated by aging (p

(54.2% and 79.2%) in Type C and 223 (71.7% and 75.0%), respectively. The
maturation rate of immature oocytes was significantly different with Type D
and Type A (71.7% vs. 51.0%, P

14.06, respectively. There was a significant inverse relationship between
increasing BMI and lower serum hCG levels in the SC group. However, there
was no relationship between BMI and percent mature oocytes. Because of the
difference in luteal support between the two groups, strict comparisons of
pregnancy rates were not possible, however there were no significant difference
in pregnancy rates.
Subcutaneous
hCG (n¼215)
Intramuscular
hCG (n¼159)
p-value
Age 34.87 35.78 0.08
BMI 25.10 25.91 0.17
AMH 2.61 2.76 0.70
Total oocytes retrieved 15.21 14.06 0.19
Number mature 11.04 10.28 0.26
Percent mature 76.28% 74.37% 0.57
CONCLUSIONS: SC-hCG is as effective as IM-hCG for final follicular
maturation in IVF cycles. There was a significant inverse relationship between
increasing BMI and lower serum hCG levels in the SC-hCG group.
However, there was no relationship between BMI and percent mature oocytes.
P-566 Wednesday, October 21, 2015
THE CORRELATION OF ANTI-MULLERIAN HORMONE CON-
CENTRATIONS TO OOCYTE MATURITY AT
RETRIEVAL. M. Bustillo, a C. Alford, a I. Collazo, a K. O. Pomeroy. b,a
a South Florida Institute for Reproductive Medicine, Miami, FL; b The World
Egg Bank, Phoenix, AZ.
OBJECTIVE: Determine if Anti-Mullerian hormone levels correlate to the
number of mature ooctyes or the percent of mature oocytes at the time of
oocyte retrieval.
DESIGN: Retrospective analysis of 2459 cycles of IVF from 2010 to 2014.
MATERIALS AND METHODS: Spearman’s rank correlation was done
on all IVF cycles where the AMH value was equal to or less than 11.10
ng/ml to determine correlation between total number of mature ova and
percent of mature ova. Cycles where the number of collected mature ova
were less than 6 were classified as poor cycles and an ROC curve was generated
to identify the cutoff value of AMH for ‘‘poor cycles.’’
RESULTS: There was a moderate correlation of AMH value to the total
number of mature oocytes retrieved (Spearman’s rho ¼ 0.578; p

emain unclear (1). Studies exploring this topic, however, do not address
more clinically significant outcomes such as clinical pregnancy rate (CPR)
or live birth rate (LBR). Therefore, the objective of this study was to determine
whether patients with PCOS have better pregnancy outcomes following
ICSI versus IVF.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Data from the 2011-2012 SART registry
were analyzed. First fresh autologous cycles in women under 40 with
the diagnosis of PCOS were included in the study. All other diagnostic
indications for IVF were excluded from this study including male factor.
Groups were further stratified based on the insemination technique: conventional
IVF or ICSI. Cycles using split IVF/ICSI were excluded. Cycles
were divided into two groups: cleavage stage embryo transfer (ET) and
blastocyst stage ET. Main outcomes measured were the number of two
pronuclear (2PN) embryos, LBR, and miscarriage rate. Secondary outcomes
included implantation rate (IR), CPR and the percentage of cycles
that did not result in 2PN embryos. Data were analyzed using two-sided
Welch’s t-test.
RESULTS: 2217 cycles reported use of conventional IVF and 2462 cycles
reported use of ICSI. In the IVF group, the number of 2 PN oocytes, CPR, and
LBR were statistically higher in both cleavage stage and blastocyst stage
transfer cycles compared to the ICSI group. Miscarriage rate was lower in patients
undergoing ICSI with blastocyst ET but higher in those with cleavage
ET compared to the IVF group. The percentage of cycles with no 2PN oocytes
was statistically lower in the ICSI group with blastocyst ET only (Table).
Although statistical significance was found between these two groups,
the differences were not clinically significant.
CONCLUSIONS: Contrary to previous studies, ICSI does not offer an
advantage over conventional IVF in terms of number of oocytes fertilized,
CPR, or LBR.
MATERIALS AND METHODS: Using an established and validated IVF
database from a single large urban IVF center in an insurance mandated state,
we investigated failed fertilization cases with more than 5 eggs and where

MATERIALS AND METHODS: Sperm samples were produced by
masturbation following 2-5 days abstinence and subjected to computer-assisted
sperm analysis (CASA), Kremer penetration testing, calcium response
recording (FLUOstar), PLCz immunocytochemistry (anti-human PLCz;
Cova-Lab UK), and electrophysiology studies using single-cell patch clamping(4).
DNA was extracted from whole blood samples and subjected to
exome sequencing.
RESULTS: 20% (5/25) had significant sperm pathologies detected,
including abnormal calcium response, reduced Kremer penetration and putative
CatSper / K+ channel abnormalities, most of which can be overcome by
ICSI. Significantly, over half of the men investigated (13/25) were found to
have reduced proportion and / or total relative fluorescence of PLCz.
Following appropriate counselling, 6 of these couples have subsequently undergone
ICSI and AOA using calcium ionophore A23187 (GM508 Cult Activ;
Gynemed) Average M2 oocyte fertilisation rate was 61.2% (54.5% -
70.8%), with 3 biochemical pregnancies and an ongoing intrauterine pregnancy
to date, and further treatment results awaited. Exome sequence analysis
is currently in progress.
CONCLUSIONS: This study supports our hypothesis that sperm dysfunction
commonly underlies unexplained infertility, and unexplained TFF,
which may not be predicted by standard diagnostic semen analysis. Utilising
a variety of laboratory approaches to investigate sperm calcium response,
motility and function can enable application of an individualised approach
to a future cycle of ART, including AOA where PLCz deficiencies are identified.
References:
1. Factors associated with failed treatment: an analysis of 121,744 women
embarking on their first IVF cycles. Bhattacharya S, Maheshwari A,
Mollison J. PLoS One. 2013 Dec 5;8(12):e82249.
2. The cytosolic sperm factor that triggers Ca2+ oscillations and egg activation
in mammals is a novel phospholipase C: PLCzeta. Swann K,
Larman MG, Saunders CM, Lai FA. Reproduction. 2004
Apr;127(4):431-9. Review.
3. Assisted oocyte activation following ICSI fertilization failure. Vanden
Meerschaut F, Nikiforaki D, Heindryckx B, De Sutter P. Reprod Biomed
Online. 2014 May;28(5):560-71. Review.
4. Patch clamp studies of human sperm under physiological ionic conditions
reveal three functionally and pharmacologically distinct cation
channels. Mansell SA, Publicover SJ, Barratt CL, Wilson SM. Mol
Hum Reprod. 2014 May;20(5):392-408.
Supported by: TENOVUS Scotland MRC Chief Scientist Office NHS Tayside
Royal Society UK Oxford University Medical Research Fund.
P-571 Wednesday, October 21, 2015
ARTIFICIAL OOCYTE ACTIVATION WITH CALCIUM IONO-
PHORE (A+23187) FOLLOWING ICSI FERTILIZATION
FAILURE. A. Sdrigotti, a G. J. Rey Valzacchi, b F. A. Leocata Nieto, c
V. E. Canada. d a Reproductive Medicine, Procrearte, Buenos Aires,
Argentina;
b Director, Buenos Aires, Argentina;
c Embriologyst, Buenos
Aires, Argentina; d Procrearte, Capital Federal, Argentina.
OBJECTIVE: To assess the CALCIUM IONOPHORE (A+23187) use in
patients with total failure or low fertilization rate on previous ICSI cycle/s.
DESIGN: Retrospective, case- control study.
MATERIALS AND METHODS: Patients with a history of one or more
ICSI cycle/s with low (

Program through the National Research Foundation of Korea funded by the
Ministry of Education, Science and Technology and a grant (A120080) from
the Korean Healthcare Technology R&D project, Ministry for Health, Welfare
and Family Affairs, Republic of Korea.
P-573 Wednesday, October 21, 2015
DELAYED PRONUCLEAR APPEARANCE AS AN INDICATION OF
COMPROMISED OOCYTE REPAIR CAPACITY. T. Cozzubbo,
Q. V. Neri, T. Paniza, L. Reisman, Z. Rosenwaks, G. D. Palermo. Reproductive
Medicine, Weill Cornell Medical College, New York, NY.
OBJECTIVE: To associate the delayed formation of two-pronuclei (2PN)
after ICSI with the integrity of the oocytes capability to cope and eventually
overcome varying levels of genomic error.
DESIGN: In a retrospective study, we investigated the impact of the degree
of sperm DNA fragmentation in relation to oocyte aging. We gauged the efficiency
of oocyte repair mechanisms in occasion of delayed pronuclear
appearance and consequent syngamy was assessed.
MATERIALS AND METHODS: We identified couples undergoing ICSI
treatment (n¼152) that had sperm chromatin assessment by TUNEL on their
ejaculates. TUNEL was performed by adjusting sperm concentration to 5
million, fixation with 4% PFA, exposure to mild permeabilizing solution,
and incubated at 37 C with TUNEL reaction mixture. At least 500 spermatozoa
were scored under fluorescent microscopy. Patients were categorized
by DFI level and grouped according to maternal age. Each subdivision of
DFI was evaluated in relation to clinical pregnancy outcome (presence of a
fetal heartbeat). Standard fertilization is performed 16-18 hours after ICSI
and delayed fertilization was conducted over 18 hours. To determine the significance
of a delayed normal fertilization, arguably linked to an extensive
processing and repair of the male genome following sperm nuclear unraveling,
we assessed cohorts of oocytes that had at least one oocyte with a delayed
appearance of the pronuclei in relation to a matched control.
RESULTS: In a previous study, we established in 315 ICSI cycles which
included 152 women with a mean age of 37.34yrs. In a cohort of women

RESULTS: Following AOA, fertilization rate was remarkably improved
from 21.4% to 67.8%, and was comparable to that of non-AOA (65.5%) After
transferring 32 AOA embryos, 6 (18.7%) implanted (IMP). Among AOA embryos
considered morphologically suitable for transfer, IMP embryos
showed earlier PN fading and first cell cleavage than non-IMP (P < 0.05).
Interestingly, these significant differences disappeared after 2-cell stage.
When AOA IMP embryos were compared to non-AOA IMP embryos (n ¼
34), no difference was observed in any of morphokinetic events.
CONCLUSIONS: Zygotes fertilized with the aid of activating stimuli
showed morphokinetics in early development similar to those of conventional
ICSI zygotes. After AOA, differences between embryos with or
without the ability of implantation were identified only at PN fading and
the first cleavage, but not at later developmental stages. This phenomenon indicates
that the effect of AOA on morphokinetics is transitory.
P-576 Wednesday, October 21, 2015
CORRELATION OF THE HYALURONAN BINDING ASSAY TO
FERTILIZATION IN VITRO. K. O. Pomeroy, a,b G. De Meglio, a
J. Garcia, c I. Collazo. d a South Florida Institute for Reproductive Medicine,
Miami, FL; b The World Egg Bank, Phoenix, AZ; c Andrology, South Florida
Institute for Reproductive Medicine, Miami, FL; d Embryology, South Florida
Institute for Reproductive Medicine, Miami, FL.
OBJECTIVE: Determine if the hyaluronan binding assay (HBA) can be
used to identify patients with poor fertilization in In Vitro Fertilization.
DESIGN: Retrospective analysis of 905 IVF cases from 2012 to 2014.
MATERIALS AND METHODS: The same sperm used for IVF was
analyzed for hyaluronan binding using the manufacturer’s instructions.
Less than 80% binding was used as the cut-off as recommended by the manufacturer.
100 sperm were counted (or 100 squares when there were few
sperm) and percent binding was determined.
RESULTS: Abnormal HBA results were found in 36.5% of patients. Fertilization
rates for sperm that had low binding (¼80%; n¼212) and poor fertilization (

their own controls. Primary outcomes included fertilization rates, number of
embryos that reached an appropriate stage (2Bc or greater) for trophoectoderm
biopsy (TEBx) and number of euploid embryos. T-tests of means and
paired t-tests were used for statistical analysis.
RESULTS: Average age of the patients in this study was 35.6 years +/- 5.2.
Average day 2 estradiol (pg/ml) and follicle stimulating hormone (mIU/ml)
values were 55.1 +/-56.5 and 6.75 +/- 2.87. Average total gonadotropin
dosage (IU) and estradiol at trigger were 3648.6 +/- 144.6 and 2789 +/-
1418.3, respectively. Approximately 19.8 +/- 9.3 oocytes on average were
retrieved with a maturation percentage of 81%. Insemination fertilization
rates were 69% +/- 23.8 and 75% +/- 13.8 (p-value of 0.28) for the ICSI
cohort. The percentage of embryos available for TEBx for the insemination
cohort was 32% and 38% for the ICSI cohort, with a p-value of 0.15 and RR
0.84 (0.652-1.07). 52.8% of embryos biopsied resulting from ICSI and 47%
of embryos from standard insemination were euploid, and paired t-test analysis
showed no significance between the two groups, p ¼ 0.07.
CONCLUSIONS: Standard insemination techniques and ICSI showed no
difference in fertilization rates, and no difference in percent of chromosomally
competent embryos. In the absence of standard indications for ICSI,
there does not appear to be any benefit or harm in the use of ICSI for
increased development of chromosomally competent embryos.
P-579 Wednesday, October 21, 2015
MULTIPLE PRONUCLEAR ANALYSIS BY IMMUNOFLUORES-
CENCE STAINING IN HUMAN EMBRYOS DERIVED FROM
PATIENTS WITH POSITIVE ANTI-CENTROMERE ANTI-
BODIES. M. Tokoro, H. Ohno, N. Aoyagi, M. Sonohara, M. Tsuiki,
K. Ishihara, Y. Funagayama, Y. Kida, N. Fukunaga, Y. Asada. Asada Ladies
Clinic Medical Corporation, Nagoya, Japan.
OBJECTIVE: Anti-centromere antibody (ACA) is an anti-nuclear antibody
(ANA), and specifically recognizes the centromere. Recently, several
studies have reported that ACA have effects on oocyte maturation and embryo
cleavage. Moreover, the rate of multiple pronuclear (MPN) formation
is higher in patients with ACA. However, the reason for the high rate of
MPN formation in ACA patients is not clear. In this study, we examined
the cause of MPN formation in embryos derived from ACA patients, by performing
immunofluorescence staining of MPN embryos and immature oocytes
of ACA patients.
DESIGN: Retrospective, nested case-control study.
MATERIALS AND METHODS: A total of 1290 patients from our clinic
were tested for ANA before oocyte retrieval from August 2014 to February
2015. After ANA testing, patients were classified according to 3 groups;
ACA patients (with only ACA), non-ACA patients (with ANA not included
ACA) and non-ANA patients (without ACA and any other ANA). The rate of
MPN formation after ICSI was compared in the 3 groups. MPN embryos with
over 3 pronuclei (PN) and immature MI oocytes were used for research after
informed consent was obtained from ACA patients. The embryos and oocytes
were stained with H3K9me2 antibody to detect the female chromosome.
Ethics Committee approval was obtained for the study.
RESULTS: Among tested patients, 366 patients were positive for ANA,
and 12 of 366 (3.3%) patients were positive for ACA. The average MPN formation
rate per oocyte retrieval was 51.3% in ACA patients, 3.3% in non-
ACA patients and 4.2% in non-ANA patients, significantly higher in ACA
patients vs other groups (P0.05). Additionally,
44.4% SNT blastocysts showed euploid chromosomes following aCGH
test compared to 53.5% euploidy rate in the non-SNT control group
(p>0.05).
CONCLUSIONS: Using the current nuclear transfer-membrane fusion
system, SNT seems to interfere with normal fertilization of human oocyte,
however, it did not impose adverse impact to the pre-implantation embryonic
development and its chromosomal complements.
P-581 Wednesday, October 21, 2015
DOES LASER ASSISTED HATCHING ON THAWED EMBRYO AF-
TER VITRIFICATION IMPROVE IMPLANTATION
RATE? S. A. Hebisha, a A. I. Ahmed, b H. N. Sallam, c M. S. Omran. d
a Lecturer of Gynecology, Alexandria University - Faculty of Medicine, Alexandria,
Egypt; b Obstetrics and gynecology MFM division, Perinatology
research/NICHD/NIH/DHHS, Detroit, MI; c Gynecology, Alexandria University
- Faculty of Medicine, Alexandria, Egypt; d Alexandria University -
Faculty of Medicine, Alexandria, Egypt.
OBJECTIVE: To evaluate the impact of laser assisted hatching on the implantation
rate of vitrified-thawed embryos transferred after IVF/ICSI
DESIGN: Prospective controlled study.
FERTILITY & STERILITY Ò
e305

MATERIALS AND METHODS: One hundred and eighty IVF/ICSI patients
undergoing frozen (vitrified) thawed embryo transfer. All embryos
were frozen on day three by vitrification using the open technique. Patients
were divided into two groups: embryos from group A were subjected to laser
assisted hatching of half thickness of one quarter of the zona two hours after
thawing and two hours before transfer, while embryos from group B patients
were transferred two hours after thawing without assisted hatching.
RESULTS: Implantation rate was significantly higher in group A
compared to group B ( 47.2 % vs 23.4 % respectively; P¼ 0.0041). Pregnancy
rate was also significantly higher in group A compared to group B (61.1 % vs
41.1 % respectively; P ¼ 0.031).
CONCLUSIONS: Laser assisted hatching of half thickness of a quarter of
the zona pellucida increases the implantation and pregnancy rates after the
transfer of thawed embryos vitrified using the open technique.
Effect of LAH on implantation rate.
Group A Group B P value
Total number of cases 90 90
Number of embryo transferred 288 252
Implanted embryos 136/288 59/252
Implantation rate 47.22% 23.41% 0.0041*
Pregnancies 55 37
Pregnancy rate 61.1% 41.1% 0.031*
* Statistically significant.
P-582 Wednesday, October 21, 2015
SPERM DNA DAMAGE IMPAIRS EMBRYO DEVELOPMENTAL
POTENTIAL AND INDUCES EPIGENETIC CHANGES IN THE RE-
SULTING MOUSE OFFSPRING CONCEIVED THROUGH
ICSI. Y. Li, H. Wang, S. Zhang, X. Huang. Reproductive Medical Center,
The First Affiliated Hospital of Wenzhou Medical University, Wenzhou,
China.
OBJECTIVE: To evaluate the effects of sperm DNA damage on fertilization
and embryonic development, and the question whether sperm DNA damage
could introduce epigenetic changes was also investigated.
DESIGN: Sperm DNA damage was triggered by freezing-thawing process,
fertilization were performed by intracytoplasmic sperm injection
(ICSI), in vitro developmental progression was observed and recorded.
Meanwhile, methylation status of differentially methylated region (DMRs)
for selected imprinted genes, a paternally imprinted gene H19 and a maternally
imprinted gene SNRPN, was analyzed in midgestation mouse fetuses.
MATERIALS AND METHODS: B6D2F1 (C57BL/6DBA/2) strain
mice were used as oocytes and semen donors, sperm was freeze-thawed
for ten times, then DNA damage extent was evaluated by sperm chromatin
dispersion (SCD) assay. Oocytes (N¼524) were fertilized and in vitro developmental
progression till to blastocyst formation was observed at different
time stage. Afrer embryo transfer, midgestation mouse fetuses were
collected, followed by DNA extraction, bisulfite-converted, and then amplified,
cloned, sequenced and finally the methylation status of DMRs of H19
and SNRPN was established by DNA methylation analysis.
RESULTS: (1) The treated spermatozoa retained their fertilization potential
and no statistically decrease existed in pronucleus formation (treated
89.1% vs. control 91.5%, P > 0.05), however, sperm DNA damage effectively
blocked fertilized oocytes from reaching 2-cell embryo stage and
decreased the formation rate of morula and blastocyst in vitro (treated
56.5% vs. control 85.3%, P < 0.05).(2) The DMRs for these two imprinted
genes displayed an abnormal methylation status in the fetuses derived
from DNA-damaged spermatozoa. For H19 DMR, there was a substantial increase
in methylation level compared with control (treated 69.9% vs.control
52.5%, P ¼0.0375), showing aberrant hypermethylation of the H19 DMR.
Oppositely, SNRPN DMR showed a considerably reduction of methylation
level, also reaching statistically significance (treated 31.5% vs.control
53.0%, P ¼ 0.0451).
CONCLUSIONS: Sperm DNA damage impairs in vitro embryo developmental
potential, and it could induce epigenetic changes in the resulting F1
mouse offspring, reinforcing the observation that the aberrant epigenetic
modification is a contributory factor in the roles of sperm DNA damage
played, which may associate the declined embryonic development.
Supported by: Department of Health of Zhejiang Province(2014KYB340).
P-583 Wednesday, October 21, 2015
GRANULOCYTE COLONY STIMULATING FACTOR (G-CSF)
LEVEL IN FOLLICULAR FLUID IS A PROGNOSTIC FACTOR
FOR EMBRYO DEVELOPMENTAL POTENTIAL IN IN-VITRO
FERTILIZATION CYCLES. N. M. Chimote, a N. M. Nath, b
B. N. Chimote. b a Embryology/Endocrinology, Vaunshdhara Clinic and Assisted
Conception Centre, Nagpur, India; b Vaunshdhara Clinic and Assisted
Conception Centre, Nagpur, India.
OBJECTIVE: Local production of G-CSF in the human ovary, its expression
in follicular luteal granulosa cells (GCs) as well as presence of higher
concentration of GCSF in follicular fluid (FF) than in serum implies a potential
autocrine/paracrine role of G-CSF within the follicular microenvironment.
Therefore the objective of this study was to evaluate if FF-GCSF
levels have a predictive value on the potential of embryo to develop to blastocyst
stage.
DESIGN: Prospective study of fresh non-donor, no male factor conventional
IVF cycles (n¼44, mean age¼ 30.32 2.70 years) carried out from
July 2014 to December 2014. Standard controlled ovarian stimulation with
r-FSH and antagonist protocol was followed. Fertilization, cleavage and blastocyst
formation rates were assessed. All cycles involved day5 blastocyst
transfer.
MATERIALS AND METHODS: For each patient FF-GCSF levels were
estimated in fluid pooled only from lead follicles (R 18 mm in diameter)
from which oocytes had been retrieved. Levels were reported as a measure
of total protein content of FF. Cycles were divided into Low FF-GCSF group
(n¼22, % 10.33 pg/mg protein) and High FF-GCSF group (n¼22, >10.33
pg/mg protein) depending on the 50th centile value.
RESULTS: Fertilization rate (71.33 0.03 vs. 50.91 0.03%, p lt
0.0001), Cleavage rate (68.53 0.03 vs. 49.09 0.02%, p lt 0.0001) and
blastocyst formation rate (47.55 0.03 vs. 26.13 0.03%, p lt 0.0001)
were significantly higher in High FF-GCSF group compared to Low FF-
GCSF group despite non-significant differences in the total number of eggs
retrieved (286 vs. 222, p¼0.13) in the two groups. Pearson r values for correlation
of FF-GCSF with fertilization, cleavage and blastocyst formation
rates were 0.44, 0.40 and 0.36 respectively. An ROC cutoff of gt12.05 ng/
mg protein increased the likelihood of embryos with good developmental potential
(AUCROC¼ 92.98%, Sensitivity¼ 90.91%, Specificity¼ 72.73%).
Although clinical pregnancy rates were higher in high compared to low
FF-GCSF group, the difference was not significant. However, since live birth
data has yet to be calculated for all patients involved in this study, it would be
interesting to observe the differences in live birth rate.
CONCLUSIONS: Endogenously produced Granulocyte Colony Stimulating
Factor (GCSF) level in pooled follicular fluid (FF) strongly correlates
with fertilization, cleavage and blastocyst formation rate and may be a predictive
marker of embryo developmental potential in IVF cycles.
P-584 Wednesday, October 21, 2015
DOES THE TIMING OF BLASTULATION PROGNOSTICATE RISK
OF ANEUPLOIDY? C. R. Juneau, M. D. Werner, J. M. Franasiak,
E. J. Forman, K. H. Hong, T. Molinaro, R. T. Scott. RMA, NJ, NJ.
OBJECTIVE: It has been established the rate of aneuploidy is higher in
embryos which blastulate more slowly (day 6) relative to those which blastulate
within the normal time range (day 5). Interpreting these data is complicated
by the fact that later blastulation is associated with increasing maternal
age and declining ovarian reserve. Thus, it is unknown if, within a single
cohort of embryos from a single treatment cycle, those embryos which blastulate
on day 6 have an increased risk of being aneuploid. This study seeks to
address that question.
DESIGN: Retrospective cohort.
MATERIALS AND METHODS: Patients in their first IVF cycle at a single
center from 2010-2014 using comprehensive chromosome screening
(CCS) were reviewed. Those patients who had a portion of their cohort biopsied
on day 5 and the remaining biopsied on day 6 were included. Blastocyst
assessment was made in the mid-afternoon of day 5 and subsequently on the
morning of day 6, 15 hours later. Blastocysts were deemed appropriate for
biopsy if there was evidence of cells herniating through an artificial opening
in the zona made on day 3 of development. The rate of aneuploidy between
day 5 and day 6 blastocysts was compared using a paired Wilcoxon signed
rank test for non-parametric data. Data were stratified by the Society of Assisted
Reproductive Technologies (SART) age groups, and the aneuploidy
rates between day 5 and day 6 blastocysts were compared using a c2 test
of proportions. Logistic regression was employed to control for confounders,
e306 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

and a generalized estimating equation (GEE) model was created to control
for patient specific differences.
RESULTS: 2143 blastocysts from 274 patients met inclusion criteria. The
mean patient age was 34.3 3.8 years. 1563 blastocysts were biopsied on
day 5 and 580 blastocysts were biopsied on day 6. In the entire population,
when controlling for basal antral follicle count, follicle stimulating hormone,
age, and IVF stimulation type using a GEE analysis, the odds of aneuploidy
were increased in day 6 blastocysts (OR 1.38, 95% CI 1.01-1.89). However,
in the paired analysis, the rate of aneuploidy per patient on day 6 was similar
to that on day 5 (35.5% versus 29.9%, p¼0.2).
CONCLUSIONS: Given the observed difference in the rate of aneuploidy
between day 5 and day 6 is negated when the patient serves as their own control
in this paired analysis, it is likely the risk of aneuploidy is intrinsic to the
patient and not related to the time of biopsy. Thus, embryos that blastulate
later maintain a potential for euploidy and should be screened.
P-585 Wednesday, October 21, 2015
ANEUPLOIDY RATES IN EMBRYOS PRODUCED BY FERTILE
COUPLES. D. Wells, a K. Ravichandran, b M. Alper, c J. Jain, d
A. Penzias, e C. A. Benadiva, f P. Colls, b M. Konstantinidis, b S. Munne. b
a Reprogenetics, Oxford, United Kingdom; b Reprogenetics, Livingston, NJ;
c Boston IVF, Waltham, MA; d Santa Monica Fertility, Santa Monica, CA;
e Boston IVF / Harvard Medical School, Waltham, MA; f University of Connecticut,
Farmington, CT.
OBJECTIVE: To determine whether the high aneuploidy rates routinely
seen in embryos of patients undergoing infertility treatments are also typical
of couples without fertility problems.
DESIGN: Examination of aneuploidy rates observed during preimplantation
genetic diagnosis (PGD) and preimplantation genetic screening (PGS)
cycles.
MATERIALS AND METHODS: 96 fertile couples underwent PGD for a
single gene disorder. Additionally, PGS was employed in order to enhance
the likelihood of transferring a chromosomally normal embryo. A total of
597 blastocyst stage embryos, derived from these couples, were biopsied
and subjected to gene testing and also comprehensive chromosome screening
using microarray-CGH (aCGH). During the same period of time 31,137 embryos
were produced by couples undergoing assisted reproductive treatments
(ARTs). Once again, embryos were subjected to aCGH at the blastocyst stage
to assist in the selection of viable embryos for transfer. Information for the
study was extracted from the eIVF database (Dallas, TX).
RESULTS: The aneuploidy rate for blastocysts from patients of proven
fertility, undergoing ART in order to generate embryos for PGD of a gene disorder,
was found to be 49% (mean maternal age 33.5). This compares to an
aneuploidy rate of 47% for embryos from infertile/subfertile patients undergoing
PGS (mean age 34.7). These aneuploidy rates are not significantly
different (P¼0.30, Chi square with Yates correction).
CONCLUSIONS: It has long been known that aneuploidy is extremely
common in human preimplantation embryos generated during ARTs. However,
it is unknown whether such abnormalities represent part of the infertile
pathology of the parents. This study assessed a large number of embryos using
a well-validated chromosome screening method, providing robust data on
aneuploidy rates. No difference in the incidence of aneuploidy was observed
in embryos from typical infertile/subfertile patients compared to those from
patients who had previously conceived naturally. Thus, it can be concluded
that in the great majority of cases the high frequency of aneuploidy in embryos
produced using ART is not related to the patient’s underlying infertility.
As all patients in the current underwent ART, we were unable to address the
question of whether the methods used for infertility treatment influence the
incidence of aneuploidy.
P-586 Wednesday, October 21, 2015
A MITOCHONDRIAL D LOOP VARIANT ASSOCIATED WITH
REDUCED RISK OF EMBRYONIC ANEUPLOIDY. M. Olcha,
X. Tao, Y. Wang, T. Xing, Y. Zhan, J. M. Franasiak, R. T. Scott,
N. R. Treff. RMA, NJ, NJ.
OBJECTIVE: Mitochondria are integral to oocyte and embryo energy production,
spindle formation, and chromosomal segregation. Single nucleotide
variants within the mitochondrial genome have been associated with susceptibility
to many disease processes, including autoimmune and inflammatory
processes. These variants could influence functions essential to early embryo
development. The aim of this study was to assess if any association existed
between rates of embryonic aneuploidy and sequence variants in the mitochondrial
genome.
DESIGN: Retrospective genetic association study.
MATERIALS AND METHODS: All patients underwent IVF at a single
center with comprehensive chromosomal screening. 469 patients were identified
as either being young with atypically high embryonic aneuploidy rates
or older with low aneuploidy rates. Patient DNAwas obtained from an on-site
repository and used for analysis. Mitochondrial DNA was amplified with Takara
LA PCR KitsÔ and mitochondria specific primers. Whole genome
sequencing was accomplished using an Ion PGMÔ system utilizing Ion
318Ô Sequencing Chips in order to achieve 20x depth reads. Individual variants
were identified by comparing sequence reads to the Cambridge reference
sequence. Patients were also assigned a haplogroup using the
HaploFind software (https://haplofind.unibo.it). Individual variants and
mitochondrial haplogroup frequencies were calculated and compared between
the two study populations using logistical regression and chi squared
contingency tables.
RESULTS: Sequencing revealed 1,475 SNPs amongst 469 tested patients.
A variant at position 16,390 (G>A) was significantly more common in patients
with low aneuploidy rates (p.0.0008), which survived multiple test
correction. Additionally, 443 individuals were assigned into 1 of 9 known European
haplogroups (H, I, J, K, T, U, V, W, X) and analyzed in this study. The
remaining patients were grouped as other. Haplogroup H contained the
largest proportion of patients (41.4%). No association was found between
aneuploidy rate and haplogroup inheritance.
CONCLUSIONS: Although prior studies suggested an association between
haplogroup variants and the incidence of oocyte chromosomal errors
the present study found no association with a significantly larger sample size.
However, with the availability of sequence data for the entire mitochondrial
genome a significantly associated single nucleotide variant was identified in
patients with lower risk of embryonic aneuploidy. The variant is located
within the D-loop, a non-coding region of the mitochondrial genome
involved with replication and transcription. It is possible that this variant confers
a reproductive advantage given its distinct location. Additional studies
need to be performed in order to further investigate the role of this variant
in embryo development.
P-587 Wednesday, October 21, 2015
TOO SLOW? WE DON’T THINK SO. OUTCOMES RELATIVE TO
EMBRYO DEVELOPMENT IN A BUSY OOCYTE CRYOPRESER-
VATION (OC) PROGRAM. S. Druckenmiller, F. Licciardi, P. Labella,
M. Clarke-Williams, N. Seta, D. H. McCulloh, N. Noyes. NYU Medical
Center, New York, NY.
OBJECTIVE: OC is now a mainstream ART, frequently used to bank
donor oocytes or defer fertility. To date, >2,000 babies have been born as
a result of OC. Thus, we analyzed the specifics of post-fertilization (fert) embryo
development in order to glean outcome differences when using autologous
(Aut; older) vs. donor (OD; younger) frozen-thawed oocytes. We also
compared OC blastocyst (BL) formation rates (BFR) to those of fresh-oocyte
cycles.
DESIGN: Retrospective and case control.
MATERIALS AND METHODS: For OC, 154 Aut (139 pts) & 53 OD (no
commercial-bank oocytes) thaw cycles completed from 2004-2015 were reviewed.
Cycle data was mined for: oocyte maturity, survival, 2-pronuclear
(2PN) fert, development, embryo implantation (EI) & pregnancy. When assessing
development, the highest stage & grade achieved before embryo
transfer (ET) was used. BL were graded using Gardner’s criteria. When evaluating
EI & pregnancy rates relative to embryo stage, only single ETs &
multi-embryo ETs with same-stage embryos were considered. BFR in 286
age-matched fresh Aut & 186 fresh OD cycles from 2011-2014 were
computed & compared to OC thaw outcomes. Fisher’s exact was used for
stats.
RESULTS: For OC, 1,098 (7/cycle) Aut (mean age 383y; range 25-44y)
& 411 (8/cycle) OD (mean age 275y; range 22-33y) oocytes were thawed,
survived & achieved 2PN fert. The Table shows post-fert development. OC
BFRs were not different when Aut (n¼428 ; 39%) & OD (n¼173; 42%) rates
were compared; however, within the BL-Stage groups, more BL expanded to
Stage 3 & 4 in OD than Aut cycles (p< .02). Conversely, a greater % of BL
only reached Stage 2 in Aut vs. OD thaws (p< .01). When comparing EI &
pregnancy rates between Aut & OD relative to the day (D) of BL ET (i.e.
D-5 vs. D-6/7), no statistical difference between ETs of similarly-staged
BL formed on D-5 (24/53 {45%} vs. 16/28 {57%};p¼0.4) or D-6/7 (1/3
{33%} vs. 1/1 {100%};p¼1) was noted; however, within the Aut & OD
groups, EI was significantly greater with ETs of BL >Stage 1 (p< .04).
FERTILITY & STERILITY Ò
e307

BFRs for fresh Aut (959/1539; 59%) & OD (2578/3658; 71%) cycles were
significantly higher than in age-matched frozen Aut & OD cycles (p28% peaked at morula or Stage-1 BL, both considered early ‘‘usable’’ embryos
resulting in reasonable, albeit lower EI & pregnancy rates. When
comparing BL maturity in Aut vs. OC cycles, OD oocytes achieved >BL
expansion with higher EI & pregnancy rates, most likely reflecting younger
oocyte age.
Embryo development & pregnancy outcomes of OC thaws using Autol vs. OD
oocytes.
Autol OC
(n¼1098)
OD OC
(n¼411)
No. 2PN-Zygotes with No 81 (7%) 39 (9%) 0.2
Division (1-cell)
No. 2PN-Zygotes with
433 (39%) 139 (34%) 0.04
Cleavage Arrest
No. Morula (highest stage 156 (14%) 60 (15%) 0.9
achieved)
No. BL - Stage 1 75 (18%) 22 (13%) 0.2
No. BL - Stage 2 181 (42%) 33 (19%)

Using these SRM assays and aCGH, we performed blastocoel protein
profiling and cytogenetic assessment on 14 embryos donated to research.
Subsequently, we conducted statistical analysis using logistic regression to
reveal any correlations between blastocoel protein profile, embryo
morphology, ploidy status, gender, patient’s age.
RESULTS: Using levels of a metabolic protein and the detection of a nuclear
protein as predictors and embryo’s chromosomal status as class label,
the statistical model was able to predict whether an embryo was euploid or
chromosomally abnormal. This was achieved with 100% accuracy within
the small cohort of embryos investigated. Statistical analysis did not show
any association between blastocoel protein profile and any of the other parameters
considered.
CONCLUSIONS: The application of logistic regression analysis to the
data obtained suggests that it may be possible to distinguish chromosomally
normal embryos from those affected by aneuploidy using a targeted proteomics
approach applied to blastocoel fluid. However, due to the small size
of the sample population investigated and the retrospective nature of the analysis,
further work will be essential to determine sensitivity and specificity of
this promising method of preimplantation aneuploidy detection. If confirmed
in larger studies, this minimally invasive proteomic approach could offer a
novel methodology for the prediction of embryonic viability and developmental
competence.
Supported by: Institutional funding.
P-591 Wednesday, October 21, 2015
SYMMETRY AT THE 4-CELL STAGE USING TIME-LAPSE
IMAGING IS CORRELATED WITH EMBRYO
ANEUPLOIDY. C. C. Shenoy, Z. Khan, C. Coddington, J. Jensen,
G. S. Daftary, E. A. Stewart, D. Morbeck. Mayo Clinic, Rochester, MN.
OBJECTIVE: Embryo selection is important for optimal ART outcomes.
Embryo selection has improved over time and the advent of time-lapse
(TL) monitoring offers increased information to aid in embryo selection.
Nonetheless, TL timing parameters alone do not provide high sensitivity
for determining embryo viability or ploidy. We sought to compare euploid
prediction using TL timings with traditional morphometric parameters that
are easily measured using a TL system.
DESIGN: Embryos undergoing preimplantation genetic screening (PGS)
that were cultured using TL monitoring were examined in this retrospective
study.
MATERIALS AND METHODS: All embryos from 2012-2015 undergoing
PGS with trophectoderm biopsy were included. The distance between
the second and first polar body was determined (PBD). Zona pellucida thickness
(ZPT) was measured at the pronuclear stage and at the 2-cell stage in
four locations and averaged. Blastomere area was assessed at the 2- and 4-
cell stages. Symmetry at the 2-cell stage was determined by percent difference
between blastomeres (2cSY). Symmetry at the 4-cell stage was the
percent difference between the smallest and largest blastomeres (4cSY). T-
test was used to compare group means.
RESULTS: Embryos (n¼182) from 21 patients were analyzed. Fouty five
percent were euploid. Patient age ranged from 22-43 (avg¼34). The only variable
that differed significantly between euploid and aneuploid embryos was symmetry
at the 4-cell stage. Aneuploidy rates were 45.5% in the lowest symmetry
quartile and 70.5% in the quartile with the most asymmetry at the 4-cell stage.
CONCLUSIONS: Poor symmetry at the 4-cell embryo stage is more predictive
of aneuploidy than TL parameters, suggesting that embryo selection
models using time-lapse parameters should incorporate cleavage-stage
morphology.
Morphologic and TL Differences Between Euploid and Aneuploid Embryos.
Euploid Aneuploid p value
PBD (mm) 43.2 24.8 43.6 26.8 0.91
ZPT at PN stage (mm) 18.0 2.7 18.3 2.4 0.45
ZPT at 4-cell stage (mm) 17.9 2.6 17.9 2.4 0.81
2cSY (% difference) 11.3 7.5 10.0 8.2 0.28
4cSY (% difference) 26.6 11.4 31.3 13.4 0.01*
t2 (h) 26.4 3.7 26.8 2.8 0.41
t5 (h) 51.2 8.3 52.2 6.9 0.39
tsb (h) 101.8 9.4 103.7 7.9 0.15
tb (h) 108.9 9.7 110.7 8.9 0.27
cc2 (h) 11.1 3.1 11.6 2.9 0.20
s2 (h) 2.2 4.2 1.1 2.4 0.06
P-592 Wednesday, October 21, 2015
FUNCTIONAL CHARACTERIZATION OF CULTURED HUMAN
GRANULOSA CELLS IN SERUM FREE CULTURE
SYSTEM. Y. Wu, a D. F. Albertini, b Q. Wang, a D. H. Barad, c
V. A. Kushnir, d E. Lazzaroni-Tealdi, a N. Gleicher. c a Center for Human
Reproduction, New York, NY; b Center for Human Reproduction & University
of Kansas Medical Center, New York, NY; c Center for Human Reproduction
& Foundation for Reproductive Medicine, New York, NY; d Center for
Human Reproduction & Wake Forest University, New York, NY.
OBJECTIVE: In vitro culture of human granulosa cells (GCs) is an important
research technique for reproductive cell biology and endocrinology
studies. However, traditional culture protocols, involving supplementation
of serum to culture medium, result in luteinization of GCs, and changes their
cell functions during culture. Establishment of a GC culture system that does
not cause luteinization would, therefore, be important.
DESIGN: Prospective laboratory study.
MATERIALS AND METHODS: GCs were collected from follicular
fluid after oocytes retrievals. After washed twice by DPBS to remove
blood contamination,GCs were seeded into 12-well plates at density of
10x105/ml in DMEM/F12 containing 10% FBS, 2mg/ml of HSA, 2
mM glutamine and 1x Insulin-Transferin-Selenium X and cultured for
eight hours at 37 C with 5% CO2 to allow cell attachment. Then the culture
medium was replaced by serum free medium (DMEM/F12 with same
supplementation but without FBS). After overnight culture, medium was
replaced once more. GCs were then cultured for another 96 hours
with or without 50ng/ml FSH supplementation. To evaluate GC functions
after culture, we examined aromatase and FSH receptor (FSHR)
mRNA expression by real-time PCR, cell proliferation by MTT assay,
apoptosis by DAPI staining and estradiol (E2) production by analyzing
medium hormone concentration after adding testosterone (1mM) to the
medium.
RESULTS: After 96 hours culture, FSH treatment significantly induced
aromatase and FSHR expression in GCs (P

test for numerical variables and chi square test for categorical variables were
employed as statistical analysis. P-value lower than 0.05 was considered as
statistically significant.
RESULTS: The incidence of embryos that were generated from fresh
donated oocytes 73.28% (192/262) and 26.72% (70/262) from thawed
donated oocytes. PNf asyn rate was 14.5% (38/262) being slightly more
frequent in thawed oocytes (17.14% vs 13.54%) without any significant difference
(p¼0.42). The average duration between the disappearance of the
first pronuclei and the second was 0.33h (IC: 0.17-0.34). The top quality embryo
rate (A category from ASEBIR) in those embryos showing PNf asyn
was 34.21% (13/38), and in synchronical was 48,86%(86/176); p¼0.10. In
Embryos with asynchronical fading, tPNf were significantly later than in
those with synchronical (24.664.16 vs 26.564.75, p

component analysis (PCA) followed by a partial least square discrimination
analysis (PLS-DA), combined with positive correlation to the blastocyst
group.
RESULTS: The PLS-DA analysis demonstrated a significant difference
between groups in which seven metabolites were differently expressed.
The differently expressed metabolites were phosphocholine arachidonic
C36:2 (Corr. 0.8588), lyso phosphocholine arachidonic C18:0 (Corr.
0.83102), GCKR** (Corr. 0.81856), phosphocholine arachidonic C36:3
(Corr. 0.79307), phosphocholine arachidonic ether C30:2 (Corr. 0.74401),
Glutamine (Corr. 0.74384), and Glutamine/Glycine (Corr. 0.7373).
CONCLUSIONS: The day three culture medium quantitative secretomics
provides important information concerning the embryo developmental
competence, therefore may be an useful tool for the selection of embryos
to be culture until day five improving the implantation and single pregnancy
rates.
Supported by: Coordination for the improvement of Higher Level or Education
Personnel - (Capes-Brazil).
P-597 Wednesday, October 21, 2015
MITOCHONDRIAL DNA CONTENT AS A VIABILITY SCORE IN
HUMAN EUPLOID EMBRYOS: LESS IS BETTER. A. Diez Juan, a
C. Rubio, b C. Marin, a S. Martinez, c P. Diaz-Gimeno, d M. Riboldi, e
N. Al-Asmar, f D. Valbuena, g C. Simon. h a Product Innovation, Igenomix,
Rocafort, Spain; b Igenomix SL; FIVI/INCLIVA, Paterna, Spain; c Product
Innovation, Igenomix, Paterna, Spain; d Fundacion IVI / INCLIVA, Paterna
(Valencia), Spain; e Igenomix Brasil, Sao Paulo, Brazil; f IviGen, Miami,
FL; g Igenomix S.L., Paterna, Spain; h FIVI/INCLIVA, Valencia University,
Stanford University, Paterna, Spain.
OBJECTIVE: To investigate the clinical relevance of mitochondrial
mtDNA content as a viability score in human euploid embryos.
DESIGN: Retrospective analysis of mtDNA content of transferred euploid
embryos.
MATERIALS AND METHODS: Setting: Reproductive genetics laboratory.
Patients: Single embryo transfer in 270 patients who underwent preimplantation
genetic screening (205 day-3 blastomere biopsies, and 65 day-5
trophectoderm biopsies), and 10 patients with double embryo transfer
(male-female). Samples: DNA from single blastomere or trophectoderm biopsy
amplified using the Sureplex DNA amplification system. Euploid embryos
were transferred at the blastocyst stage and an aliquot of their
amplified DNA was used for this study. Relative amounts of nuclear DNA
(nDNA) and mtDNA were determined by quantitative real-time (RT) PCR.
The ratio of mtDNA/nDNA was classified as the mitochondrial score (Ms),
and used as an indicator of the mitochondrial copy number per cell. MtDNA
copy number was compared using a non-parametric test, Wilcoxon’s rank
sum test and Shapiro-Wilk test were used to normalize. Ms scores are presented
based on quartiles.
RESULTS: High mtDNA copy number in euploid embryos is indicative of
lower embryo viability and implantation. Using the normalized mtDNA content,
we created the mitochondrial score or Mitoscore (Ms). Day-3 embryos
with less than 34 (MsA) had an implantation rate (IR) of 59% (n¼51); those
with 34 to 52 (MsB) had an IR of 44% (n¼52); those with 52 to 97 (MsC) had
an IR of 42% (n¼50); and those with levels > 97 (MsD) had an IR of 25%
(n¼52). Embryos with Ms>160 (n¼22) never implanted. Day-5 with less
than 18.19 (MsA) had an IR of 81%; those with 18.19 to 24.15 (MsB) had
an IR of 50% (n¼16); those with 24.15 to 50.58 (MsC) had an IR of 62%
(n¼16); and those with levels > 50.58 (MsD) had an IR of 18% (n¼17). Embryos
with levels > 60 (n¼7) never implanted.
CONCLUSIONS: An increased amount of mtDNA in euploid embryos is
related to poor implantation potential and may be indicative of reduced metabolic
fuel during oocyte maturation. We are implementing Ms in our PGS
platform to prospectively-analyze its clinical relevance.
P-598 Wednesday, October 21, 2015
WITHDRAWN
P-599 Wednesday, October 21, 2015
PROFILING OF GENDER-SPECIFIC GENE EXPRESSION IN FE-
MALE AND MALE PRIMORDIAL GERM CELLS IN
MICE. T. Kono, A. Sakashita, Y. Kawabata, Y. Jincho, H. Kobayashi.
Bioscience, Tokyo University of Agriculture, Tokyo, Japan.
OBJECTIVE: After complete migration of primordial germ cells (PGCs)
into the genital ridges by E13.5 in mice, they undergo a gender-specific fate:
Male and female PGCs undergo mitotic arrest and meiosis, respectively.
However, differentiation of PGCs on a gender-specific basis remained poorly
understood. The aim of this study was to gain further insight into the genderspecific
features of PGCs in mice.
DESIGN: We performed RNA-Seq analysis of E13.5 female and male
PGCs by using a next-generation sequencer.
MATERIALS AND METHODS: PGCs of E13.5 fetuses were isolated using
a FACSAria II Cell Sorter (BD Bioscience). Total RNA of 1 104 PGCs
was isolated, and cDNA synthesis and pre-amplification were performed using
10 ng of total RNA. Sequencing libraries were created by NEBNext Ultra
DNA Library Prep Kit for Illumina, according to the library construction protocol
(New England BioLab). Indexed libraries were pooled and sequenced
using an Illumina Hiseq 2500 Sequencer. ChIP-seq analysis was performed
for H3K4me3 and H3K27me3. RNA-Seq reads were aligned to the mouse
genome (mm10, Genome Reference Consortium Mouse Build 38) by using
a CLC Genomics Workbench (CLC bio). Aligned reads were subsequently
assembled into transcripts guided by reference annotation (mm10, UCSC
gene annotation).
RESULTS: We identified gender-specific transcripts: 651 and 428 were
specific expressed (>2-fold and P

and significance differences between implantation rates of HIGH and LOW
embryos to the previous analysis (p¼0.019, OR¼1.72CI95% 1.10-2.70).
CONCLUSIONS: Our study presents,to our knowledge, the largest set of
transferred embryos after time lapse analysis using Eeva. It has demonstrated
that embryo selection using the classification provided by Eeva is related
with reproductive outcome. The observed relationship with the implantation
potential reflects a direct link between the parameters provided by the automatic
system and embryo quality and can be used for embryo selection in a
clinical setting. Moreover, our multivariable analysis demonstrate that the
relationship between EEVA and implantation potential is robust and independent
of other clinical variables such oocyte quality or embryo morphology.
P-601 Wednesday, October 21, 2015
TIME LAPSE KINETIC MARKERS ENABLE EARLY IDENTIFICA-
TION OF DEVELOPMENTALLY DELAYED EUPLOID
BLASTOCYSTS. B. Hixon, a J. C. Parks, a J. M. Stevens, b
W. B. Schoolcraft, c M. Katz-Jaffe. c a National Foundation for Fertility
Research, Lone Tree, CO; b Fertility Labs of Colorado, Lone Tree, CO; c Colorado
Center for Reproductive Medicine, Lone Tree, CO.
OBJECTIVE: The introduction of time lapse (TL) technology into the IVF
laboratory has allowed for a more in depth and uninterrupted study of embryo
development. A viable blastocyst requires successful completion of specific
developmental events and all 23 pairs of chromosomes. Research investigating
TL data have indicated that competent blastocysts are identifiable
by early cleavage stage TL kinetic markers. This study examined embryonic
development and TL kinetic markers of euploid blastocysts, and their implantation
outcome.
DESIGN: Research Study.
MATERIALS AND METHODS: Normally fertilized zygotes from infertility
patients (mean maternal age ¼ 37.0 years) were cultured in the EmbryoScopeTM
(Vitrolife) to the blastocyst stage. A trophectoderm biopsy was
performed on either D5 or D6 of development for comprehensive chromosome
screening using qPCR (RMA-NJ) or aCGH (Illumina), to identify
euploid blastocysts (n¼272). Biopsied blastocysts were vitrified to be
warmed at a later date for a frozen embryo transfer. Statistical analysis
included Student’s t-test and Chi-square test for independence, significance
at P

CONCLUSIONS: While ovarian stimulation can negatively impact uterine
receptivity (3), this study addresses the effects of stimulation more
directly on implantation of euploid embryos. Use of more gonadotropin
and retrieval of more oocytes were both associated with decreased IR. Higher
gonadotropin doses could indicate a poorer ovarian response or that excessive
use of gonadotropin is detrimental. Higher numbers of oocytes retrieved
could indicate a more robust ovarian response or aggressive stimulation/
retrieval. Higher IR occurred when FhMG was larger, suggesting that LHlike
activity improves oocyte/embryo quality.
References:
1. Hodes-Wertz, et al. Fertil. Steril. (2015) 103:947.
2. Hong et al., Fertil. Steril. (2014) 102:41.
3. Shapiro et al., Fertil. Steril. (2011) 96:344
EMBRYO CULTURE
P-604 Wednesday, October 21, 2015
DIPEPTIDE GLUTAMINE FUNCTIONS AS AN ORGANIC OSMO-
LYTE IN MOUSE PREIMPLANTATION EMBRYOS BUT DOES
NOT SUPPORT BLASTOCYST HATCHING AT THE SAME RATE
AS INDIVIDUAL COMPONENT AMINO ACIDS. J. E. Swain. a,b a Fertility
Lab Sciences, Englewood, CO; b Colorado Center for Reproductive
Medicine, Lone Tree, CO.
OBJECTIVE: Dipeptide glutamine is included in human embryo culture
media to reduce ammonia production. However, dipeptides have not been
examined to determine if they can fulfill known physiologic roles of individual,
non-dipeptide amino acids. Our objective was to examine whether dipeptide
alanyl-glutamine (ala-glu) can act as an organic osmolyte in mammalian
embryos as efficiently as its component individual amino acids.
DESIGN: Prospective experimental study.
MATERIALS AND METHODS: Thawed 1-cell mouse embryos were
cultured in groups of 10 in 500ul in 1 of 4 media treatments for 96h in 6%
CO2, 5% O2, 89% N2. Positive controls (Pos Con) consisted of amino acidfree
Human Tubal Fluid (HTF; 280mOsm). Treatment media included HTF
with elevated NaCl to increase osmolality (320mOsm) and either 1mM dipeptide
ala-glu or 1mM alanine + 1mM glutamine (ala+glu). Negative controls
(Neg Con) consisted of 320mOsm with no amino acids. Blastocyst development
was assessed and sizes determined using Cronus software. Data were
collected over 5 replicates and analyzed using ANOVA and Tukey analysis.
RESULTS: Pos Con resulted in 79.0% 5.9 blastocyst development while
Neg Con media resulted in significantly lower rates of blastocyst formation
(20.1% 7.3), p

patients. In contrast, the arrest rate between day 3 and the blastocyst stage
increased dramatically with age. This supports the use of extended culture
to optimize embryo selection. The fact that the blastulation rates were quite
high in even the oldest patient group suggests that clinicians and embryologists
should not forego extended culture out of concern that there will be a
lack of blastulation. This allows the older high risk age groups access to
advance diagnostics such as CCS.
Embryonic performance in extended culture by age.
Age

second mitosis. Time is calculated by hours after insemination. Data were
analyzed using ANOVA and c2 test.
RESULTS: Euploidy is not correlated with the morphokinetic parameters
used, either in BB group or TB group (Table 1). Interestingly, multinucleation
at 2-cell stage did not increase aneuploidy rates in either Day 3 or
Day 5 biopsies. Euploidy rate was significantly higher in the TB group
(33%) compared with BB group (16%, p¼0.005).
CONCLUSIONS: Our results suggest that euploidy (from either BB or
TB) cannot be predicted with the selected morphokinetic parameters using
time-lapse microscopy. Future investigation should focus on different morphokinetic
parameters to predict ploidy status.
Table 1. The Mean Differences of Morphokinetic Parameters within Groups.
Biopsy stage
Trophectoderm
Biopsy
Trophectoderm
Biopsy
Blastomere
Biopsy
Blastomere
Biopsy
Parameters /ploidy Aneuploidy Euploidy p-value Aneuploidy Euploidy p-value
tPB2 3.61.5 3.21.2 0.14 4.83.4 4.73.3 0.81
tPNa 8.51.9 8.12.1 0.40 10.83.9 10.53.5 0.63
tPNf 24.53.7 23.63.1 0.27 24.66.5 24.24.6 0.69
tP2 11.24.8 11.14.9 0.49 7.86.5 7.55.8 0.67
tSB 103.79.7 101.56.6 0.26 99.98.9 100.611.6 0.75
Multinucleation
at 2 cell stage
25% 39% 0.24 37% 33% 0.62
Supported by: The authors wish to thank Dr.MH. Fakih, Dr.FN. Shamma,-
Dr.A.Hammoud and Dr. S.Dogan for data mining support.
P-610 Wednesday, October 21, 2015
RECOMBINANT ALBUMIN AS A POTENTIAL ALTERNATIVE TO
BLOOD-DERIVED ALBUMIN FOR USE IN BLASTOCYST CUL-
TURE: A SIBLING EMBRYO STUDY. M. Murakami, a
D. K. Gardner, b S. Mizumoto, a K. Tanaka, a A. Egashira, a T. Kuramoto. a
a Kumamoto Women’s Clinic, Fukuoka, Japan; b University of Melbourne,
Parkville, Australia.
OBJECTIVE: Replacing human serum albumin (HSA) with lower
amounts of recombinant human albumin (rHA) during embryo culture yields
good-quality blastocysts. In this study, the efficacy of using completely
defined media containing rHA for all ART protocols was evaluated using a
sibling embryo model to minimize experimental variation between patients.
Further, we examined the effects of the albumin source on pregnancy rates
following vitrification.
DESIGN: Prospective sibling embryo study.
MATERIALS AND METHODS: Patients undergoing elective embryo
cryopreservation between October 2014 and April 2015 were evaluated.
Forty-one patients (aged 35.5 0.6 years) who underwent oocyte
retrieval after hyperstimulation and IVF/ICSI treatment using rHA-containing
defined media and had R4 2PN oocytes at 18 h after insemination
were included in the study. In total, 362 sibling 2PN oocytes from
these patients were randomized for side-by-side culture in G1/G2 media
containing either 0.5 mg/mL rHA or 5 mg/mL HSA. All good-quality
blastocysts (R3BB) were vitrified by day 6. Morphokinetics were evaluated
by time-lapse microscopy in certain patients. Data was analyzed
for a single ET of vitrified blastocysts performed by the end of March
2015.
RESULTS: Percentages of good-quality day 3 embryos (6- to 10-cell embryos
with %25% fragmentation) and good-quality blastocysts were
similar in rHA (55.2% and 18.8%, respectively) and HSA (56.9% and
26.5%) groups. The time taken to develop into full blastocysts (with the
blastocoel completely filling the embryo) from insemination was not
different between rHA (108.6 5.6 h) and HSA (100.9 3.1 h) groups.
There were 12 and 11 cycles of vitrified blastocyst ET in the rHA and
HSA groups, respectively. All blastocysts survived after warming (1 embryo/cycle).
Clinical pregnancy rate/ET was 75.0% and 45.5%, and
ongoing pregnancy rate/ET was 75.0% and 18.2% (P < 0.05) in rHA
and HSA groups, respectively.
CONCLUSIONS: We demonstrated the feasibility of using chemically
defined ART systems with rHA to yield good-quality blastocysts, and
subsequently, high pregnancy rates. Studies involving optimization of
rHA concentration and long-term follow-up studies with more participants
are required to validate the efficacy of the procedures. Using
rHA will help eliminate variation and risks associated with blood-derived
products.
P-611 Wednesday, October 21, 2015
DOES THE TRANSITION FROM STANDARD INCUBATOR TO A
TIME-LAPSE IMAGING CHAMBER IMPACT EMBRYONIC
DEVELOPMENT AND NEONATAL OUTCOMES? R. W. Goldberg,
P. K. Gill, J. M. Goldberg, N. Desai. Obstetrics & Gynecology / Women’s
Health Institute, Cleveland Clinic, Beachwood, OH.
OBJECTIVE: Time lapse imaging chambers offer a unique environment
for uninterrupted culture while still allowing visualization of embryo development.
The altered culture environment may influence cell division and progression
to the blastocyst stage. Ability to identify abnormal cleavage
patterns as well as specific kinetic data with time lapse systems may further
aid in embryo selection and ultimately pregnancy outcomes. This study compares
embryonic development, clinical and neonatal outcomes after transition
from standard box type incubators to time lapse.
DESIGN: This study retrospectively analyzed IVF cycle data from 2011
when embryo culture was performed in Forma standard incubators (SI) to
2013 when all patient embryos were exclusively cultivated in EmbryoScope
time lapse imaging chambers (TL). For analysis the data was further stratified
by age and restricted to patients using autologous oocytes and having a fresh
transfer.
MATERIALS AND METHODS: Zygotes were individually cultured in
Global medium (LifeGlobal; Guilford, CT) with 10% SPS (SAGE; Trumbull,
CT) at 37o C with 6% CO 2 . In the SI group, dishes were removed once a day
for embryo evaluation. Embryos in the TL group underwent continuous uninterrupted
culture under low oxygen tension. Embryo development was
monitored by viewing time lapse videos. Clinical pregnancy was determined
by the presence of a fetal heart on ultrasound. Implantation rate was calculated
based on gestational sacs. Cycle data and neonatal outcomes were
compared using chi square analysis and the Student’s T-test. P-values of
37
TL
N¼222
p-value
Standard
N¼123
TL
N¼92
p-value
Embryos Transferred 2.06 2.00 0.332 2.69 2.33 0.0014
0.60 0.52
0.80 0.83
Clinical Pregnancy (%) 53.6 56.3 0.564 30.1 34.8 0.465
(126/235) (125/222)
(37/123) (32/92)
Implantation (%) 35.2 42.7 0.0192 15.7 21.0 0.113
(170/483) (190/445)
(52/331) (45/214)
Blastocyst Formation (%) 48.7 62.9

Mono media is a single formulation. Mono formulations require less manipulation
and are less expensive. Both formulations have attained good outcomes,
but an adequately powered assessment including usable
blastulation rates (USBR), ploidy risk, and SIR is lacking.
DESIGN: Paired RCT.
MATERIALS AND METHODS: Patients with normal ovarian reserve
were recruited. A paired design allowed each patient to serve as their own
control, eliminating many confounding variables seen in prior studies. After
confirming fertilization, patients’ zygotes were randomized (1:1 ratio) to
either Seq media (Quinn’s Advantage Cleavage Medium, Sage then Blast
Assist, Origio) or Mono media (Continuous Single Culture; Irvine Scientific).
Each culture system used separate incubators. Assessed endpoints for
all embryos included USBR, blastulation timing (arrest vs day 5 vs 6) and
ploidy status. Paired euploid blastocyst transfers, one from each group,
were performed. DNA fingerprinting of concepti was used as needed to
link each embryo to a definitive outcome. SIR was defined as the presence
of a fetal heart beat at 8-9 weeks gestation. Statistical analysis performed
via McNemar’s Chi Square and Wilcoxon sum rank tests.
RESULTS: 186 patients had their 2PN embryos (N¼2257) randomized to
each culture system. Seq media had a higher blastulation rate then Mono media
(p¼0.001, 55.5% vs. 46.0%). No differences were found in the day of
blastulation (p¼0.4063) or in the aneuploidy rate (p¼0.5518). Of the 168 patients
who had euploid blastocysts suitable for transfer, 126 completed a
paired embryo transfer and 42 had a SET. Amongst the SETs, the SIR was
equivalent (p¼1.0). Of the 126 double embryo transfers, 36 patients had
one fetal heart beat at discharge, however there was no statistical difference
in the likelihood of implantation between groups (p¼0.8642).
CONCLUSIONS: This is the first randomized controlled trial to systematically
examine paired euploid transfers of sibling zygotes cultured in in Seq
vs. Mono media. This study demonstrates that the usable blastocyst rate is
greatest after culture in Seq media in comparison to a Mono formulation;
however no difference exists in SIR.
Supported by: Irvine scientific provided Mono media.
P-613 Wednesday, October 21, 2015
SUPPLEMENTING SINGLE STEP CULTURE MEDIA WITH INSU-
LIN FOR CONTINUOUS UNINTERRUPTED IN VITRO CULTURE
OF HUMAN EMBRYOS AND MONITORING THE OUTCOME:
PROSPECTIVE RANDOMIZED CLINICAL TRIAL. M. Fawzy, a
A. Alaboudy, a M. Sabry, b M. Gad, a H. Morsy, a H. Kasem, a
F. Alaboudy, a E. R. Othman, c H. Abdelghaffar, b A. M. Metwalley, d
M. Abdel-Rahman, b S. Roshdy. b a Ibnsina IVF Center, Sohag, Egypt; b Sohag
Faculty of Medicine, Sohag, Egypt; c Assiut University, Assiut, Egypt; d IVF
Lab Director, Bahrain, Bahrain.
OBJECTIVE: To compare embryologically and clinical outcomes between
two different culture strategies: Uninterrupted culture in single step
IVF culture media, versus uninterrupted culture in the same medium with insulin
supplementation.
DESIGN: Prospective Randomized Clinical Trial.
MATERIALS AND METHODS: We recruited 117 patients’ Oocytes presented
to Ibnsina IVF Center, Sohag, Egypt, between September 2014 and
March 2015 to this study. Mature Oocytes (1354) underwent Intracytoplasmic
sperm injection (ICSI). We randomly assigned the injected Oocytes
into two groups (sibling Oocytes, 677 each). Group I Oocytes underwent
ICSI and cultured in insulin free single step media (GLOBAL TOTALÒ,
LIFEGLOBALÒ, Canada). Also, incubated in (7% Co2, 5% O2 and 88%
N2) for uninterrupted culture from day 0 to day 5. Group II Oocytes underwent
ICSI then cultured in insulin supplemented single step media
(GLOBAL TOTALÒ, LIFEGLOBALÒ, Canada). We supplemented the culture
media with insulin to reach a concentration of 50 ng/ml (Sigma I9278)
under the same gas phase for uninterrupted culture protocol. We randomized
the transfer of the resulting embryos between the two groups by closed envelope
method. Outcome measures included: Fertilization rate, top quality embryos
and compactionrate at day 3, blastocyst rate and quality at day 5, embryo
utilization and finally pregnancy rate. We used Chi-square test for comparing
ourdata.
RESULTS: The recruited patients were similar regarding mean age, BMI,
the dose of FSH/HMG used, number of oocytes collected and number of cycle
days. There was no difference in fertilization rate (group II 72.6% and a
group I 73%). There was no difference in top quality embryos at day 3 with a
trend toward better compaction of group II. We documented a significantly
higher blastocyst formation rate in group II versus group I (66% versus
44%, respectively. P value < 0.01). We also recorded a higher top quality
blastocyst (better expansion, intact inner cell mass and more trophectoderm
layers) in the group II (64% versus group I 41% P value

and 3 x Zand-air 100C units were used as air filtration. Chi square and student’s
t-tests were used as appropriate. There were no lab protocol changes
during the study period, besides the in-duct air system.
RESULTS: 289 ART cycles were analyzed, 163 cycles pre and 126 cycles
post in-duct air filtration installation. There were no differences between the
two groups in the mean age of the patient, the number of oocytes retrieved,
percentage of mature oocytes per patient, number of fertilized oocytes, or
number of embryos transferred. However, significant increases were seen in
the number of positive pregnancies, ongoing pregnancy rate, and viable blastocyst
development rates. Air quality VOC testing demonstrated improved air
quality after the installation of the in-duct air system (0.0 ppm) as compared
with previous VOC readings prior to installation (0.3 ppm).
Freestanding air
filtration units
alone
In-duct air
filtration
units
p value
Mean age of patient (years) 30.5 30.5
Number of patients (n) 163 126
Mean no. eggs retrieved 12.6 13.3 p¼0.3231
Mean no. mature eggs 8.77 9.77 p¼0.0583
Mean no. fertilized eggs 7.28 7.67 p¼0.4128
Mean no. embryos 1.88 1.79 p¼0.0749
transferred
Mean no. viable blastocysts 1.75 3.50 p¼0.0001
First bhCG (%) 47.2% 63.5% p¼0.0059
Clinical pregnancy rate 45.4% 55.6% p¼0.0868
(%)(gestational sac)
Ongoing pregnancy rate (%)
(fetal heart tones)
34.5% 55.6% p¼0.0003
CONCLUSIONS: Air quality in the IVF laboratory has been shown to have
an impact on embryo development and resultant pregnancy outcomes. In an
urban, multi-office, multiple story building, air quality is often difficult to
control. The results of this study show that even multiple types and numbers
of freestanding air filtration units may not be enough to create an ideal air
quality environment within the IVF laboratory. The observations seen in
this study suggest that the installation of an in-duct, positive pressure air filtration
system does increase viable blastocyst development and pregnancy rates.
P-616 Wednesday, October 21, 2015
PREDICTING BLASTOCYST FORMATION RATE: AN AUTO-
MATIC CELL TRACKING SYSTEM AIDS IN THE SELECTION
AMONG GOOD MORPHOLOGY EMBRYOS. N. Basile, a
I. Cabanes, a M. Testillano, a D. Cernuda, a J. A. Garcia-Velasco, a
M. Meseguer. b a IVI, Madrid, Spain; b Clinical Embryology, Valencia, Spain.
OBJECTIVE: To quantify, by multivariable analysis, the blastocyst formation
rate according to the three predictive categories provided by Eeva (Early
Embryo Viability Assessment).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients undergoing IVF cycles using
their own or donated oocytes. Embryos were cultured in standard incubators
including multiple Eeva systems. The Eeva test utilizes an automatic cell
tracking software to classify embryos into three categories (HIGH-ME-
DIUM-LOW) according to their probability of becoming a blastocyst. For
that aim the system relies on an algorithm based on the variables P2 ¼ t3-
t2 (time to 3 cell - time to 2 cell) and P3 ¼ t4-t3 (time to 4 cell - time to 3
cell). In order to quantify the blastocyst formation rate according to the
Eeva categories, a logistic regression analysis was performed taking in
consideration possible confounding factors: embryo morphology according
to ASEBIR (Spanish Association of Biologists; A-B-C-D), source of oocytes
(own or donated), and number of oocytes.
RESULTS: A total of 494 patients generated 3596 embryos. The overall
blastocyst formation rate was 59.3 % (1347/2269). When categorizing according
to Eeva, we found significant differences in the overall blastocyst formation
rate between LOW vs. MEDIUM (p< 0.001, OR¼ 1.964 CI95% 1.550-
2.489) and LOW vs. HIGH (p< 0.001, OR¼ 3.743 CI95% 2.724-5.143).
Development to ‘‘optimal’’ blastocyst was also analyzed with significant differences
between LOW vs. MEDIUM (p< 0.001, OR¼ 1.634 CI95% 1.248-
2.140) and LOW vs. HIGH (p< 0.001, OR¼ 3.053 CI95% 2.295-4.061). The
only confounding factor presenting a significant effect was embryo
morphology according to ASEBIR. The correlation between Eeva categories
and blastocyst formation rate differs between good quality embryos (A-B) and
poor quality embryos (C-D). Taking this in consideration, we still found significant
differences between LOW vs. MEDIUM (p¼ 0.001, OR¼ 1.554
CI95% 1.207-1.999) and LOW vs. HIGH (p< 0.001, OR¼ 2.505 CI95%
1.792-3.502) for the overall blastocyst formation rate but correlation was
reduced in 21% and 33% respectively. For ‘‘optimal’’ blastocyst formation
rate, no significant differences were observed between LOW vs. MEDIUM
(p¼ 0.110, OR¼ 1.263 CI95% 0.948-1.682) but we did observe significant
differences between LOW vs. HIGH (p< 0.001, OR¼ 1.950 CI95% 1.437-
2.647). Once again correlation was reduced in 23% and 36% respectively.
CONCLUSIONS: Eeva categories are strongly correlated with blastocyst
formation rates and the prediction is significantly affected by day 3 embryo
morphology. Therefore the best strategy, among good quality embryos, is the
combination of both: morphology and morphokinetics.
P-617 Wednesday, October 21, 2015
EFFECT OFARTIFICIAL OOCYTE ACTIVATION IN INTRACYTO-
PLASMIC SPERM INJECTION USING TESTICULAR SPERMATO-
ZOA ON SIBLING OOCYTES. C. Takahashi, S. Mizuta, R. Nishiyama,
K. Yamaguchi, K. Kitaya, H. Matsubayashi, T. Ishikawa. Reproduction
Clinic Osaka, Osaka, Japan.
OBJECTIVE: Artificial oocyte activation (AOA) has been proposed as a
suitable means to overcome the problem of failed or impaired fertilization after
intracytoplasmic sperm injection (ICSI). To analyze with calcium ionophore
after ICSI using testicular spermatozoa improves fertilization,
embryonic development and pregnancy outcome in patients with obstructive
azoospermia (OA) or non-obstructive azoospermia (NOA).
DESIGN: Prospective clinical analysis on sibling oocytes.
MATERIALS AND METHODS: This prospective study was performed
between October 2013 and April 2015. All patients involved gave written
consent, and institutional review board approval was granted. This study includes
22 OA and 47 NOA couples. We excluded the couples using only
immotile spermatozoa for ICSI. Retrieved oocytes were incubated in culture
medium (Universal IVF Medium: UIM) for 2 hours at 37C and 6% CO₂ and
were underwent ICSI with motile testicular spermatozoa. When eight or more
metaphase M (MII) oocytes were available, AOA was performed on half of
the sibling MII oocytes. After ICSI, oocytes were incubated in UIM for 30
minutes, and exposed to 10mM of calcium ionophore A23187 for 15 minutes.
The oocytes were then washed and placed in UIM. Two pronuclei (2PN) oocytes,
blastocysts development, good-quality blastocysts, biochemical pregnancies,
and clinical pregnancies rates were compared between two groups.
RESULTS: In terms of OA couples, there were no significant difference in
2PN oocytes, blastocysts development, and good-quality blastocysts rates
(73.0%, 56.1%, and 47.8% with AOA and 66.0%, 41.2%, and 45.7% without
AOA, respectively). For NOA couples, 2PN oocytes with AOA (74.0%) was
significantly higher than those without AOA (61.2%). Blastocysts development,
and good-quality blastocysts rates for NOA couples were 53.8% and
38.4% with AOA and 56.7% and 40.5% without AOA, respectively (no significant
differences).
CONCLUSIONS: AOA with calcium ionophore showed favorable effect
on fertilization rate in patients with NOA but OA. The sperm source was
strongly affect the fertility potential or clinical outcomes. Severe male factor
infertility, especially NOA, could be an indication for application of AOA.
P-618 Wednesday, October 21, 2015
AN EVALUATION OF CONTINUOUS HUMAN EMBRYO CULTURE
USING THE WOW DISH. H. Watanabe, N. Fukunaga, K. Nakayama,
M. Shimomura, H. Tsuji, H. Kitasaka, F. Tamura, Y. Konuma,
M. Kojima, Y. Asada. Asada Ladies Clinic Medical Corporation, Nagoya,
Japan.
OBJECTIVE: The WOW dish (LinKIDTMculture dish,DNP) 25 microwells
that allows group culture under a single drop of medium. Through its
design it is possible to manage embryos separately whilst in group culture.
There are several reports (SUGIMURA et al, 2013) suggesting that the
WOW dish improved bovine embryo culture results. Therefore, we investigated
whether the WOW dish is suitable for continuous human embryo culture
without exchange of culture medium.
DESIGN: Retrospective study.
MATERIALS AND METHODS: The study consisted of three experimental
groups. ‘‘15ul drop culture’’ group (control) was the standard embryo
culture method with exchange from single medium to single medium at Day3
FERTILITY & STERILITY Ò
e317

and Day5 using normal 35mm dish. The other 2 groups were ‘‘15ul drop
continuous culture’’ and ‘‘WOW dish continuous culture’’ without exchange
of culture medium. A total of 143 pronuclear embryos from 12 patients who
consented to post-thaw culture were studied. Pronuclear stage frozen embryos
were thawed and cultured across the 3 groups for up to 7days (n¼47, 48, 48).
We compared the incidence of good blastocysts (R 3BB,Gardner) and blastocyst
formation rate at Days 5, 6 and 7 in the 3 groups.
RESULTS: In the control group, blastocyst formation rate at Days 5, 6 and
7 was 46.8%, 57.4% and 59.6%, respectively. The incidence of good blastocysts
was 25.5%, 44.7%, and 48.9% (n ¼ 47) respectively. In the ‘‘15ul drop
continuous culture’’ group, blastocyst formation rate at Days 5, 6 and 7 was
45.8%,54.2% and 54.2%. The incidence of good blastocysts was
14.6%,31.3%,33.3% (n ¼ 48). In the ‘‘WOW dish continuous culture’’ group,
blastocyst formation rate at Days 5, 6 and 7 was 52.1%,58.3% and 58.3%.
Incidence of good blastocysts was 25.0%,43.8% and 47.9% (n ¼ 48). Between
each observation time interval, there was no significant differences
in both the incidence of good blastocysts and the blastocyst formation rate.
CONCLUSIONS: These results suggest that the WOW dish is suitable for
continuous culture without exchange of culture medium compared to the
15ul drop control method.
P-619 Wednesday, October 21, 2015
COMPARISON OF CONTINUOUS AND SEQUENTIAL CULTURE
MEDIA ON BLASTOCYST UTILIZATION AND IMPLANTATION
RATES USING SIBLING OOCYTES. A. E. Fritz, K. Gehrke,
D. Kim, M. Goering. Center for Advanced Reproductive Medicine, Department
of Obstetrics and Gynecology, University of Kansas School of Medicine,
Overland Park, KS.
OBJECTIVE: The broad availability of commercial embryo culture media
has improved performance and consistency in human ART laboratories, however
the variety of media compositions from different manufacturers and culture
conditions between laboratories has left a lack of consensus across the
field. The aim of this study was to provide a carefully controlled comparison
of human preimplantation embryo development in two distinct culture media
formulations, continuous (Global, Life Global Group) versus sequential
(Quinn’s Advantage, Sage).
DESIGN: Retrospective, randomized cohort study.
MATERIALS AND METHODS: A total of 247 fresh non-donor ICSI cycles
(2841 oocytes) from January 2014 through April 2015 were included in
the analysis. Study inclusion required cohorts of at least six sibling oocytes to
be randomly divided into one of two different culture media, continuous
(n¼1451) or sequential (n¼1390). All sibling oocytes and embryos were
treated identically regardless of media formulation, including protein supplementation
(5mg/mL SPS) and media refreshment on D1 and D3. Culture
occurred in Planer mini-benchtop incubators using premixed gas or Thermo
cabinet incubators with 6% CO 2 and 5% O 2. Manufacturer target post-equilibration
pH ranges were carefully monitored and varied only slightly between
formulations (7.25-7.35 +/-0.05). Rates of fertilization, D5 and D6
blastocyst conversion, embryo transfer (D5), cryopreservation, implantation,
and clinical pregnancy were compared.
RESULTS: The rate of fertilization post-injection was higher in the continuous
media (80%) versus the sequential media (73%; p

monitoring, but 50 mL prevents changes in media contained in the SAFE Sens
sensor for weeklong testing.
Osmolality Values.
SAFE Sens
Day 035 mL Oil
268 SAFE Sens Day
335 mL Oil
Culture DishDay 265 Culture Dish Day
0 5.5 mL Oil
3 5.5 mL Oil
Control Culture Dish 268 Control Culture Dish
Day 011 mL Oil
Day 3 11 mL Oil
292 SAFE Sens Day
735 mL Oil
283 Culture DishDay
75.5 mL Oil
270 Control Culture Dish
Day 7 11 mL Oil
Supported by: Thanks to Blood Cell Storage Inc. for use of their evaluation
equipment.
P-622 Wednesday, October 21, 2015
RANDOMIZED COMPARISON OF EMBRYO DEVELOPMENT IN
CLOSED TIME-LAPSE PHOTOGRAPHY SYSTEM WITH TRADI-
TIONAL STANDARD EMBRYOLOGY CULTURE WITH DAY-3 EM-
BRYO TRANSFERS. Y. Wu, a E. Lazzaroni-Tealdi, a Q. Wang, a
D. F. Albertini, b D. H. Barad, c V. A. Kushnir, d N. Gleicher. c a Center for Human
Reproduction, New York, NY; b Center for Human Reproduction & University
of Kansas Medical Center, New York, NY; c Center for Human
Reproduction & Foundation for Reproductive Medicine, New York, NY;
d Center for Human Reproduction & Wake Forest University, New York, NY.
OBJECTIVE: To prospectively compare a time-lapse photographic system
(EmbryoscopeÒ, UnisensFertilitech, Vitrolife, Aarhus, Denmark) to
the traditional standard manual embryology in the management of human
embryos during donor oocyte cycles with day-3 embryo transfers (ET).
DESIGN: Prospectively randomized open label study (PRS).
MATERIALS AND METHODS: This study had two components: In (1)
we compared embryo quality in 76 embryos from 7 oocyte donor cycles during
randomized parallel culture in our standard culture system (SC) and an
EmbryocopeÒ culture system (EC). After three day culture, embryos were
evaluated by our center’s standard morphology grading. In (2), we parallel
cultured 37 embryos from 4 donor cycles in SC and EC dishes, respectively,
in a standard incubator for 3 days before grading to determine whether culture
dishes used in EC and SC may be responsible for outcome differences.
All embryos were produced by ICSI and only normally fertilized embryos
were selected for the study.
RESULTS: The table demonstrates that the SC system produced better
quality embryos than the EC system (P 0.05).
Phase I: Culture system
test (n¼76, N¼7)
Phase II: Culture dish
test in standard
incubator(n¼37, N¼4)
Embryo
quality
n¼number of embryos; N¼number of patients.
EC
system
SC
system
817
333
268
P value
Good (%) 55.86.4 81.24.1 0.005
Fair (%) 36.88.5 7.74.1 0.01
Poor (%) 7.35.7 11.04.7 0.62
Embryo EC dish SC dish P value
quality
Good (%) 65.08.6 66.03.3 0.96
Fair (%) 16.79.6 11.45.9 0.67
Poor (%) 23.35.1 27.33.9 0.56
CONCLUSIONS: In oocytes donors (i.e. likely good prognosis patients),
undergoing day-3 embryo transfer, SC produced embryos of better quality
than EC. Since this difference does not appear to be consequence of differences
in culture dishes, the observed discrepancy in outcomes has to be
due to differences in culture conditions. Since here reported results are based
on limited study size and, therefore, do not include pregnancy outcomes,
they should be considered preliminary. They, however, do raise concerns,
and should induce the industry to perform PRSs, utilizing embryo randomization
rather than patient randomization, to assure that EC systems do not
negatively affect IVF outcomes in comparison to standard embryology
practice.
Supported by: We acknowledge the support of UnisenseFertilitech, Vitrolife,
which loaned CHR the instrument. Intramural funds from The Center for
Human Reproduction and grants from The Foundation for Reproductive
Medicine paid for installation fees and reagents.
OVARIAN STIMULATION
P-623 Wednesday, October 21, 2015
COMPARISON OF STANDARD GNRH ANTAGONIST PROTOCOL
AND LUTEAL PHASE ESTRADIOL/GNRH ANTAGONIST PRIM-
ING PROTOCOL IN THE POOR RESPONDERS. A. Erdem, a
M. F. Mutlu, b M. Erdem, a I. Mutlu, c I. Guler. a a Gazi University Faculty
of Medicine, Ankara, Turkey; b Koru Hospital, Ankara, Turkey; c NovaArt
IVF Center, Ankara, Turkey.
OBJECTIVE: To compare in vitro fertilization (IVF) outcomes between
standard GnRH antagonist protocol and luteal phase estradiol/ GnRH antagonist
priming protocol in the poor responders.
DESIGN: Prospective randomized trial.
MATERIALS AND METHODS: One hundred and five poor responders
according to ESHRE Bologna criteria whose ages were between 25 to 45
were included to the study. Based on computer generated randomization
52 patients used luteal phase estradiol/ GnRH antagonist priming protocol
and 53 patients used standard GnRH antagonist protocol prospectively. Patients
with luteal phase estradiol/ GnRH antagonist priming protocol took
0.1 mg transdermal estradiol on alternate days for 3 times starting from
7th day after ovulation in the previous cycle. In addition, on the second
day after application of patch daily subcutaneous cetrorelix 0.25 mg was
started and continued for 3 days. Both of two groups underwent controlled
ovarian hiperstimulation starting on day 3 of menstural cycle with flexible
antagonist protocol Primary outcomes of the study were the number of oocytes
retrieved, clinical pregnancy, implantation and cycle cancellation rates.
Secondary outcomes were peak estradiol levels, the number of mature oocytes,
fertilization rates. Student’s t-test was used for continuous variables,
and the chi-square test was used for categorical variables.
RESULTS: The baseline characteristics as age, body mass index, antral
follicle count, basal FSH, basal E2, the number of prior IVF attempts and
duration of infertility were similar between the two groups. The mean number
of oocytes retrieved (3.7 2.8 vs. 3.8 2.8), the number of mature oocytes
(2.6 2.2 vs. 3 2.3), fertilization rates (61.1% 37.8% vs. 71%
31.3%) and the number of embryos transferred (1.6 0.6 vs. 1.7 0.6) were
also similar. The cancellation rate was not different between groups (15.1%
vs 11.1%). There were no significant differences between groups in terms of
the implantation (8.4% vs 8.8%), clinical pregnancy (13.2% vs. 13.3%) and
ongoing pregnancy rates (9.4% vs. 4.4%) per started cycle.
CONCLUSIONS: Luteal phase estradiol/ GnRH antagonist priming protocol
has no effect to improve the IVF outcomes in terms of improved cycle
cancellation and pregnancy outcome as compared to standard GnRH antagonist
protocol in patients with poor response to gonadotropins after IVF.
P-624 Wednesday, October 21, 2015
LUTEAL PHASE LUTEINIZING HORMONE (LH) LEVELS AFTER
GONADOTROPIN-RELEASING HORMONE AGONIST (GNRH-A)
TRIGGER AND PROBABILITY OF CONCEPTION. C. B. Bartels,
B. L. Maslow, E. Anspach, C. A. Benadiva, J. Nulsen, L. Engmann. University
of Connecticut Health Center, Farmington, CT.
OBJECTIVE: It has been proposed that low luteal phase luteinizing hormone
(LH) levels following gonadotropin-releasing hormone agonist
(GnRH-a) trigger induce abnormal corpus luteum function, which may lower
conception rates. This study aimed to evaluate whether luteal phase serum
LH profile predicts probability of conception after GnRH-a trigger.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: We included all women

continuous variables. Logistic regression models were built to test significant
findings in the univariate analysis.
RESULTS: 375 cycles were included in this study. 255 (68%) were
conception cycles. There were no significant differences in baseline characteristics
between conception and non-conception cycles. There was no significant
difference in use of GnRH-a alone and GnRH-a+HCG trigger in
conception compared to non-conception cycles (p¼0.55).Median LH levels
were 1.6, 65.3, 0.3, 0.1 and 0.2 on trigger +0, +1, +5, +9 and +16 days respectively.
There were no significant differences in median LH levels at any of the
time points between those who received GnRH-a alone and those who
received GnRH-a+HCG trigger. There were also no significant differences
between conception and non-conception cycles in any time points apart
from the day of the pregnancy test. The higher LH levels on the day of pregnancy
test remained significant in the regression model (p¼0.007), even
when controlling for type of trigger.
Luteal phase luteinizing hormone (LH) levels in conception and nonconception
cycles.
Time
Conception
Cycles
LH IU/L
(MedianIQR)
N¼ 255
Non-conception
Cycles
LH IU/L
(MedianIQR)
N¼ 120
p-value
Day of Trigger 1.62.1 1.51.7 0.56
Day after Trigger 67.250.5 63.249.7 0.68
Trigger + 5 days 0.30.4 0.30.4 0.83
Trigger + 9 days 0.10.3 0.10.2 0.85
Trigger + 16 days 0.20.3 0.10.1 0.001
CONCLUSIONS: Luteal phase LH levels are generally low in both
conception and non-conception cycles after GnRH-a trigger. This is the first
study to demonstrate that luteal phase LH levels following GnRH-a trigger do
not significantly predict the probability of conception and therefore its measurement
is not warranted.
P-625 Wednesday, October 21, 2015
WITHDRAWN
P-626 Wednesday, October 21, 2015
COMPARISON OF HORMONAL LEVELS IN POLYCYSTIC OVARY
PATIENTS USING SHORTAND LONG ANTAGONIST PROTOCOLS
DURING CONTROLLED OVARIAN STIMULATION IN IVF-ET
CYCLES. M. Thakur, a A. Bolnick, a O. Abuzeid, b R. Raju, c J. Dai, a
E. E. Puscheck, a M. P. Diamond, d M. I. Abuzeid. e a Division of Reproductive
Endocrinology and Infertility, Department of Obstetrics and Gynecology,
Wayne State University/ Detroit Medical Center, Detroit, MI; b OBGYN,
Dearborn, MI; c Hurley Medical Center/Michigan State University Co, Flint,
MI; d Georgia Regents University, Augusta, GA; e Hurley Medical Center,
Flint, MI.
OBJECTIVE: The usefulness of gonadotropin-releasing hormone (GnRH)
antagonist for the inhibition of premature luteinization during ovarian hyperstimulation
has been studied. GnRH antagonist has been shown to offer
increased safety contrasted with GnRH agonist cycles, with no significant
difference in birth rate. However some studies suggest that longer than 5
days of GnRH antagonists may have a negative impact on the endometrium
and in turn on pregnancy outcome. We set out to study the affect of different
timing of the initiation of GnRH antagonist on hormonal levels in patients
with polycystic ovary syndrome (PCOS) undergoing IVF. We also evaluated
effect of length of GnRH antagonist use on pregnancy outcomes.
DESIGN: Prospective trial (ISRCTN69937179).
MATERIALS AND METHODS: Patients with PCOS undergoing IVF
were prospectively randomized to Group 1, initiating GnRH-antagonist on
day 1 of stimulation and group 2, with GnRH antagonist initiated on day
5. Estradiol (E2) and progesterone (P4) were measured during follicular
phase on cycle day 2-3, 5-6, 7-8 and daily or every other day thereafter.
Serum E2 and P4 were measured on the day of embryo transfer and every
third day thereafter during luteal phase (1st, 2nd, 3rd sample). Analysis of
hormonal values was performed to determine differences in the groups based
on day of antagonist initiation and we also studied the effect of the length of
GnRH used on pregnancy outcomes.
RESULTS: 140 patients with PCOS were recruited with Group 1 (n¼69)
and Group 2 (n¼71). Embryo transfer (ET) was performed in 122 women.
Eighteen patients (9 in each group) were excluded as they did not have ET.
Mean days of GnRH-antagonist used in Group 1 was 10 2 days (range
7-16 days) and for Group 2 was 6 2 days (range 3-15 days) [p % 0.0]. There
were no differences noted in hormonal levels in either the follicular or luteal
phase regardless of the group. In addition there were no significant differences
in the clinical pregnancy rate (68.3 % vs 56.5%) and first trimester
miscarriage rate (7.3 % vs 8.6%) between Group 1 and Group 2, respectively.
CONCLUSIONS: Our data shows that amongst PCOS patients undergoing
IVF-ET there was no difference in hormonal levels during the follicular
and luteal phases regardless of when GnRH antagonist was started. Contrary
to previous reports we did not observe any negative impact on clinical pregnancy
rate in this pilot study.
P-627 Wednesday, October 21, 2015
THE EFFICACY AND SAFETY OF 100 MCG CORIFOLLITROPIN
ALFA IN ART CYCLES FOR PATIENTS WITH HIGH SERUM
ANTI-MULLERIAN HORMONE LEVELS. T. Lee, a,b H. Chen, b
C. Huang, b M. Lee. a,b a Department of Obstetrics and Gynecology, Chung
Shan Medical University Hospital, Taichung, Taiwan; b Department of Infertility
Clinic, Lee Women’s Hospital, Taichung, Taiwan.
e320 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

OBJECTIVE: The selection of 100mcg or 150mcg corifollitropin alfa in
IVF/ICSI cycles depends on the body weight of the patients with good prognosis.
A higher percentage of excessive ovarian response and consequent
cancellation is found in corifollitropin alfa compared to daily FSH with
GnRH antagonist protocols. However, the excessive ovarian response in corifollitropin
alfa protocol did not jeopardize the ongoing pregnancy rate and
might be predicted by serum anti-Mullerian hormone levels. This analysis
tried to prove the concept that 100mcg corifollitropin alfa is adequate for
all patients with a high AMH level (>3.5 ng/ml).
DESIGN: This is a nested case-control analysis of the medical records in a
prospective cohort trial.
MATERIALS AND METHODS: A total of 121 patients with an AMH
level >3.5 ng/ml were recruited into this analysis. Forty-nine patients
received 100mcg corifollitropin alfa and 72 patients underwent daily FSH
(150 IU-225 IU) in the initial 5 days. The main outcome was the oocyte
retrieval number, ongoing pregnancy rate, and ovarian hyperstimulation
rates.
RESULTS: The estradiol (23961181 vs. 30031864 pg/ml) and progesterone
levels (0.920.54 vs. 1.220.84 ng/ml) on the day of hCG injection
were significantly lower in corifollitropin alfa group compared to daily FSH
group. The oocyte retrieval number (13.36.3 vs. 14.37.5), fertilization
rates (74.715.6 vs. 74.719.1%), day3 good embryo rates (53.532.6
vs. 58.725.3%), and pregnancy rates (28/48¼58.3% vs. 31/70¼ 44.3%)
are similar between the two groups. The OHSS rate is 2.04% (1/49) and
2.78 (2/72) for corifollitropin alfa and daily FSH group, respectively. All
were moderate OHSS and paracentesis was not necessitated.
CONCLUSIONS: The 100mcg corifollitropin alfa is effective and safe in
IVF/ICSI cycles for patients with high AMH levels. Further studies are
needed to investigate the efficacy of 100mcg corifollitropin alfa for patients
with weight above 90 Kg.
References:
1. Fatemi HM, Doody K,Griesinger G, et al., High ovarian response does
not jeopardize ongoing pregnancy rates and increases cumulative pregnancy
rates in a GnRH-antagonist protocol. Hum Reprod, 2013;28:442-
452.
2. Polyzos NP, Tournaye T, Guzman L, et al., Predictors of ovarian
response in women treated with corifollitropin alfa for in vitro fertilization/
intracytoplasmic sperm injection. Fertil Steril, 2013;100: 430-
437.
Supported by: Grant from Chung Shan Medical University Hospital CSH-
2014-C-012.
P-628 Wednesday, October 21, 2015
MINIMAL STIMULATION IN VITRO FERTILIZATION (IVF) WITH
A FREEZE-ALL APPROACH GIVES ACCEPTABLE LIVE BIRTH
RATES IN AN OVERALL POOR PROGNOSIS
POPULATION. B. G. Reed, M. Ezzati, S. N. Babayev, V. Libby,
B. Carr, O. Bukulmez. UT Southwestern Medical Center, Dallas, TX.
OBJECTIVE: Minimal stimulation IVF typically utilizes daily clomiphene
citrate and 3 low doses of gonadotropin injections on stimulation
days 5, 7, and 9. Minimal stimulation cycles are planned with the intention
to freeze all available embryos exclusively at blastocyst stage. After reasonable
embryo accumulation, frozen embryo transfer (FET) is performed.
There is a paucity of published data on the outcomes of minimal stimulation
IVF with a freeze-all protocol in patients with diminished ovarian reserve
(DOR) and/or a prior history of traditional IVF failures.
DESIGN: Retrospective observational study.
MATERIALS AND METHODS: Institutional Review Board approval
was obtained and the data for all initiated minimal stimulation IVF cycles between
2012-2014 was entered into a database. Patient and cycle characteristics,
pregnancy and live birth data were reviewed and analyzed.
RESULTS: A total of 54 patients initiated 147 cycles (102 stimulations,
and 45 FETs). Age, body mass index (BMI), and antimullerian hormone
(AMH) levels and prior history of failed traditional IVF attempts were suggestive
of an overall poor prognosis group (Table 1). Cancellation rates for
minimal stimulation and FET were 24% and 22% respectively. Stimulation
cycle cancellation reasons included premature ovulation (13), poor response
(8), early follicular growth (1), and non-compliance (2). FET cycle cancellations
were mostly due to either inadequate endometrial development (6) or
inability to be suppressed with a GnRH agonist (3). The methods of fertilization
were conventional IVF in 46 cycles and ICSI in 35 cycles. Despite being
a poor prognosis population and having a high cancellation rate, the cumulative
live birth rate per patient was 24%. The live birth rate per FET was 37%.
Of the women who have not yet achieved a live birth, 32% still have remaining
embryos to transfer. Among many cycle parameters, female age was a
strong predictor of obtaining at least one frozen blastocyst.
CONCLUSIONS: Since there is no data available on minimal stimulation
using a freeze-all approach in poor prognosis patients, this information is
important for patient counseling. Minimal stimulation cycles for embryo
accumulation at blastocyst stage result in acceptable live birth rates in a
poor prognosis population.
Table 1: Demographics and clinical outcome data in completed minimal
stimulation cycles.
Values are expressed
as either percentage
or meanSD. Range
Age (y) 37.894.1 (28-45)
BMI 25.954.6 (18.1-38.4)
AMH (ng/ml) 0.550.67 (0-3)
Patients with prior unsuccessful 44 % (24/54) -
traditional IVF
Number of retrievals per patient 1.690.99 0-4
Number of FETs per patient 0.720.63 0-2
Biochemical pregnancy rate 60% (21/35) -
per FET
Clinical pregnancy rate per FET 48.6% (17/35) -
Live birth rate per FET 37% (13/35) -
Cumulative live birth rate
per patient
24% (13/54) -
P-629 Wednesday, October 21, 2015
VERY LOW LEVEL OF ANTI-MULLERIAN HORMONE (AMH) IN
A LARGE COHORT OF ASSISTED REPRODUCTIVE TECHNOL-
OGY (ART) PATIENTS. J. Rodriguez-Purata, a M. Luna, a
E. Cervantes, a J. A. Lee, a M. C. Whitehouse, a A. B. Copperman, b
B. Sandler. a a Reproductive Medicine Associates of New York, New York,
NY; b RMANY-Mount Sinai, New York, NY.
OBJECTIVE: The counseling and management of women with low AMH
levels presents a significant challenge where either cycle cancellation or poor
response is foreseen. Prior clinical impression suggests that an extremely low
level of AMH can serve as an alert for a clinician to dissuade a patient from
utilizing certain fertility treatments. The aim of this study was to evaluate
ART laboratory outcomes from patients with extremely low AMH levels.
DESIGN: Retrospective.
MATERIALS AND METHODS: All patients scheduled for a fresh IVF
cycle with ‘‘extremely low’’ (Group A: %0.2) or ‘‘low’’ AMH levels (Group
B: 0.2 - %0.5; Group C: %1 ng/ml) between January 2009 to March 2015
were included. Main outcome measures were total retrieved oocytes and cycle
cancellation rates. Secondary outcomes analyzed were age, day 3 FSH,
basal AFC, number of follicles greater than 14mm at surge, peak estradiol,
cumulative amount of gonadotropins (GND) and days of stimulation.
RESULTS: Five hundred and forty four patients underwent 1647 cycles
(Table 1). The rate of cycle cancellation prior to VOR was significantly
higher in Group A patients (44.2%) when compared to Group B and C patients
(20.8%; 10.0%). In patients that reached VOR, the number of oocytes
retrieved per cycle was significantly lower in Group A (5.13.0) compared to
Groups B and C (6.43.5; 8.64.7). All secondary variables were statistically
different between groups except the average days of stimulation
required.
CONCLUSIONS: AMH is one of the best available tools for the detection
of low ovarian reserve and its role as a clinical test is robust. To our knowledge,
this is one of the largest studies to examine laboratory outcomes in
extremely low AMH level patients. The results demonstrated that patients
with very low AMH levels require a higher amount of gonadotropins than patients
with merely low levels. Although clinicians should inform patients the
potential adverse realities of seeking treatment, our study demonstrated that
patients should not be excluded from pursing an IVF cycle as they still
respond, fairly, to stimulation. With further research, a more personalized
stimulated treatment regimen incorporated genomic and phenotypic variables
will lead to accurate prognostic information and optimal treatment strategies.
FERTILITY & STERILITY Ò
e321

Outcomes in patients with Low and Very Low AMH levels
ANOVA Chi SquareA vs. B Chi SquareA vs. C
Cycles 437 557 653 1647
Age 38.54.5 39.04.0 38.53.9 NS NS NS
Day 3 FSH 8.87.4 7.54.7 7.64.0 p

d Calculated as the number of 2 pronuclei oocytes divided by the total number
of oocytes retrieved.
e Visit 8 corresponds to Day 14 +/- 2 days after embryo transfer.
f Live birth data were collected by each study site after the end-of-study visit.
g Calculated using the total number of neonates for the denominator.
Supported by: This study was sponsored by Ferring Pharmaceuticals, Inc.
P-632 Wednesday, October 21, 2015
EGG YIELD VS. EGG QUALITY. C. Chang, J. M. Linn, D. B. Shapiro,
A. A. Toledo, M. W. Best, Z. Nagy. Reproductive Biology Associates, Atlanta,
GA.
OBJECTIVE: To assess the correlation between donor oocyte yield and
oocyte quality using a cryo-banking model.
DESIGN: Retrospective study.
MATERIALS AND METHODS: Cryopreservation of donor oocytes
derived from 543 egg retrievals was performed using minimum volume vitrification.
All donors were treated with recFSH/GnRH antagonist/GnRH
agonist trigger. Oocytes were warmed for each matched recipient independently
(1334 recipient cycles, total of 8344 oocytes warmed). Outcome
data was grouped by oocyte yield; I)

TRANSCRIPTIONAL COMPARISON OF NATURAL AND STIMULATED
IVF CYCLES.
NATURAL
CYCLE
ANTAGONIST
CYCLE
AGONIST
CYCLE
p-value
Age 33.12.5 34.23.4 32.74.5 a:

P-638 Wednesday, October 21, 2015
THE USE OF COENZYME Q10 AND DHEA DURING COH AND IVF
CYCLES IN PATIENTS WITH DECREASED OVARIAN RESERVE
(DOR). I. Gat, a S. Blanco Mejia, b H. Balakier, a C. L. Librach, a
A. Claessens, b E. A. J. Ryan. b a Create Fertility Centre, Toronto, ON, Canada;
b Toronto West Fertility Center, Etobicoke, ON, Canada.
OBJECTIVE: Treatment of patients with diminished ovarian reserve
(DOR) is one of the biggest challenges in assisted reproductive therapeutics.
Dehydroepiandrosterone (DHEA) and Coenzyme Q10 (CoQ10) are supplements
that have been purported to have a beneficial effect in these patients.
Our objective was to compare the effect of combining DHEA and CoQ10
supplementation with DHEA alone in COH and IVF cycles in patients
with DOR.
DESIGN: Clinical retrospective study.
MATERIALS AND METHODS: We extracted data from patients charts
treated by DHEA (25mg tid) with/without CoQ10 (600mg daily) in a private
infertility clinic between Feb. 2006 to June 2014. Pre-stimulation parameters
analyzed included age, BMI, day 3 FSH and antral follicular count (AFC).
Ovarian response parameters analyzed included total dose of gonadotropins,
peak serum estradiol (E2), follicles number > 16 mm on day of triggering
and fertilization rate for IVF cycles. Clinical outcomes analyzed included
clinical and ongoing pregnancy rates per cycle initiated.
RESULTS: 797 COH cycles and 253 IVF cycles were included. Of these,
330 COH cycles involved both DHEA and CoQ10 (D+C) and 467 cycles of
DHEA (D) alone; 78 IVF cycles involved D+C and 175 D. In both COH and
IVF groups, AFC was significantly higher with D+C compared to D alone
(7.45.7 vs. 5.94.7 and 8.26.3 vs. 5.25, respectively, p 16 mm on the
trigger day during COH cycles (3.32.3 vs. 2.92.2, respectively,
p¼0.01), but no difference was observed in IVF cycles (5.53.5
vs.5.94.6, p¼0.4). There was lower gonadotropin consumption during
D+C IVF cycles compared with D cycles (3,4141141 IUs vs.
3,8771143 IUs respectively, p¼0.032) without difference in the COH comparison.
For COH cycles there was no difference in clinical or ongoing pregnancy
rates in the D+C vs. D groups (8.1 vs. 10.9, p¼0.2 and 6.2 vs. 8.2,
p¼0.3, respectively). Similarly, there was no difference regarding clinical
or ongoing pregnancy rates in the D+C vs. D groups in the IVF cycles
(25.1 vs. 29.5, p¼0.5 and 21.1 vs. 23.1, p¼0.7, respectively).
CONCLUSIONS: Combined DHEA and CoQ10 supplementation significantly
increases the AFC compared to DHEA alone, which lead to a higher
ovarian responsiveness during both COH and IVF, but without a difference in
pregnancy rate.
P-639 Wednesday, October 21, 2015
EARLY GNRH-ANTAGONIST INITIATION DOES NOT IMPROVE
PREGNANCY RATES OF PCOS CASES UNDERGOING IVF/ICSI:
A PROSPECTIVE RANDOMIZED STUDY. C. S. Atabekoglu, a
E. Kocbulut, a E. G. Pabuccu, b B. Ozmen, a B. Berker, a M. Sonmezer. a a Obstetrics
and Gynecology, Ankara University School of Medicine, Ankara,
Turkey; b Ufuk University School of Medicine, Ankara, Turkey.
OBJECTIVE: Is to assess the effect of early GnRH antagonist initiation on
the hormonal environment and pregnancy outcomes of Polycystic Ovary
Syndrome (PCOS) patients undergoing in vitro fertilization cycles due to unexplained
infertility.
DESIGN: Prospective randomized, pilot study.
MATERIALS AND METHODS: Twenty-four women participated in a
prospective randomized trial. Diagnosis of PCOS was based on Rotterdam
Criteria (1). Inclusion criteria were: age < 38 year, no more than two previous
failed IVF attempts, body-mass index between 18-29 kg/m2, no endometriosis
or previous poor response to ovarian stimulation. Following 1 month
of oral contraceptive pill usage, all participants were stimulated with daily
recombinant FSH at doses ranging from 150 IU to maximal of 200 IU according
to ovarian reserve and/or body mass index values, starting on day 2 of the
cycle. Patients received GnRH-antagonist, starting either on day 2 (n ¼ 12,
early antagonist group) or on day 6 of stimulation (n ¼ 12, flexible antagonist
group). Primary outcome was to compare the serum progesteron (P) levels on
the day of hCG administration between groups. Secondary outcome was to
compare implantation and clinical pregnancy rates.
RESULTS: Demographic characteristics of the groups were similar in
terms of age, BMI, previous history and ovarian reserve. Serum P concentrations
were not differed among groups both during early follicular phase and
on the day of hCG. Other outcome measures including mature oocyte yield,
fertilization-implantation and clinical pregnancy rates were also comperable
among groups (p

with 1,2 versus 3,0, and the luteal phase discomfort was rated as 1.2 vs. 2.5
respectively with the patient friendly protocol (corifollitropin alpha+GnRHagonist-triggering)
as compared with the conventional protocol (recFSH+hCGtriggering)
(from 1- low, to 5 - severe).
CONCLUSIONS: This is the first study to compare two different stimulation
regimens in the same donor to avoid inter-patient variation. Taking into
account the low risk of ovarian hyperstimulation syndrome (OHSS), the
lower degree of physical distress during the luteal phase as well as the similar
pregnancy rates and efficacy, the use of corifollitropin alpha combined with
GnRH-agonist for triggering appears to be the ideal standard in oocyte donors.
P-641 Wednesday, October 21, 2015
LOWER MULTIPLE PREGNANCY RATES WITH LETROZOLE
VERSUS CLOMIPHENE IN 16,001 IUI CYCLES OVER 10
YEARS. J. Toner. Atlanta Center for Reproductive Medicine, Atlanta, GA.
OBJECTIVE: To evaluate whether pregnancy and multiple pregnancy outcomes
are different in cycles stimulated with letrozole versus clomiphene citrate.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All 16,001 IUI cycles at our clinic using
letrozole or clomiphene citrate for ovulation induction or enhancement from
2004 to 2014 were included. Cycles with concomitant gonadotropin use were
excluded. hCG to trigger ovulation was optional. Cycles were subdivided by
infertility diagnosis and female age.
RESULTS: Over all diagnoses and female ages, pregnancy rates were
slightly higher with clomiphene than letrozole (17.4% vs. 15.1%), but multiple
pregnancy rates were significantly higher with clomiphene (twins/preg:
7.5% vs. 4.7%; triplets/preg: 1.3% vs. 0.2%). These effects were also seen
across diagnoses: PCO, male, pelvic, endometriosis, unexplained.Unexpectedly,
clomiphene’s higher pregnancy rates relative to letrozole vanished over
age 40, where letrozole had a small advantage (8.2% vs. 5.6%).
CONCLUSIONS: Our large experience with both letrozole and clomiphene
for many types of infertility patients suggests that letrozole produces
fewer multiple pregnancy rates and more pregnancies over age 40 relative to
clomiphene.
P-642 Wednesday, October 21, 2015
COMPARISION OF LUTEAL ESTRADIOL PATCH AND ORAL
CONTRACEPTIVE PILLS IN ANTAGONIST CYCLES FOR PRE-
TREATMENT OF THE NORMAL RESPONDER. N. Pereira, a
A. C. Petrini, b J. Lekovich, a I. Kligman, b Z. Rosenwaks. b a The Ronald
O. Perelman and Claudia Cohen Center for Reproductive Medicine, New
York, NY; b Weill Cornell Medical College, New York, NY.
OBJECTIVE: To investigate the impact of pre-treatment with luteal phase
estradiol (E 2 ) patch compared to both combined oral contraceptive pills
(OCPs) and no pretreatment on ovarian stimulation response in patients undergoing
fresh in vitro fertilization (IVF) - embryo transfer (ET) cycles.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: All patients undergoing fresh IVF-ET
cycles between January 2008 and June 2013 at our center were analyzed
for inclusion. Exclusion criteria included polycystic ovarian syndrome,
oocyte donors, age > 40 and known poor response to ovarian stimulation.
Demographic characteristics recorded were age, gravidity, parity, BMI (kg/
m 2 ), and infertility diagnosis. Ovarian stimulation parameters recorded
were as follows: total days of ovarian stimulation, total dosage of gonadotropins
administered (IU), peak estradiol (E 2 ) level (pg/mL), peak endometrial
stripe (mm), total number of oocytes retrieved, and total number of mature
oocytes. For each ET cycle, the total number of embryos transferred, clinical
pregnancy rate, biochemical pregnancy rate, spontaneous miscarriage rate,
and live birth rate was recorded. Student’s t-tests and Chi-square tests were
used to compare means and percentages, respectively. Statistical significance
was set at P < 0.05.
RESULTS: 4080 patients met inclusion criteria. There was no difference
in the baseline demographics of the two groups. Differences in the ovarian
stimulation parameters are highlighted in the table below. Patients in the
OCP group underwent ovarian stimulation longer than the E2 patch group
i.e.,10.7 (1.63) days vs. 9.92 (1.94) days (P < 0.001). As a result, the
former group required higher doses of gonadotropins (2750.6 1297.1 IU)
compared to the latter group (2550.1 1270.2 IU; P¼0.003). Patients in
the OCP group also had higher E2 levels on the day of and the day after
hCG trigger comapred to the E2 patch group. There were no differences in
pregnancy outcomes after ET when comparing all three groups.
CONCLUSIONS: Compared to E 2 patch pre-treatement, OCPs increase
the total stimulation days, total gonadotropins administered, E2 level on
the day of trigger, while slightly decreasing the total number of mature oocytes
in the normal responder. The overall clinical pregnancy and live birth
rates remain unaffected by E 2 patch, OCP or no pretreatment.
P1: E2 patch-Ant vs. OCP-Ant; P2: E2 patch-Ant vs. No E2 patch/OCP-Ant.
Parameter
E2 Patch-Ant
(n¼1057)
OCP-Ant
(n¼1035)
No E2/OCP-Ant
(n¼1988) P1 P2
Age (years) 36.1 (2.73) 35.9 (3.91) 36.3 (3.08) 0.15 0.08
Basal AMH (ng/mL) 0.98 (0.13) 0.97 (0.17) 1.00 (0.14) 0.41 0.73
Total stimulation days 9.92 (1.94) 10.7 (1.63) 9.89 (2.03)

CONCLUSIONS: BRCA status does not appear to correlate with response
to controlled ovarian stimulation, oocyte yield, and cancellation rate in patients
undergoing ovarian stimulation. Prospective investigation in a larger
cohort is needed.
P-644 Wednesday, October 21, 2015
THE TIMING OF ADMINISTRATION OF LETROZOLE SIGNIFI-
CANTLY AFFECTS THE OOCYTE RECOVERY RATE IN BREAST
CANCER PATIENTS UNDERGOING CONTROLLED OVARIAN
STIMULATION FOR FERTILITY PRESERVATION. C. Diaz-Garcia,
a J. Domingo, b A. Romero, a M. Martinez, c J. M. Rubio, d
J. A. Garcia-Velasco, c A. Pellicer. a,e a Woman’s Health Area, La Fe University
Hospital, Valencia, Spain; b IVI-Las Palmas, Las Palmas de Gran Canaria,
Spain; c IVI Madrid, Madrid, Spain; d La Fe University Hospital, Valencia,
Spain; e IVI-Valencia, Valencia, Spain.
OBJECTIVE: Diminished ovarian response to controlled ovarian stimulation
(COH) in breast cancer (BC) patients has been reported by different authors,
although others do not find such differences. COH protocols in patients
suffering from BC have some distinctive features: aromatase inhibitors are
commonly used to avoid an increase in estradiol (E2) levels. To what extent
this is a consequence of using modified stimulation protocols remains unclear.
The aim of this study was to quantify the effect of two different protocols
of COH on the number of oocytes retrieved in BC patients undergoing
COH for fertility preservation.
DESIGN: Observational multicentric prospective cohort study.
MATERIALS AND METHODS: Patients were allocated to receive two
different stimulation protocols: group A-patients started COH using letrozole
5mg/d and hrFSH was added from the third day of stimulation.
Group B-patients started COH using hrFSH and letrozole was added
from the third day of stimulation to prevent the rise of estradiol (E2)
levels. All patients started stimulation during the early follicular phase,
GnRH antagonist was administered when the leading follicle reached
14 mm, and final oocyte maturation was triggered with 0.2 mg of
GnRH agonist. E2 levels the day of triggering, the total number of oocytes
retrieved and the proportions of MII oocytes were compared between
groups. A GLM-based multivariate regression analysis was
applied to control for potential confounders.
RESULTS: Patients in group A (n¼44) and group B (n¼257) had similar
age and antral follicle count (AFC). Duration of stimulation was similar between
groups. Patients in group A yielded higher estradiol level the day of
ovulation triggering and higher numbers of total and MII oocytes retrieved
(Table 1) when compared to patients in group B. The mean differences in
oocytes retrieved (5.2-95%CI: 2.5-7.9) and MII oocytes retrieved (2.9-95%
CI: 0.9-5.0) remained significant after adjustment for doses of rhFSH used
during stimulation (total oocytes: 4.7-95%CI: 1.9-7.5 and MII: 2.5-95%CI:
0.4-4.7).
CONCLUSIONS: The use of letrozole before administration of rhFSH decreases
the number of mature oocytes retrieved after COH in BC patients undergoing
fertility preservation. On the other hand, the estradiol pick at
ovulation triggering is higher when letrozole is administered after rhFSH.
The total dose of rhFSH used during COH acts and effect modifier.
Baseline Characteristics of the patients and COH results.
Group A Group B p-value
Age (years) 32.6 (3.4) 33.7 (3.9) n.s.
AMH (pM)
27.0 (17.0-43.0) 25.7 (13.9-35.3) n.s.
Duration of COH (days) 9.3 (1.5) 9.3 (3.8) n.s.
Starting dose of rhRSH (IU) 248.3 (59.2) 173.1 (82.0) p

analysis in integrating available literature evidence to support dose selection
of recFSH.
Simulated RecFSH dose-response at clinical relevant doses based on the model
based meta analysis.
recFSH Dose 100 IU 150 IU 200 IU 250 IU 300 IU
Predicted Mean 7.16 9.99 10.6 10.83 10.94
Oocytes Count
(95 % CI)
(4.7 – 8.3) (8.8 – 10.7) (9.7 – 11.3) (9.9 – 11.5) (9.9 – 11.7)
References: Table Note: 10,000 simulations conducted incorporating
parameter uncertainty.
Supported by: Merck & Co., Inc., Kenilworth, NJ, USA.
P-647 Wednesday, October 21, 2015
LUTEAL-PHASE OVARIAN STIMULATION VERSUS CONVEN-
TIONAL OVARIAN STIMUATION IN PATIENTS WITH NORMAL
OVARIAN RESERVE TREATED FOR IVF: A LARGE RETROSPEC-
TIVE COHORT STUDY. N. Wang, a Y. Wang, b Y. Kuang. c a Department
of Assisted Reproduction, Shanghai Ninth People’s Hospital, Shanghai Jiaotong
University School of Medicine, Shanghai, China; b Reproductive Endocrinology,
Shanghai, China; c Shanghai Ninth People’s Hospital, Shanghai,
China.
OBJECTIVE: To systematically assess the efficiency and security of the
luteal-phase ovarian stimulation (LPS) strategy for infertility treatment by
comparing with conventional ovarian stimulation protocols.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients with normal ovarian reserve
taking ovum pick-up (OPU) cycles between April 2012 and September
2013 from three ovarian stimulation protocols were enrolled (727 cycles
from the LPS protocol, corresponding to 708 patients; 830 cycles from the
mild treatment protocol, corresponding to 745 patients, and 1385 cycles
from the short-term protocol, corresponding to 1287 patients). Main outcomes
were number of mature oocytes retrieved, number of top-quality embryos
obtained, implantation rate, pregnancy rate, live birth and ongoing
pregnancy rate, and the neonatal outcomes.
RESULTS: Compared with mild treatment protocol, the LPS was associated
with higher number of mature oocytes retrieved per OPU cycle (10.97.6 vs.
3.73.0, P0.05), live birth and
ongoing pregnancy rate (44.4% vs. 41.7%, P>0.05) in the LPS and mild treatment
protocols, respectively. Comparisons between the LPS protocol and
short-term protocol indicated that the LPS could achieve more mature oocytes
and top-quality embryos per cycle (10.97.6 vs. 9.15.5, 4.64.3 vs.
3.73.1, respectively, both P
95%th percentile for the study cohort i.e., > 13 days. Patients with history
of poor response to ovarian stimulation or with PCOS were excluded. Non
GnRH-antagonist-based cycles were excluded. Demographic characteristics
included age, gravidity, parity, body mass index (kg/m 2 ) and infertility diagnosis.
COS parameters recorded were total days of GnRH-antagonist administration,
total dosage of gonadotropins administered (IU), peak E 2 level (pg/
e328 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

mL), number of mature oocytes retrieved, fertilization rate (%) number of
embryos transferred, and day of ET. Clinical pregnancy, biochemical pregnancy,
spontaneous miscarriage, and live birth rates in patients undergoing
COS < 13 days and > 13 days were estimated. Wilcoxon rank sum test
and student’s t-test were utilized for continuous variables. Chi-square (c2)
test with Mantzel-Hansel correction was used for categorical variables.
Odds ratios (OR) with 95% confidence intervals (CI) for live birth were
calculated. Statistical significance was set at P 13 days, respectively.
Overall, there were no differences in demographic characteristics. Patients in
the > 13 day group had more total antagonist days and total gonadotropins
administered. COS < 13 days was associated with increased odds of clinical
pregnancy 1.76 (95% CI 1.33 - 2.34) and live birth 1.70 (95% CI 1.33 - 2.34),
which remained unchanged when adjusting for total antagonist days and total
gonadotropins administered. Patients in the > 13 day group who had live
births were younger (34.6 4.91 vs. 38.2 4.72; P < 0.001).
CONCLUSIONS: Our findings suggest that COS < 13 days is associated
with increased odds of clinical pregnancy and live birth. In patients undergoing
COS > 13 days, younger age is associated with greater odds of live birth.
Comparison of Demographics, COS response and IVF-ET outcomes.
Parameter >13 days (n¼240) P
Age (years) 37.6 (4.57) 37.5 (4.99) 0.67
BMI (kg/m2) 23.0 (6.32) 23.4 (6.68) 0.34
Total antagonist days 4.14 (1.44) 5.85 (2.34)

OBJECTIVE: Women of reproductive age with DOR may have regular
menses but respond poorly to stimulation and/or have suboptimal fecundity
compared to those of similar age. We sought to evaluate DOR patients
engaged in IVF treated with either a luteal estradiol/gonadotropin-releasing
hormone antagonist (antGnRH) (Estradiol-Priming protocol (EPP)) or with
an oral contraceptive pill (OCP) daily micro-dose leuprolide acetate (Microflare)
protocol (MFP). Our goal was to use age, baseline hormone values,
number of injections, and outcome to optimize choice of ovarian stimulation
protocol.
DESIGN: Retrospective.
MATERIALS AND METHODS: Patients who were identified as having
DOR who underwent an IVF utilizing a MFP or an EPP were included.
DOR was defined by: 1) history a canceled IVF; 2) poor response to stimulation
(

significantly higher than for the isolated markers (age ¼ 0.67, AFC ¼ 0.74,
AMH ¼ 0.75, FSH ¼ 0.61). The cut-off values and false positive rates for
obtaining detection rates of 50% and 80% are presented in Table 1.
CONCLUSIONS: The formula combining age, FSH, AMH, and AFC is
more accurate in predicting POR than individual markers. More studies are
still needed before recommending its use in clinical practice.
CONCLUSIONS: IVF protocols that include letrozole yield comparable
fertilization rates and an improved oocyte yield in BRCA-positive women.
The addition of letrozole to the BRCA-positive patient’s IVF protocol should
be considered regardless of cancer or estrogen-receptor status. Further
studies are needed to determine whether these findings hold true in a larger
cohort and to investigate pregnancy outcomes.
Cut-off values and false positive rates (FPR) to predict poor ovarian response.
Detection rate ¼ 50% Detection rate ¼ 80%
Cut-off FPR Cut-off FPR
Age R35 22% R33 57%
AFC %4 13% %8 42%
AMH %1,0 18% %1,5 43%
FSH R7,0 24% R4,3 72%
Formula R0,5 10% R0,37 27%
P-655 Wednesday, October 21, 2015
LETROZOLE PROTOCOLS ARE ASSOCIATED WITH IMPROVED
OOCYTE YIELD IN BRCA POSITIVE PATIENTS. A. P. Melnick,
E. M. Murphy, G. L. Schattman, Z. Rosenwaks. The Ronald O. Perelman
and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medical
College, New York, NY.
OBJECTIVE: To determine whether letrozole-based protocols improve
ovarian response in BRCA-positive women.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients with BRCA mutations undergoing
IVF for fertility preservation (breast cancer patients) or preimplantation genetic
diagnosis (no history of cancer) were included. Patients with breast cancer
who had received prior chemotherapy were excluded. Patients were stratified
into two groups: those who received letrozole continuously throughout their cycle
and those who received conventional GnRH agonist- or antagonist-based
protocols without the addition of letrozole. Outcomes assessed included total
and mature oocyte yield, oocyte maturity rate, fertilization rate, and cancellation
rate. Statistical analysis included student’s t-test, chi-square and Fisher’s
exact tests. P

EMBRYO TRANSFER
Treatment outcomes.
P-657 Wednesday, October 21, 2015
G-HRT
protocol
Previous
protocol
P-value
PITUITARY SUPPRESSION BEFORE FROZEN EMBRYO TRANS-
FER BENEFICIAL FOR THE IDIOPATHIC REPEATED IMPLAN-
TATION FAILURE PATIENTS. X. Yang, R. Huang, X. Liang.
Reproductive Medicine Centre, The Sixth Affiliated Hosptial of Sun Yat-
Sen University, Guangzhou, China.
OBJECTIVE: We were interested in whether long-term GnRHa pretreatment
combine with hormone replacement therapy (G-HRT)could improve
pregnancy outcomes in idiopathic repeated implantation failure(RIF) patients.
DESIGN: 18 idiopathic RIF patients undertaken G-HRT protocol were
included in this retrospective self-control study, compared clinical data
with their 49 previous embryo transfer cycles.
MATERIALS AND METHODS: 18 idiopathic RIF patients undertaken
G-HRT Protocol. The first injection, leuproreline acetate 2.5 mg i.m.,
was administered during the early follicular phase of the menstrual cycle
for G-HRT patients. Twenty eight days later, 1.875 mg of leuproreline acetate
was administered, and 15 days later the HRT protocol was started.
Estradiol valerate was commenced orally for at least 10 days, progesterone
intramuscular injection 40mg twice daily were added when the endometrial
thickness is > 7mm. FETwas performed on the fourth day of administration
of progesterone for day 3 embryo transfer or on the sixth day for blastocyst
embryo transfer. The same doses of estrogen and progesterone were
continued until a serum beta human chorionic gonadotropin(hcg) assay
was performed 14 days after FET. If the assay was positive, HRT was
continued until 10 weeks of gestation, and the patient was followed with ultrasonography
to determine fetal viability until approximately 7 weeks of
gestation. Pregnancy rate was defined by positive assay of hcg 14 days after
FET. The Clinical pregnancy was defined by the presence of an gestational
sac and a live fetus on TVS at 7 weeks of gestation. Implantation rate was
defined as number of gestational sacs observed divided by the number of
embryos transferred. In their previous cycles, HRT was performed identically
as above description but without GnRHa pretreatment. Or embryos
were transferred after ovulation with appropriate luteal support in Natural
cycle.
RESULTS: The data showed that after RIF, patients with advanced age
result in significantly promotion of reproductive outcome in G-HRT protocol,
the pregnancy rate, clinical pregnancy rate , implantation rate and
on-going pregnancy rate were 0.67,0.61,0.44 and 0.33, compared with
same parameters in their previous cycle were 0.17,0.11,0.07 and0.04,respectively.
Similar number of embryo were transferred with same endometrium
thickness.
CONCLUSIONS: Hormonally controlled endometrial preparation with
prior GnRHa suppression could be used for patients who have had RIF of
IVF-ET treatment despite having morphologically optimal embryos and
may be useful for increasing receptivity of the endometrium in these patients.
Endometrium thickness on 10.840.52 11.390.35 0.42
progesterone day (mm)
E2 level on progesterone 148.214.87 759.8142.7 0.0037
administration day
(pg/ml)
p Level on progesterone 19.312.58 40.554.93 0.0042
administration day
(ng/ml)
Mean No. of embryo 1.780.13 2.040.10 0.15
transferred
Pregnancy rate 0.67 0.117 0.167 0.0537 0.0012
Implantation rate 0.6111 0.12 0.1163 0.049 < 0.0001
Clinical pregnancy rate 0.44 0.098 0.071 0.033

quality (n¼78), were more likely to be euploid than non-freeze quality blastocysts
(n¼369) (64.1% vs., 43.0%, respectively; P

hatching with a differentiated inner cell mass and trophectoderm. All blastocysts
were collapsed prior to freezing. Vitrifcation and warming were performed
using Irvine Scientific freeze and thaw kits and Cryolocks. Patients
who used donor eggs were excluded from this study. The outcomes of interest
were positive hCG and positive clinical pregnancy rates (defined as the presence
of fetal activity at 7 weeks).
RESULTS: A total of 253 of the 354 (71%) FET patients had a positive
hCG compared to 61 of the 94 (67%) of euploid FET patients. Of the FET
patients, 182 (of 354) developed fetal cardiac activity at their 7 week ultrasound,
resulting in 51% clinical pregnancy rates. In comparison, 35 (of 94)
patients who transferred euploid embryos developed fetal cardiac activity, resulting
in 53% clinical pregnancy rate. Statistical analysis by t-Test demonstrated
no significant difference between standard frozen embryo transfer and
transferring frozen euploid embryos.
CONCLUSIONS: Vitrified blastocyst that reach freeze quality have no
significant difference in positive hCG or clinical pregnancies compared
to transferring vitrified euploid embryos. These results may be indicative
that biopsy may impact the ability of the embryo to implant or develop.
Also, the criteria of the embryos that are biopsied is not as consistent as
what is classified as ‘‘freeze quality’’. This flexibility in grading may be
detrimental to the embryo’s ability to survive the biopsy and subsequent
transfer.
P-662 Wednesday, October 21, 2015
AVOIDING CANCELLATION OF EMBRYO TRANSFER: EM-
BRYOS WITH POOR MORPHOLOGY ON DAY 5 YIELD PREG-
NANCIES AND LIFE BIRTHS. G. Koustas, H. Smith, C. Sjoblom.
Obtetrics and Gynaecology, University of Sydney/ Westmead Fertility
Centre, Sydney, Australia.
OBJECTIVE: The objective of this study was to assess the occurrence
and outcome of clinical pregnancies and live birth following transfer of
embryos on day 5 of development with deviant morphological parameters.
DESIGN: Retrospective analysis of fresh cycles with resulting poor quality
embryos on day 5 for single embryo transfer (SET) and double embryo
transfer (DET) between January 2010 and January 2015.
MATERIALS AND METHODS: This study included patients having conventional
IVF and/or ICSI in a total of 5019 fresh non-donor transfer cycles
conducted from January 2010 to January 2015. Embryos were assessed on
day 5 of development based on morphology. A poor embryo was identified
as having inadequate compaction and/or poor or absent inner cell mass
(ICM) and trophectoderm (TE), in accordance with the grading system
defined by Gardner and Schoolcraft (1999).
RESULTS: Out of 5019 fresh cycles, we identified 367 cycles where the
embryos transferred had been assessed as poor. There was a significant difference
in age between patients with poor quality embryos compared to
the overall average age of patients during this period. The proportion of
IVF/ ICSI and number of embryos transferred was found to be significantly
higher in the poor embryo group (P

among women aged 35-39 allows the physician to choose between either
fresh or FET according the patient’s needs. A very low ongoing pregnancy
rate in women R40 suggests that donor oocytes cycles should be highly recommended
for this age group.
P-664 Wednesday, October 21, 2015
DOES THE DEGREE OF EXPANSION AT TIME OF FROZEN EM-
BRYO TRANSFER (FET) AFFECT OUTCOMES OF SINGLE
THAWED EUPLOID EMBRYO TRANSFERS
(STEET)? N. M. Sachdev, a D. H. McCulloh, b J. Grifo. c a Obstetrics and
Gynecology, New York University Langone Medical Center Fertili, New
York, NY; b New York University Fertility Center, New York, NY; c NYU Langone
Medical Center, NY, NY.
OBJECTIVE: To evaluate if the degree of blastocyst expansion following
rewarming of euploid embryos in a frozen embryo transfer cycle affects on
pregnancy outcomes.
DESIGN: Retrospective Cohort Study.
MATERIALS AND METHODS: Patients undergoing a frozen euploid
embryo transfer from June 2013 to October 2014 were included. Preimplantation
genetic screening using array comparative genomic hybridization
designated embryos to be euploid or aneuploid. All embryos had undergone
trophectoderm biopsy followed by vitrification. Following our standard FET
protocol, vitrified euploid embryos were rewarmed on the day of transfer.
Embryologists documented degree of expansion at time of transfer for all
frozen embryo transfers. Patients with all causes of infertility, and both autologous
and donor cycles were included. Primary outcome included pregnancy,
defined by presence of a gestational sac, and clinical pregnancy, defined by
live birth or fetal heart beat through cycle day 63. Statistical analysis was
done using relative risk and Chi Square.
RESULTS: A total of 228 embryo transfers were included in the study.
The mean age of patients at time of embryo freezing included in the study
was 36.20 +/- 4.59 (range 21-45). There were 143 expanded, 64 partially
expanded and 21 collapsed blastocysts at time of transfer. The overall
pregnancy rate was 66.2% and clinical pregnancy was 60.1%. There is
a higher rate of clinical pregnancies in embryos that were expanded at
time of transfer (Table 1). The collapsed embryo group had similar pregnancy
rates but was noted to have a lower clinical pregnancy rate.Blastocysts
were biopsied and vitrified on day 5 (166,72.8%) or on day 6
(62,27.2%). No difference was noted on its affect on pregnancy or clinical
pregnancy rates.
unknown whether hatching at this point is associated with poor embryo survival.
This study sought to determine whether single embryo transfers (SETs)
of euploid embryos that hatched during the warming process were associated
with lower pregnancy rates (PRs).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients underwent trophectoderm biopsy
with qPCR-based comprehensive chromosome screening (CCS) from
June 2011 to January 2015. Only patients who underwent SET of a frozen
euploid embryo were included. All embryos were graded according to the
Gardner scale both prior to freeze by vitrification and after thaw prior to
transfer. Patients were segregated into two groups: A) FETs of embryos
that hatched from expansion 4 or 5 to expansion 6 during the warming process;
and B) FETs of embryos expansion 4 or 5 that remained the same after
re-warming. Contingency tables were generated for PR and clinical PR based
on embryo hatching stage and the PR and clinical PR were computed. Statistical
analysis was calculated by fisher exact test with significance at p< 0.05.
RESULTS: Couples underwent 262 euploid frozen SETs of oocytes aged
23.5-44.3 yo. Embryos that had hatched prior to biopsy to grade 6 at transfer
(n¼86) experienced PRs (0.76, 0.65-0.84 95% CI) that were not significantly
different from those (n¼176) that remained grade 4 or 5 (0.78, 0.71-0.84 95%
CI, p¼0.75). Clinical PRs from euploid SET in embryos that had hatched
from grade 4 or 5 prior to biopsy to grade 6 (0.65, 0.54-0.75 95% CI) were
not significantly different from those that remained grade 4 or 5 (0.66,
0.59-0.73 95% CI, p¼0.89).
Embryo Transfer Outcomes.
Blast 4/5 Embryos at Freeze At Transfer Blast 6 vs 4/5
Embryo Transfer Count 176 86
Pregnancy 137 65
Pregnancy Rate 0.78 0.76 0.88 [0.46-1.70]
Clinical Pregnancy 117 56
Clinical Pregnancy Rate 0.66 0.65 0.94 [0.53-1.69]
CONCLUSIONS: Extended culture has been improved and is more
commonly applied in practice, consequently increasing the prevalence of
blastocyst cryopreservation. Embryos hatching from their zona pellucida
during the re-warming process experienced comparable PR and clinical
PRs to those hatching prior to cryopreservation. There is no cause for concern
when upon rewarming an unhatched embryo, complete escape from the zona
pellucida occurs. Further studies utilizing larger subsets of patients are
needed to confirm this study’s findings.
Pregnancy Outcomes by Embryo Expansion at Time of Transfer
N
Total Pregnant (presence
of gestational sac)
Percent
Total Clinical Pregnancy (fetal
heart beat of Live Birth)
Percent
Expanded 143 106 74.12 95 66.43
Partially Expanded 64 34 53.13 32 50.0
Collapsed 21 11 64.70 10 47.62
Total 228 151 137
P value 0.0047 0.0392
CONCLUSIONS: There is a higher chance of implantation and ongoing
pregnancies that lead to live birth in embryos that are re-expanded at time
of transfer following the thaw of a euploid embryo.
P-665 Wednesday, October 21, 2015
DOES COMPLETE HATCHING AT THE TIME OF THAW NEGA-
TIVELY AFFECT PREGNANCY RATES? J. Gingold, a J. Rodriguez-
Purata, b M. C. Whitehouse, b B. Sandler, b A. B. Copperman. c,a a Obstetrics,
Gynecology and Reproductive Science, Icahn School of Medicine at Mount
Sinai, New York, NY; b Reproductive Medicine Associates of New York, New
York, NY; c Reproductive Medicine Associates, New York, NY.
OBJECTIVE: After re-warming for frozen embryo transfers (FET),
embryonic expansion and hatching frequently often observed in biopsied embryos,
even when they were previously unhatched at time of vitrification. It is
P-666 Wednesday, October 21, 2015
ECONOMIC IMPLICATIONS OF THE SART GUIDELINES ON
EMBRYO TRANSFER: HEALTHCARE DOLLARS SAVED BY
REDUCING IATROGENIC TRIPLETS. M. S. Lee, a,b B. T. Evans, b,c
M. D. Hornstein. a,b a Obstetrics and Gynecology, Brigham and Women’s Hospital,
Boston, MA; b Harvard Medical School, Boston, MA; c Harvard Combined
Orthopaedic Residency Program, Massachusetts General Hospital,
Boston, MA.
OBJECTIVE: Since SART first released guidelines in 1998 on number of
embryos to transfer, there has been a consistent decrease in the number of embryos
transferred per cycle and the percentage of higher order multiple (HOM)
gestations resulting from ART in the United States. The purpose of this study
was to estimate the total national cost savings resulting from reductions in
FERTILITY & STERILITY Ò
e335

HOM gestations since the publication of the initial SART guidelines on embryo
transfer (ET).
DESIGN: Descriptive cost analysis and three-point estimation.
MATERIALS AND METHODS: A systematic review of the literature was
conducted to estimate the hospital costs of singleton and triplet pregnancies
and subsequent delivery. National data reported to the CDC from 1996 to
2012 were reviewed. The trends in ET and reductions in resulting HOM gestations
were examined. The number of HOM deliveries prevented following
the introduction of SART guidelines was estimated. The total costs were estimated
based on direct hospital charges for associated obstetric and perinatal
conditions/complications, and projected to 2014 US dollars.
RESULTS: A singleton gestation (including pregnancy, delivery and up to
one year of neonatal care) was estimated to cost between $17,112-24,212. A
triplet gestation was estimated at $190,788-453,935. Comparable estimates
of singleton and triplet gestations demonstrated the latter to be between 11
and 27 times as costly. The percentage of HOM gestations amongst all
ART pregnancies decreased from 11.4% in 1997 to 2.0% in 2012, with the
sharpest year-to-year decline of 20.3% occurring from 1998-1999, the year
following the publication of the initial SART guidelines. Similarly, the number
of liveborn HOM infants secondary to ART has decreased from 59.8 per
1,000 fresh non-donor cycles in 1997 to 13.2 in 2012. From 1998 to 2012, the
cumulative number of prevented HOM deliveries was estimated to be 13,512.
This corresponds to an estimated net total savings in direct hospital-related
costs of $4.27B (range $2.35-5.85B, 2014 dollars).
CONCLUSIONS: Iatrogenic HOM gestations represent a substantial economic
burden to our healthcare system. Changes to the practice of ET
following the publication of the initial SART guidelines in 1998 have resulted
in a dramatic decrease in the HOM rate. The associated cumulative
cost savings to the US healthcare system are estimated to be over $4B.
Supported by: No financial support. We are indebted to Drs. Dmitry Kissin
and Sara Crawford at the CDC for their invaluable assistance and support.
P-667 Wednesday, October 21, 2015
FROZEN BLASTOCYST TRANSFER IN NATURAL CYCLE VS
HORMONE REPLACEMENT THERAPY CYCLE. A. Biryukov, a
V. Apryshko, a,b I. Zorina, a E. Osina, a N. Dmitrieva, a S. Yakovenko. a,c a Altravita
IVF clinic, Moscow, Russian Federation; b Faculty of Biology, Moscow
State University, Moscow, Russian Federation; c Faculty of Physics,
Moscow State University, Moscow, Russian Federation.
OBJECTIVE: The introduction of vitrification in assisted reproduction
technology leads to significant increase in embryo survival rates by up to
95-100%. Frozen blastocyst transfer (FBT) treatment can be scheduled in a
natural ovulatory cycle or in a hormone replacement therapy (HRT) in which
the endometrium is prepared with estrogen and progesterone supplementation.
Natural cycle FBT is easy to implement and can be offered to women
that have a regular menstrual cycle and are proven to ovulate. Conversely,
HRT is an option for anovulatory patients and offers flexibility for the doctor
by controlling the timing of the cycle. In this research the hypothesis is that
there is no significant difference in efficiency between natural FBT and HRT
FBT.
DESIGN: Retrospective analysis of 500 natural FBT cycles and 500 HRT
FBT cycles. Clinical pregnancy, miscarriage and live births rates were
compared. Cycles with donor embryo transfers were excluded from this
research.
MATERIALS AND METHODS: Embryos were cultured in Global total
media at 7.3% CO2, 36.6 C for 5-6 days and were vitrified using CryoTech
vitrification method at blastocyst stage. Only AA, AB, BA and BB grades
(Gardner’s system) of blastocysts were frozen. After thawing, blastocysts
were cultured for 120 minutes prior to transfer in order to make sure of survival.
The average number of transferred blastocysts was 1.4 for natural cycle
and 1.3 for HRT. The average age of patients was 34.8 for natural cycle and
35.1 for HRT. Statistical differences between the values were made using
analysis of variance (ANOVA).
RESULTS: Clinical pregnancy rates for natural cycle FBT were greater
than HRT FBT (43.7% vs 39.8%). However this was not statistically significant
(p¼0.26). Miscarriage rates for natural cycle FBT were greater than
HRT FBT (33.7% vs 36.4%). However this was not statistically significant
(p¼0.2). Live birth rates for natural cycle FBT were lower than HRT FBT
(38.7% vs 44.3%). However this was not statistically significant (p¼0.14).
CONCLUSIONS: This retrospective cohort study found no significant
difference in clinical pregnancy, miscarriage and live birth rates between
natural cycle FBT and HRT FBT. At present our success rates for natural
cycle vs HRT FBT suggest that if a patient has a regular menstrual cycle
then they should be offered a natural cycle FBT. This avoids the use of
multiple medications, reduces the cost for patients, and simplifies the
treatment.
P-668 Wednesday, October 21, 2015
DEMOGRAPHIC CHARACTERISTICS ASSOCIATED WITH ELEC-
TIVE SINGLE EMBRYO TRANSFER IN IVF: WHO CHOOSES JUST
ONE? E. M. Munch, a K. M. Summers, a G. Ryan, a J. D. Kapfhamer, a
B. Collura, b G. D. Adamson. c a Reproductive Endocrinology and Infertility,
University of Iowa Hospitals and Clinics, Iowa City, IA; b RESOLVE,
McLean, VA; c PAMF Fertility Physicians of Northern California, Saratoga,
CA.
OBJECTIVE: Younger patient age has been found in previous IVF studies
to be associated both with desire for twins and the choice of single embryo
transfer; we sought to examine this and other demographic and geographic
parameters in relation to elective single embryo transfer (eSET).
DESIGN: Descriptive analysis of online survey results from a crosssectional
sample of U.S. community women.
MATERIALS AND METHODS: An online survey, advertised through
RESOLVE (the National Infertility Association) was conducted over 5
weeks in 2014. Interested participants were screened for gender and cycle
eligibility and gave consent by acknowledging an online privacy statement.
Inclusion criteria were age < 40 and the completion of at least one IVF cycle
with embryo transfer. Participants were asked to identify their age, race,
income, highest education completed, and insurance coverage in the survey.
The outcome of interest was the election of single embryo transfer (eSET,
defined as multiple embryos available for transfer but electing a single embryo
transfer) versus multiple embryo transfer (MET) in the first IVF cycle.
We examined geographic characteristics by US Census region and by state
IVF access quartile, as previously published. 1 All variables, including age
and income, were categorical, and X 2 was used to compare proportions
among groups.
RESULTS: Of 888 participants, 587 met age and cycle inclusion criteria.
Participants who chose eSET tended to be younger than those choosing MET,
with 25% of participants under 29 choosing eSET, compared to 12% of those
aged 35-39 (p¼0.014). Education level, race, income, and insurance
coverage for infertility did not differ between eSET and MET groups. There
was no association between eSET or MET with regard to accessibility to IVF
in the participant’s state. When comparing patterns of eSETand METaccording
to US Census region, patients from the Midwest were significantly more
likely to choose MET over eSET (91% for the Midwest vs an average of 80%
for other census regions, p¼0.003).
CONCLUSIONS: Even in patients

DESIGN: Retrospective cohort study of fresh first IVF cycles started May
2012 to November 2013.
MATERIALS AND METHODS: Among 1186 cycles, 857 had a day 3 ET
and 329 a day 5 ET; day 5 ET was based on a clinical algorithm that included
day 3 availability of R3 embryos with at least 8-cells with

alance between the potential cryo-damage to the embryos from the vitrification
procedure and improved endometrial receptivity. FET does not increase
miscarriage rate.
IMPLANTATION
P-673 Wednesday, October 21, 2015
EMBRYO S-PHASE LENGTH ANALYSIS ON FIRST AND SECOND
CELL CYCLE AND ITS RELATIONSHIP WITH REPRODUCTIVE
OUTCOME. J. A. Aguilar, a E. Munoz, b A. Galan, c Y. Motato, c
M. Ojeda, b V. Garcia, c M. Meseguer. d a IVF Laboratory, IVI Vigo, Vigo,
Spain; b IVI Vigo, Vigo, Spain; c IVI Valencia, Valencia, Spain; d Clinical
Embryology, Valencia, Spain.
OBJECTIVE: to analyse the length of the s-phase (synthesis of DNA), in
the first cell cycle (ECC1) and in each blastomere of the second cell cycle
(ECC2) of known implantation data (KID) embryos, and to relate them
with reproductive outcome.
DESIGN: Observational retrospective study of 1679 transferred embryos
from 940 ICSI patients from our egg donation program between January
2011 and January 2014 in IVI Vigo and IVI Valencia, cultured in Embryoscope.
Sperm samples under 1 million/ml were used as an exclusion
criteria.
MATERIALS AND METHODS: Only non-multinucleated embryos
which either failed to implant or fully implanted were included in the study.
The s-phase was calculated as the period while nuclei were visible, being
ECC1 S-phase¼tPNfading-tPNappearance, and ECC2 S-phase¼
t2MONO1(f)- t2MONO1(a). ANOVA test and c2-test were performed
when applicable to assess the influence of the length of S-phase in the implantation
rate.
RESULTS: S-phase in both cell cycles were calculated in 904 KID embryos
out of 1679, and therefore were analyzed. 32,74% (n¼296) fully implanted
(KID+), and 67,25% (n¼608) failed to implant (KID-). The
average length of s-phase in the ECC1 in the KID+ embryos was longer
than in KID-, 15,50h v.s. 14,38h, and slightly shorter in ECC2, 8,35 in
KID+ v.s. 8,60h in KID-. 46,1% (n¼112) of the embryos implanted when
the difference between both s-phases was greater than 7.96h, being 29,1%
for those where the difference is between 5,96h-7,95h; 23,1% for those between
3,61h-5,95h; and 30% for those below 3,6h.
CONCLUSIONS: Results show that the s-phase ECC1, nearly double the
length of s-phase ECC2, and the greater the difference between both s-phase,
the greater the embryo implantation, suggesting that the replication in the
first cell cycle may implicate particularities related to the genome combination,
and that those embryos which do not implant may not complete the
replication of its genome during the first cycle producing a larger reduction
in the duration of the second.
P-674 Wednesday, October 21, 2015
REPEATED IMPLANTATION FAILURE RELATES TO CIRCULA-
TORY ABNORMALITIES. D. A. Pattinaja, M. E. Spaanderman,
C. Ghossein-Doha, R. J. van Golde. Obstetrics & Gynaecology Department,
Maastricht University Medical Centre, Maastricht, Netherlands.
OBJECTIVE: A diminished pre-pregnant plasma volume as marker of
reduced cardiovascular reserves may influence embryo implantation by circulatory
redistribution at the expense of uterine perfusion. In this pilot study
we tested the hypothesis that recurrent implantation failure in in-vitro-fertilization
(IVF) patients is associated with decreased plasma volume and
increased uterine arteries resistance as proxy measures for circulatory anomalies.
DESIGN: In this cross-sectional pilot study we included patients who had
at least 2 embryo transfers after IVF treatment (n¼10) and a control group
consisting of women with a history of uncomplicated reproduction (n¼10).
Both groups were assessed for mean arterial blood pressure (MAP), plasma
volume and the pulsatility index (PI) of both uterine arteries.
MATERIALS AND METHODS: Blood pressure was measured at three
minute intervals during a half hour in sitting position. Plasma volume was
measured by the iodine 125-labeled albumin indicator dilution method and
expressed in ml/ body surface area (m2). Resistance of the uterine arteries
was obtained by measuring the pulsatility index by transvaginal doppler
flow ultrasonography. Data was analyzed using the Mann Whitney U test.
A p-value below 0.05 was considered statistically significant.
RESULTS: The median duration of subfertility in the study group is 39
months [IQR 32-51]. The conventional diagnoses of subfertility were male
factor (n¼6; one women combined with endometriosis and one women combined
with cycle irregularity), unexplained subfertility (n¼3) and tubal pathology
(n¼1). Baseline characteristics were comparable between both
groups (Table 1). Plasma volume was lower in the IVF group (1313 [IQR
1189-1406] ml/m2) compared to the control group (1468 [IQR 1362-1551]
ml/m2). Furthermore the pulsatility indices of the uterine arteries were higher
in the IVF group compared to the control group with 3.5 [IQR 3.0-3.8] vs 2.1
[IQR 1.6-2.7] for the right branch and 3.2 [IQR 2.7-3.7] vs 2.2 [IQR 1.5-2.5]
in the left branch respectively.
CONCLUSIONS: Repeated implantation failure after IVF relates to
diminished plasma volume along with increased uterine vascular resistance.
We speculate that reduced plasma volume relates to circulatory redistribution
at the expense of the uterine perfusion, which in turn negatively affects reproductive
performance.
Baseline characteristics and circulatory variables, presented as median with
interquartile range.
Control
group(n¼10)
Study group
(n¼10)
p-
value
Age (years) 32 (30-33) 31 (29-35) 0.80
Height (metres) 1.7 (1.6-1.7) 1.6 (1.6-1.7) 0.35
Weight (kilograms) 60.6 (56.6-65.3) 67.5 (57.8-75.0) 0.17
Body mass index (kg/m2) 21 (20-24) 23 (21-29) 0.08
Body surface area (m2) 1.7 (1.7-1.8) 1.8 (1.6-1.9) 0.82
Parity 2 (1-3) 0 (0-0) 0.02
Mean arterial pressure 88 (81-108) 88 (82-94) 0.87
(mmHg)
Plasma volume per body 1468 (1362-1551) 1313 (1189-1406) 0.02
surface area (ml/m2)
PI right uterine artery 2.1 (1.6-2.7) 3.5 (3.0-3.8)

47.8%, p

are generated, the implantation rates are independent of female reproductive
age. The application of CCS appears to be useful in the identification of blastocysts
with high implantation potential in women of all reproductive ages.
Implantation rates in each age group after vitrified euploid blastocysts ET.
FET
cycles (n)
Total
number of
blastocysts
transferred
(n)
Mean
number of
blastocysts
transferred
(n)
Heart
Beat# (n)
Implantation
rates (%)
Donor 126 202 1.6 111 55%
42y 14 17 1.2 11 65%
P-679 Wednesday, October 21, 2015
EMBRYO CYTOPLASMATIC WAVE AND EVENNESS OF BLASTO-
MERES CHARACTERIZATION AT THE END OF CELL CYCLE
AND THEIR RELATION WITH CLINICAL
OUTCOME. J. A. Aguilar, a E. Munoz, b M. Ojeda, b E. Taboas, b
M. Perez, b M. Meseguer. c a IVF Laboratory, IVI Vigo, Vigo, Spain; b IVI
Vigo, Vigo, Spain; c Clinical Embryology, Valencia, Spain.
OBJECTIVE: Early cell cycle events have recently been studied as
possible markers for embryo selection and related with implantation rate.
The aim of this research is to characterize the cytoplasmic wave and the evenness
of blastomeres at the end of 2 cells and 4 cells stage in embryos from
fresh donated oocytes with known implantation data.
DESIGN: Observational retrospective study conducted in IVI Vigo, between
January 2011 and January 2014. 275 transferred embryos, cultured
in an Embryoscope incubator in a 37 C, 6% CO2 and 20% O2 atmosphere,
from 171 oocyte donation cycles were analyzed.
MATERIALS AND METHODS: Cytoplasmic wave was annotated at its
beginning and its end, and its duration was calculated as CWduration¼
tCWend- tCWinitiation. CWduration were categorized and the frequency
of embryos was also registered. The symmetry of the embryo blastomeres
were assessed at the end of the 2cells, and 4 cells stage, in order to avoid
the change in their volume due to the embryo dynamic behaviour. Only those
embryos with which either failed to implant or fully implanted (KID+) were
included in the study. ANOVA test, t-test and c2-test were performed when
applicable to assess the influence of the continuous and categorical variables
in the implantation rate.
RESULTS: 275 embryos were analyzed. CW was always present, 19.2%,
of embryos showed a CWduration shorter than 1.5h, 58,5% between 1,5-3h,
19.6% between 3-4,5h and 2,54% longer than 4.5h. No significant differences
were found between these categories and embryo implantation
(p¼0.13)In KID+ embryos blastomere evenness was more frequent 33.8%
vs 8% (p¼0.26) (KID-) at the end of the 2cells stage. Differences were
greater at the end of the 4cells stage, and statistically significant 37.09%
vs 4,72 (p¼0.025).
CONCLUSIONS: The majority of embryos present a CW grouped between
1,5h and 3h, although it seems not to be related to embryo implantation
The unevenness of the blastomeres at the end of the 4 cells stage is negatively
related to embryo implantation, while it seems not to be at the end of the 2
cells stage. Although numbers from these observations are still reduced,
we suggest to annotate the latter embryo morphological characteristic
following our descriptions, due to the continuous modification of embryo
blastomeres during the cell cycles, enhancing the annotation at the end of
the 4 cells stage, and considering for the future elaboration of embryo selection
algorithms.
P-680 Wednesday, October 21, 2015
HOW WIDE IS THE UTERINE IMPLANTATION
WINDOW? D. H. McCulloh, a C. McCaffrey, b J. Grifo. c a Obstetrics
and Gynecology, New York University Fertility Center, New York, NY;
b NYU Fertility Center, New York, NY; c NYU Langone Medical Center,
New York, NY.
OBJECTIVE: The period of time when embryos implant following natural
cycles with intercourse or following in vitro fertilization is quite
diverse spanning more than one week. Diverse times of implantation
(TIs) are consistent with at least two different models: 1) the uterus is
receptive to implantation for a long period of time following its onset or,
2) the uterus is receptive to implantation for a brief period with diverse
times of onset. Our objective was to determine whether the window of uterine
receptivity is wide (model 1) or narrow (model 2) and to determine just
the window’s width.
DESIGN: Retrospective Analysis.
MATERIALS AND METHODS: Human Chorionic Gonadotropin
(hCG) levels and fetal sacs were assembled for patients who became pregnant
and where all detected fetal sacs delivered. TIs were estimated by
extrapolating the log(hCG) levels on cycle days 28 and 35 to a time
when the hCG was10 mIU/mL per fetal sac. Although we refer to this
time as TI, it, strictly speaking, lags behind implantation with a delay.
Distributions for two TIs were compared: singletons versus twin pregnancies
following fresh, day 3 embryo transfers. The method to determine the
width of the uterine window of receptivity relied upon narrowing of the TI
distribution that occurs when twins implant (Our method estimates only
one ‘‘averaged’’ TI when twins implant at diverse times. Narrowing is
analogous to standard errors with 2 samples being smaller than the standard
deviation.).
RESULTS: TIs for singletons averaged 7.6 +/- 2.1 days (N ¼ 231) after
oocyte retrieval. TIs for twins averaged 8.1 +/- 1.8 days (n ¼ 89 twin
pregnancies). Standard deviation of the distribution of TIs estimated of
the width of the distribution of TIs. The distribution for twins was
0.86 X the width for singletons. When twins implant simultaneously,
the distribution of TIs is expected to be as wide as the distribution of
TIs for singletons. The twin distribution was significantly narrower
than this expectation. Twins implanting randomly throughout the distribution
of times seen for singletons would be expected to lead to a
twin distribution narrowed to 0.71 X the singleton TI distribution. The
twin distribution was significantly wider than this expectation. Modelling
the implantation of twins within a defined interval led to estimates that
the width (standard deviation) of the uterine window of receptivity is
0.6 times as wide as the standard deviation of the singleton implantation
distribution (1.26 days).
CONCLUSIONS: The standard deviation (width) of the window of uterine
receptivity of 1.26 days (0.6 x the width of the singleton distribution) is
consistent with a restricted window of uterine receptivity. Therefore, we
believe that Model 2 is correct, indicating that there is a variable time of onset
with a restricted period of uterine receptivity. This suggests that as many as
20-40% of embryos that are capable of implantation may fail to implant simply
due to asynchrony between embryo and uterus.
P-681 Wednesday, October 21, 2015
DOES ENDOMETRIAL INJURY IMPROVE PREGNANCY OUT-
COMES IN OVUM DONATION CYCLES? E. Szlit Feldman, a
E. A. Salama, a A. Torno, b L. Ferle, b G. Arruguete, c A. Sdrigotti. d a Procrearte
Director, Buenos Aires, Argentina; b Gynecologist - Procrearte, Buenos
Aires, Argentina; c Procrearte, Buenos Aires, Argentina; d gynecologist and
obstetrician. fellow in reproduct, Buenos Aires, Argentina.
OBJECTIVE: To explore the possibility that local endometrial injury
(LEI) improves the clinical outcomes in ovum donation recipients.
DESIGN: Retrospective cohort analysis.
Main results.
Study Group
(n¼66)
Control
Group(n¼104)
p Value
Donor age (min-max) 24,3 (21-30) 24,8 (21-30) 0.5
Recipients age
43,5 (33-55) 41,85 (33-55) 0.13
(min-max)
Severe Male factor 1/66 (1.5%) 4/106 (3.7%) 0.69
Number of embryos 2,01 2,01 NS
transfered
Implantation Rate( %) 32/133 (24%) 43/210 (20.47%) 0.5
Clinical Pregnancy (%) 24/66 (36%) 31/104 (29.8%) 0.47
Multiple pregnancy
Twins
3/24 (12.5%) 8/31 (25.8%) 0.37
e340 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

MATERIALS AND METHODS: A total of 170 ovum donation cycles
were included from 2012 to 2014 in a private fertility center: the study group
included 66 patients who underwent the local endometrial injury; the control
group included 104 patients without the experimental treatment. Oocyte donors
were 21-30 years old with proven fertility, and recipients were women
with indication of ovum donation due to diminished ovarian reserve or
advanced maternal age. The local injury was performed once on day 20-22
of the previous menstrual cycle. The endometrial scratch was done by vacuum
aspiration with a Frydman Catheter and a 5 cc. syringe on the uterine
wall until a sample of tissue was removed through the tube. Statistical analysis
was performed using the SPSS. p< 0.05 was considered statistically significant.
RESULTS: There were no demographic differences between the groups.
The difference between implantation, clinical pregnancy and multiple pregnancy
rates was not statistically significant.
CONCLUSIONS: Our results show that local injury of endometrium by
aspiration did not improve implantation, pregnancy and multiple pregnancy
rates in ovum recipients. To our knowledge this is the second study (1) presented
in order to evaluate the influence of LEI on ovum donation recipients.
The benefit of LEI is controversial in cycles of IVF in patients with recurrent
implantation failure. The advantage of using oocyte donor cycles in the analysis
of LEI is that it eliminates the potential bias of oocyte quality on pregnancy
outcome. A prospective randomized study will be conducted soon in
order to elucidate the controversies of results when using this procedure in
different studies.
Reference:
1. Dain L, Ojha K, Bider D, Levron J, Zinchenko V, Walster S, Dirnfeld
M. Effect of local endometrial injury on pregnancy outcomes in
ovum donation cycles. Fertil Steril. 2014 Oct;102(4):1048-54.
P-682 Wednesday, October 21, 2015
MECHANICAL ENDOMETRIAL INJURY DOES NOT INCREASE
SERUM LEVELS OF PRO-IMPLANTATION
CYTOKINES. A. P. Melnick, a I. Ramer, a E. M. Murphy, a
Z. Rosenwaks, a S. Witkin, b S. D. Spandorfer. a a The Ronald O. Perelman
and Claudia Cohen Center for Reproductive Medicine, Weill Cornell Medical
College, New York, NY; b Obstetrics and Gynecology, Weill Cornell Medical
College, New York, NY.
OBJECTIVE: Mechanical endometrial injury via endometrial biopsy or
hysteroscopy in the menstrual cycle prior to in vitro fertilization (IVF) has
been proposed as a way to increase implantation rates by improving endometrial
receptivity. Specifically, endometrial injury may induce an inflammatory
response with upregulation of cytokines, adhesions molecules, and growth
factors critical to implantation. We have previously demonstrated an association
between cytokine levels and IVF outcome. The goal of this study was to
determine whether mechanical endometrial injury affects serum levels of
pro-implantation cytokines in patients undergoing IVF with autologous
endometrial coculture (AECC).
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: Patients with a history of prior IVF
failure undergoing IVF with autologous endometrial coculture at our center
were included. All patients underwent luteal phase endometrial biopsy
in preparation for IVF. Biopsy samples were either utilized for IVF in the
consecutive menstrual cycle or were frozen and thawed for use in a future
cycle. Cycles were stratified into two groups by biopsy timing: biopsy
performed in the cycle prior to IVF and biopsy performed more than
one cycle prior to IVF. Serum samples were obtained from day of IVF cycle
start and levels of cytokines implicated in implantation were determined
by quantitative ELISA performed by blinded personnel. Primary
outcome was levels of IL-1b, IL-6, IL-11, IL-17, IGF-1, IGF-2, TNF-a,
IL-1RA, IFN-g, BDNF, NT-3, and NT-4. Statistical analysis included
Mann-Whitney U, chi-square, and Fisher’s exact tests. P140 pg/mL, Progesterone > 10 ng/mL at the day of transfer),
with good blastocyst (3BB or more with Gardner’s classification), were
included in the study. Pregnancy test (serum hCG), serum copper, zinc and
ceruloplasmin concentrations were measured 16 days after the first date of
progesterone replacement. Hundred women with pregnant without miscarriage
at 10 weeks of gestation (group P) and 176 women without pregnant
(group NP) were compared. Student’s t test was used for comparison between
two groups. Cutoff line was determined by receiver-operator characteristic
(ROC) curve, and Chi-squared test was used for 2x2 contingency table. Statistical
significance was defined as P

(embryos were biopsied day 5, vitrified day 6). A logistic regression analysis
was used to determine a correlation of elapsed time in culture from biopsy to
FET and PR.
RESULTS: A total of 362 cycles met criteria for inclusion (SDBV¼325
and NDBV¼37). PR observed for SDBV was 75.2% and for NDBV was
70.3%. No correlation was found between time in culture from biopsy to
FET and pregnancy outcomes (p>0.05).
CONCLUSIONS: Vitrification is an excellent tool in the IVF laboratory,
enabling better clinical flexibility in planning a patient’s IVF protocol.
This study is the first of its kind to analyze the impact of delayed vitrification
of biopsied embryos. We found no correlation between time in culture to
successful ET. Embryologists can be reassured that prolonging culture of
embryos following biopsy does not negatively impact implantation.
P-685 Wednesday, October 21, 2015
PREDICTABILITY OF BLASTOCYST, EUPLOIDY AND IMPLAN-
TATION RATE WITH EARLY MORPHOKINETICS
PARAMETERS. E. Rocafort, A. Leza, M. Guijarro, L. Medrano,
B. Ramos, M. Fernandez, J. Aizpurua. IVF Spain, Alicante, Spain.
OBJECTIVE: To determine if early morphokinetic parameters , such as
P2 (time from 2 to 3 cells) and P3 (time from 3 to 4 cells), are correlated
with the likelihood of blastocyst formation, euploidy and implantation
rate.
DESIGN: Unicentric and restrospective (Sept.2013 - April 2015). A total
of 76 patients were included to a private clinic Preimplantation Genetic
Screening (PGS) program. 740 embryos were cultured into
EevaÔ. EevaÔ is an automated time-lapse system that provides on Day
3 of embryo culture a prediction (High/Medium/Low) depending on P2
and P3 morphokinetic parameters. Euploid embryos with Known Implantation
Data (KID) and their implantation depending on EevaÔ prediction
were studied.
MATERIALS AND METHODS: All embryos were cultured and biopsied
on Day 5-6 of embryo development and screened by Next Generation
Sequencing. All transfers were differed and under HRT. Embryos selected
for transfer were based on comprehensive-chromosomal-screening results
and morphology criteria. Implantation was calculated using known implantation
confirmed by ultrasound at 6-7 weeks.
RESULTS: In 740 embryos, 29.8% were High, 20.5% Medium and 49.7%
Low. Blastocyst formation rates for Eeva High, Medium and Low embryos
were 80.9% (178/220), 72.4% (110/152) and 40.2% (148/368), respectively.
The difference in blastocyst formation rates between Eeva High, Medium and
Low embryos was statistically significant. Euploidy rates were 35.5% (78/
220) in High embryos, 31.6% (48/152) Medium and 18.5% (68/368) Low
with significant differences from High and Medium to the Low group
(p< .0001). However, no significant difference was found between Blastocyst
Euploidy rates for Eeva High, Medium and Low (43.8% (78/178), 43.6% (48/
110) and 45.9% (68/148), respectively). A total of 62 KID euploid blastocyst
were analyzed, being their implantation rate for High, Medium and Low of
52.6% (20/38), 33.3% (3/9) and 13.3% (2/15), respectively. The difference
in KID euploid blastocyst between High and Low was statistically significant
(p¼0.009).
CONCLUSIONS: Based on our results, P2 and P3 are correlated with blastocyst
formation, euploidy and implantation rates. However, these parameters
are not useful in euploidy predictability when embryo culture is
performed until Day 5. Automated time-lapse systems may be useful in combination
with PGS in terms of selecting the blastocyst with the highest potential
among euploid embryos. This preliminary results may indicate that
implantation rates do not only depend on the ploidy of the embryo as a factor
for a successful pregnancy.
P-686 Wednesday, October 21, 2015
REVISITING THE PREDICTIVE VALUE OF UTERINE ARTERY
PULSATILITY INDEX FOR UTERINE RECEPTIVITY. H. Zaki, a
E. Geneidi, b C. Coulam. c a Director Ganin Fertility Center, Cairo, Egypt;
b Radiology, Faculty of Medicine, Cairo, Egypt; c Fertility & Cryogenics
Lab, Downers Grove, IL.
OBJECTIVE: Key factors for the process of implantation include endometrial
thickness, embryo competence, hormonal milieu and uterine blood
flow.There have been conflicting reports assessing the usefulness of doppler
ultrasound for predicting pregnancy outcome, part of the problem is the interaction
of all key factors. Now that substantial enhancement in embryo selection
with comprehensive chromosome screening has given the chance of
transferring single euploid embryo. Again, the substantial improvement in
embryo freezing with vitrification methods, allows eliminating the negative
effect of supernatural levels oestrogen achieved during ovarian stimulation
upon implantation.There is a renewed focus upon assessing other key factors
such as uterine blood flow. The objective of this study is to assess the role of
pulsatility index (PI) in the process of implantation.
DESIGN: Prospective Study.
MATERIALS AND METHODS: We evaluated 83 patients under the age
of 40 years of age, undergoing thawed single euploid embryo transfer. All
embryos are the result of long protocol of controlled ovarian stimulation,
when embryos reach blastocyst at day 5 or day 6, embryo biopsy were
done and the embryos were vitrified. Comprehensive Chromosome
Screening (CCS) were done using q-PCR platform.Frozen cycles were performed
using oral hormonal replacement therapy with oral 17 beta estradiol
valerate. Progesterone injections of 50 mg were taken when the endometrium
thickness exceeds 7.5 mm and had a trilaminar pattern. Ultrasound doppler
measurements of Pulsatility Index(PI) were done in both uterine arteries during
the last day before starting the progesterone injections. Pregnancy was
diagnosed by positive pregnancy test two weeks after embryo transfer and
fetal heart detection 3 weeks after pregnancy test. T-test was used and statistical
significance was established on p 0.3) were recruited
for the study during actual ICSI cycles. Intervention(s): Patients
self-administered sildenafil citrate (Viagra), vaginal suppositories (100 mg/
once daily) for 7days, starting from the day of ovulation triggering till the embryo
transfer day (ET D5). Endometrial thickness and spiral artery RI & PI
were assessed using trans-vaginal color-pulsed Doppler 2D & 3D ultrasound
on the sub-endometrial zone in two sessions, (day of triggering and day of
embryo transfer). Main Outcome Measure(s): Endometrial thickness and
RI & PI plus reconstructed 3D Doppler images were the primary outcomes
while the implantation rate and pregnancy rate were the secondary outcomes.
RESULTS: Improvement of endometrial thickness and uterine blood flow
with significant reduction of RI & PI were detected in 15 (68%) out of 22 patients.
Implantation rate and pregnancy rate were higher in patients with positive
effect in comparison to the other patients with no effect; (26% vs 7%) &
(40% vs 14%) respectively.
CONCLUSIONS: Vaginal sildenafil might be an interesting therapeutic
option during ICSI cycles as it may improves the uterine perfusion especially
with women with a thin endometrium. Further studies are needed on sildenafil
to determine whether there are other mechanisms for improvement of implantation
rather than its direct effect on the endometrium.
Reference: Thin endometrium.
e342 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

P-688 Wednesday, October 21, 2015
EFFECT OF DURATION OF PROGESTERONE SUPPLEMENTA-
TION BEFORE THE TRANSFER OF VITRIFIED-WARMED
BLASTOCYSTS ON IMPLANTATION RATES IN FROZEN
EMBRYO TRANSFER (FET) ESTROGEN REPLACEMENT
CYCLES. S. H. Anderson, a,b D. Brasile, a,b E. S. Verrecchio, a
H. Pearlstein, a W. P. Haberstroh, a M. J. Glassner. a,b a Main Line Fertility
Center, Bryn Mawr, PA; b Obstetrics and Gynecology, Drexel University College
of Medicine, Philadelphia, PA.
OBJECTIVE: There have been few studies evaluating the optimal timing
of progesterone (P4) supplementation prior to transfer of embryos in FET cycles.
The objective of this study was to determine whether starting P4 supplementation
five days or six days before transfer of vitrified-warmed
blastocysts results in higher implantation rates in estrogen replacement
cycles.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: A total of 229 estrogen replacement FET
cycles were studied. All blastocysts were vitrified-warmed using the same
protocol, and only expanding good or excellent quality blastocysts were
warmed and transferred. Blastocysts were transferred on the day of warming.
Patients were prepared for transfer with oral, intramuscular (IM), and/or
transdermal estradiol to maintain a serum estradiol of 200 pg/ml. After endometrial
thickness of >8 mm and tri-laminar pattern were attained, patients
started P4 in the evening, either five or six days prior to blastocyst transfer.
Patients were given 50 mg of IM P4 every-other-evening, and 400 mg vaginal
P4 daily until at least 8 weeks of pregnancy. The Mann-Whitney non-parametric
test was used to determine if starting P4 supplementation five vs six
days before FET had a statistically significant effect on the blastocyst implantation
rates.
RESULTS: There was no difference in maternal age between the group
of patients that started P4 five days before FET (mean age¼32.3)
compared to the group that started P4 six days before FET (mean
age¼31.8). Based on the non-parametric statistical test, the mean implantation
rate of vitrified-warmed blastocysts was higher (p-value¼0.03)
when P4 was started five days before transfer (36.6%; N¼117) compared
to the implantation rate when P4 was started six days before transfer
(26.3%; N¼112).
CONCLUSIONS: Starting P4 supplementation five days before the transfer
of vitrified-warmed blastocysts resulted in higher implantation rates
compared to starting P4 six days before transfer in estrogen replacement
FET cycles. Prospective, randomized studies should be performed to further
pinpoint the duration of P4 supplementation before FET, as well as the route
of administration, that optimizes pregnancy outcomes.
P-689 Wednesday, October 21, 2015
TO EVALUATE THE PREGNANCY RATE AFTER ENDOMETRIAL
SCRATCHING IN COUPLES WITH UNEXPLAINED INFERTILITY
IN OVULATION INDUCTION AND IUI CYCLES. R. Mahey, T. Goel,
M. Gupta, G. Kachhawa, A. Kriplani. Obstetrics & Gynaecology, All India
Institute of Medical Sciences, New Delhi, New Delhi, India.
OBJECTIVE: Embryo implantation is the rate-limiting step for success of
IVF and IUI cycles and is affected by various hormonal and biological
markers. Endometrial scratching done in same cycle or previous cycles has
been shown to increase endometrial receptivity thus making capable of
receiving and adhering the embryo. The aim of present study was to compare
the pregnancy rates during IUI cycles with or without endometrial scratching
in follicular phase in couples with unexplained infertility.
DESIGN: A prospective randomized study.
MATERIALS AND METHODS: Sixty four couples with unexplained
infertility were recruited during March 2014 to March 2015. Patients were
randomized into two groups by computer generated random table. Patients
in group I (32 women, 78 cycles) underwent endometrial scratching on
day 8 of the same menstrual cycle after receiving 7 days of ovulation induction
with clomiphene citrate 50mg (day 2 to day 6) and Inj hMG 75 IU on day
6,7. Group II(32 women, 81 cycles) received ovulation induction (CC+hMG)
without endometrial scratching. Transvaginal USG was done to monitor the
follicular growth and endometrial thickness from day 8 till at least one follicle
reached 18mm diameter.Primary outcome was measured by pregnancy
rate and secondary outcome was measured by abortion rate and ongoing
pregnancy rate(>12wks).
RESULTS: Both groups were comparable in regard to mean age, BMI,
duration and type of infertility. There was no difference in the dose of clomiphene
and gonadotropin requirement in two groups. The mean endometrial
thickness in group I was 5.82.3mm on day 8 and 8.73.4mm on day of
hCG trigger. The respective values in group II were 6.11.8mm (day 8)
and 8.52.3mm (day of hCG trigger) (p value-0.9). The pregnancy rate in
group I was 18.7% (6/32) as compared to 9.3% (3/32) in group II (p value-
0.43). The abortion rate was 1/6 (16.7%)in group I and 0 in group II. The
ongoing pregnancy rate was 5/6(83%) in group I and 2/3 (66.7%)in group
II with the one being an ectopic pregnancy in group II. There were no multiple
pregnancies.
CONCLUSIONS: Endometrial scratching is associated with better pregnancy
rates, implantation rate and ongoing pregnancy rates however, further
trials with large sample size are required to establish its role in IUI cycles.
P-690 Wednesday, October 21, 2015
RECURRENT ECTOPIC PREGNANCY HISTORY IS ONE OF THE
RISK FACTORS OF INTRAUTERINE IMPLANTATION FAILURE
IN WOMEN UNDERGOING FROZEN-THAWED EMBRYO TRANS-
FER CYCLES. T. Du, H. Chen, Q. Lyu, Y. Kuang. Reproduction Department,
Shanghai Ninth People’s Hospital, Shanghai, China.
OBJECTIVE: To assess the influence of ectopic pregnancy history on intrauterine
implantation outcome in patients undergoing frozen-thawed embryo
transfer(FET) treatments.
DESIGN: Retrospective cohort study.
MATERIALS AND METHODS: A total of 7,118 patients undergoing
10,043 FETs in the period between September 2013 and April 2015 were
included and classified into two groups: implanted group (n¼4424 FETs)
and non-implanted group (n¼5619 FETs).Endometrial preparation and
FETs were performed in ordinary practice.
RESULTS: Among all subjects, 784 patients had ectopic pregnancy once
before (corresponding to 1082 FETs), 284 patients had twice or more
ectopic pregnancies before (corresponding to 390 FETs). The total times
of ectopic pregnancies (0[range 0-3] vs. 0[range 0-3]), the age
(31.294.05 vs. 32.834.95 years) and the duration of infertility (3[range
0-24] vs. 3[range 0-21] years) were significant lower in implanted group
compared with non-implanted group, whereas the total number of embryo
transferred (2[range 1-2] vs. 2[range 1-3]) and the BMI (21.703.14 vs.
21.662.96 kg/m2) was significantly higher. No differences were detected
regarding the endometrial thickness on embryo transfer (ET) day, estradiol
and progesterone levels on ET day when comparing patients in two groups.
After adjusting confounding factors including the age, BMI, uterine disorders,
duration of infertility, different stage of the embryos transferred, total
number of embryos transferred and times of ectopic pregnancy, the risk of
implantation failure significantly increased for age, uterine disorder history
and the duration of infertility, the adjusted odds ratios were 0.938 (95%
confidence interval [CI]: 0.929-0.947), 0.835 (95% CI: 0.741-0.940) and
0.983 (95% CI: 0.968-0.998), respectively. Compared with those who
had no ectopic pregnancy history, the risk of implantation failure significantly
increased for the patients had twice or more ectopic pregnancies previously,
the adjusted odds ratios was 0.783 (95% CI: 0.637-0.962). The
patients transferred blastula embryo had a significantly increased odds
for implantation compared with patients transferred cleavage stage embryo,
so were the patients with thicker endometrium on ET day and larger total
number of embryo transferred, the adjusted odds ratios were 2.053 (95%
CI: 1.772-2.380), 1.063 (95% CI: 1.045-1.081) and 1.854 (95% CI:
1.629-2.111), respectively.
CONCLUSIONS: Recurrent ectopic pregnancy is one of the risk factors of
intrauterine implantation failure. The declined intrauterine implantation rate
to some extent indicates an altered endometrium, which may play an important
role in the pathogenesis of ectopic pregnancy.
P-691 Wednesday, October 21, 2015
CAN OOCYTE RECIPIENT OUTCOME BE PREDICTED BY
OOCYTE DONOR AGE? E. Cater, a L. Jenner, a A. Campbell, b
S. Fishel. c a Embryology, CARE Fertility, Nottingham, United Kingdom;
b Director of Embryology, CARE Fertility, Nottingham, United Kingdom;
c C.E.O, CARE Fertility, Nottingham, United Kingdom.
OBJECTIVE: It is widely accepted that maternal age is one of the major
determining factors in ART success. It is therefore a consideration that in
oocyte recipient cycles, the age of the donors may also be an influential factor.
The literature is however not conclusive; some articles describing
FERTILITY & STERILITY Ò
e343

predictive power in oocyte donor age for recipient outcome, some showing
no difference and others describing a reduction in outcome from very young
donors. This study aims to establish if oocyte donor age can predict the recipient
outcome.
DESIGN: Standard oocyte recipient cycles from four private ART clinics
between January 2008 and December 2013 were retrospectively reviewed
and analysed for number of oocytes retrieved, maturity, fertilisation, achievement
of an embryo transfer (ET), clinical pregnancy rate/ET (CPR), implantation
rate (IR) and live birth rate/ET (LBR), based on oocyte donor age.
MATERIALS AND METHODS: 1229 cycles were divided into 8 groups
based on oocyte donor age of 2 year intervals; 20-21(n¼18), 22-23(n¼37),
24-25(n¼82), 26-27(n¼111), 28-29(n¼148), 30-31(n¼238), 32-
33(n¼266), 34-35(n¼329).Significance was determined using a Z-test of
two proportions; a significance value of 0.05 and two tailed hypothesis.
RESULTS: No significant difference was apparent in embryological
parameters assessed up to ETalong with CPR, IR and LBR within the groups;
a decrease in the three latter was seen in groups 28-29 and 34-35. Combining
the two upper age groups and comparing to the two lower to increase the n
number also did not show significance.CPR; 41.2% (7/17), 41.9% (13/31),
36% (27/75), 36.9% (38/103), 30.8% (44/143), 39.5% (88/223), 38.4%
(93/242), 33.4% (100/299) respectively according to the above age groups.
IR; 26.5% (9/34), 28.8% (17/59), 25.5% (38/146), 23.5% (47/200), 21.8%
(60/275), 28.3% (121/427), 24.9% (118/473), 22% (128/582) respectively.
LBR equalled the CPR up until the age of 25, after which miscarriages
occurred. LBR; 34.9% (36/103), 26.6% (38/143), 34.5% (77/223), 33.1%
(80/242), 27.8% (83/299) respectively.
CONCLUSIONS: No statistical difference was seen in outcome parameters
of oocyte recipient cycles based on oocyte donor age. It should be noted
that the 8 groups have an uneven number of cycles included which may affect
data analysis. Those with a greater number of cycles are likely to be more
reflective of the true result. As one of the largest studies so far published,
it is reassuring to note that the age range provided by the HFEA guidelines
shows no significant variation in the incidence of live Birth.
P-692 Wednesday, October 21, 2015
MID-SECRETORY EUTOPIC ENDOMETRIUM IN INTRAMURAL
FIBROIDS AND SEVERE ENDOMETRIOSIS: RELEVANCE TO
FERTILITY. L. Aghajanova, J. Irwin, L. C. Giudice. Obstetrics, Gynecology
and Reproductive Sciences, University of California San Francisco,
San Francisco, CA.
OBJECTIVE: Data are conflicting on the expression of endometrial receptivity
markers in women with intramural uterine fibroids and the effect of on
fertility. We aimed to investigate the mid-secretory phase (MSE) endometrial
transcriptome of women with intramural fibroids and compare them to controls
and those with severe endometriosis.
DESIGN: In silico and laboratory-based study.
MATERIALS AND METHODS: Well-annotated endometrial tissue samples
were obtained through the UCSF/NIH Human Endometrial Tissue Bank.
MSE samples were from 8 women with no uterine/endometrial pathology, 4
with intramural fibroids (no submucosal component) and 8 with severe endometriosis.
Purified total RNAwas subjected to microarray analysis with Gene
1.0 ST Affymetrix platform. Data were analyzed with GeneSpring and Ingenuity
Pathway Analysis. Menstrual cycle phase was assigned by endometrial
histology, estrogen/progesterone levels and bioinformatics methods. Microarray
analysis validation was performed with Fluidigm array and real-time
PCR.
RESULTS: Intramural fibroid MSE samples clustered separately from the
control or endometriosis MSE. Comparison of differentially regulated genes
revealed dysregulation of 1496 genes (989 up and 506 down) in endometrial
samples from women with intramural fibroids vs. controls, 244 genes (139 up
and 105 down) in severe endometriosis vs. controls and 1936 (1232 up and
704 down) genes in fibroid vs. endometriosis samples. Comparison of the
gene lists above with the 238 Endometrial Receptivity Aarray (ERA) genes
(Diaz-Gimeno et al, 2011) showed that only 8 and 1 genes were dysregulated
beyond the ERA 3-fold threshold in samples from women with intramural
fibroids or severe endometriosis respectively vs. controls. IGF-1 and nitric
oxide signaling were the top regulated pathways in the fibroid group, and
planar cell polarity and CRH signaling were the top regulated pathways in
the endometriosis group, while comparison of the two diseases revealed
involvement of hypoxia and oxidative stress pathways.
CONCLUSIONS: While ERA receptivity genes are minimally affected in
the setting of intramural uterine fibroids and severe endometriosis, other
genes are markedly different vs. normal controls, which may contribute to
the known poor pregnancy outcomes in these populations. Further studies
are needed to validate or refute these observations.
Reference:
1. Diaz-Gimeno, Horcajadas JA, Martınez-Conejero JA, et al. A genomic
diagnostic tool for human endometrial receptivity based on the transcriptomic
signature. Fertil Steril 2011; 95(1):50-60.
Supported by: NIH NCTRI P50HD055764 (LCG).
LUTEAL PHASE SUPPORT
P-693 Wednesday, October 21, 2015
IMPACT OF SHORT LUTEAL PHASE ON NATURAL
FERTILITY. N. M. Crawford, K. Chantala, A. Steiner. University of
North Carolina, Chapel Hill, NC.
OBJECTIVE: To determine the impact of a short luteal phase on fecundability.
DESIGN: Prospective, time-to-pregnancy study.
MATERIALS AND METHODS: Women, 30-44 years old with no history
of infertility, who were trying to conceive for less than 3 months, were
enrolled and followed until pregnancy. Each day women recorded bleeding,
ovulation predictor test results, intercourse, and pregnancy test results for
up to 4 months while attempting to conceive. For each cycle in which a
woman did not conceive, the length of the luteal phase was determined
by calculating the number of days from the first positive ovulation test to
the last day of the menstrual cycle. We defined a short luteal phase as
%11 days in length. We subsequently examined the probability of
conceiving in a cycle based on presence or absence of a short luteal phase
in the preceding non-conception cycle. Discrete time models were created
to calculate fecundability ratios (FR) adjusting for maternal age. A FR

Yuden index was applied for identifying the best cut-off value of the progesterone
(P4) level at day 16 in recognizing the 12th ongoing pregnancy status.
Using the best cut-off value of the P4 level at day 16, we categorized our subjects
into ‘‘with CLR (corpus luteum rescue)’’ and ‘‘without CLR’’. Logistic
regression with age and BMI adjustment was utilized to evaluate the association
between the duration of LPS and 12th week ongoing pregnancy in each
group.
RESULTS: A total of 158 (57.9%) women were confirmed as pregnancy at
day16. Among them, 56 (35.4%) women had LPS until 7-12th week, and 102
(64.6%) women had LPS 20.6ng/ml) and 38 (24.1%) women without CLR (P4

Summary of the main findings.
Vaginal
progesterone
capsules Oral dydrogesterone RR (95% CI)
Participants /
Studies
Authors’ Interpretation
Quality of the
evidence
Ongoing pregnancy 20% 25% (21-31%) 1.24 (1.02-1.51) 1,611 / 5 Dydrogesterone is better Moderate
Clinical pregnancy 27% 34% (30-39%) 1.24 (1.08-1.41) 2,286 / 6 Dydrogesterone is better Moderate
Miscarriage 23% 17% (11-26%) 0.75 (0.49-1.14) 438 / 5 No evidence of difference Low
Dissatisfaction 26% 3% (1-10%) 0.10 (0.02-0.39) 430 / 1 Dydrogesterone is better High
Vaginal progesterone gel Oral dydrogesterone
Ongoing pregnancy 27% 27% (23-31%) 0.97 (0.83-1.13) 1,735 / 2 No relevant difference High
Clinical pregnancy 31% 29% (25-34%) 0.95 (0.82-1.09) 1,735 / 2 No relevant difference High
Miscarriage 11% 9% (5-15%) 0.81 (0.48-1.35) 523 / 2 No evidence of difference Low
Dissatisfaction 18% 5% (3-8%) 0.26 (0.16-0.42) 822 / 1 Dydrogesterone is better High
OBJECTIVE: To identify, appraise and summarize the evidence from randomized
controlled trials (RCTs) comparing oral dydrogesterone with progesterone
for luteal-phase support (LPS) in women undergoing IVF/ICSI.
DESIGN: Systematic review and meta-analysis.
MATERIALS AND METHODS: Record screening, study selection, data
extraction, and evaluation of the risk of bias were performed independently
by two authors. The comparison between dydrogesterone and progesterone
were summarized as risk ratio (RR) and the precision of the estimates was
assessed by the 95% confidence interval (CI).
RESULTS: Our electronic search was performed in Mar-17-2015 and 7
studies were included in our quantitative analysis: 5 studies compared oral
dydrogesterone with vaginal progesterone capsules; 1 study compared oral
dydrogesterone with vaginal progesterone gel; and 1 study has three arms:
oral dydrogesterone, vaginal progesterone capsules and vaginal progesterone
gel, being considered in both comparisons. We didn’t find any study
comparing oral dydrogesterone with intramuscular or oral progesterone.
The main results are reported in the table.
CONCLUSIONS: The available evidence from RCTs suggests that oral
dydrogesterone for LPS provides at least similar reproductive outcomes
and less dissatisfaction than vaginal progesterone capsules or gel.
Supported by: Conselho Nacional de Desenvolvimento Cientıfico e Tecnologico
(CNPq); S~ao Paulo Research Foundation (FAPESP).
P-698 Wednesday, October 21, 2015
THE ADDITION OF GNRH AGONIST FOR LUTEAL PHASE SUP-
PORT IN OVUM DONATION CYCLES. P. Casanova, a E. Szlit Feldman,
b G. J. Rey Valzacchi, b L. A. Blanco, c C. A. Carrere, b A. Torno, d
M. A. Rodriguez Kubrusli, a J. I. Mannara, d V. E. Canada. e a Gynecologist
Procrearte, Buenos Aires, Argentina; b Procrearte Director, Buenos Aires,
Argentina; c Reproductive Medicine, Caba, Argentina; d Procrearte, Buenos
Aires, Argentina; e Procrearte, Capital Federal, Argentina.
OBJECTIVE: To evaluate the effect of 0,1mg triptorelin in a single injection
administered to ovum donation recipients the 6th day after donor oocyte
retrieval on pregnancy outcomes.
DESIGN: Prospective trial.
MATERIALS AND METHODS: This study involved 78 ovum donation
cycles between August 2014 and February 2015, in a private fertility center.
Patients were divided into two groups. Group A (Study group): 43 patients
received one dose of 0,1mg Triptorelin (GonapeptylÒ Ferring) on
the day 6th after donor oocyte retrieval in addition to regular luteal phase
support (vaginal progesterone 600-800 mg/day and oral 17 B estradiol 6-
8 mg/day). Group B (Control group): 35 patients received only regular
phase support. The study group was conformed according to patients’
agreement to participate. Embryos were transfered at cleaving stage
(48-72hs) with Frydman Catheter. Patients with at least one good quality
embryo (class I-II) were included. Severe male factor were excluded
(WHO criteria).Statistical analysis was performed using SPSS. p

supplementation over the period 01.11.2013 to 30.06.2014. Previously, the
patients were enrolled as a vaginal p. group between 01.11.2012 and
01.11.2013. In the combined p. group, IM progesteron 100 mg/day and
vaginal progesteron 90 mg/day were administered from embryo transfer
day to bhCG day (a total of 10 days). 90 mg of vaginal progesteron gel
was given twice daily in the vaginal p. group. Luteal phase support was
continued with vaginal progesteron up to 10 weeks of gestation for pregnant
women.
RESULTS: The clinical pregnancy rate per embryo transfer (ET), ongoing
pregnancy rate/ETand live birth rate/ETwere significantly higher in the combined
p. group than in the vaginal p. group (Table). There was no difference in
the serious side effects between the groups.
CONCLUSIONS: This study reveals a new perspective in the luteal phase
support. Our results indicate that, short-term, intensive luteal phase support
significantly improved live birth rates in intracytoplasmic sperm injection cycles.
Demographic features and cycle outcomes of the groups.
Combined p.
gr.(IM+Vag. p)
n¼386
Vaginal p. gr.
n¼251
P value
Age (y) 305 30.94.6 0.04
Total gonadotrophin 2286979.9 2295.8948 NS
dose (IU)
No. of MII oocytes 8.45.1 9.45.9 NS
Estradiol level on hCG 2549.11768.7 2925.42063.7 0.049
day (pg/ml)
Estradiol level on OPU 1660.31189.8 1839.21413.9 NS
day(pg/ml)
Progesterone level on OPU 7.954.4 8.275.6 NS
day (ng/ml)
Estradiol level on ET 1546.71290.6 1900.71512 0.003
day(pg/ml)
Progesterone level on ET 46.728.2 50.843.4 NS
day(ng/ml)
Clinical pregnancy rate/ET 43.2 28.5 0.001
Ongoing pregnancy rate/ET 29.1 20.2 0.001
Live birth rate/ET 28.6 20.2 0.001
EARLY PREGNANCY
P-700 Wednesday, October 21, 2015
DO ELEVATED TSH LEVELS PREDICT EARLY PREGNANCY
LOSS IN ART PATIENTS? J. Rodriguez-Purata, a J. Gingold, b
M. C. Whitehouse, a J. A. Lee, a A. B. Copperman. c a Reproductive Medicine
Associates of New York, New York, NY; b Icahn School of Medicine at Mount
Sinai, New York, NY; c RMANY-Mount Sinai, New York, NY.
OBJECTIVE: Infertility related to ovulatory dysfunction is commonly
associated with thyroid disorders such as hypothyroidism. A number of
studies have observed increased miscarriage rates in patients with hypoand
hyperthyroidism, but the basis of this association remains unclear. While
the manufacturer-listed normal range TSH is

P-702 Wednesday, October 21, 2015
WOMEN’S EXPERIENCE POST MISOPROSTOL TREATMENT
FOR FIRST TRIMESTER MISCARRIAGES. B. Zhou. OB GYN,
Houston Methodist Hospital, Houston, TX.
OBJECTIVE: To report the outcomes and patient perspectives of the medical
management of nonviable pregnancies before 12 weeks gestation.
DESIGN: Retrospective cohort analysis.
MATERIALS AND METHODS: The study included women with first
trimester miscarriages who underwent treatment with vaginal misoprostol
from 1/2011 through 12/2014 at Houston IVF Clinic. Gestational age were
calculated by certain menstrual dates or date of embryo transfer and
compared with ultrasound criteria. Pregnancies were considered nonviable,
if no cardiac activity was detect by a CRL greater than 7mm, no fetal pole
detected when mean sac diameter was greater than 25mm, or if the pregnancy
displayed abnormal growth. Non-viability was confirmed with serial ultrasound
scans. Once non-viability was confirmed, patients were given misoprostol
800mcg vaginally and followed up 24 hours post placement. The
patients then filled out a post treatment questionnaire, which assessed onset
of pain, bleeding, passage of tissues, and their overall experience. The primary
outcome measured was whether or not patients would recommend misoprostol
as a treatment to peers in the future. Means and standard deviations
were calculated.
RESULTS: 83 underwent medical treatment with misoprostol for their first
trimester miscarriage. Five patients were excluded because of incomplete
completion of the questionnaire. Complete miscarriage was induced without
the need for surgery in 75 women (96%). Surgical dilation and curettage
(D&C) was performed in 3 patients (4%). Women reported average onset
of pain was 2.9 hours after miso placement, average pain rating was 7/10
and average duration of pain was 7.4 hours. Onset of bleeding after miso
placement was 4.7 hours and 54% (42/78) perceived the bleeding as heavy,
and 91% (71/78) reported noticing passed products of conception. Among
those who returned patient questionnaires, 91% participants (71/78) indicated
they would recommend medical management to peers, while 3% (3/
78) indicated they would undergo surgery next time, and 5% (4/78) were undecided.
40% (31/78) of participants have had a D&C prior to this study. The
major reason participants recommended misoprostol over D&C was that misoprostol
was more simple, fast, and not as painful as expected.
CONCLUSIONS: The medical management of first trimester miscarriage
on the outpatient basis with misoprostol is effective and highly recommended
by patients who experienced it.
P-703 Wednesday, October 21, 2015
PRE-PREGNANCY LOW TO MODERATE ALCOHOL INTAKE
AND RISK OF SPONTANEOUS ABORTION. A. J. Gaskins, a
J. W. Rich-Edwards, b P. Williams, c T. Toth, d S. A. Missmer, e
J. E. Chavarro. f a Department of Nutrition, Harvard T.H. Chan School of Public
Health, Boston, MA; b Connors Center for Women’s Health and Gender
Biology, Brigham and Women’s Hospital and Harvard Medical School, Boston,
MA; c Harvard T. H. Chan School of Public Health, Boston, MA; d Massachusetts
General Hospital; e Brigham and Women’s Hospital and Harvard
Medical School, Boston, MA; f Harvard School of Public Health, Boston,
MA.
OBJECTIVE: To examine the relationship between pre-pregnancy alcohol
intake and risk of spontaneous abortion (SAB).
DESIGN: Prospective cohort study.
MATERIALS AND METHODS: Our prospective cohort study included
27,580 pregnancies reported by 17,929 women with no history of SAB in
the Nurses’ Health Study II between 1990 and 2009. Pre-pregnancy alcohol
intake over the past year was assessed in 1989, 1991, and every 4 years thereafter
using a validated questionnaire. Pregnancies were self-reported with
case pregnancies lost spontaneously at less than 20 weeks of gestation and
comparison pregnancies ending in ectopic pregnancy, induced abortion, or
live birth. Multivariable log-binomial regression models with generalized
estimating equations were used to estimate the relative risks (RRs) and
95% confidence intervals (CIs) adjusting for age, year, BMI, smoking status,
physical activity, history of infertility, marital status, race, caffeine intake,
and multivitamin use.
RESULTS: Incident spontaneous abortion was reported in 4,326 (15.7%)
pregnancies. Pre-pregnancy alcohol intake was not associated with pregnancy
loss. Compared to women who did not consume alcohol, the multivariable
RRs (95% CIs) for increasing categories of alcohol intake were 1.04
(0.97, 1.12) for 0.1-1.9 g/day, 1.02 (0.94, 1.11) for 2-4.9 g/day, 1.01 (0.92,
1.10) for 5-9.9 g/day, and 0.98 (0.88, 1.09) for R 10 g/day (p-trend¼0.45).
Women who consumed R2 servings of beer per week prior to pregnancy had
a 9% (95% CI 1, 17%) lower risk of pregnancy loss compared to women
consuming

P-705 Wednesday, October 21, 2015
PRECONCEPTION LOW DOSE ASPIRIN TREATMENT IM-
PROVES CLINICAL PREGNANCY AND LIVE BIRTH IN WOMEN
WITH HIGHER SYSTEMIC INFLAMMATION. L. Sjaarda, a
E. Mitchell, a S. L. Mumford, a R. Radin, a N. J. Perkins, a N. Galai, b
R. M. Silver, c E. Schisterman. a a NICHD, NIH, Rockville, MD; b Haifa University,
Haifa, Israel; c Unversiy of Utah, Salt Lake City, UT.
OBJECTIVE: While some studies of low dose aspirin (LDA) to
improve reproductive outcomes have found a beneficial effect, others
have found none. Recently, we reported that preconception LDA
(81mg) increased fecundability and live birth rates relative to placebo
in a subset of women with a history of one recent (prior 12 months) pregnancy
loss in the EAGeR trial. Here we aimed to evaluate the effect of
LDA, a known anti-inflammatory agent, according to inflammatory status
to explore possible mechanisms and best identify women who may benefit
from treatment.
DESIGN: Multicenter, block-randomized, double-blind, placebocontrolled
trial of 1228 women to evaluate the effect of preconception-initiated
daily LDA on clinical pregnancy and live birth in women with a history
of pregnancy loss.
MATERIALS AND METHODS: We assessed high-sensitivity C-reactive
protein (CRP), a marker of systemic inflammatory status, at baseline. Women
were stratified into tertiles of baseline CRP concentration: lower
(

were included in this study. Exclusion criteria included donor oocyte cycles,
frozen-thawed embryo cycles, pregnancies of unknown location, and treatment
with salpingectomy. LFTs, specifically alanine aminotransferase (ALT), aspartate
aminotransferase (AST), albumin and total bilirubin levels were measured
on day of MTX administration (day 1) and 7 days later (day 7). The change in
LFTs (D) between day 1 and day 7 was calculated for both single and double
dose MTX protocols. Furthermore, the change in LFTs (D) for single dose and
double dose MTX protocols was compared. Continuous variables were checked
for normality and expressed as mean standard deviation. Paired student’s t-tests
were utilized. Statistical significance was set at P

Table 1. Notch Activity and Expression of Notch Proteins and Ligands in the Decidua and Placenta
E8.5 Notch Activity Notch1 Notch2 Notch4 Dll4 Jag1 Jag2
Maternal decidua ECs
[capillaries &
ECs Decidual cells ECs,
Decidual cells
ECs ECs
[spiral arterioles]
spiral arterioles]
Ectoplacental cone TBs, TGCs TBs, TGCs TBs, TGCs TBs, TGCs
E12.5 Notch Activity Notch1 Notch2 Notch4 Dll4 Jag1 Jag2
Maternal decidua ECs
ECs ECs ECs
[capillaries]
Junctional zone Spongio-TBs Spongio-TBs
Labyrinth
Fetal ECs,
TGCs
Fetal ECs TGCs, syncytio-TBs TGCs, syncytio-TBs Fetal ECs TGCs
capillaries and Dll4 activates Notch1 in fetal placental vessels. In the developing
placenta at E8.5, Dll4 may activate Notch2 and Notch4 in TBs, while
Jag2 may activate Notch2 and Notch4 in TGCs in the mature placenta. Taken
together these results suggest Notch signaling dynamically regulates
decidual angiogenesis and placentation.
Supported by: Robert Wood Johnson Foundation.
P-711 Wednesday, October 21, 2015
PREGNANCY-ASSOCIATED PLASMA PROTEIN A (PAPP-A): A
BIOMARKER FOR THE AID IN RISK STRATIFICATION OF
NONVIABLE PREGNANCY. R. J. Batson, a B. B. Mills, b Z. Nagy, c
W. Roudebush. d a Biomedical Sciences, University of South Carolina School
of Medicine Greenville, Greenville, SC; b Obstetrics & Gynecology, Greenville
Health System, Greenville, SC; c Reproductive Biology Associates, Atlanta,
GA; d Biomedical Sciences, University of South Carolina School of
Medicine, G, Greenville, SC.
OBJECTIVE: Pregnancy-associated plasma protein A (PAPP-A) is a
placenta-derived glycoprotein produced by trophoblastic cells, which rises
gradually during the early weeks of a viable pregnancy. Its biological function
is unknown but measuring serum levels has proven useful in a number
of abnormal obstetrical conditions. Currently, serum quantitative bhCG
levels and ultrasound are utilized for following early pregnancies and, ultimately,
separating viable from nonviable pregnancies (miscarriages and ectopics);
however, the diagnosis is often delayed due to limitations of these
tests. The objective of this study is to compare PAPP-A levels in human
sera obtained from women with viable intrauterine pregnancies to those
with nonviable pregnancies following assisted reproductive technology.
DESIGN: Retrospective cohort study in patients undergoing in vitro fertilization
and embryo transfer (IVF-ET) at a tertiary fertility clinic.
MATERIALS AND METHODS: Women with positive bhCG measurements
14 days after IVF transfer werecategorized into two groups: (a)
nonviable pregnancy, including biochemical pregnancy (bhCG
5-50 mIU/mL) and pregnancies with bhCG >50 mIU/mL which did not double
within 72 hours, and (b) viable pregnancy (bhCG >50 mIU/mL which at
least doubled within 72 hours). Serum PAPP-A levels were measured via an
ultrasensitive enzyme-linked immunosorbent assay (Beckman Coulter,
Chaska, MN). PAPP-A levels between 4 weeks-4 days and 6 weeks-3 days
were compared for viable and nonviable pregnancies.
RESULTS: Twenty-five pregnant women undergoing IVF-ET were followed
for this study, 7 resulted with a biochemical pregnancy, 5 with an
ectopic pregnancy, and 13 with a viable intrauterine pregnancy. Three or
four serum samples from each patient (n¼97 samples) were assayed for
PAPP-A. The levels of PAPP-A ranged from 0.60 ng/mL to 3.3 ng/mL
with analytical and functional sensitivities of 0.2 ng/mL and 1.25 ng/mL with a sensitivity of 69.2 and a specificity
of 100.0 at a gestational age of 5 weeks-4days.
CONCLUSIONS: From gestational week 5 onwards in normal pregnancies,
PAPP-A was significantly increased in normal viable pregnancies
over nonviable pregnancies. In nonviable pregnancy, PAPP-A appeared
to remain low for the entire observation period. Pregnancy-associated
plasma protein A (PAPP-A) appears to be a biomarker for nonviable pregnancy.
P-712 Wednesday, October 21, 2015
THE RELATIONSHIP BETWEEN INITIAL HCG LEVELS AND
DEVELOPMENT OF PREECLAMPSIA IN PREGNANCIES
FOLLOWING IN VITRO FERTILIZATION. M. Erdem, a
A. Najafaliyeva, a I. Guler, a A. Erdem, a N. Bozkurt, a M. Oktem, a
M. F. Mutlu. b a Obstetrics and Gynecology, Gazi University Faculty of Medicine,
Ankara, Turkey; b Koru Hospital, Ankara, Turkey.
OBJECTIVE: To evaluate whether initial levels of hCG in pregnant patients
after an IVF-ICSI cycle is related with development of preeclampsia.
DESIGN: Retrospective case-control study.
MATERIALS AND METHODS: IVF and obstetric records of 417 patients
who had a live birth after IVF from 2005 to 2012 were evaluated. Serum betahCG
levels on day 12 and 14 after embryo transfer were compared between
patients who were treated with ICSI and fresh or frozen/thawed embryo
transfer cycle and had a singleton live birth that was complicated with preeclampsia
or not.
RESULTS: A total number 168 patients were included into the analysis. In
32 patients, hCG levels were measured other than day 12 or 14 and were
excluded from the study. In 136 patients day 12 and day14 hCG levels
were measured and included to the study. Twenty-five (18%) patients had
preeclamsia. Demographic data of patients were comparable between each
group. Day 12 and day 14 beta hCG values were significantly lower in preeclampsia
group (137, 17 72,0 IU/l and 254.1 158,7 IU/l, respectively) as
compared to control group (205,1 99,5 and 457.5 267,3 IU/l, respectively)
(p¼0.001 and p¼0.003, respectively).
CONCLUSIONS: Initial plasma levels of hCG were inversely correlated
with development of preeclampsia in pregnancies following embryo transfer.
Our results support the theory of impaired invasion of trophoblasts and
abnormal placentation in the main pathophysiology of preeclampsia.
P-713 Wednesday, October 21, 2015
ASSOCIATION BETWEEN LEPTIN AND PREGNANCY
LOSS. T. Plowden, a S. Zarek, a E. Schisterman, a L. Sjaarda, a
R. M. Silver, b N. Galai, c A. DeCherney, a S. L. Mumford. a a NICHD,
NIH, Bethesda, MD; b Unversiy of Utah, Salt Lake City, UT; c Haifa University,
Haifa, Israel.
OBJECTIVE: Previous research in both mouse and human models suggests
that leptin plays an important role in reproductive health, although
the exact mechanism of action is unclear. Our objective was to explore if
maternal leptin levels are associated with early pregnancy loss.
DESIGN: Prospective cohort study of a multi-center, block-randomized
placebo-controlled trial of pre-conception low dose aspirin to assess its
impact on reproductive outcomes in healthy, fertile women who have had
1 or 2 prior pregnancy losses. Leptin was measured in serum drawn at baseline.
Women who became pregnant during the study were followed prospectively
throughout their pregnancy.
MATERIALS AND METHODS: A total of 1194 women had measured
leptin levels and were included in this analysis. The women were classified
into low (0.007-11.3ng/ml), middle (26.3-97.4 ng/ml) and high
(11.4-26.2 ng/ml) tertiles, with the middle tertile serving as the reference
group. Pregnancy status was established using daily first-morning urine
collection and spot urine clinic pregnancy tests at monthly visits. Chemical
pregnancy loss was defined as positive hCG without clinical evidence of
FERTILITY & STERILITY Ò
e351

pregnancy (n ¼56) and clinical pregnancy loss as a loss following ultrasound
confirmed pregnancy (n¼133). RR and 95% CIs for pregnancy loss were estimated
using generalized linear models adjusted for age and the probability of
confirmed pregnancy using stabilized inverse-probability-weights.
RESULTS: Among women who became pregnant during this study
(n¼771), 279 were in the low tertile, 275 were in the middle tertile and 217
were in the high tertile. After adjusting for age, those in the low and high leptin
tertiles did not have a higher risk of chemical pregnancy loss compared to the
reference middle tertile group (low tertile: RR 0.89, 95% CI 0.48, 1.65; high
tertile: RR 1.13 95% CI 0.61, 2.09). Additionally, there was no difference in
clinical pregnancy loss among the groups (low tertile: RR 0.88, 95% CI
0.61, 1.28; high tertile: RR 1.02, 95% CI 0.69, 1.46) compared to the middle
tertile. Similar results were noted after adjusting for body mass index.
CONCLUSIONS: Preconception leptin levels were not associated with an
increased risk of chemical or clinical pregnancy loss among women with a
history of prior pregnancy losses. Our data suggests that leptin levels are
not associated with early pregnancy loss.
Supported by: Intramural Research Program, DIPHR, PRAE, NICHD,
NIH.
P-714 Wednesday, October 21, 2015
C-REACTIVE PROTEIN AND PREGNANCY LOSS: RESULTS
FROM THE EFFECTS OF ASPIRIN IN GESTATION AND REPRO-
DUCTION (EAGER) TRIAL. S. L. Mumford, a L. Sjaarda, b
R. Silver, c R. Radin, d E. Mitchell, b E. Schisterman. e a NICHD, NIH, Rockville,
MD; b NICHD, Rockville, MD; c University of Utah, Salt Lake City, UT;
d NIH, Rockville, MD; e Eunice Kennedy Shriver National Institute of Child,
Rockville, MD.
OBJECTIVE: A relationship between systemic inflammation and pregnancy
loss has been reported. Therefore, we determined whether baseline inflammatory
status, as indicated by C-reactive protein (CRP), was associated
with pregnancy loss.
DESIGN: Secondary analysis of the EAGeR Trial, a multicenter, blockrandomized,
double-blind, placebo-controlled clinical trial to evaluate the effect
of preconception-initiated daily low dose aspirin on reproductive outcomes
in women with a history of pregnancy loss.
MATERIALS AND METHODS: Participants were attempting pregnancy,
aged 18-40 years, with one to two prior pregnancy losses and no history
of infertility or other gynecologic disorders. We assessed highsensitivity
CRP, a marker of systemic inflammatory status, at baseline prior
to randomization to LDA or placebo. Women were allocated to tertiles
based on their CRP concentration at baseline (12 missing CRP): low
(n¼279,

embryo transfer number or eSETwas directly correlated with choosing eSET,
with higher numbers of channels being associated with higher rates of eSET
(p

from off-site work, as shown in the latest 2014 survey. In addition, most survey
responses showed satisfaction with their current jobs and optimism on
the future reproductive laboratory job market.
CONCLUSIONS: Reproductive lab professionals have experienced an
overall steady increase in salaries relative to national labor wage and most
other clinical laboratory wage benchmarks. They also maintain an optimistic
outlook on their future job market. Potential factors and impacts on these
trends warrant further investigation.
P-720 Wednesday, October 21, 2015
FERTILITY-RELATED SMARTPHONE APPLICATION USE
AMONG PATIENTS SEEKING TREATMENT FOR
INFERTILITY. M. Lanham M. A. Christensen. University of Michigan,
Ann Arbor, MI.
OBJECTIVE: To determine how women presenting with infertility to a subspecialty
clinic are using fertility-related smartphone applications ("apps").
DESIGN: Cross-sectional observational study of women presenting as
new patients to an reproductive medicine clinic.
MATERIALS AND METHODS: Paper surveys were offered to 310
consecutive, English-speaking women who presented as new patients to an
outpatient fertility clinic. IRB approval was obtained prior to initiating the
study. The response rate was 89.7% (278/310).
RESULTS: Of those patients completing the survey, the most common
age range was 30 to 34 years (35.0%), with 88.0% of participants’ ages falling
between 25 and 39 years. The majority of patients (81.5%) had an
annual household income greater than fifty-thousand dollars. Thirty-two
percent of women had been attempting pregnancy between 6 and 12
months, while 56.5% had been attempting pregnancy for over 12 months.
Eighty-eight percent of participants were actively trying to become pregnant,
68.0% had regular periods, and 97.4% owned a smartphone. The
vast majority of patients owned either an iPhone (63.5%) or an Android device
(35.0%). The participants who were trying to become pregnant were
more likely to be tracking their periods (odds ratio [OR] 7.0, 95% confidence
interval [CI] 3.2-15.4). Of those trying to become pregnant, those
with regular periods were more likely to be tracking their periods (OR
3.2, 95%CI 1.4-7.7) and those who were tracking periods were more likely
to have fertility apps installed on their smartphone (OR 23.7, 95%CI
5.5-102.6). Of those who were trying to become pregnant and tracking their
periods, age under 35 years was associated with an increased likelihood of
using an app to track periods compared to other tracking methods such as
basal body temperature or ovulation predictor kits (OR 2.8, 95%CI 1.5-
5.3). Out of a total of 37 reported fertility apps, the top three most popular
by reported usage were "Fertility Friend Mobile Smart Ovulation Tracker
Calendar" (used by 10.4% of participants), "My Days - Period & Ovulation"
(9.0%) and "Period Tracker" (7.9%). The top three most desired functionalities
for a fertility app, chosen from a list of 10 options, were an
appointment reminder (92.2% were interested), a way to send cycle data
to a physician (85.1%), and access to medical records (82.1%). The least
desired function was delivery of information on healthy activities (47.2%).
CONCLUSIONS: Smartphones are nearly ubiquitous in the population of
women presenting to the fertility clinic in this study. Women who are under
age 35, trying to become pregnant, and have regular cycles are more likely to
use apps to track their periods than other women. There is demand for
increased interaction with providers through a digital medium and for integration
of consumer health apps with clinical medicine. The possibilities
are numerous and might include, for example, remote clinical monitoring
during cycles of stimulated ovulation. Although the specific benefits or harms
of fertility apps remain to be determined the potential is large.
Reference: Study data were collected and managed using REDCap electronic
data capture tools hosted at the University of Michigan.
Supported by: Michigan Institute for Clinical & Health Research grant
support (CTSA: UL1RR024986).
SEXUALITY
P-721 Wednesday, October 21, 2015
THE PROSPECT OF CHILDBEARING AND FAMILY FORMATION
IN THE EYES OF CHINESE YOUNG LESBIANS: A QUALITATIVE
STUDY. I. P. Lo C. Chan. The University of Hong Kong, Hong Kong,
Hong Kong.
OBJECTIVE: This research aims at exploring Chinese young lesbians’
attitudes and intention towards the use of assisted reproductive technologies
(ART) and the ways in which they construct the meanings of childbearing
and family in a culture where homosexuality still remains
stigmatized.
DESIGN: This is a qualitative study. Twelve lesbian-identified Chinese
women in Hong Kong who aged between 18 and 35 were recruited through
local LGBT organizations. In-depth semi-structured interviews were conducted
in April 2015.
MATERIALS AND METHODS: Based on grounded theory, this study
investigated Chinese young lesbians’ lived experiences with regard to regulatory
social practices and processes that frame their perceptions of ART,
childbearing, and family formation. Participants were invited to share their
desire and motivation for childbearing and the prospective use of ART, as
well as their coping strategies in a heterosexist society where same-sex marriage,
civil partnership, and ART are not legally allowed. Each interview
lasted around 90 minutes. All interviews were audio-recorded and transcribed
for coding.
RESULTS: The majority of respondents reported that one of the major obstacles
in achieving parenthood was the perceived negative costs of ‘‘being
outed’’ in the public by conceiving children and becoming parents. Even
though they would like to have their own children, they felt reluctant to
disclose their sexual orientation particularly to their parents since being homosexual
would be considered a shame to the family. Most of them also
worried that their children born as a result of ART would face discrimination
due to their non-normative family formation and structure. In this regard, it
was found that they would internalize the feelings of shame and guilt about
their inevitable use of unconventional means of conception. Homophobic
messages in society had probably led them to believe that their sexual identity
was incompatible with reproduction, which is not only practically unachievable
but also socially censured in Hong Kong.
CONCLUSIONS: This study highlights that the nature of involuntary
childlessness among Chinese young lesbians is by no means only a medical
issue but also a psychosocial and ethical issue which is tightly linked with
gender and heterosexual family norms in the Hong Kong Chinese context.
It signifies a major step forward to understand local lesbians’ attitudes and
intentions towards childbearing as well as the role of parental and social pressure
on their fertility plan. It sensitizes mental health practitioners to the
emotional burden of Chinese lesbians in relation to their fertility plan. Public
education is needed to understand the needs and wants of lesbians and to
create social space for same-sex relationships and families to be visible
and legitimized.
Supported by: This research is Supported by the Small Project Funding
from University Research Committee of the University of Hong Kong [Project
No.: 201409176248].
MENTAL HEALTH
P-722 Wednesday, October 21, 2015
AS BOTH PERSON AND A PROFESSIONAL: CULTIVATING
COMPASSION SATISFACTION AMONG HEALTHCARE PRACTI-
TIONERS SPECIALIZED IN ASSISTED REPRODUCTIVE TECH-
NOLOGIES (ART). H. Chan, a S. Wong, b M. Tam. c a Queen Mary
Hospital, Hong Kong, Hong Kong; b The University of Hong Kong, Hong
Kong, Hong Kong; c Department of Social Work and Social Administratio,
Hong Kong, Hong Kong.
OBJECTIVE: Healthcare practitioners in ART setting work under high
pressure environment with heavy caseloads and patients facing a lot uncertainty
and uncontrollability. This study aims at evaluating a professional
development program which was develop to cultivate compassion satisfaction
(CS), which refers to the pleasure derived from doing health care
work effectively, and enhancing their ability to receive gratification from
caregiving.
DESIGN: This is a quasi-experimental study. Twenty-five healthcare practitioners
from 11 licensed ART clinics in Hong Kong, including obstetricians
and gynecologists, nurses, social workers and embryologists, attended a
professional training course on infertility counseling. Effectiveness of the
program was evaluated before and after the implementation of the program.
MATERIALS AND METHODS: The professional development program
was composed of nine three-hour weekly sessions, including components
such as self-reflective activities, mindfulness, compassionate meditation
and personal reflection on vulnerabilities and resilience, life and death, as
e354 ASRM Abstracts Vol. 104, No. 3, Supplement, September 2015

well as pain and suffering. All of them were invited to complete a self-administered
questionnaire before (T0) and after (T1) the course, which contained
Professional Quality of Life (ProQol) measuring CS, compassion fatigue
(CF), and other validated measures related to psychological wellbeing.
RESULTS: Participants showed significant improvement in CS after
attending the course (Pre-course: 33.94 + 4.4; Post-course: 36.35 + 3.82,
t¼-3.29, p

RESULTS: Correlational analyses showed positive associations between
anxiety and all of the predicted risk variables. To assess the unique
effect of the minority-specific stressor, a multiple regression model was
used. Results showed that, after controlling for the effects of general
stressors, perceptions of heterosexist bias were associated with higher
levels of anxiety.
Table 1: Effect of General and Minority-Specific Stressors on Anxiety.
Anxiety
Step 1: General stressors .21*
Length of time trying to conceive -0.06
Cost of treatment 0.18
Perceived lack of control 0.39*
Step 2: Minority-specific stressors .13*
Perceived heterosexist bias 0.33*
Note: *p< .05
b
R 2 Change
CONCLUSIONS: In the context of fertility treatment, sexual minorityspecific
stressors appear to uniquely contribute to anxiety for lesbian women
above and beyond the effect of general stressors. Changes in practice that
reduce perceptions of heterosexist discrimination, or interventions to help
women cope with such experiences, may help optimize the experiences
that lesbian women have in treatment settings and reduce psychological
distress.
P-726 Wednesday, October 21, 2015
MOTHERHOOD AFTER AGE 50: LONG-TERM FOLLOW UP OF
PHYSICAL AND MENTAL WELL-BEING OF WOMEN WHO
BECAME MOTHERS THROUGH OOCYTE
DONATION. E. Davenport, H. Burks, R. Paulson. Obstetrics and Gynecology,
University of Southern California, Los Angeles, CA.
OBJECTIVE: To compare long-term mental and physical health outcomes
of women who became mothers in their 50s to outcomes of younger women
who also became mothers through oocyte donation.
DESIGN: A descriptive study using validated surveys.
MATERIALS AND METHODS: Participants were former infertility patients
at an academic IVF center who became pregnant at any age through
oocyte donation, and who had a live birth of one or more children who are
now adolescents age 11 to 18. Participants completed a demographic survey
and the SF-36-v2 Health Survey, a multi-purpose survey to assess mental and
physical well-being. The SF-36v2 scores for physical and mental well-being
were calculated for each participant, and median scores were calculated for
each age group:

OBJECTIVE: To evaluate parent and donor-conceived child characteristics
in families headed by single mothers (SM) compared with same-sex female
couples (SSC) and heterosexual couples (HC).
DESIGN: Cross-sectional study of 216 parents (n ¼ 85 SM, n ¼ 72
SSC, n ¼ 59 HC). Participants were recruited from fertility clinics,
donor programs, and parenting groups. The 216 children (4-17 years
old; M age ¼ 7.56 years, SD age ¼ 3.82 years) were conceived via
IVF (31.0%) or IUI (69.0%) with donor sperm (83.4%), donor egg
(10.6%), or donor sperm and donor egg (6.0%). Participating families
were from the U.S. (77.4%), Canada (14.3%), Europe (7.4%), and
Australia (0.9%).
MATERIALS AND METHODS: Parents completed an online survey assessing
parental well-being (Center for Epidemiological Studies - Depression
- 10 items; Radloff, 1977), parent-child relationship satisfaction
(Marital Opinion Questionnaire; Huston & Vangelisti, 1991), parent-child
communication climate, which facilitates positive, open communication on
various topics (Revised Family Communication Pattern Instrument - Conversation;
Ritchie & Fitzpatrick, 1990), and child psychosocial adjustment
(Strengths and Difficulties Questionnaire; Goodman, 1997). Covariates
included parent age, family income, multiple birth status, child’s sex and
age. ANCOVAs were used to test differences in outcomes across family
types.
RESULTS: After accounting for covariates, parents in SM, SSC and HC
families had similar levels of parental well-being (Msm ¼ 5.76, Mssc ¼
6.20, Mhc ¼ 5.61; clinical cutoff: 10.00), parent-child relationship satisfaction
(Msm ¼ 4.30, Mssc ¼ 3.80, Mhc ¼ 4.29), and child emotional (Msm ¼
2.89, Mssc ¼ 4.07, Mhc ¼ 3.10; normal range: 0.00 - 5.00), behavioral (Msm
¼ 4.22, Mssc ¼ 4.17, Mhc ¼ 4.95; normal range: 0.00 - 7.00), and prosocial
adjustment (Msm ¼ 8.54, Mssc ¼ 8.66, Mhc ¼ 8.36; normal range: 6.00 -
10.00). SSC families had a more open parent-child communication climate
(Mssc ¼ 4.42) than SM families (Msm ¼ 4.28) and HC families (Mhc ¼
4.23; F(2, 205) ¼ 3.46, p ¼ .033).
CONCLUSIONS: Families headed by single mothers should not be
perceived as differing substantially from traditional nuclear families. SM
families appear to be functioning well.
References:
1. Goodman R. The strengths and difficulties questionnaire: a research
note. J Child Psychol Psychiatry 1997;38:581-586.
2. Huston TL, Vangelisti AL. Socioemotional behavior and satisfaction in
marital relationships: a longitudinal study. J Pers Soc Psychol
1991;61:721-733.
3. Radloff LS. The CES-D scale: a self-report depression scale for
research in the general. Appl Psych Meas 1977;1:385-401.
4. Ritchie LD, Fitzpatrick MA. Family communication patterns:
measuring interpersonal perceptions of interpersonal relationships.
Comm Res 1990;17:523-544.
Supported by: University of Minnesota (UMN) Agriculture Experiment
Station, UMN Grant-in-Aid, UMN College of Education & Human Development
Research Development Investment Grant.
P-729 Wednesday, October 21, 2015
MENTAL HEALTH DISORDERS IN INFERTILE WOMEN: PREVA-
LENCE, PERCEIVED EFFECT ON FERTILITY, AND WILLING-
NESS FOR TREATMENT FOR ANXIETY AND
DEPRESSION. H. S. Hoff, a N. M. Crawford, b J. E. Mersereau. c a Reproductive
Endocrinology and Infertility, University of North Carolina, Chapel
Hill, NC; b University of North Carolina, Chapel Hill, NC; c UNC, Chapel
Hill, NC.
OBJECTIVE: To evaluate whether women with infertility identify with
being anxious or depressed, assess their perception that mental health disorders
affect fertility, and determine their willingness to undergo treatment.
DESIGN: Prospective, observational study.
MATERIALS AND METHODS: An electronic survey was sent to all
new infertile female patients over a period of 2 years at UNC Fertility.
Informed consent was obtained prior to survey initiation. The proportion
of respondents affected by anxiety and depression was evaluated using
the NIH PROMIS, a validated 4 question scale, and we assessed their
views about mental health disorders. Summary statistics were used to
evaluate the population cohort. Data was analyzed using student’s t-
test and Person’s chi square for continuous and categorical variables,
respectively.
RESULTS: A survey response rate of 43% was achieved (414/959).
Overall, the cohort was young (

(SC), Hypomania (MA), and Social Introversion (SI). Cohorts were segregated
by status of cycle obligation (‘‘Complete-Oocytes Retrieved’’; ‘‘Incomplete-Dropout’’).
A random-number generator was used from a larger subset
to isolate the ‘‘Complete-Oocytes Retrieved’’ group.
RESULTS: Sixty oocyte donors were evaluated and MMPI was deemed
valid and acceptable based on program criteria. Of the MMPI taken by
both who completed (n¼45) and did not complete (n¼15) donation cycles,
no statistical difference between the 13 personality characteristics was
observed. A difference in the PA category was detected, although it did not
reach significance.
CONCLUSIONS: The MMPI test does not prospectively predict donor
compliance to protocols and obligations in an ovum donation program.
Donors demonstrated relatively consistent personality traits. Donors who
scored the highest on the ‘‘hysteria’’ and ‘‘paranoia’’ scale were less likely
to complete the process. These characteristics are common in individuals
who display hypervigilance in social interactions, and have a tendency to
be apprehensive to interpersonal communication and resist authorities’ direction.
Although the MMPI did not find a differentiation in donors’ personality
traits, future studies that evaluate alternative personality or psychopathology
could assist in identification of appropriate donors who will be compliant
with all participation requirements.
P-731 Wednesday, October 21, 2015
EFFECTIVENESS OF A SELF-HELP INTEGRATIVE BODY-MIND-
SPIRIT INTERVENTION (I-BMS) IN REDUCING INFERTILE
WOMEN’S ANXIETY DURING THEIR IN VITRO FERTILIZATION
(IVF) TREATMENT RESULT AWAITING PERIOD. C. Chan, a
S. Wong, a M. Tam. b a The University of Hong Kong, Hong Kong, Hong
Kong; b Department of Social Work and Social Administratio, Hong Kong,
Hong Kong.
OBJECTIVE: This study aims at evaluating the effectiveness of a series of
self-help techniques developed based on the Body-mind-spirit intervention
(I-BMS) intervention model to fulfill the psychosocial needs of infertile
women during the most distressful treatment stage, which is the result awaiting
period after embryo transfer.
DESIGN: This is a quasi-experimental study. 20 women who underwent
IVF treatment were recruited and randomly assigned into intervention
group (n¼10) and control group (n¼10). The study adopted the clinical
framework of I-BMS model, which aims at maintaining a dynamic and
harmonious balance between physical, psychosocial and spiritual wellbeing.
MATERIALS AND METHODS: Participants in the intervention group
attended a 3-hour I-BMS workshop organized by a registered social
worker in educating the self-help techniques, and received a self-help exercise
book containing self-help exercises, spiritual proverbs and journal
writing for home practice on a 14-day basis. They were asked to practice
the self-help techniques and report their subjective well-being on a daily
basis during the result awaiting period after embryo transfer. Meanwhile,
participants in control group received educational materials on healthy
diet for home reading.All participants were invited to complete a selfadministered
at three time points: on the day of recruitment (T0), on
the day of embryo transfer (T1), before pregnancy test (T2). The scales
used include Importance of Childbearing Index, Chinese Kansas Marital
Satisfaction Scale (C-KMS) and Chinese State-Trait Anxiety Inventory
(C-STAI).
RESULTS: By using ANOVA, participants in the intervention group
showed significant reduction in the importance in childbearing (T1: 26.90
+ 4.09; T2: 24.30 + 3.98, F¼2.10, p

OBJECTIVE: While the Uterine Fibroid Symptom and Quality of Life
questionnaire (UFS-QOL) is a validated and helpful tool in determining
response to treatment, it does not allow for the texture and intensity of fibroid
related experiences to be conveyed. The purpose of this study is to qualitatively
identify and characterize challenges that impact quality of life and
emotional well-being of women with symptomatic fibroids.
DESIGN: Qualitative semi-structured interviews, health literacy assessments,
and demographic surveys.
MATERIALS AND METHODS: Women with symptomatic uterine
fibroids were recruited from an urban academic medical center and community-based
organizations in a large Midwest metropolitan area. Participants
completed in-depth, one-on-one interviews, a health literacy assessment
and a demographic survey. Interviews were transcribed verbatim and uploaded
to NVivo version 10 for data management and thematic coding. Three
coders identified major themes and subthemes using a grounded theory
approach.
RESULTS: Sixty women completed the study for a total of 35 hours of interviews
yielding 1,357 transcribed pages. The k across coders was 0.94. The
mean age of participants was 43.0 6.8 (mean SD). 61.7% of participants
self-identified as African-American, 25.0% as Caucasian, 8.3% as Hispanic
and 5.0% as Asian. 68.3% of the participants had at least a 4-year college degree
and 55.0% of the women had a total annual household income of less
than $75,000. Two of the major themes identified were impact of fibroids
on work and impact on social life. Work impact examples include women
feeling distressed due to fear of soiling their clothes and furniture. They
also felt frustration and anxiety because heavy bleeding and pain symptoms
frequently led to missed days of work. Women’s social lives were also
impaired by having symptomatic fibroids. Women reported feeling lonely
and socially isolated due to staying at home to avoid embarrassment caused
by unexpected pain or bleeding and expressed guilt about missing out on major
social events and disruption of relationships. Women also expressed
shame about their appearance, difficulty planning for social events, and
limited behavioral coping strategies as a result of having to work around
the symptoms of their fibroids.
CONCLUSIONS: Symptomatic fibroids greatly impair women’s work
and social lives in ways that cannot be fully captured in a survey. These challenges
impact the quality of life in women largely by limiting their work productivity
and social engagement with loved ones. The emotional distress
reported by women in this qualitative study suggests the need for the inclusion
of mental health professionals in improving clinical care and wholehealth
outcomes for women suffering from these prevalent tumors.
Supported by: NIH WRHR Program K12HD050121; RWJ Foundation;
NMH; Evergreen Foundation (EEM).
P-734 Wednesday, October 21, 2015
DONOR EGGS: ON ICE, STILL NICE? J. C. Patel, a
D. L. Cunningham, a M. Y. Fung, b D. A. Sheehan, a M. K. Connerney, a
K. J. Go. b a Embryology, IVF New England, Lexington, MA; b IVF New England,
Lexington, MA.
OBJECTIVE: The vitrification and thawing of mature human eggs are
among the newest of assisted reproductive technologies (ART), and present
the opportunity for creating donor egg banks. Given some of the advantages
to the recipient patient of using vitrified eggs (VE), e.g., greater selection of
donors, lower cost, and more convenience with no need for cycle synchronization
with the donor, ART laboratories may encounter VE cycles more
frequently, requiring development of expertise in using VE. To learn how
VE compare with fresh eggs (FE) and identify the technical differences in using
each egg type, we compared clinical outcomes for recipients of VE and
FE from July 1, 2012 through December 31, 2014. VE were purchased from a
single egg bank in this study.
DESIGN: Retrospective data analysis of IVF cycles of recipients of FE or
VE from a single VE bank. Statistical analyses were conducted by Student’s
t-test and Mann-Whitney U test.
MATERIALS AND METHODS: Recipient cycles of FE or VE were
compared for average number of eggs per cycle, fertilization rate, #embryo
transfers, average # embryos transferred, pregnancy rate,and chance for the
recipient to have supernumerary embryos for cryopreservation. FE were
inseminated conventionally or by ICSI; VE received ICSI exclusively. Assisted
hatching was used electively on cleavage stage embryos (CSE)from FE
but on all VE CSE. Embryologists were trained in the specific thawing protocol
of VE obtained from a single egg bank.
RESULTS: 80 FE and 168 VE cycles were analyzed, with 1595 and 1171
eggs, respectively, available for the recipients (averages of 19.9 FE vs 7 VE
per recipient). Fertilization rates of 76.18.5% and 84.18.0%)of inseminated
FE and VE, respectively, were achieved(significant, p¼0.0011) leading
to averages of 1.2 (FE) and 1.3 (VE) embryos per transfer (no statistical difference)and
49/75 FE and 91/158 VE clinical pregnancies per ET (not significant,
chi-squared¼1.2699, p¼0260).72.5% of FE recipients had frozen
embryos after ET compared to 51.2% of VE.
CONCLUSIONS: Despite the almost 3-fold higher number of eggs available
per treatment cycle for FE vs. VE, comparable fertilization rates, application
of elective single embryo transfer, and clinical pregnancy rates per ET
were obtained. FE patients had an advantage in more frequently having supernumerary
embryos for freezing from their cycles. VE can provide more
cost-effective and convenient treatment to patients but require specific
training in the thawing protocol prescribed by the egg bank to realize this
advantage.
P-735 Wednesday, October 21, 2015
THE PSYCHOSOCIAL IMPACT OF INFERTILITY AMONG
WOMEN SEEKING FERTILITY TREATMENT. W. D. Winkelman, a
P. P. Katz, b J. F. Smith, c T. Rowen. a a Obstetrics, Gynecology and Reproductive
Medicine, University of California, San Francisco, San Francisco, CA;
b Department of Medicine, University of California, San Francisco, San Francisco,
CA; c Department of Urology, University of California, San Francisco,
San Francisco, CA.
OBJECTIVE: To identify factors that are associated with increased psychosocial
impact on women seeking infertility treatment.
DESIGN: Cross sectional study of women seeking infertility treatment
who presented for an initial intake exam.
MATERIALS AND METHODS: Responses to written questionnaire that
included items on the sexual, personal, marital, and social impact of infertility
using a previously validated scale. Respondents also classified the underlying
cause of infertility as female factor, male factor, concurrent male
and female factor or unexplained infertility. Multivariate regression
Table 1: Impact of infertility among respondents for select demographic traits, adjusted model,
Characteristic
Sexual impact
score, mean (p)
Personal impact
score, mean (p)
Marital impact
score, mean (p)
Social impact
score, mean (p)
Perceived Infertility Male factor only 22 (ref) 58 (ref) 27 (ref) 55 (ref)
Etiology
Combined 15 (0.09) 67 (0.1) 29 (0.68) 40 (0.2)
Female factor only 33 (

analyses were used to identify factors independently associated with
increased impact.
RESULTS: A total of 809 women met the inclusion criteria, of which 396
(49%) completed the questionnaire. Most participants (75.5%) reported having
no prior children. The majority (58.8%) attributed infertility to only female
factors, 30.4% to a combination of male and female factors, 7.3% to
only male factor, and 3.5% reported that the etiology had not been determined.
The actual cause of infertility from the medical records was statistically
different from the perceived cause of infertility, p

MATERIALS AND METHODS: Study participants were 152 female
infertility patients who were enrolled in an IVF clinical trial at a fertility
clinic in the United States. To measure anxiety, depression, grief, and
maternal identity centrality, all women completed self-report questionnaires
at the outset of treatment in advance of the start of a first cycle of IVF. Data
were collected between October 2012 and June 2014.
RESULTS: Mean anxiety and depression scores for the clinical sample
were significantly lower than for a community sample (p < .001). Mean grief
scores for the clinical sample were significantly lower than for a control sample
(p < .01), although grief symptoms were endorsed with greater frequency
than either anxiety or depression. Predicted interaction effects between
maternal identity centrality and grief on the presence of anxiety and depression
were not Supported. While hypotheses involving maternal identity centrality
were not confirmed, a statistically significant positive relationship was
found between maternal identity centrality and grief.
CONCLUSIONS: The findings of this investigation demonstrated that
clinical symptoms are not reported in the majority of women beginning
IVF treatment. Instead, more women report the experience of grief, which
suggests a normative psychological process in response to loss. The significant
positive relationship found between maternal identity centrality and
grief implicates the role of development and identity in shaping psychological
response to infertility. These findings support approaching subclinical
distress in treatment-seeking women through the grief paradigm and explaining
adverse psychological response to infertility as resulting, in part, from the
normative desire for motherhood as a means of identity fulfillment.
Reference:
1. Levin, K., Samstag, L., & Freeman-Carroll, N. (2014). Psychological
distress in women presenting for first-time in vitro fertilization: Relationships
among maternal identity centrality, grief, and psychopathology.
Available from ProQuest Dissertations and Theses databases
(UMI No. pending).
P-739 Wednesday, October 21, 2015
INTEGRATING DONOR CONCEPTION INTO IDENTITY:
PARENT-CHILD RELATIONSHIPS AND IDENTITY DEVELOP-
MENT IN DONOR-CONCEIVED ADOLESCENTS. J. Slutsky, a
V. Jadva, b T. Freeman, b S. Persaud, a W. Kramer, c M. Steele, a
H. Steele, a S. Golombok. b a Center for Attachment Research, The New
School for Social Research, New York, NY; b Centre for Family Research,
University of Cambridge, Cambridge, United Kingdom; c The Donor Sibling
Registry, Nederland, CO.
OBJECTIVE: This study explores the experiences of donor-conceived adolescents
connecting to same-donor offspring (individuals conceived using
the same donor), and whether the quality of parent-child relationships relates
to how donor-conceived adolescents, informed of their genetic origins in
early development, integrate donor conception into their sense of identity.
DESIGN: During in-home visits, qualitative and quantitative data were
collected from adolescents conceived by sperm donation on their experiences
of being donor conceived, connecting to same-donor offspring, and their relationships
with their parent(s) and siblings.
MATERIALS AND METHODS: Twenty-three adolescents completed
questionnaires and a semi-structured interview including the Friends and
Family Interview (FFI) (Steele & Steele, 2005; Steele et al. 2009), designed
to explore the adolescents’ relationship with their parent(s), siblings, and
same-donor offspring. Participants also completed a questionnaire designed
to evaluate the extent to which donor conception factors into their sense of
identity. Participants were members of the Donor Sibling Registry, an American-based
registry that facilitates contact between same-donor offspring,
their parents, and donors.
RESULTS: Participants, (16 female, 7 male) ages 12 to 19 (mean 14.5
years), had all made contact with at least one same-donor offspring; none
had located their donor. Just over half (13, 56 %) felt ‘neutral/indifferent’ about
their donor conception, 8 (32%) felt ‘positive/interested’, and 2 felt ‘negative/
avoidant’. The importance of knowing the donor varied greatly from those who
felt it was ‘unimportant’ (5, 23%) to those who felt it was ‘extremely important’
(5, 20%), with the remainder seeing it as ‘somewhat important’. Data
will be presented on the FFI and its relationship to donor conception identity.
CONCLUSIONS: This study presents the first in-depth data from donorconceived
adolescents on their experience of being donor conceived and contacting
same-donor offspring, their familial relationships, and their incorporation
of donor conception into their identity. These results enable greater
understanding of the factors that affect identity development in donorconceived
individuals.
References:
1. Steele, H. & Steele, M. (2005). The construct of coherence as an indicator
of attachment security in middle childhood: the friends and family
interview In: Kerns, K.A. & Richardson, R.A. (Eds.) Attachment in
middle childhood (Chapter 7). New York: Guilford Press.
2. 2.Steele, H., Steele, M. & Kriss, A. (2009). The friends and family
interview (FFI): coding guidelines. Unpublished manuscript.
Supported by: This study was Supported by the Wellcome Trust [097857/
Z/11/Z].
P-740 Wednesday, October 21, 2015
PATIENT PERSPECTIVES ON IVF SUCCESS AND LIKELIHOOD
OF MULTIPLE GESTATIONS. S. J. Miller, a K. D. Schoyer, b
E. M. Wozniak, b J. Davis, a J. Sandlow, b E. Y. Strawn, c K. E. Flynn. b
a OB/GYN, Medical College of Wisconsin Affliated Hospital, Wauwatosa,
WI; b Medical College of Wisconsin, Milwaukee, WI; c Medical College of
Wisconsin, Mequon, WI.
OBJECTIVE: To describe couples’ perceptions of fertility treatment success
rates and chance of multiples.
DESIGN: Longitudinal, prospective, mixed-methods study of patients and
their partners who presented for care with a reproductive endocrinologist
(REI) at an academic medical center.
MATERIALS AND METHODS: Both members of 37 couples (2 same-sex
female) separately completed surveys and qualitative interviews before their
initial consultation with an REI and up to 5 additional times over the subsequent
12 months. We present means (SD) and ranges of perceived chances
(%) of success and multiples with in vitro fertilization (IVF) from survey
data pre-consult and at 12 months. For couples with diagnoses of diminished
ovarian reserve (DOR) or anovulation, we present means (SD) of perceived
chances of success specific to the treatment they were planning next, based
on interview data at 2 months, after testing and diagnoses. The study was primarily
qualitative in nature and not powered to make statistical comparisons.
RESULTS: The Table shows the perceived chance (%) of IVF success
and multiples for all respondents (patients and partners) stratified by patient
age. There was little difference pre-consult and 12 months later. At 2
months, couples with a diagnosis of DOR who were planning IVF with
donor oocytes perceived their chance of success to be 72.9% (12.9). Those
with DOR planning ovulation induction with intrauterine insemination
(IUI) perceived their chance of success to be 23.0% (22.8). Couples with
anovulation planning ovulation induction with IUI perceived their success
to be 38.7% (26.2).
CONCLUSIONS: The very wide ranges of perceived likelihood of success
with IVF and other diagnosis-specific treatments confirm the need for patient/partner
education. Additionally, there is significant potential for education
regarding options to minimize the risk of multiple births with IVF.
Table.
Mean (SD)
Range
Patients
Patients 35-40 years old
Prior to consult with REI 12 months later Prior to consult with REI 12 months later
Perceived likelihood of success with IVF 43.1% (23.2)
1.0-90.0
Perceived likelihood of multiples with IVF 30.8% (20.3)
3.0-78.0
41.2% (20.4)
25.0-60.0
32.2% (22.4)
3.0-85.0
50.3% (30.0)
1.0-90.0
32.8% (21.8)
4.0-60.0
36.6% (21.1)
11.0-86.0
29.8% (21.5)
5.0-70.0
FERTILITY & STERILITY Ò
e361

AUTHOR INDEX
Aballa, T., O-19
Aballa, T. C., O-24
Abdalmageed, O. S., P-48, P-568
Abd El Aal, D. M., P-161
Abdelaleem, A. A., O-228, P-211
Abdelaziz, M., O-6
Abdelghaffar, H., P-233, P-613
Abdel-Hamid, S., P-48
Abdelmagied, A. M., O-69, O-259, P-137, P-211
Abdel-Rahman, M., P-613
Abdel-Rahman, M. Y., P-233
Abdo, G. A., P-403
Abdulwahab, F. M., P-283
Abedi Asl, Z., O-156
Abozaid, T. I., P-107, P-695
Abuelhasan, A. M., O-69, O-259, P-211
Abu-Elmagd, M., P-112, P-555
Abusamaan, M. S., P-113, P-115
Abu-Soud, H., P-215, P-282
Abu-Soud, H. M., O-143
Abuzeid, M. I., O-70, P-81, P-107, P-609, P-626,
P-695
Abuzeid, O., O-70, P-81, P-107, P-626, P-695
Acacio, B., O-34, P-516
Acharya, C. R., O-38, O-41, O-118, P-568
Acharya, K. S., O-38, O-41, O-118, P-568
Achour-Frydman, N., O-159
Acton, A. J., O-52
Adamowicz, M., P-516
Adamson, G. David., O-216, P-659, P-668, P-716,
P-717
Adamu, A., P-4
Adashi, E. Y., P-89
Adda-Herzog, E., P-311
Adedji-Fajobi, T., O-212
Adir, M., O-136
Agarwal, A., O-111, P-101, P-111, P-112, P-114,
P-116, P-119, P-381, P-387, P-451, P-534,
P-544, P-545, P-552, P-555, P-559
Agarwal, N., P-157
Agbo, C., P-413
Aghajanova, L., P-29, P-692
Agrawal, S., P-31
Aguilar, J. A., O-45, P-593, P-673, P-679
Aguirre, M., P-183, P-192, P-327, P-349
Ahmad, A. K., P-64
Ahmad, G., P-101, P-114, P-116, P-119, P-451,
P-552
Ahmady, A., P-412
Ahmed, A. I., P-484, P-581, P-637
Ahmed, H., P-228
Ahn, J.-H., P-543
Ainsworth, A., P-98
Ait-Ahmed, O., O-239
Aitken, R. J., P-117, P-123
Aizpurua, J., O-128, O-173, P-685
Akbas, H., O-117
Akin, N., O-162, P-33, P-475, P-634
Akopians, A. L., P-247, P-473
Akutsu, H., P-407
Alaboudy, A., P-233, P-613
Alaboudy, F., P-613
Alama, P., P-12
AlAnsari, A. A., O-111
Al-Aref, I., P-174
Al-Asmar, N., P-597
Alazami, A. M., P-283
Albahlol, I., P-333
Albert, C., P-256
Albertini, D. F., O-151, P-253, P-592, P-622
Albertsen, H. M., O-175
Alegretti, J. R., P-351
Alexander, C., P-240, P-490
Alexander, C. J., O-260
Alfarawati, S., O-252
Alford, C., P-566
Al-Hassan, S., P-283
Al-Hendy, A., O-6, O-73, O-74, O-75, O-76,
O-182, O-185, O-186, O-211, P-113, P-115
Al-Hussaini, T. K., O-228
Ali, M. K., P-161
Alikani, M., P-323
Alkabra, M., P-283
Alkruaya, F. S., P-283
Alkudmani, B., P-200, P-443
Alkusayer, G. M., P-150
Allen, R., P-14
Allon, R., P-704
Allshouse, A. A., O-201
Al Ma’mari, N., P-302
Alohali, A., O-1
Alper, E., P-634
Alper, M., P-585
AlRumaihi, K. R., O-111
Al-Safi, Z. A., O-274, P-425
AlSaid, S., O-111
Alshahrani, S., P-409
Altman, E., O-94
Alur, S., O-54
Alvarez, J. P., O-120, P-473
Alvarez Sedo, C., O-36, P-487, P-515
Alvero, R., O-258
Alviggi, C., P-630
Amand, G., P-418
Amano, N., P-320, P-561
Amdani, S., P-570
Amo, A., P-285
Ampeloquio, E., O-123, P-489
Amrane, S., O-60, P-348
Anahory, T., O-247, P-360
Anchan, R. M., O-188, P-162
Anderson, A. R., O-5
Anderson, K. N., P-727, P-728
Anderson, M., O-170
Anderson, R. E., P-507, P-541
Anderson, S., P-508
Anderson, S. H., P-688
Andreucci, S., P-296
Andriani, L., P-421
Anduaga Marchetti, I. A., P-415
Anspach, E., P-255, P-479, P-624
Antaki, R., O-132, P-309
Anthony, J. T., P-337
Antoine, Y., P-182
Antoniassi, M. P., P-99, P-539
Antunes, D. M. F., P-271
Ao, A., P-506
Aono, N., P-57, P-536, P-575
Aoyagi, N., P-579
Aparicio-Ruiz, B., P-600
Apryshko, V., P-523, P-663, P-667
Apter, D., O-100, P-371
Aquino, A. P., P-261, P-351
Arafa, M. M., O-111
Arai, G., P-391
Arav, A., O-62
Arbo, E., O-129, P-305
Arbona, C., O-235
Archer, D. F., O-181, P-191
Arheart, K. L., P-651
Ariza, M., P-41
Arju, R., P-422
Armant, D., O-89, O-221
Armenti, E. M., O-193
Armstrong, A., O-39, O-268
Arora, R., P-196
Arredondo, F., P-8, P-345, P-419
Arruguete, G., P-681
Arthur, R., P-196
Arvas, A., P-645
Arvizu, M., O-271
Asada, H., P-21, P-144
Asada, Y., O-64, P-579, P-618
Asemota, O., O-273
Asemota, O. A., P-47
Ashcraft, L., P-402
Ashraf, M., O-70, P-107, P-695
Aslan, K., P-167
Assidi, M., P-111, P-381
Assou, S., O-239
Aston, K., P-46, P-541
Ata, B., P-167, P-250
Atabekoglu, C. S., P-639
Ates, S., P-229
Attar, E., O-51
Attia, G., O-265, P-651
Auer, H., O-47
Ausin, I., P-331
Austin, C. M., O-251
Avci, B., P-167, P-250
Avendano, C., P-414, P-415
Avila, J., P-9
Avril, C., P-305
Awonuga, A. O., P-215
Ayaz, A., P-111, P-112, P-559
Ayers, J., P-440
Ayers, J. W., P-609
Aygun, M., O-117
Azab, M., O-237
Azuma, Y., P-170
Babayev, S. N., P-186, P-218, P-400, P-628
Baccarelli, A. A., O-136
Bae, J., P-149, P-187
Bahceci, M., P-303
Baillargeon, J.-P., O-253
Baillet, S., P-254
e362 Author Index Vol. 104, No. 3, Supplement, September 2015

Baird, D., O-77, P-423
Baird, G., O-234
Baker, V., O-39, O-141, O-268
Baker, V. L., P-26, P-550
Bako, I. G., P-4
Balaban, B., O-162, P-33, P-475, P-634
Balasch, J., P-427
Baldwin, M., P-569
Balthazar, U., O-5
Banchereau, J., O-122
Bankowski, B., O-44
Banks, N., P-15, P-287
Bar, H., P-278
Baracat, E. C., O-149
Barad, D. H., O-151, P-253, P-428, P-439, P-592,
P-622
Barash, O., O-246
Baratz, A. Y., P-196, P-557
Barbato, V., P-464
Barberet, J., P-360
Barberry, C., O-23
Barbosa, C. P., P-179
Barbosa, M. W. P., P-697
Bar-Chama, N., O-112, O-164, O-204
Bardos, J., O-121, O-245, P-131
Bareh, G., P-432, P-483
Barfield, W., P-109
Barnett, R., O-244
Barnhart, K. T., O-88, P-26
Baroni, R. H., O-26
Barragan, M., O-47
Barratt, C. L.R., P-570
Barrera, A. D., P-483
Barrett, C., P-569
Barrett, E. S., P-299, P-377
Barretto, T., O-236
Barriere, P., O-129, P-305
Barrionuevo, M., O-193
Barritt, J., P-240, P-247, P-473, P-490, P-500
Barros, B., P-261, P-351
Bartels, C. B., P-624
Bartmann, A. K., O-159
Bartolomei, M. S., O-212, O-226, P-556
Bartolucci, A., P-134, P-354, P-479
Baruffi, R., O-131, O-270
Bashook, P., P-104
Basile, N., P-616
Bastu, E., O-51, O-127, P-205
Batcheller, A. E., P-94, P-438
Bates, G., P-220, P-279
Batmaz, G., P-229
Batson, R. J., P-711
Battello, N. V., P-414
Batzofin, D., O-84
Bauer, C., O-95
Bauer, J. D., P-317, P-403
Baumgarten, S. C., O-14
Bayer, A. H., O-12
Bayles, A., P-294
Baysal, B., O-127
Beall, S., P-321
Beauchamp, C., P-252
Bedairy, M. H., P-333, P-347
Bedaiwy, M. A., P-150, P-155, P-701
Bedard, J., P-511
Bedoschi, G., P-458, P-462, P-471
Begum, S., P-163
Behr, B., P-134, P-288, P-354, P-550
Bekheirnia, M., O-208
Belan, M., O-253
Belisle, S., O-85
Bell, E., O-154
Bellerose, H., O-166
Bello, S., O-154
Belloc, S., O-247
Bellver, J., O-148
Belo, A. S., P-261
Beltsos, A. N., P-288, P-508
Benadiva, C., P-479
Benadiva, C. A., P-255, P-288, P-508, P-585,
P-624
Benages, C. A., P-7
Benard, J., P-460, P-476
Bender, J., P-707
Bendigeri, T., P-163
Bendikson, K., P-108
Bendikson, K. A., P-88
Bennett-Toomey, J. N., O-101
Benoit, J., O-132, P-309
Berga, S. L., P-19
Berger, S. L., P-556
Bergeron, M., P-158
Bergh, C. M., O-83
Bergh, P. A., O-83
Berglund, A., P-224
Berkeley, A. S., O-198
Berker, B., P-639
Berkkanoglu, M., P-435, P-653
Berkowitz, K., P-280
Berman, J. M., P-113, P-115
Bernardi, L. A., O-77, O-179, P-423
Bernstein, L. R., O-161, P-270
Berro, R., P-419
Berry, T., O-76
Bertolla, R., O-24, P-99, P-539
Best, M. W., P-632
Bhagavath, B., P-139
Bhatt, D., O-275
Bhattacharya, S. M., O-11, P-72
Bhusane, K., P-163
Bialobrzeska, D., P-36, P-40
Bianchi, E., P-199
Bianco, B., P-179
Bieniek, J. M., O-171, O-199, P-133
Biesiada, J., P-94
Bildik, G., O-162, P-33, P-475, P-634
Biley, Y., P-82
Biryukov, A., P-667
Bishop, C. V., O-130
Bishop, L. A., P-421
Bisignano, A., O-196, P-14
Bissonnette, F., O-132, P-297, P-309
Bissonnette, L., P-193
Blanchard, A., O-203
Blanchette Porter, M. M., P-83
Blanco, L. A., P-698
Blanco Mejia, S., P-638
Blazek, J., P-239, P-507
Bleess, J., O-107
Blesson, C. S., P-50
Blum, K., P-529
Bochkovsky, S., P-82
Boehnlein, L. M., P-346
Boekelheide, K., P-199
Boggino, C., P-636
Bohrer, C., O-134, P-269, P-511
Boivin, J., O-106
Bolkas, M., P-323
Bolnick, A., O-89, P-81, P-626
Bolt, A., P-663
Bond, K. R., O-97, O-99, P-129
Bonetti, T., P-159
Boostanfar, R., P-308
Boots, C. E., O-15, P-78
Borges Jr., E., P-275, P-276, P-277, P-281, P-596
Borghi, C. M., O-36, P-296, P-636
Borini, A., O-176, O-252, P-103, P-151
Bormann, C. L., O-138, P-564, P-669
Borras, A., P-427
Bosch, E., P-656
Bose, G., P-353, P-398
Botes, A., O-84
Botting, B. J., O-93
Boudoures, A., O-15
Bouet, P. E., P-309
Bouknight, J. M., P-220
Boulet, S., O-263, P-221, P-236, P-326, P-501
Boulet, S. L., P-633
Boylan, C. F., P-509
Boynukalin, F. K., P-303
Bozdag, G., O-67, P-441
Bozkurt, N., O-68, P-712
Brachtchenko, J., P-557
Brackett, N., O-24
Brackett, N. L., O-19
Bradford, A. P., O-274, P-425
Brady, P. C., P-343
Braga, D. P. A. F., P-275, P-276, P-277, P-281,
P-596
Brakta, S., O-6, O-74, O-182, O-211
Brannigan, R., O-171
Brasile, D., P-688
Braun, S., P-464
Braverman, J., O-151
Bray, M. A., P-334
Brewer, A., P-238
Bringer, S., P-182
Bristow, S. L., O-168, O-196, P-14
Britten, J., O-4
Britten, J. L., O-184, P-142
Broce, M., P-359
Bronet, F., P-41
Brower, M., P-500
Brown, L., P-15
Brown, M. B., O-115, O-241, P-298, P-452
Brown, S. G., P-570
Browne, A. S., O-5
Brunet, C., O-248, P-193, P-254
Bry, H., P-418
B.S., G., P-353
Buchanan, S., P-104
Buchheit, K., O-256
Buckbinder, J., P-88
Buckett, B., P-297, P-302
Buckett, W., P-263, P-506
Buck Louis, G., O-154
Buck Louis, G. M., O-114, O-219, O-225
Bukulmez, O., P-186, P-628
Bulun, S. E., O-179
Bulut, H., P-435, P-653
Burks, C. A., P-51
Burks, H., P-88, P-726
Burnett, B., P-8
Burns, J. S., P-97
Bush, M., P-519
Bustillo, M., P-566
Butler, S. A., P-352
FERTILITY & STERILITY Ò
e363

Butler, V. L., P-197
Butts, S., O-212, O-226, O-255, P-58
Butz, A., P-306
Buyalos, R. P., P-678
Buyru, F., O-51, O-127, P-205
Buyuk, E., O-273, O-276, P-47, P-338
Buzas, B., P-436
Cabanes, I., P-616
Cabanillas, S., O-235
Cabey, R., O-193
Cabey, R. E., P-508
Cabral, E. C., P-275, P-276, P-277
Cadieux, G., P-402
Cai, L., P-27, P-265, P-406, P-672
Cakmak, H., P-273
Calafat, A. M., O-96, O-223
Calhoun, A. R., O-97
Calis, P., P-441
Calle, A., P-123
Camargo, M., O-24, P-99, P-539
Camillo, J., P-257, P-596
Camlibel, T., P-645
Campbell, A., P-691
Can, S., P-205
Canada, V. E., P-571, P-698
Canas, M. T., O-131
Cao, Y., O-43
Capalbo, A., P-520, P-630
Cardozo, E. R., O-150
Cardozo, K., O-24, P-99
Carignan, C. C., O-223
Carlson, N., O-274
Carlson, N. E., P-425
Carlsson, M., O-172
Carneiro, M. M., P-148, P-376
Carnethon, M., O-77, P-423
Carnevali, O., O-176, P-103, P-151
Carney, S. M., P-509
Carr, B., P-186, P-400, P-628
Carrell, D., P-541
Carrell, D. T., P-46
Carrere, C. A., P-698
Carusi, D. A., P-706
Carvalho, A. T., P-148
Carvalho, V., O-24, P-99
Casals, G., P-427
Casanova, P., P-698
Casson, P. R., O-258
Castells, M., O-256
Castillon, G., O-148
Cataldo, N. A., P-366
Catapano, G., P-464
Cater, E., P-691
Catherino, W. H., O-4, O-184, P-142
Cavagna, M., O-131, O-270
Cearsolo, A., P-331
Cebi, Z., P-645
Cedars, M., P-29, P-64, P-273
Cedenho, A. P., P-99
Celestine, C. K., P-370
Celia, G., P-227
Celik, C., P-303
Celik, H. T., P-62
Celik, S., P-303
Cengiz, C., O-208
Centola, G. M., O-203
Cernuda, D., P-616
Cervantes, E., O-146, P-5, P-629, P-652, P-675
Cevher, F., O-68
Cevik, O., P-195, P-430, P-431
Cha, H.-J., P-543
Cha, J.-H., P-543
Chaffin, C. L., O-161, P-270
Chakraborty, P., P-157, P-243, P-398, P-401,
P-531
Chakravarty, B., P-157, P-243, P-353, P-398,
P-401, P-531
Chan, C., O-189, P-424, P-721, P-731, P-732
Chan, C. H. Y., P-723
Chan, H., P-722, P-723, P-732
Chan, J., O-197, P-447
Chan, P. J., P-432, P-483
Chang, C.-C., O-155, P-237, P-632, P-719
Chang, C.-H., P-32
Chang, E., P-433
Chang, H.-M., P-3
Chang, J., O-90, P-501
Chang, S., O-58, P-69, P-338
Chang, T.-C., P-719
Chang, W., P-240, P-490, P-508
Chantala, K., P-693
Chantilis, S. J., P-241, P-245, P-259, P-480, P-658
Chapa, C. R., P-405
Charles, C., P-160, P-168
Charo, L. M., O-7, P-2
Charron, M., O-273, P-47
Chattopadhaya, R., P-401
Chattopadhyay, R., P-243, P-353, P-398, P-531
Chatzicharalampous, C., P-334
Chauhan, A., P-163
Chauhan, S., P-239, P-322, P-361
Chavarro, J. E., O-205, O-209, O-271, P-703
Chavez, V., P-374
Chavez-Badiola, A., O-63, P-580
Chawla, S., P-650
Chazenbalk, G. D., O-49, O-53, O-120
Chen, A. A., P-134
Chen, C.-D., P-32
Chen, C.-H., P-455, P-461
Chen, D., P-499
Chen, H., P-690
Chen, H.-H., P-627
Chen, H.-W., P-166
Chen, M., P-727, P-728
Chen, M.-J., P-416, P-446
Chen, S., P-79
Chen, S. H., O-44, P-14
Chen, S.-U., P-32, P-694
Chen, X., P-181
Chen, Y.-F., P-416, P-446
Chen, Z., O-114
Cheng, J.-C., P-3
Chenoz, L., P-311
Cherala, G., O-98
Chernyak, E., P-37
Chetani, M., P-157
Chettier, R., O-175
Cheung, S., P-378, P-386
Chi, H., P-267, P-344
Chian, R.-C., P-27, P-265, P-406, P-672
Chiang, Y.-C., P-514, P-522
Chiang, Y.-T., P-32
Child, T., P-590
Chiles, K. C., O-5
Chiles, K., P-373
Chimote, B. N., P-59, P-583
Chimote, N. M., P-59, P-583
Chin, H. B., P-448, P-450
Chinthala, S., P-162
Chiware, T. M., P-440
Cho, S., P-146
Cho, W., P-180
Cho, Y., P-149, P-178, P-187
Choi, D., P-154
Choi, J., P-154
Choi, J. M., P-472
Choi, Y., P-433, P-572
Cholkeri-Singh, A., O-72, P-202
Chong, W., O-19
Chosich, J., O-274
Chosich, J. D., P-425
Choudhary, M., P-513
Chow, V., P-538
Christensen, G. L., P-94
Christensen, M. A., P-720
Christianson, M. S., O-10, O-39, O-258, O-268,
P-1, P-468
Christman, G. M., O-258
Christofolini, D. M., P-179
Christy, A., O-78, O-258, P-330
Chu, B., O-163
Chuan, S. S., P-310
Chulet, S., P-185
Chung, H., P-65, P-102
Chung, K., P-108, P-339, P-453, P-677
Chung, M., P-671
Chung, P., O-60, P-248
Chuong, F. S., O-58, P-468
Chwalisz, K., O-181
Ciapponi, A., P-296
Cil, A., P-268
Cil, A. P., O-117
Cilingir, O., P-430
Cilingir, O. T., P-195, P-431
Cimadomo, D., P-630
Cipolla, M., O-276
Claessens, A., P-638
Clapp, M. A., P-67
Clarke, N. J., P-453
Clarke, R., O-249, P-358
Clarke, R. N., P-492
Clarke-Williams, M., P-587
Clemons, J., O-214
Coates, A., P-498
Cobo, A., P-256, P-258, P-314, P-485
Cocuzza, M., O-26, P-535
Coddington, C., O-107, P-591
Coello, A., P-256, P-258
Coetzee, K., P-435, P-653
Cohen, J., O-91
Cole, A., P-548
Collado, D., P-12
Collazo, I., P-284, P-397, P-566, P-576
Collins, G., P-412
Collins, R., O-32, P-503
Colls, P., O-35, O-63, P-585
Collura, B., O-216, P-659, P-668, P-716, P-717
Comtet, M., P-476
Conaghan, J., P-134
Conceicao, C., P-379
Confino, E., P-733
Connaughton, H., P-117
Connell, M. T., O-61, O-187, P-194
Connerney, M. K., P-734
Connor, J. J., P-727, P-728
Considine, R. V., O-52
Conti, M., P-273
e364 Author Index Vol. 104, No. 3, Supplement, September 2015

Convissar, S. M., O-14
Conway, D. A., P-294
Cooney, L., P-477
Copperman, A. B., O-57, O-80, O-112, O-121,
O-146, O-160, O-164, O-204, O-245, P-5,
P-8, P-130, P-131, P-345, P-368, P-419,
P-442, P-629, P-652, P-665, P-675, P-684,
P-700, P-730
Cordeiro, C. N., P-468
Cordeiro, F. B., P-257, P-276, P-277, P-596
Corrado, J., O-70
Correa, L. F., O-266
Corselli, J., P-432, P-483
Cosar, E., P-146
Coscia, A., P-636
Coskun, S., P-283
Costantini, L., O-257
Costantini, M., P-660
Coulam, C., P-686
Coutifaris, C., O-3, O-144, O-212, O-226, O-258,
P-306
Coutinho, A., P-252
Coward, K., P-570
Cox, J., O-4, O-184, P-142
Cox, J. M., P-325
Cozzubbo, T., O-2, O-110, P-546, P-560, P-573
Craig, L. B., O-50, O-170, O-227, O-254
Crawford, N. M., O-224, P-693, P-729
Crawford, S., O-90, P-109, P-236, P-326
Creus, M., P-427
Crowe, R., O-217
Cruz, M., P-12
Csokmay, J., O-78, P-341
Cui, Z., P-534, P-555, P-559
Cunningham, D. L., O-81, P-734
Czuprenski, E., P-512, P-518
Dabaja, A., P-216
Dabaja, A. A., O-22, O-213, P-456
Da Broi, M. G., P-110
Daftary, G., O-107
Daftary, G. S., O-266, P-591
Dahan, M., O-85, P-413
Dai, J., P-81, P-626
Dalloul, M., P-160, P-168
Dambaeva, S., O-157, P-128
Daneshmand, S., P-183, P-192, P-327, P-349
Daneyko, M., O-80, P-368, P-730
Daniel, C. E., O-250
Danisman, N., P-63
Danovitch, Y., P-497
Danzer, H., P-240, P-473, P-490, P-500, P-677
Daris, M., P-158
Darmon, S. K., P-253, P-428, P-439
Darwish, A. M., P-48
Dasmahapatra, P., P-163
Daubert, M., P-80
Davenport, D. L., P-542
Davenport, E., P-726
Davidson, K., P-241
Davidson, M., P-724
Davie, J., P-295, P-307
Davies, M. C., O-93
Davila, A., P-405
Davis, J., O-264, P-740
Davis, L., P-210
Davis, O. K., P-348, P-420
Davis, P., P-294
Dean, N., O-132
Dean, R., O-25
Debrock, S., P-136
de Carvalho, C. V., P-159
De Chavez, P. D., O-77, P-423
DeCherney, A., O-61, O-78, O-140, O-142, O-187,
O-191, O-220, P-6, P-15, P-194, P-287,
P-321, P-328, P-330, P-341, P-382, P-394,
P-399, P-713
Declercq, E., O-37, O-42
De Gheselle, S., O-55, P-249
de Haydu, C., O-22, P-456
Deibert, C., P-383
Deibert, C. M., O-171
Deimling, T. A., P-141
Delaney, A. A., O-266
Delaroche, L., P-193
Delgado, A., O-45
de los Santos, J., P-485
de los Santos, M., P-485, P-600
De Meglio, G., P-576
de Melo, A. A., P-257, P-596
DeMichele, A., O-7, P-2
Demick, J. L., O-203
Demir, B., O-67, P-350, P-699
Demiral, I., O-51, O-127
Demirci, U., O-188
Demko, Z., P-519
Denes, F. T., O-26
Deng, J., P-710
Denomme Tignanelli, M. M., O-46
De Paz, C. C P., P-110
Desai, N., O-251, P-553, P-611
Dessapt, A., P-418
De Sutter, P., O-55, P-249
Detti, L., O-13, P-91
Devine, K., O-61, O-187, P-84, P-287, P-321,
P-328, P-330, P-382, P-394, P-421
Dewailly, D., O-129
Dhanjani, D. G., O-199
D’Hooghe, T., P-136
Di, C.-G., P-457
Dia, F., P-280
Diamond, M., O-6, O-74
Diamond, M. P., O-73, O-75, O-76, O-92, O-182,
O-185, O-186, O-201, O-211, O-258, P-81,
P-113, P-115, P-215, P-626
Diaz-Garcia, C., P-449, P-644
Diaz-Gimeno, P., P-597
Dickler, M., P-458
Dieamant, F., O-131, O-270
Diez Juan, A., P-597
DiGiovanni, L., O-197
Dilbaz, B., P-375
Dilbaz, S., P-375
DiMattina, M., P-227, P-677
Dimitriadis, I., O-200
Ding, J., P-385
Ding, X., O-49
Diop, H., O-37, O-42
Dmitrieva, N., P-663, P-667
Do, L., P-197
Dodds, W., P-127
Dodge, L., P-301
Dodge, L. E., P-340
Dokras, A., O-169, P-58
Dolinko, A. V., P-469
Domar, A. D., O-106, P-301
Domchek, S. M., O-197
Domingo, J., P-644
Domingues, T. S., P-261, P-351
Donahue, G., P-556
Dong, F., O-8
Dong, J., P-27, P-406
Dong, S., P-100
Donjacour, A., P-242
Doody, K., P-400
Douglas, N. C., O-189, P-424, P-710
Doyle, J., P-482
Doyle, N., P-399
Drevet, J., P-123
Drewlo, S., O-89, O-221
Driggers, P., O-58, P-84
Druckenmiller, S., O-123, O-231, P-489, P-567,
P-587
Druschel, C., O-154
Du, H., P-367
Du, T., P-690
Dubaut, J. P., O-8
Dudley, P., P-677
Duhamel, A., O-129
Duke, C. M. P., O-22, O-213, P-216, P-456
Duke, J., O-214
Duke, M., O-146, P-684
Dumesic, D. A., O-49, O-53, O-120
Dumitriu, B., O-142
Dun, E. C., O-213, P-216
Duncan, F. E., P-269, P-422
Duncan, J., P-738
Dundee, J. A., P-83
Dunn, R., P-239, P-322, P-361
Duong, J., O-8
Du Plessis, S. S., P-101, P-114, P-116, P-544,
P-552
Durairajanayagam, D., O-111, P-381, P-387
Dural, O., O-51, O-127, P-205
Duros, S., P-460
Duval, K., O-253
Dwan, P. G., O-81
Dye, T., P-299, P-377
Dyson, M. T., O-179
Dzidic, N., P-516
Eaton, J., P-48
Eaton, J. L., O-40, O-119, O-261
Eberlin, M. N., P-275, P-276, P-277
Eckel, R. E., O-274, P-425
Edmonds, J. W., P-279
Edwards, T. L., P-145
Efymow, B., P-477
Egashira, A., P-610
Eid, M. E., P-687
Eid, S., O-102
Eisenberg, E., O-92, O-201, O-258
Eisenberg, M., O-114, P-550
Eisenberg, M. L., O-203
Eisermann, J., P-284, P-397
Eken, M., P-195, P-430, P-431
Elam, L., O-73, O-185, O-186
Elam, L. A., O-75, O-76
El Assal, R., O-188
ElBardisi, H., O-111
Elbareg, A. M., P-204
El-Baz, M. A. H., O-228
Eleswarapu, S. V., O-22, P-456
El Hachem, H., O-132, P-309
El-Husseini, H., O-74
Elias, R., P-649, P-707
Elias, R. T., P-135
El-khayat, W., O-66
Ellen, J., P-540
FERTILITY & STERILITY Ò
e365

Elliot, M. C., P-332
Elliott, T., O-155
Elnashar, I., O-228, P-211
El Sadek, M., O-66
Eltsova, E., P-10
Emerson, G., P-650
Emirdar, V., P-230, P-462, P-471
Engmann, L., O-255, P-255, P-479, P-624
Entezami, F., P-182, P-193
Er, T.-K., P-147
Erdem, A., O-68, P-623, P-635, P-712
Erdem, E., P-645
Erdem, M., O-68, P-623, P-635, P-712
Erdinc, A. S., P-23
Erdogan, D., P-558
Eren, F., P-52
Ergin, M., P-52, P-60, P-63
Erkilinc, S., P-23
Ermilova, I., P-663
Ersoy, A. O., P-62
Ersoy, E., P-62
Escudero, T., O-35, P-495
Esfandiari, N., P-83
Eskew, A. M., P-332
Esteves, S., P-535
Eum, J., P-433
Evans, B. T., P-666
Evans, E. A., O-166
Evans, J. P., P-274
Evans, S., O-105
Evans-Hoeker, E. A., P-45, P-231
Ezzati, M., P-186, P-400, P-628
Fabregues, F., P-427
Fadiel, A., O-276
Fadiloglu, E., P-375
Fadiloglu, S., P-375
Falcao Jr, J. A., P-148
Falcone, T., O-251, P-174, P-198, P-203
Fanchin, R., O-159, P-305, P-311
Fang, C., O-133, P-457
Fang, L., P-3
Farghaly, T. A., O-69, O-228, O-259, P-211
Farhadifar, R., P-356
Farland, L. V., O-232, P-49, P-343, P-356, P-469
Farrington, P., O-175
Faulkner, N., P-307
Faustmann, T., O-100, P-371
Fawzy, M., P-233, P-613
Fazleabas, A., P-127
Fedder, J., O-202
Fedick, A., P-505
Feinberg, E., P-704
Feinberg, R. F., P-509
Feldman, G., P-7
Feldman, R., O-181, P-58
Feliciano, M., P-373
Feng, C., P-75, P-121
Feng, R., O-144
Feng, X., P-177
Ferguson, K., P-538
Ferigolo, P., P-539
Ferle, L., P-681
Fernandes, R., P-179
Fernandez, M., O-128, O-173, P-685
Fernandez Gallardo, E., P-136
Ferrando, M., P-314
Ferrer, A., O-47
Ferriani, R., P-110, P-697
Ferriani, R. A., P-654
Ferrieres-Hoa, A., O-239, P-254, P-360, P-444
Ferrieres-Hoa, A. F., O-248
Feskov, O., P-396
Feskov, V., P-396
Feuer, S., P-242
Ficicioglu, C., P-437
Fierro, M. A., O-14
Figueira, R. C. S., P-281
Fino, M., P-25
Fino, M. E., O-104, O-231, P-77
Fiorentino, I., P-464
Fisch, B., O-237
Fischer, E., P-519
Fishel, S., P-691
Fisseha, S., P-736
Fiszbajn, G., O-36, P-636
Flaws, J. A., O-10
Fletcher, N. M., O-13, P-91, P-113, P-115, P-215
Florensa, M., P-520
Flowers, L., P-501
Flyckt, R., P-174, P-198
Flynn, K. E., P-740
Fode, M., O-28, P-537
Foer, D., O-256
Folger, S., P-109
Fontenot, G. K., O-29
Foong, S., O-215
Ford, J. B., O-96
Forman, E., P-526
Forman, E. J., O-180, O-244, P-223, P-527, P-584,
P-606
Foster, K., P-86
Fothergill, A., P-448, P-450
Fox, J. H., P-706
Fragouli, E., O-91, O-252
Franasiak, J. M., O-31, O-135, O-137, O-180,
O-244, P-223, P-370, P-527, P-584, P-586,
P-594, P-595, P-606, P-612, P-660, P-715
Franca, U., P-564
Franciosi, F., P-273
Franco Jr, J. G., O-131, O-270
Frankel, R., P-11
Fraser, B., P-117
Frattarelli, J. L., O-196
Fredrickson, J., P-98
Freeman, T., O-103, P-737, P-739
Friebel, T. M., P-447
Friedman, L., P-104
Fritz, A. E., P-619
Fritz, R., O-89, O-221
Fru, K. N., P-325
Fuehrer, D., O-107
Fujii, M. G., O-149, P-718
Fukuda, A., P-56, P-563
Fukunaga, N., O-64, P-579, P-618
Funabiki, M., O-192, P-320, P-561
Funagayama, Y., P-579
Fung, J. L., P-120
Fung, M. Y., O-81, P-734
Furuya, K., P-459
Gable, E., P-377
Gabriele, D. A., P-494
Gabrielsen, J., O-205
Gad, M., P-233, P-613
Gada, R., P-245, P-259, P-480, P-658
Gaines, T., P-1
Gajbhiye, R., P-163
Gala, A., O-247, O-248, P-254, P-360, P-444
Galai, N., O-140, O-220, P-6, P-705, P-713
Galal, A. F., P-648
Galan, A., P-256, P-673
Galiana, Y., P-520
Galkina, E., P-319
Galliano, D., O-262
Ganguly, N., P-82
Gantt, P. A., P-359
Ganzabal, T., P-331
Gao, J., O-181
Gaona, M., O-84
Garcia, B. A., P-556
Garcia, D., O-153
Garcia, J., P-576
Garcia, J. E., P-468
Garcia, V., P-673
Garcia-Velasco, J. A., P-41, P-616, P-644
Gardner, D. K., O-152, P-610
Garg, D., P-264, P-709
Gargiulo, A. R., O-256
Garner, F., P-183, P-192, P-327, P-349
Garnsey, H., P-269, P-526
Garrido, N., O-262
Garrisi, M., O-163
Garzo, G., P-495
Gaskins, A. J., O-205, O-209, P-703
Gat, I., O-236, O-238, P-638
Gates, D., P-308
Gauthier-Fisher, A. S., O-236, O-238
Gavrilova-Jordan, L., O-182, O-185
Gayet, V., O-129
Gebhart, M. B., O-82
Gehrke, K., P-619
Gemzell-Danielsson, K., O-100
Geneidi, E., P-686
Genro, V., O-159
Gentry, A. L., P-45
George, J., O-122
Gerami-Naini, B., O-188
Gerkowicz, S. A., P-651
Gerson, M., O-204
Ghadir, S., O-196, P-240, P-490, P-495
Ghanem, M., P-333, P-347
Ghant, M., P-138, P-143, P-733
Gharagozloo, P., P-117, P-123
Ghassabian, A., O-154
Gheyas, F., P-646
Ghosh, B., P-398, P-401, P-531
Ghosh, S., P-157, P-353, P-398, P-401, P-531
Ghossein-Doha, C., P-674
Ghuge, A., P-163
Gibbons, W. E., P-50, P-565, P-607
Gibson-Corley, K. N., P-547
Giles, J., O-148
Gill, P. K., P-611
Gingold, J., O-146, O-164, O-204, P-368, P-442,
P-665, P-700
Ginsburg, E., O-17, O-232, P-49, P-343, P-469
Gioacchini, G., O-176, P-103, P-151
Giorgi, V. S I., P-110
Giorgini, E., O-176, P-103, P-151
Girao, M. B., P-159
Giudice, L. C., P-692
Giuliani, E., P-127
Givens, C., P-519
Glass, K., O-215, P-196, P-557
Glassner, M. J., P-688
Gleason, K., P-499
Gleicher, N., O-151, P-253, P-428, P-439, P-592,
P-622
e366 Author Index Vol. 104, No. 3, Supplement, September 2015

Globus, S. T., O-57
Glujovsky, D., P-296
Go, K. J., O-81, P-734
Godfrey, E. M., O-257
Goel, T., P-689
Goering, M., P-619
Gold, M., O-168
Goldberg, J., O-194
Goldberg, J. D., O-166
Goldberg, J. M., O-251, P-198, P-203, P-611
Goldberg, R. W., P-611
Goldberg-Strassler, D., O-193, P-508
Goldfarb, J. M., O-38, O-41, O-118
Goldfarb, S., P-458
Goldman, K. N., O-12, O-104, O-123, O-198,
O-231, P-25, P-422
Goldman, M. B., P-120
Goldman, R. H., P-49, P-669
Goldstein, M., O-109, P-373, P-546
Golombok, S., O-65, O-103, O-105, P-737, P-739
Gomes, A., P-718
Gomes, A. P., O-149
Goncalves, S. P., O-149, P-718
Gonzalez, F., O-52, O-267
Gonzalez, M., P-331
Gonzalez-Fernandez, R., P-9
Gonzalez-Foruria, I., P-427
Goodall, N.-N., O-163
Goodman, L. R., O-251, P-174, P-198, P-203
Goodrich, D., P-511
Gopal, D., O-37, O-42, P-292
Gordon, J., P-227
Gordon, K., P-308
Gordon, T. T., P-512
Gorman, J., P-470
Goswami, M., P-353, P-531
Goswami, S. K., P-243, P-353, P-401, P-531
Goswami, S., P-458
Goto, H., P-285
Goyal, A., P-174
Gracia, C., O-92, O-197, P-445, P-447, P-477
Graham, J., P-238, P-252, P-482
Grainger, D. A., O-8
Grantz, K. L., O-219
Grazi, R., O-275, P-264, P-709
Green, K. A., O-135, P-594, P-595, P-715
Greenberg, S. A., P-210
Greenblatt, E., P-196
Greene, A. F., O-56
Greene, N., O-260, P-313, P-316
Griesinger, G., P-336
Griffin, D. K., O-48, P-491, P-676
Grifo, J., O-35, O-44, O-91, O-123, O-196, O-198,
P-77, P-82, P-422, P-478, P-489, P-493,
P-499, P-504, P-510, P-524, P-525, P-578,
P-603, P-664, P-680
Grifo, J. A., O-34
Grimstad, F., P-86, P-335
Grober, E. D., O-171, O-199, P-133
Grogan, T. R., O-53
Gronier, H., P-476
Gross, J., O-106
Grossman, L. C., O-189, P-266, P-424
Grunert, G., P-322
Grunert, G. M., P-239, P-361
Grunfeld, L., O-245, P-130, P-131, P-652
Grynberg, M., P-460, P-476
Gualtieri, R., P-464
Guan, Y., P-308, P-646
Guedikian, A. A., O-120
Gueye, N.-A., P-198
Guijarro, M., O-128, O-173, P-685
Guillen, A., O-148
Guillen, J., O-195
Gulati, S., P-398
Guler, I., O-68, P-623, P-635, P-712
Gultomruk, M., P-303
Gunes, S., P-545
Gunes, Z. K., O-44
Gungor Ugurlucan, F., P-205
Guo, H., P-173
Guo, M., P-105
Guo, Y., O-59, P-44, P-70, P-670
Gupta, M., P-198, P-689
Gupta, S., P-101, P-111, P-112, P-114, P-116,
P-119, P-381, P-451, P-544, P-552, P-559
Gurtcheff, S. E., P-294
Gustofson, R. L., O-125
Gutierrez-Adan, A., P-123
Gutmann, J. N., P-8, P-345, P-419
Guven, S., O-188
Guvenir, H., P-350
Guzel, Y., O-162, P-33, P-475, P-634
Gwinnett, D., O-250
Ha, T. Kim., P-400
Haberman, S., P-709
Haberstroh, W. P., P-688
Hacker, M. R., P-301, P-315, P-340
Haddad, B., P-418
Haddad, G., O-34
Hade, E. M., P-34
Haden, S., P-738
Hadjiliadis, D., P-374
Hahn, K. A., P-17
Haji, A., P-404
Hakim, L., P-548
Halder, S. K., O-75
Hall, S. J., P-199
Halow, N. G., O-101
Haltas, H., P-61
Hamamah, S., O-239, O-247, O-248, P-182,
P-193, P-254, P-305, P-360, P-444
Hamer, J., O-36
Hammes, S., O-54
Hammes, S. R., P-139
Hammond, K. R., P-366
Han, M., P-149, P-187
Han, O., P-38
Han, T., P-399
Hancock, K., P-707
Hanna, C., O-95, O-98
Hannam, T., O-190
Hansen, K. R., O-50, O-170, O-254, O-255, O-258
Hanson, B. M., P-227
Hanson, H., P-46, P-541
Hao, C., P-66
Hao, J., P-44
Haouzi, D., P-182, P-193
Haque, I. S., O-166, O-167, O-194
Harada, T., P-170
Harden, S. A., P-45
Harkins, G., P-141
Harlev, A., P-101, P-116, P-552
Harris, A., P-280
Harris, M., O-274
Harris, M. A., P-425
Harrity, C., P-426, P-574
Hartman, M., P-287
Hartman, T. J., P-120
Hartmann, K., P-145
Harutunian, A., P-507
Hasegawa, E., P-407
Hasegawa, H., P-232
Hashimoto, S., P-285
Hasson, J., P-302
Haswell, C., O-264
Hatch, E. E., P-17, P-22, P-384
Hattori, H., P-536
Haunschild, C., O-7, P-2
Hauser, R., O-96, O-136, O-209, O-223, O-271,
P-97
Haviland, M., P-315
Hawa, N. S., P-404
Hawkins Bressler, L., O-77
Hayashi, T., O-192, P-320, P-561
Hayden, R. P., O-207
Hayward, B., O-155
Hazlett, W. D., P-615
He, A. W., O-255
Healy, M. W., O-25, O-61, O-187, P-194, P-328
Hebert, J., O-70, P-695
Hebisha, S. A., P-484, P-581, P-637
Heindryckx, B., P-249
Heiser, P. W., P-631
Helal, A. S., P-333, P-347
Heller, B., P-724
Hellmers, A., P-661
Helwa, I., O-182
Hennebold, J. D., O-101
Henriquez, S., P-651
Hernandez, J., P-9
Herraiz, S., P-449
Hershlag, A., O-44, O-163, P-323, P-497
Hesla, J., O-34
Hesley, G., P-137
Hickman, C., O-250
Hijazi, H., P-283
Hill, D., P-500
Hill, D. L., O-120, P-240, P-247, P-473, P-490
Hill, M., P-287
Hill, M. J., O-61, O-187, P-194, P-328, P-330,
P-341
Hill, W. D., O-6
Hillis, L., O-81
Hinckley, M., O-246
Hindoyan, R. H., P-339
Hines, R. S., O-82
Hipp, H., O-90
Hipp, H. S., P-633
Hirshfeld-Cytron, J., P-724
Hirshfeld-Cytron, J. E., P-51
Hixon, B., P-329, P-601
Ho, S.-M., P-94
Hoang, L., O-37, O-42
Hobeika, E., O-60, P-649
Hodes-Wertz, B., O-104, P-493, P-504, P-603
Hodis, H. N., P-88
Hoeger, K., O-54, P-299, P-377
Hoff, H. S., O-16, P-729
Hoffman, D. I., P-508
Holden, J. P., P-530
Holder, S., O-226
Holland, A. C., O-82
Holley, S. R., P-725
Holmes, R., P-238
Holoch, K. J., P-198
Holzer, H., P-263
FERTILITY & STERILITY Ò
e367

Homel, P., P-264
Homer, M. V., O-7, P-2
Hong, K. H., O-31, O-244, P-584, P-594, P-595,
P-606, P-612, P-715
Hopeman, M. M., P-445
Horne, A. W., P-701
Hornstein, M. D., P-666, P-669
Hotaling, J., P-541
Hotaling, J. M., P-46
Hourvitz, A., P-87, P-89
Howard, B., O-102
Howards, P. P., P-448, P-450
Howles, C. M., P-336
Hsu, J. W., P-50
Hsu, J. Y., P-266
Huang, C.-C., P-627
Huang, H., P-100, P-105, P-190, P-417
Huang, J.-Y., P-188
Huang, R., P-657
Huang, W., P-156, P-177
Huang, X., P-66, P-582
Huang, Y., P-417
Huang, Y.-H., P-530
Hubbard, J., O-47
Huberlant, S., P-360
Hubert, G., P-678
Huddleston, H., P-64
Hudson, C., P-183, P-192, P-327, P-349
Huffman, C. S., O-79
Hughes, J., P-15
Hughes, M., P-512, P-517, P-518
Huleihel, M., O-237
Humane, A. C., P-163
Humm, K. C., P-291
Hunter Cohn, K., P-345
Hur, J., P-71
Hurd, W. W., O-40, O-119, O-261, P-48, P-568
Hurst, B. S., P-332
Hu-Seliger, T., P-8, P-419
Hutchinson, A. P., P-135, P-649
Hwang, K., O-21, O-234, P-199
Iaconelli Jr., A., P-275, P-276, P-277, P-281
Iba, Y., P-251
Ibrahim, E., O-19, O-24
Ibrahim, Y., P-315
Ida, M., P-56
Ikuma, S., P-226, P-324, P-369, P-388
Ilagan, Y., O-178
Iles, R. K., P-352, P-620
Iliodromiti, S., O-9
Ilioi, E. C., O-65
Ilnitsky, S., P-696
Imbalzano, M., O-226
Imrie, S., O-105
Imudia, A., P-201
Inal, H. A., P-23, P-38, P-61
Inal, Z. O., P-38
Ince, U., O-162
Ingles, S., P-339
Inoue, M., P-285
Insua, M. F., P-314
Intasqui, P., P-99, P-539
Irani, M., O-275, P-264, P-709
Irani, S., O-275
Irwin, J., P-692
Isaacs, J. D., P-197
Isaacson, K., P-209
Isaka, K., P-407
Ishihara, K., P-579
Ishikawa, T., O-27, O-113, P-390, P-549, P-617,
P-683
Islas, C. A., P-405
Israel, M. A., P-719
Ito, H., P-407
Ivani, K., O-246
Iwahashi, K., P-262
Iwahata, T., P-391, P-532
Iwaki, Y., O-192, P-320, P-561
Iwata, K., P-251
Jacobs, I., O-93
Jacobson, M. H., P-448
Jacoby, V., P-137
Jadhav, P., P-646
Jadva, V., O-65, O-105, P-737, P-739
Jahoor, F., P-50
Jain, G., P-185
Jain, J., P-585
Jain, R., O-181
Jalas, C., P-494, P-505
James, A. N., O-25
James, D., O-229
Jamieson, D., P-109
Jamieson, D. J., O-90, O-263, P-236, P-326, P-633
Jang, H., P-433
Jarmuz, P., P-569
Jaroudi, S., O-163, P-508, P-590
Jarvi, K., O-236
Jarvi, K. A., O-171, O-199, P-133
Jasani, S., P-51, P-724
Jasulaitis, S., P-724
Jean-Denis, F., O-253
Jeng, G., P-501
Jenkins, J., P-334, P-336
Jenner, L., P-691
Jensen, C. F., O-28, P-537
Jensen, J., O-95, O-98, O-107, P-591
Jeon, J., P-65, P-102
Jeong, H. J., P-671
Jeong, J. H., P-562
Jeong, K., P-65, P-102
Jha, A., O-11, P-72
Ji, D., O-43
Jianini, B. T., P-110
Jimenez, C., P-331
Jimenez, J., P-367
Jin, H., P-670
Jin, S., O-92, O-258
Jincho, Y., P-599
Jindal, S., O-273
Jindal, S. K., P-47, P-338
Jo, J.-D., P-267
Jo, M., P-154
Joelsson, L. S., P-224
Joergensen, N., O-205
Johnson, D., P-240, P-490
Johnson, L., O-169
Johnson, L. N. C., O-197
Johnson, M. D., P-92
Johnson, M. R., P-284
Johnstone, E. B., P-46
Johnston-MacAnanny, E. B., P-19
Jones, C., P-570
Jones, J. M., P-346
Jones, M. E., O-93
Jones, T., P-108
Jordan, A., O-163, O-193, P-677
Jorge, S., O-58
Jou, G., P-32, P-694
Juarez, L., P-724
Julka, N., O-275
Juneau, C. R., O-31, O-137, O-180, O-244, P-370,
P-584, P-594, P-595, P-606, P-660, P-715
Jungheim, E., P-293
Jungheim, E. S., P-43
Juvet, T., P-133
Jwa, J., P-312
Jwa, S., P-312
Kachhawa, G., P-689
Kadoch, I.-J., P-225, P-297
Kahn, B. E., P-542
Kahraman, S., O-117
Kahyaoglu, I., O-67, P-350, P-699
Kaing, A., P-500
Kalem, M. N., P-38
Kalgi, B., O-275, P-264
Kaliappan, S., P-264
Kalinchenko, S., P-10
Kalinina, E., P-663
Kalmbach, K., P-271, P-272
Kalra, B., P-445
Kamel, M. A., O-69, O-259, P-211
Kamga-Ngande, C., P-225
Kang, B.-M., P-180
Kang, H. H., P-562
Kang, S. M., P-411
Kanninen, T., O-158, P-126
Kanninen, T. T., P-125
Kan-Ool, L., P-557
Kant, G., P-185
Kao, A.-P., P-166
Kao, C.-N., P-29, P-64
Kapfhamer, J. D., O-216, P-659, P-668, P-716,
P-717
Kaplanaoglu, I., P-350
Kaplanoglu, I., P-558, P-699
Karaca, N., P-229
Karacan, M., P-645
Karahasanoglu, A., O-117
Karakas, N., P-52, P-60
Karande, V., P-615
Karita, M., O-192, P-320, P-561
Karmon, A. E., O-150, P-35
Karvir, H., O-57, O-160, P-345
Kasapoglu, I., P-167
Kasem, H., P-233, P-613
Kaser, D. J., P-669, P-706
Kashanian, J. A., O-171
Kaskar, K., P-607
Kassira, S., P-220
Kastury, R. D., P-410
Kathiresan, A., O-260
Kathrins, M., P-408, P-481, P-528, P-533
Katilius, J., O-257
Katsoula, E., P-640
Katz, P. P., P-735
Katz-Jaffe, M., O-32, O-46, O-48, O-125, O-165,
O-174, P-234, P-289, P-329, P-438, P-503,
P-589, P-601, P-676
Kavoussi, S. K., P-346
Kawabata, Y., P-599
Kawwass, J. F., O-90, P-633
Ke, Z., P-190
Keefe, D. L., O-168, P-271, P-272, P-422
Keller, M., O-96, O-271
Kellogg, G. R., O-168
Kenigsberg, D., O-193
Kenigsberg, S., O-236
e368 Author Index Vol. 104, No. 3, Supplement, September 2015

Kennedy, E., O-215
Ketterson, K., P-498
Keyhan, S., O-38, O-41, O-118, P-48, P-568
Khalafalla, K., O-111
Khan, I., P-609
Khan, O., O-28, P-537
Khan, S., O-143, P-282
Khan, Z., O-107, O-266, P-591
Kharitonova, M., P-663
Khater, M. K., O-74
Kholy, A. M., P-333
Khruapenkova, T., P-663
Kida, Y., O-64, P-579
Kiehl, M., P-519
Kieslinger, D. C., O-55
Kilburn, B., O-221
Kilburn, B. A., O-89
Kilpatrick, S. J., P-316
Kiltz, R., O-193
Kiltz, R. J., O-34
Kim, B. Y., P-562
Kim, C.-H., P-180
Kim, D., P-619
Kim, H., P-180, P-380
Kim, H.-Y., P-543
Kim, J., P-268, P-465
Kim, J. H., P-411
Kim, J. W., P-671
Kim, J. Y., P-562
Kim, K.-H., P-365
Kim, M., P-572, P-671
Kim, M. J., P-562
Kim, M.-L., P-149, P-178
Kim, R., P-572
Kim, S., O-114, P-267, P-344, P-380
Kim, T., P-71
Kim, T. H., P-562
Kim, Y., P-71
Kim, Y. S., P-562
Kim, Y.-J., P-543
Kimura, T., P-459
Kingsland, C., P-288
Kinloch, M., P-150
Kirienko, K., P-523
Kirkpatrick, G., P-538
Kiskis, E., P-477
Kissin, D., P-501
Kissin, D. M., O-90, O-263, P-109, P-221, P-236,
P-326, P-633
Kitajewski, J., P-710
Kitasaka, H., O-64, P-618
Kitaya, K., O-27, P-390, P-549, P-617, P-683
Kitchen, J., P-512
Kjelland, M. E., P-404
Klausen, C., P-150, P-701
Klebanoff, M. A., P-34
Klepacka, D., O-165
Klepukov, A., P-663
Klett, D., O-22
Kligman, I., P-454, P-642, P-707
Kline, M., O-189, P-424
Klipstein, S., P-615
Knight, A. N., P-470
Knopman, J. M., O-245, P-442
Ko, J.-J., P-365, P-572
Kobayashi, H., P-599
Kobayashi, J., P-232
Kobayashi, M., O-240, P-42, P-232
Kobayashi, R., P-563
Kobayashi, T., P-391, P-532
Kobori, Y., P-391, P-408, P-481, P-532, P-533
Kocbulut, E., P-639
Kocer, A., P-123
Koch, L., P-452
Kofinas, J., P-77, P-567
Kofinas, J. D., P-82, P-578
Kogan, P., P-547
Kohan-Ghadr, H.-R., O-89, O-221
Kohlmeier, A., P-440
Koizumi, K., P-459
Kojima, J., P-407
Kojima, M., P-618
Kokeguchi, S., P-24
Kolb, B., O-34
Kolp, L. A., P-468
Komiyama, J., P-393
Kondapalli, L. A., O-32, O-125, P-503
Kono, T., P-599
Konstantinidis, M., O-44, O-163, P-508, P-580,
P-585
Konuma, Y., P-618
Kopelman, A., P-159
Korrick, S. A., P-97
Kort, J., P-496
Kostelijk, E. H., O-55
Kotelchuck, M., O-37, O-42, P-292
Kounogi, S., O-64
Koustas, G., P-662
Kovacs, P., P-436
Kovalak, E. E., P-229
Kovalevsky, G., P-429, P-509
Kovanci, E., P-239, P-322, P-361, P-565
Kramer, W., P-737, P-739
Kramer, Y. G., P-271
Krawetz, S., O-201
Krieg, A., P-86
Krieg, S., P-86
Krikun, G., O-178, P-146
Kriplani, A., P-689
Krisher, R. L., O-56, P-260, P-269, P-329
Krishnamoorthy, K., O-265
Kroener, L., P-247, P-500
Kuang, Y., O-33, P-16, P-647, P-690
Kuchibhatla, M., P-80
Kudesia, R., P-37, P-67, P-69
Kuehl, T. J., P-357
Kuji, N., P-407
Kulkarni, A., P-326
Kumar, A., P-445, P-678
Kumar, D., P-299
Kumar, N., O-168, O-196, P-14, P-310, P-497
Kumasawa, K., P-459
Kumbak, B. A., O-117
Kuo, P.-L., P-188
Kuokkanen, S., O-18
Kuramoto, T., P-610
Kuroda, S., P-459
Kus, C., O-154
Kushnir, V. A., O-151, P-253, P-428, P-439,
P-592, P-622
Kusunoki, H., P-222, P-388
Kuwahara, A., P-312
Kuwahara, M. K., P-372
Kwak, S., P-267, P-344
Kwak-Kim, J., O-157, P-128
Kyono, K., P-57, P-536, P-575
Kyrou, D., P-640
Labarta, E., P-656
Labella, P., O-123, O-231, P-489, P-567, P-587
LaBrie, S., P-402
Lachgar, H., P-297
Lagunov, A., O-190
Lai, T.-H., P-189
Laknaur, A., O-75, O-185
Lamaita, R. M., P-148
Lamb, D. J., O-208
Lambalk, C. B., O-55
Landis, J., O-31, O-135, P-269
Landis, J. N., O-137, P-511
Lange, A., O-223, P-35
Lanham, M., P-720, P-736
Lannon, B. M., P-291
Lapensee, L., O-132, P-225, P-309
Large, M., P-507
Larreategui, Z., P-314
Larsen, C., P-237
Lathi, R., O-86, P-496
Laufer, M. R., P-162
Laughlin-Tommaso, S. K., P-137
Lavery, S., O-250
Lavolpe, M., P-515
Lawson, A., P-733, P-736
Lazarin, G. A., O-166, O-167, O-194
Lazzaroni-Tealdi, E., P-592, P-622
Leach, R., P-127
Leboeuf, M., P-158
Lebovic, D. I., P-346
Le Bras, A., P-311
Lechtenberg, L., P-445
Lee, D., P-65, P-102, P-433
Lee, D. M., O-130
Lee, D. R., P-562
Lee, D.-Y., P-154
Lee, E., P-154, P-380
Lee, F.-K., P-189
Lee, H.-L., O-123, P-489
Lee, H. S., P-671
Lee, J., P-380, P-433
Lee, J. A., O-80, O-112, O-121, O-146, O-160,
O-164, O-204, O-245, P-5, P-130, P-131,
P-368, P-442, P-629, P-652, P-675, P-700,
P-730
Lee, J.-W., P-543
Lee, K., P-245, P-259, P-658
Lee, K.-A., P-365
Lee, K. L., P-241, P-480
Lee, K.-H., P-267, P-344
Lee, M. S., O-150, P-666
Lee, M. M., P-97
Lee, M.-S., P-627
Lee, S., P-71, P-149
Lee, S.-K., P-543
Lee, S.-Y., P-365
Lee, T.-H., P-627
Lee, W., P-433, P-572
Lee, W. S., P-562
Lee, Y. J., P-411
Lefebvre, J., O-132
Legro, R., O-92, O-258
Lekovich, J., O-60, O-124, O-242, P-135, P-153,
P-248, P-342, P-454, P-463, P-642, P-649,
P-707
Lemyre, M., P-158
Leocata Nieto, F. A., P-571
Leon, J., P-405
Lerner, V., P-25
Lessey, B. A., O-16, O-217
FERTILITY & STERILITY Ò
e369

Leung, P. C. K., P-3, P-701
Leung, Y.-K., P-94
Levens, E., P-287
Levin, H. I., P-710
Levin, K., P-738
Levitas, E., O-237
Levy, M., O-187
Levy, M. J., P-382
Lewis, C. M., O-170
Lewis, E. I., O-232, P-356
Leza, A., O-173, P-685
Li, B., P-363
Li, D., P-299, P-377
Li, E., P-227
Li, F., O-177, O-178, P-609
Li, G., O-59
Li, J., P-190, P-417
Li, M., P-678
Li, Q., P-364, P-367
Li, S., O-203, P-550
Li, T., P-181
Li, X., O-49, O-130, O-222, P-212
Li, Y., P-90, P-582
Liang, X., P-90, P-657
Liang, X.-Y., P-457
Libby, V., P-186, P-400, P-628
Liberty, A., P-231
Librach, C., O-238, P-200, P-443
Librach, C. L., O-236, P-196, P-557, P-638
Licciardi, F., O-163, P-310, P-587, P-603
Liebermann, J., P-51, P-134, P-238, P-288, P-354,
P-481
Lieman, H., P-67, P-69, P-338
Lien, Y. Ron., P-694
Light, A., O-54
Lim, J., P-238, P-482
Lin, H., O-133
Lin, J., O-33, P-155
Lin, W., O-17
Lin, Y., P-406
Linan, A., P-41
Lindeman, M. R., P-512
Lindheim, S. R., O-214
Lindsey, J. S., O-188, P-162
Lindstrom, M. J., P-346
Ling, Q.-D., P-189
Lingaiah, S., P-55
Link, M., P-46
Linn, J. M., P-632
Lira, J., P-376
Lisonkova, S., P-150
Liu, D., O-225
Liu, E., O-35
Liu, H., O-63, O-274, P-425, P-580
Liu, H.-C., O-233, P-118, P-420
Liu, H.-E., P-457
Liu, J., O-33, P-27, P-79, P-265, P-406
Liu, K., P-196
Liu, L., P-181
Liu, X., P-85, P-242
Liu, Y., O-182, O-230, P-417
Liu, Z., P-521
Lo, I. P. Y., P-721
Lo, K., O-236, O-238
Lo, K. C., O-199, P-133
Lo, P., P-429
Lobel, A., P-248, P-454
Lobo, R., O-189, P-266, P-424
Logan, P., O-94
Lonczak, A., P-494
Londra, L., O-87
Lorenzi, D., P-487, P-515
Lo Turco, E. G., P-257, P-275, P-276, P-277,
P-596
Louie, K., P-108, P-244
Loup, V., O-248
Loy, G., P-104
Lu, G., O-222, P-212
Lu, Q.-S., P-121
Lu, Y., P-16
Lu, Z., O-63, P-580
Lubimkina, E., P-10
Lue, J., O-185
Luense, L. J., P-556
Lujan, M. E., P-68
Lujan, S., P-528
Lukaszuk, A., P-36
Lukaszuk, K., P-36, P-40
Luke, B., O-37, O-39, O-42, O-115, O-241, O-268,
P-292, P-298, P-335, P-452
Lukes, A. S., O-181
Lukkari-Lax, E., O-100
Luna, M., O-146, P-5, P-629, P-652, P-675
Lunenfeld, E., O-237
Lv, P., P-75, P-100, P-417
Lvov, E., O-138
Lynch, C. D., O-219, P-34
Lynch, D.-M., O-256
Lynch, K. A., P-402
Lynne, C., O-19
Lynne, C. M., O-24
Lysiak, J., O-23
Lyu, Q., P-16, P-608, P-690
Ma, L., P-27, P-265
Ma, S., P-244, P-538
Maas, K., P-319
Machtinger, R., O-136
Mack, W., P-88
Mackenzie, A., P-270
Mackenzie, A. C. L., O-161
Madjid, T., P-176
Madrigal, V., O-49, O-53
Madugu, H. N., P-4
Maghen, L., O-236, O-238
Magno, E., P-238
Mahalingaiah, S., P-384
Mahey, R., P-689
Mahmood, S. N., P-404
Mahmoud, S., P-283
Mahony, M. C., O-155
Mahran, A., P-233
Mahutte, N., P-297
Mainigi, M. A., O-3, O-144, P-306, P-337
Maisel, L., P-19
Maisenbacher, M. K., O-218
Maisog, J., O-219
Maje, I. M., P-4
Majzoub, A., O-111
Mak, W., O-133, P-335
Makarem, M. H., O-69, O-259, P-211
Makinen, S., P-221
Malik, M., O-4, O-58, O-184, P-142
Malik, S., P-92
Malmsten, J., P-358
Man, L., O-229
Manau, D., P-427
Mangal, R., P-239, P-322, P-361
Manichaikul, A., O-141
Mannara, J. I., P-698
Manoharan, A., O-196
Mansour, A., O-136
Margolis, D. J., O-53
Margolis, M., P-133
Marin, C., P-597
Marin, D., P-656
Marinuzzi, J., P-148
Marquis, K., P-86
Marrs, R. P., P-495
Marsh, E. E., O-77, O-269, P-138, P-143, P-423,
P-733
Marsidi, A. M., P-255
Martikainen, H., P-221
Martin, J., P-367
Martinez, A., P-15
Martinez, C., P-577
Martinez, E., P-41
Martinez, M., P-644
Martinez, S., P-597
Martins, M. V., P-379
Martins, W., P-697
Martins, W. P., P-654
Martins Da Silva, S. J., P-570
Martins-Filho, O. A., P-148
Mas, A., O-73, O-76, O-186, O-211
Maslow, B.-S., P-479
Maslow, B.-S. L., O-122, P-624
Massaro, F. C., O-131, O-270
Massin, N., P-418
Mata, A., P-414, P-415
Mataro, D., O-153
Mather, K. J., O-52
Mathews, J., P-51
Matsubayashi, H., O-27, P-390, P-549, P-617,
P-683
Matsukawa, N., P-536
Matsumoto, H., P-563
Matsumoto, Y., P-24
Matsuura, K., P-393
Matsuzaki, T., P-459
Matyas, R. A., O-139
Mauri, A. L., O-131, O-270
Mauricio, R., O-12
Maurya, P. K., P-166
Maxwell, R., O-214
Mayordomo, E., P-449
Mazetto, R., P-261, P-351
Mazur, E., P-322, P-361
Mazur, E. C., P-239
McAvey, B., O-18, P-37
McCaffrey, C., O-91, P-478, P-524, P-603, P-680
McCalla, S., P-709
McCallie, B., O-48, P-589, P-676
McCarthy, K., O-102
McCormick, S., O-174, P-260
McCoy, R., P-519
McCulloh, D. H., O-104, O-231, P-82, P-271,
P-478, P-493, P-504, P-510, P-524, P-525,
P-578, P-587, P-603, P-664, P-680
McCullough, A., P-540
McGarvey, M., P-289
McGee, E., P-83
McGinnis, L. A., P-274
McIntire, D. D., P-400
McKinnon, C. J., P-22
McLean, M., P-220
McManaman, J. L., O-272
McQueen, D., P-704
e370 Author Index Vol. 104, No. 3, Supplement, September 2015

McReynolds, S., O-165, P-289
McWilliams, K., P-517, P-518
McWilliams, T. K., P-512, P-517, P-518
Medrano, L., O-128, O-173, P-685
Medvedovic, M., P-94
Mehr, H., P-82
Mehta, R., P-241
Meintjes, M., P-241, P-245, P-259, P-480, P-658
Melnick, A. P., O-126, P-39, P-342, P-348, P-420,
P-643, P-655, P-682
Mendiola, J., O-205
Mendola, P., O-225
Mendoza, G., P-138, P-143, P-733
Meng, H., P-265
Meng, Y., P-100
Merchenthaler, I., O-161
Merchenthaler, I. J., P-270
Mergler, R. J., P-68
Merhi, Z., O-276
Merolla, A., P-464
Merrion, K., O-218
Mersereau, J. E., P-729
Mertens, A., P-448
Mertens, A. C., P-450
Mesaros, C., O-226
Meseguer, M., O-45, P-256, P-258, P-314, P-485,
P-520, P-593, P-600, P-616, P-673, P-679
Messerlian, C., O-96
Metwalley, A., P-233
Metwalley, A. M., P-613
Metzgar, T., P-526
Metzgar, T. L., P-612, P-660
Meyer, L. R., P-615
Michau, A., P-311
Michels, K. A., P-11
Mifsud, A., P-485
Mikhail, E., P-201
Miki, T., P-226, P-324, P-369, P-388
Mikkelsen, E. M., P-384
Miller, B. T., P-8, P-345
Miller, C. E., O-72, P-202, P-631
Miller, M., P-279
Miller, S. J., P-740
Mills, B. B., P-711
Milne, P. A., P-570
Minguez-Alarcon, L., O-223, P-97
Minjarez, D. A., O-125, P-503
Minkoff, H., P-709
Mio, Y., P-132, P-251
Miorin, J., O-149, P-718
Mirisol, R., P-718
Mirisol, R. J., O-149
Mironova, A., P-663
Missmer, S. A., O-232, P-49, P-162, P-219, P-340,
P-343, P-356, P-469, P-669, P-703
Mitchell, E., P-30, P-705, P-714
Mitchell-Leef, D., P-237
Miyake, T., P-459
Miyata, A., P-391
Miyauchi, O., P-300
Mizoguchi, C., P-251
Mizumoto, S., P-610
Mizuno, S., P-563
Mizuta, S., O-113, P-390, P-617
Mneimneh, A., P-236
Moazamian, A., P-123
Moazamian, R., P-117
Mocanu, E. V., P-650
Mohamed, S. A., O-6, O-182
Mohan, A., P-185
Moley, K. H., O-15, P-78
Molinari, E., P-278
Molinaro, T., O-83, O-180, P-370, P-584
Mollamahmutoglu, L., P-350, P-699
Mollen, C., P-374
Montalvo, N., P-397
Montegriffo, E., O-100
Monteleone, P. A., O-149, P-718
Moon, J.-W., P-180
Moon, K., P-421
Moore, R., P-279
Mor, A., P-264, P-709
Moravek, M. B., P-736
Morbeck, D., P-98, P-591
Moreau, C., O-87
Mori, R., P-56
Moriguchi, H., O-116
Morimoto, Y., P-56, P-285, P-563
Morin-Papunen, L., P-55
Morris, P. L., O-21
Morrison, L. S., P-509
Morsy, H., P-613
Moschini, R. A., P-684
Moskovstev, S., P-557
Mostisser, C., O-226
Motan, T., P-696
Motato, Y., P-673
Motta, E. L. A., P-261, P-351
Mouille, B., P-203
Mounce, G., P-570
Moustafa, S., P-95, P-216
Moy, F., O-210, P-458, P-462
Mskhalaya, G., P-10
Muasher, S. J., O-38, O-41, O-118, P-48, P-568
Mucowski, S., P-88
Muenke, M., P-15
Muhsen-Alansarri, S. A., P-404
Mukerji, B., P-280
Mukherjee, T., O-245, P-5, P-130, P-131, P-368,
P-652, P-675
Mulla, Z., P-217
Mullen, T., P-295
Mullet, T., O-247, O-248, P-444
Mullin, C., P-323, P-497
Mullinax, S., P-86
Mumford, S. L., O-1, O-139, O-140, O-191,
O-220, P-6, P-11, P-18, P-30, P-321, P-325,
P-705, P-713, P-714
Mumusoglu, S., O-67
Munch, E. M., O-216, P-659, P-668, P-716, P-717
Muneyyirci-Delale, O., P-160, P-168
Munkwitz, L., P-503
Munne, S., O-34, O-35, O-36, O-63, O-91, O-193,
P-14, P-490, P-495, P-497, P-498, P-580,
P-585, P-677
Munoz, E., O-45, P-593, P-673, P-679
Munoz, M., P-12
Murakami, M., P-610
Murphy, E. M., P-39, P-348, P-420, P-643, P-655,
P-682
Murphy, M. J., O-101
Murtadi, G., O-200, P-35
Musul, B., O-210
Mutlu, I., P-623, P-635
Mutlu, L., O-177, O-178, P-95
Mutlu, M. F., O-68, P-623, P-635, P-712
Muzzey, D., O-167
Naaman, R. R., P-403
Nachtigall, L. E., O-12
Nachtigall, M., P-25
Nachtigall, M. J., O-12
Naeemi, F., O-20
Naftolin, F., O-12, O-276
Nagayoshi, M., P-222, P-226, P-324, P-369, P-388
Nagras, Z. G., P-537
Nagy, Z., O-34, P-237, P-632, P-711
Nagy, Z. P., O-155
Najafaliyeva, A., P-712
Najari, B. B., O-109
Najdecki, R., P-640
Nakajima, S. T., P-26
Nakajo, Y., P-536
Nakamura, H., P-459
Nakamura, Y., O-192, P-320, P-536, P-561
Nakaoka, Y., P-285
Nakayama, K., O-64, P-618
Nakhuda, G., P-677
Nangia, A., P-335
Naqvi, H., O-178
Nasr, A., P-54
Nassar, J., O-159
Nasseri, A., P-499
Nassr, A. A., O-69, O-259, P-211
Nastri, C. O., P-697
Natan, Y., O-62
Natarajan, L., O-7, P-2
Nath, N. M., P-583
Naumann, P. W., P-547
Naumova, A., P-663
Navarrete, G., P-259, P-480, P-658
Navarrete, G. R., P-241, P-245
Navarro, P. A., P-110, P-654, P-697
Nayar, K. D., P-185
Nazem, T. G., O-198
Needleman, D., P-356
Nehir Aytan, A., O-51
Neisani Samani, E., O-177, O-178
Neithardt, A. B., P-509
Neitzel, D., P-295, P-307
Nelson, A., P-371
Nelson, S. M., P-28
Neri, Q. V., O-2, O-110, P-122, P-378, P-386,
P-546, P-560, P-573
Neubauer, B. R., P-115, P-215
Newell, J., P-141
Ng, N. W., O-188, P-162
NICHD’s Reproductive Med Network, O-255
Niederberger, C., P-408, P-481, P-528, P-533
Nieman, L., O-184
Nieman, L. K., O-4
Nimeh, T., P-528
Nishiyama, R., O-27, P-390, P-549, P-617, P-683
Niu, W., O-59
Noble, L. S., P-217
Noblia, F., P-515
Nodar, F., P-487, P-515
Noel, M., P-29
Nogales, M., P-41
North, J., O-181
Norton, H., P-332
Norwitz, E., O-243
Notarstefano, V., O-176, P-151
Novoa Sanchez, Y., P-651
Noyes, N., O-104, O-123, O-168, O-231, P-489,
P-567, P-578, P-587
Nulsen, J., O-122, P-255, P-479, P-624
Nunes, F. F., P-148
FERTILITY & STERILITY Ò
e371

Obata, R., P-57, P-575
O’Brien, J. E., O-20, O-147
Ocali, O., P-569
Odenwald, K., P-346
O’Gradney, S., P-210
Oguz, Y., O-68
Oh, S., P-187
Ohgaki, A., P-563
Ohl, D. A., O-28, O-172, P-537
Ohno, H., O-64, P-579
Ojeda, M., P-593, P-673, P-679
Oka, N., P-57, P-575
Okada, H., P-391, P-532
Okada, K., O-21
Okada, M., P-21, P-144
Okitsu, O., P-393
Oktay, K., P-458
Oktay, K. H., O-17, O-210, P-230, P-268, P-462,
P-465, P-471
Oktem, M., O-68, P-635, P-712
Oktem, O., O-162, P-33, P-475, P-634
Okubo, T., P-300
Okuda, T., P-57, P-575
Okuyama, N., P-57, P-536, P-575
Olcha, M., O-134, P-586
Olds, S., P-621
O’Leary, T., P-238
Oleinki, T. D., P-257
Oliana, O., O-250
Oliveira, J. A., O-131, O-270
Oliveira, R., P-179
Oliveira-Pelegrin, G. R., O-131, O-270
Olson, M., P-127
Omil-Lima, D., O-234
Omran, M. S., P-484, P-581, P-637
Onalan, G., P-304
Ono, M., P-262
Ophir, L., P-87
Oppenheimer, A., O-159
Ord, T. S., O-3, O-144
Ori, A., P-590
Oron, G., P-263
Ortiz Maffei, N., P-636
Orvieto, R., O-237, P-87, P-89
Orwig, K. E., P-92
Osborne, S. E., O-13
Osina, E., P-667
Othman, E., P-233
Othman, E. R., P-613
Ouhilal, S., P-297
Ouyang, Y., O-222, P-212
Owen, C. M., P-328
Ozcan, P., P-229, P-437
Ozgur, K., P-435, P-653
Ozimek, J. A., P-316
Ozler, S., P-52, P-60, P-62, P-63
Ozmen, B., P-639
Ozsurmeli, M., O-127
Oztas, E., P-52, P-60, P-62, P-63
Pabon, D., P-485
Pabuccu, E. G., P-639
Pacheco, A., P-12
Pacheco, F., P-230
Paczkowski, M., P-357
Padovani, G. P., O-26
Padte, K., P-163
Page, D., P-15
Pagou, E., P-640
Paik, K. G., O-218
Pak, K.-A., P-543
Pakhalchuk, T., P-523
Pal, L., O-213, P-95
Palermo, G. D., O-2, O-110, P-122, P-153, P-378,
P-386, P-546, P-560, P-573
Palhares, M. B., P-654
Palmerola, K. L., P-266, P-472
Palumbo, A., P-9
Pan, Y., P-323
Paniza, T., O-110, P-122, P-546, P-573
Papadakis, M., P-332
Papanikolaou, E. G., P-640
Papier, S., O-36, P-487, P-515, P-636
Papouchis, N., P-738
Parasar, P., O-188, P-162
Pardue, S. L., O-52
Parfitt, D.-E., O-57, P-8, P-345, P-419
Parikh, T. P., O-142
Park, H., P-71, P-217
Park, I., P-180, P-267, P-344
Park, J., P-149, P-187, P-267
Park, J. H., P-671
Park, K.-S., P-365
Park, L., P-122, P-546
Park, M., P-572
Park, R., O-26
Park, S., P-65, P-102, P-187, P-392
Park, S.-Y., P-543
Parks, J. C., O-48, P-589, P-601, P-676
Parry, J., P-197
Parsons, S., O-112, O-164, O-204
Pascale, C., P-310
Pasch, L., P-725
Pasqualotto, F. F., P-281
Pasquier, M., P-418
Pastore, L., O-141
Pastuszak, A. W., O-208
Patel, A., O-4, O-184, P-142
Patel, B., O-30, P-412, P-548, P-551
Patel, J. C., O-81, P-734
Patounakis, G., O-61, O-187, P-287, P-328, P-382,
P-394, P-399
Patrizio, P., O-62, P-278
Pattinaja, D. A. P. M., P-674
Patton, A. L., O-48, P-589, P-676
Paulson, R., P-339, P-726
Payson, M., P-227
Pearlstein, H., P-688
Peavey, M., P-565, P-607
Peck, J. D., O-170, O-227, O-254
Pedro, J., P-379
Pellicer, A., O-262, P-449, P-644, P-656
Pelts, E. J., P-51
Peluso, C., P-179
Peluso, J. J., P-85
Penarrubia, J., P-427
Peng, B., P-150, P-155, P-701
Penman, A. D., O-82
Penrose, L., P-605, P-614
Penzias, A., O-243, P-291, P-315, P-585
Peralta, S., P-427
Pereira, N., O-60, O-124, O-242, O-249, P-135,
P-153, P-248, P-454, P-463, P-642, P-649,
P-707
Perez, M., O-45, P-593, P-679
Perez-Albala, S., P-600
Pericuesta, E., P-123
Perkins, K., O-263, P-236
Perkins, N. J., O-139, O-191, P-18, P-705
Perlman, J., P-709
Perreault-Micale, C., P-295, P-307
Perretti, J., P-284
Persaud, S., P-737, P-739
Peters, K., P-92
Petersen, C. G., O-131, O-270
Petersen, P., O-149, P-718
Petrini, A. C., O-124, P-642
Petrov, D., P-519
Petrozza, J. C., P-35
Pettersen, B., O-218
Phan, J. D., O-49
Phillips, S., O-132, P-252
Picou, A., P-661
Pier, B., P-279
Pietin Vialle, C., P-418
Piltonen, T., P-55
Pinkas, H., O-237
Pisarska, M. D., O-260, P-247, P-313, P-316,
P-473
Pliego, J. F., P-357
Plociennik, L., P-36, P-40
Plosker, S., P-201
Plowden, T., O-78, O-140, O-191, O-220, P-6,
P-321, P-713
Pohlmeier, A., O-102
Poizat, C., P-283
Polackwich, A. S., P-553
Polhemus, A. M., P-117, P-123
Poli, M., P-590
Pollard, J., O-18
Polotsky, A. J., O-39, O-201, O-258, O-268,
O-272, O-274, P-425
Pomeroy, K. O., P-284, P-397, P-566, P-576
Pons, J., O-47
Pool, T. B., P-346
Porcu Buisson, G., P-305
Pospisil, C., O-56
Post, M., P-499
Pouliot, M., P-158
Pourret, E., O-239
Powell, M., P-220
Prasad, N., P-401
Pratap, N. K., P-213
Prates, R., O-163, P-508
Price, T. M., P-26, P-80, P-137
Prien, S., P-605, P-614
Prins, G., P-481
Prins, G. S., P-533
Prizant, H., P-139
Proctor, C., O-267
Proctor, G., P-519
Prokai, D., P-19
Provost, M., O-38, O-41, O-118
Provost, M. P., O-40, O-119, O-261
Purandare, N. C., P-650
Purcell, S., P-241, P-245, P-259, P-658
Purcell, S. H., P-480
Puscheck, E., O-143, P-26
Puscheck, E. E., O-258, P-7, P-81, P-364, P-626
Puurunen, J., P-55
Pyle, A., P-278
Qian, Y., P-417
Qin, Y., P-241
Quach, K., O-238, P-196
Quaglieri, C., P-84
Quea, G., P-331
Quevedo, S., P-331
Quinteiro Retamar, A., O-36
e372 Author Index Vol. 104, No. 3, Supplement, September 2015

Quistorff, J., P-458
Rabinowitz, M., P-519
Raburn, D. J., P-48
Racicot, M.-H., P-252
Racowsky, C., O-136, O-138, O-193, P-356,
P-469, P-564, P-669, P-706
Radhakrishnan, G., P-184
Radin, R., P-18, P-30, P-140, P-705, P-714
Rafizadeh, M., P-526
Ragheb, A. M., P-389
Rahil, T., P-402
Rahmawati, E., P-166
Raia, M., O-166
Raju, R., P-81, P-626
Rakhila, H., P-158
Ramalingam, N., P-45
Ramer, I., O-158, P-125, P-126, P-682
Ramirez, D., P-217
Ramirez, E., O-53
Ramirez, L. B., O-138, P-564
Ramos, B., O-128, O-173, P-685
Ramos, L., P-316
Ramos, L. G., P-313
Rangarajan, S., P-200
Rappolee, D. A., P-364
Rarosi, F., P-436
Rasheed, S. M., P-233
Rauch, E., O-145
Ravichandran, K., O-44, P-585
Rebbeck, T. R., P-447
Reda, C. V., P-526, P-612, P-660
Reed, B., P-218, P-400
Reed, B. G., P-186, P-628
Reed, S., P-294
Reid, A., O-88
Reid, M., P-78
Reis, H. S., P-730
Reisman, L., O-110, P-122, P-546, P-573
Remohi, J., P-256, P-258, P-314, P-485, P-600,
P-656
Ren, H., P-538
Renzi, A., O-131, O-270
Requena, A., P-12
Resetkova, N., P-291
Revich, B., P-97
Rey Valzacchi, G. J., P-571, P-698
Rheaume, C., P-158
Ribeiro de Andrade, M., P-539
Riboldi, M., P-597
Ribustello, L., O-34, O-35, P-677
Ricard, J., P-397
Riche, D., P-197
Rich-Edwards, J. W., P-703
Richter, K. S., O-20, O-147, P-287, P-321, P-394,
P-399, P-421, P-482
Riddle, M. P., O-108
Ridha-Albarzanchi, M. T., P-404
Rienzi, L., P-520, P-630
Riley, J. K., P-43, P-293
Riley, K., P-141
Rinaudo, P., P-242
Rippon, G., O-172
Risteli, J., P-55
Robertshaw, I., P-708
Robinson, M. K., P-49
Robinson, R., P-432
Robinson, T. S., P-1
Robledo, C., O-274, P-425
Roby, K., P-86
Rocafort, E., O-128, O-173, P-685
Rodolosse, A., O-47
Rodosthenous, R., O-136
Rodrigo, L., O-148
Rodrigues, D., O-149, P-718
Rodriguez, A., O-153, O-195
Rodriguez, S., O-168, P-14, P-310
Rodriguez-Iglesias, B., P-367
Rodriguez Kubrusli, M. A., P-698
Rodriguez-Purata, J., O-80, O-121, O-146, O-245,
P-5, P-131, P-629, P-652, P-665, P-675,
P-684, P-700
Roeca, C., P-706
Rogel, S., O-128
Rogers, K., P-725
Romany, L., P-600
Romero, A., P-644
Romero, J., P-485
Rooney, K., O-106
Rosario, M., P-514, P-522
Rose, S. D., P-96
Rosen, A. M., O-44
Rosen, J., O-94
Rosen, K., P-371
Rosenberg, L., P-140
Rosenblad, A., P-224
Rosenbluth, E., O-246
Rosenwaks, Z., O-2, O-60, O-110, O-124, O-126,
O-229, O-242, O-249, P-39, P-122, P-135,
P-153, P-248, P-348, P-358, P-378, P-386,
P-420, P-454, P-463, P-492, P-546, P-560,
P-573, P-642, P-643, P-649, P-655, P-682,
P-707
Roshdy, S., P-613
Ross, L., P-108, P-453
Roth, E. W., O-269
Rothman, K. J., P-17, P-22, P-384
Rotker, K. L., O-234
Rotoli, D., P-9
Roudebush, W., P-711
Roudebush, W. E., P-602
Rowen, T., P-735
Royster IV, G. D., P-330, P-341
Rozenchan, P., P-159
Rubin, R. S., O-20
Rubio, C., O-148, P-597
Rubio, J. M., P-644
Rudick, B., P-108
Rueter, M. A., P-727, P-728
Ruiter-Ligeti, J. J., P-413
Ruman, J., P-631
Russell, D., P-514, P-522
Ryan, A., P-519
Ryan, E. A. J., P-638
Ryan, G., O-216, P-659, P-668, P-716, P-717
Ryan, G. A., P-650
Rydze, R. T., P-50
Ryu, S., P-572
Sabanegh, E., P-114, P-119, P-387
Sabanegh, E. S., P-101, P-111, P-112, P-116,
P-381, P-451, P-534, P-544, P-545, P-552,
P-553, P-559
Sabry, M., P-233, P-613
Sachdev, N. M., P-478, P-510, P-525, P-664
Sadeghi-Nejad, H., P-548
Saed, G., P-282
Saed, G. M., O-13, P-91, P-113, P-115, P-215
Saed, M. G., P-113, P-115, P-215
Saha, I., P-157
Sahin Ersoy, G., P-195, P-430, P-431, P-434
Saito, H., P-312
Saitou, S., P-232
Saji, F., P-320, P-561
Sakashita, A., P-599
Saketos, M., P-334
Sakkas, D., O-243, P-291, P-301, P-315, P-319,
P-569
Salama, E. A., P-681
Salazar Garcia, M. D., O-157, P-128
Saldivar, S., P-217
Salem, W. H., P-107
Salemi, J., P-201
Salihu, H., P-201
Sallam, H. N., P-484, P-581, P-637
Salmon, K., P-240
Samanta, L., P-101, P-111, P-112, P-116, P-381,
P-387, P-552, P-555, P-559
Sammel, M. D., O-88, O-169, O-197, O-212,
P-447
Samplaski, M. K., O-199
Samstag, L., P-738
San, A., P-367
Sanchez, M., P-210
Sanchez Sarmiento, C., P-414, P-415
Sandler, B., O-44, O-80, O-121, O-146, O-245,
P-5, P-130, P-131, P-629, P-652, P-665,
P-675, P-684
Sandlow, J., P-383, P-740
Sandlow, J. I., O-171
Sandya, R., P-213
Sanfilippo, J. S., P-92
Sankaranarayanan, K., O-170
Santamaria, X., O-235
Santanam, N., P-172
Santoro, N., O-39, O-92, O-201, O-258, O-268
Santos, R., P-405
Santos, T. G., P-257
Sanz, J., P-449
Sapienza, C., P-306
Sapra, K. J., O-114, O-219
Sarac, G. N., P-558
Sargin Oruc, A., P-38
Saribal, S., P-250
Sasaki, C., P-536
Sasaki, K., O-72, P-202
Sasamine, K., P-390
Sato, Y., P-536
Sauer, M. V., O-189, P-266, P-424, P-472
Sawant, R., P-163
Sbracia, M., P-152
Scalici, E., O-247, O-248, P-444
Scarpellini, F., P-152
Schanne, A., P-445
Schattman, G. L., O-124, O-229, P-463, P-643,
P-655
Scheib, J. E., P-727, P-728
Schenk, L., P-239, P-322, P-361
Schenkman, E., P-323
Schickler, R., P-201
Schiewe, M. C., P-507
Schisterman, E., O-139, O-140, O-191, O-219,
O-220, P-6, P-18, P-30, P-705, P-713, P-714
Schlenker, T., P-260
Schliep, K. C., P-30
Schmelter, T., P-371
Schneider, R., P-422
Schneiderman, A., O-174
Schon, S. B., P-556
FERTILITY & STERILITY Ò
e373

Schoolcraft, W. B., O-32, O-46, O-48, O-56,
O-125, O-165, O-174, O-190, P-234, P-260,
P-289, P-329, P-438, P-503, P-589, P-601,
P-676
Schor, E., P-159
Schorsch, K., P-615
Schoyer, K. D., O-264, P-740
Schreiber, C. A., P-374
Schrepferman, C. G., P-542
Schufreider, A., P-704
Schulte, M. B., P-43
Schutt, A., P-50
Schweitz, M., O-165, P-289
Schweitzer, A., O-47
Schymura, M. J., P-452
Scoccia, H., O-14
Scott, R. T., O-31, O-134, O-135, O-137, O-180,
O-244, P-223, P-269, P-370, P-494, P-505,
P-511, P-526, P-527, P-584, P-586, P-594,
P-595, P-606, P-612, P-660, P-715
Sdrigotti, A., P-571, P-681
Seal, P., P-287
Seaman, E., P-529
Segal, T., P-325
Segars, J., O-58, P-26, P-84
Seifer, B., P-146
Seifer, D. B., O-275
Sekhon, L., P-130
Selter, J. H., P-468
Seminsky, M., O-215
Senapati, S., O-88, O-212, P-306
Senbabaoglu, F., O-162
Sengoba, K., P-138, P-143, P-733
Sengul, O., O-67
Seong, S., P-178
Serafini, P. C., P-351
Sereni, E., O-176, P-103, P-151
Sergeyev, O., P-97
Session, D. R., P-633
Seta, N., O-231, P-489, P-567, P-587
Setti, A. S., P-275, P-276, P-277, P-281
Setton, R., O-126, P-39
Seungdamrong, A. M., O-92
Sevillano, G., P-485
Seybold, D. J., P-359
Seyhan Ata, A., P-33, P-634
Seymen, M. C., P-558
Sha, H., O-43
Shaaban, A., P-347
Shaaban, O. M., O-228, P-161
Shaeib, F., O-143, P-282
Shah, D. K., P-219
Shah, M., O-86, P-496
Shah, T., O-28, O-30, P-537, P-551
Shahar, A., O-62
Shalaby, S. M., O-6, O-74, O-182, O-211
Shalamova, E., P-663
Shamonki, M., P-678
Shandley, L. M., P-448
Shapiro, B. S., P-183, P-192, P-327, P-349
Shapiro, D., P-631
Shapiro, D. B., P-632
Shapiro, M., P-408
Sharara, F., P-107, P-286, P-325, P-352, P-403,
P-620
Sharara, F. I., P-317
Sharara, N. F., P-403
Sharma, P., P-196, P-200, P-443, P-557
Sharma, R., P-101, P-111, P-112, P-114, P-116,
P-119, P-381, P-387, P-451, P-534, P-544,
P-545, P-552, P-555, P-559
Sharma, S., P-157
Sharpe, A. N., P-513
Shaulov, T., O-85
Shavell, V., P-127
Shavit, T., P-263, P-302
Shawber, C., P-710
Shazly, S. A., O-259
Shazly, S. A. M., O-69
Sheehan, D. A., P-734
Shelinbarger, C. L., P-614
Shen, W., P-1
Shen, Y., P-100
Sheng, J., P-100
Shenoy, C. C., P-591
Shi, H. L., O-138
Shibasaki, S., P-536
Shigeta, M., P-262
Shim, S., P-392
Shimada, H., P-407
Shimomura, M., O-64, P-618
Shimomura, Y., P-391
Shin, D., O-30, P-433, P-551
Shin, J., P-71, P-380
Shin, P. R., O-20
Shin, S.-H., P-543
Shin, T., P-391, P-532
Shipley, S., O-20
Shlush, E., O-236, O-238, P-557
Shoupe, D., P-88
Shraga, R., O-168, O-196, P-14, P-310
Shu, Y., P-19
Shwayder, J. M., P-197
Siegersma, K., O-269
Sifer, C., P-460, P-476
Sigman, M., O-234, P-199
Sigurjonsson, S., O-218
Silber, S. J., P-15
Silva, E., P-269, P-329
Silva, L. R., P-697
Silva-filho, A. L., P-148, P-376
Silver, R. M., O-139, O-140, O-191, O-220, P-6,
P-18, P-705, P-713, P-714
Silverberg, K., P-661
Simbulan, R., P-242
Simon, A., P-519
Simon, C., O-73, O-148, O-186, O-235, P-367,
P-597
Sinaii, N., P-160, P-168
Singer, A., O-88
Singer, T., P-497
Singh, S., P-243
Singh, T., P-280
Sisti, G., O-158, P-125, P-126
Sjaarda, L., O-191, P-6, P-11, P-18, P-705, P-713,
P-714
Sjoblom, C., P-662
Skanes-DeVold, H. D., O-214
Skaznik-Wikiel, M. E., O-272
Skrip, L., P-95
Slayden, O. D., O-95, O-97, O-99, P-129
Slizewski, D., O-217
Slutsky, J., P-737, P-739
Smarr, M. M., O-225
Smigulina, L., P-97
Smith, A. D., P-28
Smith, H., P-662
Smith, J. F., O-94, P-735
Smith, K., P-46, P-541
Smith, M., P-490
Smith, R., O-23, P-260
Smith-Harrison, L., O-23
Smotrich, D., O-84
Soh, S., P-391
Soh, Y., O-152
Somova, O., P-396
Son, W.-Y., P-263
Song, S.-H., P-392
Song, Y., P-156, P-177
Sonigo, C., P-460, P-476
Sonksen, J., O-28, P-537
Sonmezer, M., P-471, P-639
Sonohara, M., P-579
Sood, A., O-107, O-213, P-216
Sota, N., P-485
Souter, I., O-200, P-35
Spaanderman, M. E. A., P-674
Spaine, D. M., P-539
Spandorfer, S. D., O-126, O-158, O-242, P-39,
P-125, P-126, P-342, P-682
Spath, K., O-252, P-590
Spector, L. G., O-241, P-298, P-452
Spencer, J., P-448
Spencer, J. B., P-450
Spiessens, C., P-136
Sroga, J. M., P-708
Srougi, M., O-26, P-535
Srougi, V., P-535
Srouji, S., P-469
Srouji, S. S., P-669
Stalzer, A., P-359
Stanczyk, F. Z., P-80, P-88, P-453
Stanford, J., O-191, P-22
Stanhiser, J., P-203
Stapleton, G., P-104
Stecher, V., O-172
Steele, H., P-737, P-739
Steele, M., P-737, P-739
Stegmann, B. J., P-308, P-646
Stein, D. E., O-121
Steinberg, M. L., O-269
Steiner, A., O-92, O-224, P-693
Steiner, A. Z., P-231
Steinkampf, M. P., P-366
Steller, C., O-72, P-202
Stelling, J., O-193, P-334
Stemm, K., P-621
Stern, J. E., O-37, O-42, O-115, P-292, P-335,
P-452
Stevens, J. M., O-56, O-125, O-174, P-289, P-329,
P-438, P-601
Stewart, E., O-107
Stewart, E. A., O-181, P-137, P-591
Stewart, J., P-248, P-342, P-454
Stillman, R., P-399
Stillman, R. J., O-147
St. Marie, P., P-402
St-Michel, P., P-297
Stobezki, R., O-210
Stocco, C., O-14
Stoppa, M., P-520
Stouffer, R. L., O-101
Strachan, G., O-102
Strashnova, A., P-523
Stratton, P., P-160, P-168
Straub, R. J., P-237
e374 Author Index Vol. 104, No. 3, Supplement, September 2015

Strawn, E. Y., O-264, P-740
Strobino, D., O-87
Strug, M., P-127
Strumbly, D., P-412
Strynar, M., O-224
Styer, A. K., O-39, O-255, O-258, O-268
Su, I., O-7, P-2, P-470
Su, Y., O-59
Su, Y.-T., P-719
Such, E., P-449
Sudhakaran, S., P-464
Sueldo, C., P-296
Sueldo, C. M., P-85
Sugino, N., P-21, P-144
Sugishima, M., P-132, P-251
Suh, C., P-380
Sukhwani, M., P-92
Sullivan-Pyke, C. S., P-710
Sulo, S., O-72
Sultan Ahamed, A. M., P-235
Summers, K. M., O-216, P-659, P-668, P-716,
P-717
Summers-Colquitt, R. B., P-346
Sun, H., P-267, P-344
Sun, L., O-225
Sun, Y., O-59, P-3, P-670
Sundaram, R., O-154, O-219, O-225
Sunderam, S., P-109
Sung, L., P-334
Sung, N., O-157, P-128
Surrey, E., O-125
Surrey, M., P-240, P-473, P-490, P-495
Sutcliffe, A. G., O-93
Suzuki, K., P-391, P-407
Suzuki, R., P-232
Swain, J. E., O-56, O-190, P-234, P-346, P-438,
P-604
Swan, S. H., O-205
Swanson, K., O-79
Swanson, S., P-557
Swerdlow, A. J., O-93
Sylvestre, C., O-132
Sylvestre-Margolis, G., O-145
Szlit Feldman, E., P-681, P-698
Taboas, E., O-45, P-593, P-679
Tada, Y., O-192, P-320, P-561
Tadros, T., P-311
Taguchi, S., O-192, P-320, P-561
Taiyeb, A. M., P-404
Takahashi, C., P-617
Takahashi, M., P-536
Takano, T., P-320, P-561
Takaya, Y., O-27, P-549, P-683
Take Kaplanoglu, G., P-558
Takemoto, Y., P-222
Takeuchi, K., P-246
Takeuchi, M., O-64
Takeuchi, T., O-113, P-57, P-536, P-575
Takiuchi, T., P-459
Tal, O., O-275
Tal, R., O-275, P-709
Talebian, S., P-442
Talevi, R., P-464
Taliadouros, G. S., P-410
Tam, M., P-722, P-723, P-731, P-732
Tamura, F., P-618
Tamura, H., P-21, P-144
Tan, L., P-134
Tan, O., P-400
Tan, S.-J., P-455, P-461
Tanacan, A., P-441
Tanaka, A., P-222, P-226, P-324, P-369, P-388,
P-407
Tanaka, I., P-226, P-324, P-369, P-388
Tanaka, K., P-610
Tang, B., O-102
Tang, Y., O-233, P-118
Taniguchi, F., P-170
Tanrikut, C., O-205, O-207, O-209, O-271
Tao, X., O-31, O-135, O-137, P-269, P-494, P-505,
P-511, P-526, P-586
Tapanainen, J., P-221
Tapanainen, J. S., P-55
Tarozzi, N., O-252
Tatpati, L. L., O-8
Tavares, A., O-26
Taylor, D., O-5
Taylor, H. S., O-177, O-178, P-146
Taylor, L., O-88
Taylor, R. N., P-19
Taylor, T. H., P-491, P-516
Tecson, V., O-163
Tejera, A., P-520
Tepper, A. B., O-83
Terakawa, N., P-170
Testillano, M., P-616
Thakore, S., P-412
Thakur, M., O-143, P-7, P-81, P-626
Thomas, A., O-53
Thomas, M., P-259, P-658
Thomas, M. A., P-94, P-708
Thomas, M. R., P-241, P-245, P-480
Thomas, S. M., O-40, O-119, O-261
Thompson, A., O-15, P-78
Thompson, K., P-526
Thompson, W., O-73
Thornton, K., O-273, P-47, P-315, P-319, P-338
Thornton, K. L., P-340
Tibaldi, D. S., P-539
Tiegs, A. W., P-493
Tiitinen, A., P-221
Tilley, B., P-241, P-245, P-259, P-480, P-658
Timur, H., P-23, P-38, P-61
Tiseo, B. C., O-26, O-205, O-209, P-535
Titus, S., O-17, O-210
Tjaden, B. L., O-8
Tjoa, M., O-102
Tokmak, A., P-52, P-60, P-62, P-63
Tokoro, M., P-579
Tokudome, M., P-246
Toledo, A. A., P-237, P-632
Toner, J., P-641
Toner, J. P., P-326
Torno, A., P-681, P-698
Toth, T., O-150, P-703
Toth, T. L., O-200, O-223
Trabucco, E., P-630
Tran, N. D., O-94
Traxler, S. A., P-374
Treff, N. R., O-31, O-134, O-135, O-137, P-269,
P-370, P-494, P-505, P-511, P-526, P-527,
P-586
Trevino, L. S., O-76
Trevisan, C. M., P-179
Trew, G., O-250
Trofimenko, V., P-541
Troshina, M., P-663
Troup, S., P-354
Truong, T. T., O-152
Tsai, E.-M., P-147
Tsai, S., P-43
Tsuiki, M., P-579
Tsuji, H., P-618
Tsukamoto, K., P-232
Tsuneto, M., P-251
Tucker, M., P-238
Tucker, M. J., O-147, P-482
Tulandi, T., P-263, P-302
Turan, V., P-230, P-462, P-465, P-471
Turki, R., P-555
Turki, R. F., P-387
Turkkani, A., P-350
Turner, T. G., P-661
Tusheva, O. A., P-497
Tvrda, E., P-451
Tyden, T., P-224
Tzeng, C.-R., P-166, P-455, P-461
Ubaldi, F., P-520
Ubaldi, F. M., P-630
Uhler, M. L., P-481, P-704
Ulker, K., O-117
Ulug, M., P-645
Umezawa, A., P-407
Unal, S., O-117
Uncu, G., P-167, P-250
Underberger, C., P-240
Upham, K. M., O-31, P-594, P-595, P-612
Uquillas, K., P-25
Urich, M., P-609
Uriondo, H., P-487
Urman, B., O-162, P-33, P-475, P-634
Utsunomiya, T., P-407
Uvarova, E., P-523
Vaast, M., P-254
Vaccari, L., O-176, P-103, P-151
Vagnini, L. D., O-131, O-270
Vaiarelli, A., P-630
Valbuena, D., P-597
Valdes, C., P-565
Valdes, C. T., P-50
Vallejo, V., O-145
van den Abbeel, E., O-55, P-249
van Golde, R. J. T., P-674
Vanijgul, C., P-239
van Rijswijk, J., O-55
Van Voorhis, B., P-219, P-326
VanWort, T. A., P-368
Vargas, M. V., P-206, P-214
Vaskivuo, T. E., P-55
Vassena, R., O-47, O-153, O-195
Vaughan, D. A., O-243, P-426, P-574
Vaughan, L. E., P-137
Vega, M. G., P-428, P-439
Vega, R. R., P-405
Veleva, Z., P-221
Velez Edwards, D. R., P-145
Vergouw, C. G., O-55
Verlinsky, O., P-523
VerMilyea, M., P-337
Vernaeve, V., O-153, O-195
Verrecchio, E. S., P-688
Victorino, A. B., P-596
Vidal, C., O-148
Vidali, A., O-151
Vigneswaran, H. T., O-234
Villamon, E., P-449
Villeneuve, K., O-190
FERTILITY & STERILITY Ò
e375

Villette, C., P-418
Viloria, T., O-45
Vincens, C., O-247, P-182, P-254
Vinning, T., P-538
Vintejoux, E., P-444
Vitek, W., O-39, O-258, O-268, P-377
Vitiello, D., O-193
Vitonis, A., P-219
Voong, C., O-197
Voronich, N., P-523
Vyas, N., P-313, P-316
Wachs, D., O-246
Wachter, K., P-621
Wactawski-Wende, J., O-1
Wagner Coughlin, C., P-354, P-495
Wald, M., P-547
Walker, C., O-73
Walker, C. L., O-76
Wallace, M. E., O-225
Wang, B., P-70
Wang, C., P-181
Wang, E. T., O-260, P-247, P-313, P-316, P-473
Wang, H., P-582
Wang, N., P-647
Wang, Q., P-592, P-622
Wang, R., P-548
Wang, S., P-514, P-522
Wang, W., O-233, P-118
Wang, X., O-84, P-44, P-457, P-556
Wang, Y., P-586, P-647
Wantman, E., O-115
Ward, K., O-175
Warinner, C., O-170
Warne, D. W., P-336
Warner, E., O-215
Warner, L., P-109
Warty, N., P-163
Watanabe, C., P-563
Watanabe, H., P-232, P-618
Watanabe, S., P-222, P-388
Waud, K., P-191
Weaver, A., P-137
Webster, W., O-257
Weckstein, L., O-246
Weedin, E. A., O-227
Weinerman, R. S., O-3, O-144, P-306
Welch, C., P-498
Welliver, C., P-540
Wellons, M., P-145
Wells, D., O-44, O-91, O-252, P-585, P-590
Werland, H., P-661
Werner, M. D., O-31, O-135, O-137, O-180,
O-244, P-223, P-370, P-584, P-594, P-595,
P-606, P-612, P-660, P-715
Wesselink, A., P-22
Wesselink, A. K., P-384
Wessels, C. E., P-605, P-614
Whalley, K., P-570
Wheeler, K., O-23
Whitcomb, B. W., P-287, P-382, P-470
Whitehouse, M. C., O-80, O-121, O-146, O-164,
O-204, P-5, P-130, P-368, P-442, P-629,
P-665, P-675, P-684, P-700
Whiting, S., P-117
Wickner, P., O-256
Widra, E., P-328
Widra, E. A., O-61
Wiehle, R., O-29
Wiemer, K., P-621
Wild, R. A., O-50
Wilkinson, T., O-250
Willey, R., P-631
Williams, C. L., O-93
Williams, H. L., P-570
Williams, L. A., O-181
Williams, L. J., O-13, P-91
Williams, M., P-452
Williams, P., O-96, O-223, P-703
Williams, P. L., P-97
Williams, S. E., P-602
Willman, S., O-246
Wilson, E. E., P-218
Wilson, L., P-279
Wincek, T., P-357
Winchester, P. D., O-267
Wing, R., P-491
Wininger, J. D., P-602
Winkel, A., P-25
Winkelman, W. D., P-735
Winston, N. J., O-14
Wise, L. A., P-17, P-22, P-140, P-384
Witkin, G., P-730
Witkin, S., O-158, P-682
Witkin, S. S., P-125, P-126
Witt, B., O-163
Witz, C. A., P-631
Wiweko, B., P-355
Wolff, E. F., O-142, P-341
Wong, J. M. K., P-736
Wong, K., P-443
Wong, S., P-722, P-723, P-731, P-732
Woo, I., P-339
Wood, E., P-397
Wood, G. J. A., P-535
Woodruff, T. K., P-269
Woodrum, D. A., P-137
Wormer, K., P-332
Wozniak, E. M., P-740
Wright, D. L., O-150
Wright, K., P-172
Wu, H., P-345
Wu, J., P-1
Wu, X., P-84
Wu, Y., P-417
Wu, Y.-G., P-592, P-622
Wun, W.-S. A., P-239, P-322, P-361, P-565
Wyatt, M., P-512, P-517, P-518
Wylie, B., O-96
Xia, M., P-27
Xiao, L., P-156, P-177
Xie, C., O-267
Xie, Y., O-154
Xing, T., P-586
Xu, F., O-98
Xu, H., P-190, P-417
Xu, J., O-170
Xu, W., O-23
Yagi, H., P-391
Yakin, K., P-634
Yaklic, J., O-214
Yakovenko, S., P-523, P-663, P-667
Yakut, H. I., P-63
Yallampalli, C., P-50
Yamagata, Z., P-407
Yamaguchi, K., O-27, O-113, P-390, P-549,
P-617, P-683
Yamaguchi, T., P-226, P-324, P-369, P-388
Yamanaka, M., P-285
Yamochi, T., P-285
Yan, Z., P-53, P-608
Yanagihori, S., P-57, P-575
Yang, J., P-16, P-385
Yang, M., P-521
Yang, N., P-433
Yang, P.-K., P-694
Yang, Q., O-76, O-185
Yang, X., P-657
Yang, Y., P-364
Yang, Z., O-33
Yango, P. L., O-94
Yankov, V. I., P-631
Yao, S., O-95, O-98
Yarnall, S., O-168, O-196, P-14, P-310
Yasa, C., O-127, P-205
Yasmin, S., P-398, P-531
Yasui, Y., P-459
Yauger, B., O-25, P-194, P-341
Ye, A., P-11
Ye, Z., O-249, P-358, P-492
Yeboah, E., P-495
Yee, B., P-288, P-354
Yee, S., O-215
Yegunkova, O., P-396
Yeh, J. S., O-38, O-40, O-41, O-118, O-119,
O-261, P-80
Yelumalai, S., P-570
Yerkes, S. E., O-80, P-730
Yerushalmi, G. M., P-87, P-89
Yesiladali, M., P-437
Yeste, M., P-570
Yeung, E., O-154
Yi, K., P-71
Yi, Y., O-222, P-212
Yih, Y.-C., P-416, P-446
Yildirim, U., P-699
Yilmaz, G., P-205
Yilmaz, N., P-23, P-38, P-61, P-62, P-63
Yilmaz, S., P-23, P-61
Yin, O., O-10
Yin, T., P-385
Yong, P., P-150
Yoon, J.-S., P-543
Yoon, S., P-433, P-572
Yoon, T., P-433, P-572
Yoon, T. K., P-562
Yoshida, A., O-240, P-42
Yoshimura, T., O-64
Young, D. O., O-165
Young, J., P-127
Young, L. K., P-320, P-561
Young, M. J., O-218
Young, N., O-142
Young, S. L., O-16, O-141, O-217
Younis, J. S., O-237
Yu, D., P-100
Yu, Y.-P., P-3
Yuksel, S., P-60
Yumoto, K., P-132, P-251
Yung, Y., P-87, P-89
Yurttas Beim, P., O-57, O-160, P-8, P-345, P-419
Yuting, Y., P-367
Zaca, C., O-176, P-103, P-151
Zacur, H. A., O-10
Zadeh, S., O-103
Zaghmout, O., O-70
Zajic, S., P-646
Zakarin Safier, L., O-189, P-424
e376 Author Index Vol. 104, No. 3, Supplement, September 2015

Zaki, H., P-686
Zaletova, V., P-10
Zamah, A. M., O-14
Zamara, C., O-131
Zaninovic, N., O-249, P-358, P-492
Zarek, S., O-140, O-142, O-191, O-220, P-6, P-30,
P-330, P-713
Zarutskie, P., P-607
Zavy, M. B., O-50
Zavy, M. T., O-50
Zbella, E. A., P-210
Zeadna, A., O-237
Zeeck, K. M., P-383
Zengin, D., O-67, P-441
Zeyneloglu, H. B., P-304
Zgodic, A., O-57
Zhan, H., P-155
Zhan, Q., O-249, P-358, P-492
Zhan, Y., O-137, P-586
Zhang, D., P-417, P-646
Zhang, F., P-670
Zhang, H., O-92, O-201, O-258
Zhang, J., O-33, O-34, O-63, P-495, P-521, P-580,
P-677
Zhang, L., P-506
Zhang, S., P-181, P-582
Zhang, W., O-15
Zhang, X., P-94, P-506
Zhang, Y., P-236, P-385
Zhao, D., O-254
Zhao, Y., O-87, O-230, P-74
Zheng, H., P-79
Zheng, Y., O-266
Zhou, B., P-565, P-702
Zhou, M., P-156, P-177
Zhou, Y., O-178
Zhu, Y., O-225
Zhylkova, L., P-396
Zimmerman, A., O-182
Zimmerman, R., P-505
Ziv-Polat, O., O-62
Zlatopolsky, Z., P-523
Zollinger, C., P-499
Zorina, I., P-667
Zozula, S., P-507
Zullo, F., P-630
FERTILITY & STERILITY Ò
e377

TOPIC INDEX
ART- In Vitro Fertilization: O-037, O-038, O-040,
O-041, O-042, O-061, O-068, O-078, O-081,
O-090, O-115, O-116, O-118, O-119, O-120,
O-127, O-129, O-153, O-169, O-187, O-216,
O-241, O-242, O-247, O-248, O-260, O-261,
O-262, O-264, O-267, O-270, P-023, P-027,
P-028, P-029, P-095, P-096, P-107, P-108,
P-109, P-157, P-196, P-211, P-220, P-221,
P-222, P-223, P-225, P-226, P-227, P-228,
P-230, P-233, P-235, P-236, P-237, P-240,
P-242, P-243, P-251, P-263, P-265, P-286,
P-287, P-288, P-292, P-293, P-294, P-297,
P-298, P-300, P-301, P-302, P-303, P-304,
P-314, P-315, P-316, P-317, P-319, P-320,
P-321, P-324, P-325, P-326, P-327, P-328,
P-329, P-330, P-332, P-333, P-334, P-335,
P-336, P-337, P-338, P-339, P-340, P-341,
P-342, P-345, P-346, P-347, P-348, P-349,
P-351, P-353, P-354, P-355, P-356, P-357,
P-358, P-364, P-385, P-388, P-396, P-398,
P-399, P-406, P-498, P-519, P-578, P-600,
P-626, P-633, P-637, P-645, P-646, P-651,
P-652
ART-Other: O-065, O-083, O-084, O-087, O-103,
O-117, O-154, O-257, O-263, P-224, P-229,
P-231, P-232, P-234, P-239, P-244, P-266,
P-289, P-291, P-295, P-296, P-299, P-305,
P-306, P-307, P-308, P-309, P-310, P-311,
P-312, P-313, P-322, P-323, P-331,
P-343, P-344, P-350, P-352, P-359, P-360,
P-361, P-407, P-543, P-601, P-638, P-719,
P-737
Cancer: O-007, O-017, O-093, P-447, P-448,
P-449, P-450, P-451, P-452, P-453, P-454
Contraception/Family Planning: O-095, O-098,
O-100, O-101, O-102, O-214, P-371, P-372,
P-373, P-374, P-375, P-376, P-377, P-378,
P-379
Cryopreservation: O-003, O-062, O-124, O-155,
O-203, P-238, P-241, P-245, P-246, P-247,
P-248, P-249, P-250, P-252, P-253, P-254,
P-255, P-256, P-257, P-258, P-259, P-260,
P-261, P-262, P-264, P-267, P-268, P-478,
P-479, P-480, P-481, P-482, P-483, P-484,
P-485, P-487, P-533
Early Pregnancy: O-059, O-085, O-086, O-088,
O-089, O-096, O-139, O-140, O-158,
O-194, O-219, O-220, O-221, O-222, P-030,
P-700, P-701, P-702, P-703, P-704, P-705,
P-706, P-707, P-708, P-709, P-710, P-711,
P-712, P-713, P-714, P-715
Embryo Biology: O-044, O-045, O-046, O-048,
O-063, O-091, O-134, O-144, O-243,
O-252, P-579, P-580, P-581, P-582, P-583,
P-584, P-585, P-586, P-587, P-589, P-590,
P-591, P-592, P-593, P-594, P-595, P-596,
P-597, P-599, P-602, P-603
Embryo Culture: O-056, O-138, O-152, O-249,
O-251, P-604, P-605, P-606, P-607, P-608,
P-609, P-610, P-611, P-612, P-613, P-614,
P-615, P-616, P-617, P-618, P-619, P-620,
P-621, P-622
Embryo Transfer: O-005, O-039, O-125, O-126,
O-145, O-146, O-147, O-149, O-150,
O-244, O-245, O-268, P-657, P-658, P-659,
P-660, P-661, P-662, P-663, P-664, P-665,
P-666, P-667, P-668, P-669, P-670, P-671,
P-672
Endometriosis: O-016, O-175, O-176, O-177,
O-178, O-179, O-180, O-266, P-146, P-147,
P-148, P-149, P-150, P-151, P-152, P-154,
P-155, P-156, P-158, P-159, P-160, P-161,
P-162, P-163, P-166, P-167, P-168, P-170,
P-172, P-173, P-174, P-176, P-177, P-178,
P-179
Endometrium: O-018, O-217, P-180, P-181,
P-182, P-183, P-184, P-185, P-186, P-187,
P-188, P-189, P-190, P-191, P-192, P-193
Environment and Toxicology: O-205, O-209,
O-223, O-224, O-225, O-226, O-228, P-092,
P-093, P-094, P-097, P-098, P-099, P-100,
P-101, P-102, P-103, P-104, P-105
Female Reproductive Endocrinology: O-004,
O-092, O-121, O-142, O-162, O-188, O-212,
O-256, O-274, P-003, P-004, P-005, P-006,
P-007, P-008, P-009, P-010, P-011, P-012,
P-013, P-014, P-015, P-016, P-017, P-018,
P-019, P-020, P-021, P-022, P-024, P-025,
P-026, P-031, P-032, P-033, P-034, P-035,
P-036, P-037, P-038, P-039, P-040, P-041
Female Reproductive Surgery: O-067, O-213,
P-194, P-195, P-197, P-198, P-199, P-200,
P-201, P-202, P-203, P-204, P-205, P-206,
P-209, P-210, P-212, P-213, P-214, P-215,
P-216, P-217, P-218, P-219
Female Reproductive Tract: O-097, O-099, O-181,
P-129, P-130, P-131
Fertility Preservation: O-022, O-104, O-215,
O-229, O-230, O-231, O-232, O-233,
O-234, P-455, P-456, P-457, P-458, P-459,
P-460, P-461, P-462, P-463, P-464, P-465,
P-468, P-469, P-470, P-471, P-472, P-473,
P-475, P-476, P-477
Fertilization: O-250, P-153, P-568, P-569, P-570,
P-571, P-572, P-573, P-574, P-575, P-576,
P-577
Genetic Counseling: O-036, O-165, O-166,
O-167, O-193, O-195, O-196, O-197,
O-198, O-204, O-218, P-367, P-368, P-369,
P-370
Imaging: O-055, O-064, O-069, O-070, O-074,
P-132, P-133, P-134, P-135, P-136
Implantation: O-066, O-122, O-156, P-673, P-674,
P-675, P-676, P-677, P-678, P-679, P-680,
P-681, P-682, P-683, P-684, P-685, P-686,
P-687, P-688, P-689, P-690, P-691, P-692
Leiomyoma: O-058, O-071, O-072, O-073, O-075,
O-076, O-182, O-184, O-185, O-186, O-211,
O-269, P-137, P-138, P-139, P-140, P-141,
P-142, P-143, P-144, P-145
Luteal Phase Support: P-693, P-694, P-695, P-696,
P-697, P-698, P-699
Male Factor: O-020, O-110, O-170, O-173, O-174,
O-200, O-271, P-380, P-381, P-382, P-383,
P-384, P-386, P-387, P-389, P-390, P-391,
P-392, P-393, P-394, P-397, P-400, P-401,
P-402, P-403, P-404, P-405, P-408, P-409,
P-410, P-411, P-412
Male Reproductive Endocrinology: P-528, P-529,
P-530, P-531
Male Reproductive Urology: O-019, O-021,
O-024, O-025, O-026, O-027, O-028,
O-029, O-030, O-109, O-111, O-112,
O-113, O-114, O-164, O-199, O-207,
O-208, O-210, P-532, P-534, P-535, P-536,
P-537, P-538, P-539, P-540, P-541, P-542,
P-544, P-545, P-546, P-547, P-548, P-549,
P-550, P-551, P-552, P-553
Menopause: O-008, O-009, O-011, P-001, P-002
Mental Health: O-105, O-106, O-107, O-108,
P-721, P-722, P-723, P-724, P-725, P-726,
P-727, P-728, P-729, P-730, P-731, P-732,
P-733, P-734, P-735, P-736, P-738, P-739,
P-740
Nursing: O-079, O-080, P-366
Obesity and Metabolism: O-015, O-171, O-201,
O-227, O-253, O-273, P-042, P-043, P-044,
P-045, P-046, P-047, P-048, P-049, P-050,
P-051
Oocyte Biology: O-043, O-047, O-133, O-135,
O-143, P-269, P-270, P-271, P-272, P-273,
P-274, P-275, P-276, P-277, P-278, P-279,
P-280, P-281, P-282, P-283, P-284, P-285
Oocyte Maturation: P-561, P-562, P-563, P-564,
P-565, P-566, P-567
Ovarian Function: O-006, O-014, O-136, O-272,
P-083, P-084, P-085, P-086, P-087, P-088,
P-089, P-090, P-091
Ovarian Reserve: O-077, O-131, O-141, O-151,
O-159, O-189, P-418, P-419, P-420, P-421,
P-422, P-423, P-424, P-425, P-426, P-427,
P-428, P-429, P-430, P-431, P-432, P-433,
P-434, P-435, P-436, P-437, P-438, P-439,
P-440, P-441, P-442, P-443, P-444, P-445,
P-446
e378 Topic Index Vol. 104, No. 3, Supplement, September 2015

Ovarian Stimulation: O-060, O-128, O-130,
O-132, O-160, O-254, O-255, P-623, P-624,
P-627, P-628, P-629, P-630, P-631, P-632,
P-634, P-635, P-636, P-639, P-640, P-641,
P-642, P-643, P-644, P-647, P-648, P-649,
P-650, P-653, P-654, P-655, P-656
Oxidative Stress: P-110, P-111, P-112, P-113,
P-114, P-115, P-116, P-117, P-118, P-119,
P-120, P-121, P-122, P-123
Polycystic Ovary Syndrome: O-010, O-013,
O-049, O-050, O-051, O-052, O-053,
O-054, O-258, O-265, O-275, O-276, P-052,
P-053, P-054, P-055, P-056, P-057, P-058,
P-059, P-060, P-061, P-062, P-063, P-064,
P-065, P-066, P-067, P-068, P-069, P-070,
P-071, P-072, P-073, P-074
Practice Management: O-057, O-259, P-716,
P-717, P-718, P-720
Preimplantation Genetic Diagnosis: O-031,
O-032, O-033, O-034, O-035, O-082,
O-123, O-148, O-163, O-168, O-246, P-489,
P-490, P-491, P-492, P-493, P-494, P-495,
P-496, P-497, P-499, P-500, P-501, P-503,
P-504, P-505, P-506, P-507, P-508, P-509,
P-510, P-511, P-512, P-513, P-514, P-515,
P-516, P-517, P-518, P-520, P-521, P-522,
P-523, P-524, P-525, P-526, P-527
Reproductive Hormones: O-001, O-012, O-161,
O-190, O-191, O-192, P-075, P-076, P-077,
P-078, P-079, P-080, P-081, P-082
Reproductive Immunology: O-137, O-157, P-125,
P-126, P-127, P-128
Sexuality: O-172
Sperm Biology: O-023, P-555, P-556, P-557,
P-558, P-559, P-560
Sperm Preparation: P-413, P-414, P-415, P-416
Stem Cells: O-094, O-235, O-236, O-237, O-238,
O-239, O-240, P-363, P-365
Testis: O-002, O-202, P-417
FERTILITY & STERILITY Ò
e379

AUTHOR AND SPOUSE/PARTNER DISCLOSURES INDEX
All speakers at the 2015 ASRM Annual Meeting and Postgraduate Courses were required to complete a disclosure form. These
disclosures were reviewed and potential conflicts of interest resolved by the Subcommittee on Standards of Commercial Support of
the Continuing Medical Education Committee. Each abstract or video author is listed below along with any relationships their
partners/spouses disclosed.
Adamson, G. D.
Advanced Reproductive Care, CEO,
Shareholder; LabCorp, Paid
consultant; Bayer, Paid consultant;
Ferring, Paid consultant; Auxogyn,
Grant recipient; Ziva, Direct
stockholder
Ahmed, H.
Celgene, Full-time company
employee
Albertini, D.
Springer Publishing, Paid consultant;
Italian Ministry of Science, Grant
recipient; State of Oklahoma,
Honoraria; TEDCO, Paid
consultant
Albertsen, H. M.
Juneau Biosciences, LLC, Company
officer; Juneau Biosciences, LLC,
Direct stockholder
Alikani, M. Reprogenetics, Direct stockholder;
Life Global, Paid consultant
Alper, M. M.
EMD Serono, Honoraria; Good Start
Genetics, Advisory Board; EMD
Serono, Honoraria; Reprosource,
Advisor-Board; Optum, Honoraria
Alvero, R.
Cooper Surgical, Paid consultant
Andriani, L.
I own non-controlling interests in
publicly traded stock of
companies in the medical field.
These securities were purchased
and are managed on a fully
discretionary basis by an SEC
registered independent investment
advisor or are owned through
pooled, investment vehicles like
mutual funds and/or exchange
traded funds and represent
substantially less than 1%
ownership interests. Investments
include: Celgene, Cerner, Costco,
Danaher, Gilead, Johnson &
Johnson, Merck, Mylan, Proctor &
Gamble, & Walgreens.
Apter, D.
Bayer, Grant recipient; Merck, Grant
recipient; Exeltis, Grant recipient;
Bayer, Merck and Exeltis,
Speakers bureau; GSK, Grant
recipient
Arbo, E.
Ferring, Full-time company employee
Archer, D. F.
AbbVie, Paid consultant; AbbVie,
Grant recipient; TherapeuticsMD,
Paid consultant; TherapeuticsMD,
Grant recipient; Bayer Healthcare,
Paid consultant; Bayer Healthcare,
Grant recipient; Agile
Pharmaceuticals, Paid consultant;
Exeltis/CHEMO France, Paid
consultant; Endoceutics, Paid
consultant; Endoceutics, Grant
recipient; TEVA/HR Pharma, Paid
consultant
Avril, C.
Merck Serono, Paid consultant;
Ferring, Paid consultant
Baker, V. L.
Good Start Genetics, Advisory Board;
Ovuline, Unpaid consultant
Barad, D. H.
U.S. Patents, DHB is a co-inventor on
a number of FMR1 gene-related
U.S. patents and still pending
patent applications, which claim
diagnostic benefits from
evaluations of the gene. One of
these patents was licensed to
Generation Medical Associates,
PLLC.; Generation Medical
Associates, PLLC, Direct
stockholder; U.S. Patents, DHB
holds patents that claim therapeutic
benefits from androgen
supplementation in women with
LFOR and hypoandrogenism and
receives licensing fees for the
patents from Fertility
Nutraceuticals, LLC.
Barnhart, K.
Bayer, Paid consultant; SPD, Paid
consultant
Barrett, C. B. ReproSource Fertility Diagnostics,
Paid consultant
Barriere, P.
Genevrier France, Honoraria; Merck
Serono France, Paid consultant;
MSD France, Paid consultant;
HRA Pharma, Paid consultant;
Ferring France, Honoraria
Bauer, J. D.
Ferring, Employee
Behr, B.
Auxogyn, Direct stockholder; Ivigen,
Direct stockholder
Bellerose, H.
Counsyl, Inc, part-time contracted
employee
Beltsos, A. N.
Merck Pharmaceuticals, Speakers
bureau; EMD Serono
e380 Author Disclosures Index Vol. 104, No. 3, Supplement, September 2015

Pharmaceuticals, Speakers bureau;
Ferring Pharmaceuticals, Speakers
bureau; Diclegis, Speakers bureau;
Good Start Genetics, Paid
consultant; Optum, Paid consultant
Benadiva, C. A.
OvaScience, Paid consultant
Bendikson, K.
Theralogix, Paid consultant
Berga, S. L. Ferring Pharmaceuticals, Attended
fundraiser at own expense;
Ferring Pharmaceuticals, Grant
recipient; Pfizer, Paid consultant;
UpToDate, Editor; Pfizer, Grant
recipient
Bergh, C. M.
MedSoftware, Company officer
Berkeley, A. S.
Merck, Direct stockholder; Pfizer,
Direct stockholder; Glaxo, Direct
stockholder; Becton, Dickenson,
Full-time company employee;
Bristol Myers, Direct stockholder
Bernstein, L. R.
Merck, Received free medication.;
Fertility Center of Maryland, Paid
consultant
Berro, R.
Celmatix, Inc, Full-time company
employee
Bhagavath, B.
Hologic, Paid consultant; Abbvie,
Paid consultant
Bisignano, A. Recombine, Company officer;
Recombine, Direct stockholder;
Recombine, Full-time company
employee
Bissonnette, F.
YAD, Direct stockholder
Blazek, J.
Genesis Genetics, Full-time company
employee
Boekelheide, K.
Boehringer Ingelheim, Grant
recipient; Tb Alliance, Paid
consultant; Drugs for Neglected
Diseases, Paid consultant;
CytoSolv, Direct stockholder
Boivin, J.
Merck & Co, Paid consultant;
Actavis, Paid consultant; EMD
Ltd, Honoraria
Borini, A.
Merck Serono GFI, Grant recipient;
Merck Serono Poland, Speakers
bureau; Unisense fertiliTech,
Participate advisory board
Brannigan, R. E.
Abbvie, Inc., A grant in support of
Northwestern University’s
Andrology fellowship was
provided to Northwestern
University, Feinberg School of
Medicine. I am the Director of the
Andrology Fellowship.; The
American Urological Association/
The Journal of Urology, I am an
Assistant Editor for The Journal of
Urology
Bristow, S. L.
Recombine, Full-time company
employee; Recombine, Stock
options
Butler, S. A.
MAP Diagnostics Ltd, Company
officer
Carlsson, M.
Pfizer Inc., Full-time company
employee; CSG INC., Full-time
company employee
Carrell, D. T.
Episona, Inc, Company officer
Castells, M.
Merck, Sanofi, Novartis, Paid
consultant
Cataldo, N. A.
CenseoHealth, Independent
contractor; GoodStart Genetics,
Paid consultant
Catherino, W. H.
Abbvie, Paid consultant; Viteava
Pharmaceuticals, Paid consultant;
Medical College of Wisconsin,
Honoraria; Recombine, Full-time
company employee; Bayer, Grant
recipient
Cedars, M. Ferring Pharmaceutical, Research
support - investigator - initiated
Celia, G.
Good Start Genetics, Inc., Providing
research samples to the company
Centola, G. M.
New England Cryogenic Center, Paid
consultant; Cryos International
USA, Paid consultant; Manhattan
Cryobank, Paid consultant;
NYMHB Fertility Services;
Dr. George Kofinas, Paid
consultant; Seattle Sperm Bank/
Phoenix Sperm Bank, Paid
consultant
Chan, C.
Level, Company officer
Chang, C.-C.
MyEggBank, Direct stockholder
Chen, A. A.
Progyny, Full-time company
employee
Chen, D.
Gilead Sciences, Direct stockholder;
Pfizer, Direct stockholder
Chen, S. H.
Ovascience, Paid consultant; Hologic,
Paid consultant; Optum, Paid
consultant; Recombine, Paid
consultant
Chettier, R.
Juneau Biosciences, Full-time
company employee; Affiliated
Genetics; Juneau Biosciences,
Direct stockholder
Cholkeri-Singh, A. Ethicon Endo Surgery, Speakers
bureau; Ethicon Endo Surgery,
Advisory Board Member; Bayer,
Speakers bureau; Bayer, Advisory
Board Member
Christman, G. M.
Abbvie Pharmaceuticals, Grant
recipient; Abbvie Pharmaceuticals,
Honoraria; Bayer Pharmaceuticals,
Grant recipient; Bayer
Pharmaceuticals, Paid consultant
Chwalisz, K.
AbbVie Inc., Full-time company
employee; AbbVie Inc., Direct
stockholder
Clarke, N. J.
Quest Diagnostics, Full-time
company employee
Coddington, C.
PG, Merck, Stock
Cohen, J.
Reprogenetics LLC, Direct
stockholder; Reprogenetics LLC,
Paid consultant; Life Global Inc.,
Paid consultant
Conaghan, J.
Irvine Scientific, Paid consultant;
Auxogyn, Paid consultant
Considine, R.
Eli Lilly Research Labs, Paid
consultant; Merck Research Labs,
Paid consultant
Copperman, A. B.
EMD Serono, Speakers bureau;
Merck, Speakers bureau; Ferring,
Speakers bureau
Corrado, J.
Good Start Genetics, Member of the
Nurse Advisory Panel
Costantini, L.
Prima-Temp, Inc, Company officer
FERTILITY & STERILITY Ò
e381

Craig, L.T. B. Ferring Pharmaceuticals, Principal
Investigator; Roche Diagnostics,
Principal Investigator
Czuprenski, E.
Genesis Genetics, Full-time company
employee
Daneshmand, S.
Actavis Inc., Grant recipient
Daubert, M.
Stealth Peptides, Paid consultant
Davie, J. Good Start Genetics, Inc., Paid
consultant
Davies, M. C.
Centre for Reproductive and Genetic
Health, 256 Grays Inn Road,
London WC1X 8LD UK, Paid
consultant
Demirci, U.
DxNow, Co-founder, Scientific
Advisor; Koek Biotech,
Co-founder, Scientific Advisor
Demko, Z.
Natera, Full-time company
employee; Natera, Direct
stockholder
Desai, N.
Fertilitech, Advisory Board
D’Hooghe, T.
WERF (World Endometriosis
Research Foundation), Board
Membership till July 2011; Bayer
Pharma, Proteomika,
MPharmaplex, Astellas, roche
Diagnostics, Paid consultant;
Ferring, Merk Serono, Merck,
Besins, Pharmaplex, Grant
recipient; Ferring, Merk Serono,
Merk, Travel/accomodations/
meeting expenses unrelated to
activities listed.Money to our
institution, University Hospitals
Leuven, Belgium
Diamond, M. P. NICHD, AbbVie, EMD Serono,
Baxter, Grant recipient; Teijin
Pharmaceuticals, Auxogyn, Paid
consultant; Advanced
Reproductive Care, Board of
Directors and Stockholder
Diaz-Gimeno, P.
Igenomix, ERA Patent authorship
Diez Juan, A.
Igenomix, Full-time company
employee
Domar, A. D.
Merck, Grant recipient;
Johnson&Johnson, Paid
consultant; Merck, Speakers
bureau; Ovascience, Paid
consultant; Ferring, Paid
consultant; Merck, Paid
consultant; UptoDate, Paid
consultant; Nora Therapeutics,
Paid consultant
Doody, K.
Merck Pharmaceutical, Paid
consultant; Ferring
Pharmaceutical, Paid consultant;
Finox Pharmaceutical, Paid
consultant; Serono
Pharmaceutical, Speakers bureau;
Good Start Genetics, Paid
consultant
Drevet, J. R.
Celloxess, Board member
Driggers, P.
Eisai, Inc., Speakers bureau; novonordisk,
Speakers bureau; Eisai,
Inc., Paid consultant; Eisai, Inc.,
Clinical Investigator; novonordisk,
Clinical Investigator
Dudley, P. S.
MyEggBank, N.A., Direct
stockholder; Attain Genetics,
LLC, Direct stockholder
Dundee, J. A.
New England Fertility Society/
Ferring Pharmaceuticals, Grant
recipient
Dunn, R. D.
Finox, PI for one of many sites for
Afolia ART Study
Dwan, P. G.
EMD SERONO, Speakers bureau;
EMD SERONO, Paid consultant
Dye, T. D.
Pfizer, Grant recipient; Humana, Inc.,
Paid consultant
Dzidic, N. Combimatrix, Full-time company
employee
Eid, S.
Teva Pharmaceuticals, Paid
consultant
Eisenberg, M. Sandstone Diagnostics, Direct
stockholder; Reprovantage, Direct
stockholder; Glow, Advisor;
EmbraceHer, Direct stockholder
Evans, E.
Counsyl, Company officer
Fanchin, R. Merck-Serono, Grant recipient;
Ferring, Grant recipient
Farrington, P. Affiliated Genetics, Direct
stockholder; Juneau Biosciences,
Company officer; Juneau
Biosciences; Affiliated Genetics,
Direct stockholder; Juneau
Biosciences, Full-time company
employee
Faulkner, N.
Good Start Genetics, Full-time
company employee
Faustmann, T. A.
Bayer Pharma AG, Full-time
company employee
Feinberg, E.
Abbvie, Data Safety Monitoring
Board; Natera, Advisory Board
Feinberg, R. F.
AbbVie, Clinical Trial; Nora, Clinical
Trial; Ferring, Clinical Trial;
Finox, Clinical Trial
Fode, M.
Eli Lilly, Paid consultant; Astellas,
Paid consultant; Menarini, Paid
consultant; Coloplast, Honoraria
Foong, S.
EMD Serono, Honoraria
Forman, E. J.
Ferring Pharmaceuticals, Speakers
bureau
Gao, J.
AbbVie, Full-time company
employee
Gardner, D. K.
Vitrolife, Grant recipient
Gargiulo, A. R.
OmniGuide, Inc., Paid consultant;
Kawasaki Robotics (USA), Inc.,
Paid consultant
Gates, D.
Merck and Co., Full-time company
employee; Covance, Full-time
company employee
Gayet, V.
Ferring, Honoraria
Gemzell-Danielsson, K.
Bayer AG; MSD/Merck; HRA
Pharma; ExelGyn; Gedeon
Richter, Honoraria; Bayer, Grant
recipient
Gharagozloo, P.
CellOxess LLC, Company officer
Gheyas, F.
Merck, Company officer
Ginsburg, E.
UptoDate, Honoraria; Springer Inc,
Honoraria; Serono: investigator
initiated study, Grant recipient
Giudice, L. ASRM, Company officer; Merck;
Pfizer, Direct stockholder
Givens, C.
Merck, Paid consultant
Gleicher, N.
Fertility Nutraceuticals, LLC, Direct
stockholder; Generation Medical
Associates, PLLC, Direct
stockholder; Fertility
Nutraceuticals, LLC, Receive
e382 Author Disclosures Index Vol. 104, No. 3, Supplement, September 2015

patent licensing fees; Generation
Medical Associates, PLLC, Patent
licensed; U.S. Patents, NG holds
patents that claim therapeutic
benefits from androgen
supplementation in women with
LFOR and hypoandrogenism.;
U.S. Patents, NG is a co-inventor
on a number of FMR1 gene-related
U.S. patents and still pending
patent applications, which claim
diagnostic benefits.
Globus, S. T.
Celmatix Inc., Full-time company
employee
Go, K. J.
Ferring Pharmaceuticals, Full-time
company employee
Godfrey, E. M.
Prima-Temp, Grant recipient; Bayer
Pharmaceuticals, Grant recipient;
Teva Women’s Health, Grant
recipient; Merck, Sharp & Dohme
Corp, Speakers bureau
Goering, M. C.
MedTech for Laboratory Solutions,
Paid consultant
Gold, M.
Recombine, Full-time company
employee; Recombine, I have
stock options.
Goldberg, J. D.
Counsyl, Inc, Full-time company
employee
Goldfarb, J. M.
Lumara Health – no longer affiliated
with teh company since 11/14,
Direct stockholder
Goldstein, M.
Therologix, advisory board
Gordon, K.
Merck & Co, Full-time company
employee
Gordon, T. T.
Genesis Genetics, Direct stockholder;
Rubicon Genomics, Paid
consultant
Grainger, D. A.
Abbvie, Speakers bureau; Shionogi,
Inc., Speakers bureau
Grantz, K. L.
Mylan, Inc., Direct stockholder
Grifo, J.
Illumina, Speakers bureau;
Ovascience, Medical Advisory
Board
Grifo, J. A.
Illumina, Speakers bureau;
Ovascience, Medical Advisor
Grober, E. D. Abbott, Paid consultant; Paladin,
Grant recipient; Eli Lilly, Paid
consultant
Gross, J.
Novartis, Full-time company
employee
Guan, Y.
Merck&Co., Inc., Full-time company
employee
Gutmann, J.
somalogic, Paid consultant
Hadjiliadis, D.
Gilead, Advisory board; Bayer,
Advisory board; Vertex, Advisory
board
Hakim, L. S.
ENDO/AMS, Speakers bureau
Hamamah, S.
Ferring, Grant recipient
Hammond, K.
Good Start Genetics, Paid consultant;
CenseoHealth, Independent
contractor
Hannam, T.
EMD Serono, Our company sells
pharmaceutical products to
patients; Organon, Our company
sells pharmaceutical products to
patients; Ferring, Our company
sells pharmaceutical products to
patients
Hansen, K. R.
Roche Diagnostics, Grant recipient;
Ferring International
Pharmascience Center US, Grant
recipient
Haque, I. S.
Counsyl, Full-time company
employee
Harkins, G.
Ethicon EndoSurgery, Paid
consultant; Intuitive Surgical, Paid
consultant
Harutunian, A.
Genesis Genetics, Direct stockholder
Hasson, J.
‘‘Temp-drop LTD’’; This is a start-up
company which manufactures
a self monitoring device which
continuously measures body
temperature in sleep., Medical
adviser. The position is unpaid.
Hayward, B.
EMD Serono, Inc., Full-time
company employee
Heiser, P. W.
Ferring Pharmaceuticals, Inc., Fulltime
company employee
Heller, B.
Pulling Down the Moon Inc.,
Company officer
Hennebold, J. D.
AbbVie Pharmaceuticals, Direct
stockholder; Abbott Laboratories,
Direct stockholder; Omnicare,
Direct stockholder; Gedeon
Richter PregLem SA, Grant
recipient
Hesley, G.
Insightec, Grant recipient
Hirshfeld-Cytron, J. E.
Duchesnay, Speakers bureau
Hornstein, M. D. WINFertility, Meidcal Advisory
Board; Up-To-Date, Author
Hotaling, J. M.
Andro360, own equity in early stage
start up company that has not
made a profit; SpermDx, own
equity in early stage start up
company that has not made a profit;
StreamDx INC, own equity in early
stage start up company that has not
made a profit
Howard, B. Teva Pharmaceuticals, Full-time
company employee
Howles, C. M.
Finox Biotech AG., Paid consultant
Hubbard, J. W.
Affymetrix Inc, Full-time company
employee
Hunter Cohn, K.
Celmatix Inc, Full-time company
employee
Hu-Seliger, T.
Celmatix, Full-time company
employee
Iles, R. K.
MAPDiagnostics Ltd, Company
officer
Israel, M. A.
Progyny, Inc., Full-time company
employee
Jacobs, I.
ABCODIA, Company officer
Jacoby, V.
Halt Medical, Grant recipient
Jadhav, P.
Merck, Full-time company employee
Jain, R.
AbbVie, Full-time company
employee
Jasulaitis, S.
Merck Pharmaceuticals, Speakers
bureau
Jenkins, J.
Finox Biotech A.G., Company officer
Kadoch, I.-J.
Clinique Ovo, Clinical director; Yad-
Tech, Shares holder
Kalra, B.
Ansh Labs, Full-time company
employee
Kamga-Ngande, C.
Clinique Ovo, Clinical director
Karvir, H.
Celmatix Inc., Full-time company
employee
FERTILITY & STERILITY Ò
e383

Katilius, J.
Prima-Temp,Inc, Paid consultant
Kellogg, G. R.
Recombine, Full-time company
employee; Recombine, Stock
Options
Ketterson, K.
Recombine, Shareholder
Kimura, T.
Takeda Pharmaceutical Co.Ltd.,
Grant recipient; Nohon Kayaku
Co.Ltd., Paid consultant; Nihon
Shinyaku Co.Ltd, Grant recipient;
Fuji Pharmaceutical Co. Ltd.,
Grant recipient; MSD Co.Ltd.,
Speakers bureau; Chyugai
Pharmaceutics Co. Ltd., Grant
recipient
Kolb, B. A. My Fertility, Company officer;
Ferring Pharmaceuticals, Speakers
bureau; EMD Serono, Paid
consultant; Good Start Genetics,
Paid consultant
Kovalevsky, G.
AbbVie, Clinical research trial
investigator; Nora, Clinical
research trial investigator; Ferring,
Clinical research trial investigator;
Finox, Clinical research trial
investigator
Krawetz, S.
Taylor and Francis, EIC of Systems
Biology in Reproductive Medicine
Krisher, R. L.
Merck Serono, Grant recipient
Kumar, A.
AnshLabs, Full-time company
employee
Kumar, D.
Hologic Inc., Although I am an
employee of the University of
Rochester, part of my salary is paid
by Hologic Inc. as I am
coordinating a study sponsored by
Hologic Inc. This study is
regarding the effectiveness of the
Novasure Global Endometrial
Ablation procedure in t
Kumar, N.
Recombine, Full-time company
employee
Kushnir, V. A.
Generation Medical Associates,
PLLC, Direct stockholder
Kyrou, D.
MSD, Honoraria
Lamb, D. J.
Cellmatrix, Paid consultant
Lannon, B.
Dyax Inc., Direct stockholder
Large, M.
Genesis Genetics, Full-time company
employee
Lazarin, G. A.
Counsyl, Full-time company
employee
Leboeuf, M.
Actavis, Speakers bureau; Bayer,
Speakers bureau; Abbvie, Speakers
bureau
Legro, R. S.
Ferring, Grant recipient; Astra
Zeneca, Grant recipient;
Euroscreen, Paid consultant;
Kindex, Paid consultant; Takeda,
Paid consultant; Clarus
Therapeutics, Paid consultant
Leone Roberti Maggiore, U. DEKA, Speakers bureau
Lessey, B. A.
Pfizer, Paid consultant; Sepal, Inc.,
Licensed technology and potential
royalty payments
Levy, M.
Donor Egg Bank, USA: President,
Company officer
Li, M.
PacGenomics, Company officer
Liebermann, J.
Sage/Origio, Paid consultant; Vivere,
Paid consultant; Irvine Scientific,
Speakers bureau
Lindeman, M. R.
Genesis Genetics, Full-time company
employee
Lindheim, S. R.
Hologic, Speakers bureau; Abbvie,
Speakers bureau; Progenity,
Speakers bureau; Bayer, Grant
recipient; Cooper Surgical,
Non-paid Advisor
Liu, K.
Ferring Canada, Grant recipient
qukaszuk, K.
Ferring Pharmaceuticals, Honraria;
Roche Diagnostics, Honoraria;
Merck, Honoraria
Luke, B.
Society for Assisted Reproductive
Technology, Paid consultant
Lukes, A. S.
Abbvie, Grant recipient; Abbvie, Paid
consultant; Glaxo-Smith Kline,
Paid consultant; Mirabilis Medica,
Paid consultant; Hologic, Paid
consultant; Bayer, Paid consultant;
Agile, Grant recipient; Sequoia,
Grant recipient; Therapeutics,
Grant recipient; Bayer, Grant
recipient; Watson, Grant recipient;
Hologic, Grant recipient; Merck,
Grant recipient; Amgen, Grant
recipient
Lukkari-Lax, E.
Bayer Oy, Full-time company
employee
Mahony, M. C.
EMD Serono, Inc., a subsidiary of
Merck KGaA, Darmstadt,
Germany, Full-time company
employee
Malik, M.
AB Sciex, Full-time company
employee
Manoharan, A. P.
Recombine, Full-time company
employee; Recombine, Stock
options
Martinez, C.
Upward Labs Holdings, Inc.
(www.glowing.com), Company
officer
Massin, N.
MSD, Honoraria; Merck, Honoraria
Mather, K.
Novo Nordisk, Grant recipient; sanofi
aventis, Grant recipient; Merck
Inc, Grant recipient; Abbott, Grant
recipient; Boehringer Ingelheim,
Paid consultant
Matsuura, K.
ZEON Corporation, Speakers bureau
McCoy, R.
Stanford University, Co-inventor on
a provisional patent filed by
Stanford University, related to
work in collaboration with Natera.
McCulloh, D. H. ReproART: Georgian American
Center for Reproductive
Medicine, Tbilisi, Georgia,
Company officer; Biogenetics
Corporation, Mountainside, New
Jersey, USA, Company officer;
NYU Fertility Center, New York,
New York, USA, Full-time
company employee
McCullough, A. Repros, Paid consultant; Repros,
Direct stockholder; Pfizer, Direct
stockholder; Pfizer, DSMB;
Antares, Grant recipient; Repros,
Grant recipient; ISSM, Honoraria;
AUA, Honoraria; Capital Region
Medical Research Fund, Grant
recipient
McWilliams, K.
Genesis Genetics, Full-time company
employee
e384 Author Disclosures Index Vol. 104, No. 3, Supplement, September 2015

McWilliams, T. K.
Genesis Genetics, Full-time company
employee
Meintjes, M.
Vitrolife AB, Paid consultant
Merrion, K. Natera, Inc., Full-time company
employee; Natera, Inc., Option to
hold stock in Natera, Inc.
Miller, C. E. Covidien, Femasys, Olympus,
Novartis, Abbvie, Intuitive
Surgical, Gynesonics, Grant
recipient; Ethicon, Covidien,
Femasys, Abbvie, Halt Medical,
Intuitive Surgical, Gynesonics,
Paid consultant; Ethicon, Smith &
Nephew, Intuitive Surgical,
Speakers bureau
Minjarez, D. A.
Ferring, Speakers bureau
Moazamian, A.
CellOxess LLC, Full-time company
employee
Moazamian, R. J.
CellOxess, Intern
Moley, K. OvaScience, Scientific Advisory
Board Member
Montegriffo, E.
Bayer HealthCare Pharmaceuticals,
Full-time company employee
Morbeck, D.
Vitrolife, Research equipment loan
Mullen, T.
Good Start Genetics, Full-time
company employee
Munne, S. Reprogenetics, Direct stockholder;
Recombine, Direct stockholder
Muzzey, D.
Counsyl Inc., Full-time company
employee
Nagy, Z. P.
My Egg Bank, Direct stockholder;
Cooper Surgical/Origio, Paid
consultant; Fertilitech, Paid
consultant; MERCK MSD,
Speakers bureau
Natan, Y.
FertileSafe Ltd., Full-time company
employee
Neitzel, D.
Good Start Genetics, Full-time
company employee
Nelson, A.
Agile; Bayer, Grant recipient;
Actavis; Bayer; Merck; Pfizer ;
Teva, Honoraria; Actavis; Agile;
Bayer; ContraMed; Merck; Teva;
MicroCHIPS Biotech; Pharmanest,
Paid consultant
Niederberger, C.
American Urological Association,
Journal section editor; update
series editor; Ferring, Grant
recipient; IBSA, Grant recipient;
NexHand, Company officer
North, J.
AbbVie Inc., Full-time company
employee; Abbott Laboratories,
Prior employee
Norwitz, E. R.
Natera, Unpaid member of Advisory
Board; Hologic, Unpaid member
of Advisory Board; Bayer
Pharmaceuticals, Named inventor
on a patent for the diagnosis of
preeclampsia purchased by Bayer
Ohl, D. A. Pfizer, Scientific Advisor; Endo,
Grant recipient; Coloplast, AMS,
Surgical Consultant and Advisory
Board
Page, D. C.
Counsyl, Scientific Advisory Board
Paik, K. G.
Natera, Inc., Full-time company
employee
Palermo, G. D.
Irvine Scientific, Royalties
Park, S. O.
CHA Gangnam Medical center, CHA
University, Full-time company
employee
Patrizio, P.
Counsyl, Honoraria; FertileSafe,
scientific advisor
Paulson, R.
Ferring Pharmaceuticals, Honoraria;
Origio, Paid consultant
Penrose, L.
Reproductive Solutions Inc.,
Company officer
Penzias, A.
OvaScience, Company Advisor;
ReproSource, Company Advisor;
Nora Therapeutics, Company
Advisor; Ferring Pharmaceuticals,
Honoraria
Peralta, S. Merck-Serono, Attendance at the
meeting and travel expenses
Petrozza, J. C.
Interlace Medical / Hologic,
Scientific Advisory Committee
Pettersen, B.
Natera, Inc., Full-time company
employee
Pohlmeier, A. M.
Teva Pharmaceuticals, Full-time
company employee
Polhemus, A. M.
CellOxess, LLC, Full-time company
employee
Pomeroy, K. O.
The World Egg Bank, Full-time
company employee
Pool, T. B.
Auxogyn, Honoraria
Porcu Buisson, G.
Ferring SA, Honoraria; Merck Serono
SA, Honoraria
Prien, S.
Reproductive Solutions Inc,
Company officer
Rabinowitz, M.
Natera, Inc., Company officer
Racowsky, C.
Life Global Group, Paid consultant;
UpToDate, Honoraria; World
Health Organization, Paid
consultant
Rafizadeh, M. J. Colgate-Palmolive Company,
Summer Internship Offer
Raia, M. H.
Counsyl (Genetic Counselor),
Full-time company employee
Reed, B. G.
Sequenom, Direct stockholder
Riboldi, M.
Igenomix Brasil, Full-time company
employee
Riche, D. M.
Merck, Novo Nordisk, Speakers
bureau
Rippon, G. A.
Pfizer Inc., Full-time company
employee
Robinson, M. K.
Thrive Rx, Paid consultant
Robinson, R. D.
AbbVie, Grant recipient
Rodriguez, S.
Recombine, Full-time company
employee
Rosen, K. A. Bayer Healthcare Pharmaceuticals
Inc., Full-time company employee
Ruman, J. Ferring Pharmaceuticals, Full-time
company employee
Ryan, A.
natera, Full-time company employee
Sadeghi-Nejad, H.
Auxilium (now Endo), Grant
recipient
Sakkas, D. Ferring, Grant recipient; Origio,
Scientific Advisory Board;
Fertilitech, Scientific Advisory
Board; Good Start Genetics, Paid
consultant; INVO Biosciences,
Direct stockholder
Saldivar, S.
Myriad Genetics, Speakers bureau;
Intuitive Surgical Inc, Paid
consultant
FERTILITY & STERILITY Ò
e385

Sammel, M. D.
Swiss Precision Diagnostics, Paid
consultant
Santoro, N.
Bayer Inc, Grant recipient;
Menogenix Inc, Stock Options
Sasaki, K.
E-medicine, Honoraria
Schattman, G. L.
Femasys, medical advisor, clinical
investigator; Theralogix, Paid
consultant; Ferring, Speakers
bureau
Schmelter, T.
Bayer Pharma AG, Full-time
company employee
Schoolcraft, W. B. OvaScience, Advisory Board
Member; Ferring, Paid consultant;
Serono, Paid consultant
Schweitzer, A. C.
Affymetrix, Full-time company
employee
Scott, R. T. Foundation for Assessment &
Enhancement of Embryonic
Competence, Inc.; Neither myself
or my program get any personal
benefit., Company officer; Ferring
Pharmaceutical, Scientific
Advisory Board
Seifer, D.
Rutgers Medical School/ MGH
licensing aggreement with
Beckman-Coulter, Co-inventor of
AMH as a method of determining
ovarian reserve; Beckman-Coulter,
Paid consultant; Women’s
Integrated Network, Paid
consultant; none, none
Shah, D.
Grand Rounds Health, Paid
consultant
Shapiro, B. S.
Actavis Inc., Grant recipient
Shapiro, D. Actavis, Speakers bureau; Merck,
Honoraria; EMD-Serono, Paid
consultant; Ovascience, Paid
consultant
Sharara, F.
MAP Diagnostics, Company officer;
Ferring Pharmaceuticals, Speakers
bureau
Shawber, C. Eisai Pharmaceuticals, Columbia
University has established
a research agreement (CU12-3625
Eisai Research Collaborative
Agreement) with Dr. Kitajewski as
the PI to develop of Notch1 decoys.
This research agreement provided
funds to me as an inventor and
my lab
Shin, D.
Endo Pharmaceutical, Speakers
bureau
Shraga, R.
Recombine, Full-time company
employee; Recombine, Stock
Options
Shwayder, J. M.
Cook Ob-Gyn, Royalties for
SonoBiopsy catheter
Sigurjonsson, S.
Natera, Full-time company employee
Silverberg, K.
Abbvie, Honoraria; Good Start
Genetics, Paid consultant; Actavis,
Paid consultant; Serono, Paid
consultant; Illumina, Paid
consultant; Myriad Genetics, Paid
consultant; Finox, Grant recipient;
Auxogyn, Grant recipient
Simon, A.
Natera, Inc., Full-time company
employee
Simon, C. Equipo IVI, Direct stockholder;
Igenomix SL, Direct stockholder;
OVASCIENCE, Paid consultant;
TEVA; EXCEMED; MSD;
Ferring, Honoraria
Slayden, O. D.
Bayer Pharma AG, Berlin, Grant
recipient
Soh, Y. M.
Vitrolife, Grant recipient
Sood, A.
Global Center for Resiliency and
Wellbeing, Owner
Stanczyk, F. Z. Merck & Co., Paid consultant;
TherapeuticsMD, Paid consultant;
Noven Pharmaceuticals, Paid
consultant; Enteris Biopharma,
Paid consultant; AbbVie, and
Agile Therapeutics, Paid
consultant
Stecher, V.
Pfizer Inc, Full-time company
employee
Stegmann, B. J.
Merck & Co., Full-time company
employee
Steinkampf, M. P.
AbbVie Pharmaceuticals, Speakers
bureau
Stewart, E. A.
AbbVie, Astellas,Bayer,
GlaxoSmithKline, Gynesonics,
Welltwigs, Viteava, Paid
consultant; UpToDate, Honoraria
St-Michel, P.
MD Serono, Paid consultant; Ferring
Canada, Grant recipient
Strachan, G.
Teva Pharmaceuticals, Full-time
company employee
Stratton, P.
Alllergan has provided a grant to my
institution (NIH) for a study on
which I am the medically
responsible investigator. I have no
direct of personal profit from this
grant., Grant recipient
Strumbly, D.
Janssen Pharmaceuticals, Contract
Employee
Sung, L.
Ferring, Grant recipient
Surrey, E.
AbbVie Laboratories, Advisory
board, speaker’s bureau, grant
recipient
Swain, J. E.
Irvine Scientific, royalties from
a previously designed culture
medium
Swerdlow, A. J.
GlaxoSmithKline, Direct stockholder
Tan, L.
Progyny (formerly Auxogyn), Fulltime
company employee; Progyny
(formerly Auxogyn), Direct
stockholder
Tang, B. Teva Pharmaceutical, Full-time
company employee
Taylor, R. N. AbbVie, Paid consultant; Ferring,
Paid consultant
Taylor, T. H. CombiMatrix, Full-time company
employee
Thomas, M. A.
Medicines 360, Grant recipient;
Agile, Grant recipient; Merck,
Grant recipient; Bayer, Grant
recipient
Tjoa, M. L.
Teva Pharmaceuticals, Full-time
company employee
Toth, T. L.
Good Start Genetics, medical
advisory board
Treff, N. R.
EMD Serono, Grant recipient
Truong, T. T.
Vitrolife AB, Grant recipient
e386 Author Disclosures Index Vol. 104, No. 3, Supplement, September 2015

Tulandi, T.
Actavis Inc, Honoraria; AbbVie
Canada, Honoraria
Valbuena, D.
Igenomix S.L., Full-time company
employee
Vaskivuo, T.
Roche Diagnostics, Speakers bureau;
Siemens Diagnostics, Speakers
bureau
VerMilyea, M.
Irvine Scientific, Paid consultant;
Progyny (formerly Auxogyn), Paid
consultant
Wang, R. AMS, Paid consultant; Coloplast,
Paid consultant; Men MD, Paid
consultant; Eli Lilly, Speakers
bureau
Ward, K. Affiliated Genetics, Inc.; Juneau
Biosciences, Direct stockholder;
Affiliated Genetics, Inc., Full-time
company employee; Affiliated
Genetics, Inc., Direct stockholder;
Juneau Biosciences, Company
officer; Juneau Biosciences, Fulltime
company employee
Warne, D. W. Finox, Paid consultant; PregLem,
Paid consultant
Webster, W.
Prima-Temp, Inc, Company officer
Welch, C. Reprogenetics, Full-time company
employee
Wellons, M.
Pfizer, advisory board
Wells, D. Reprogenetics, Direct stockholder;
Illumina, Paid consultant; Ferring,
Honoraria; Recombine, Direct
stockholder; Merck Serono,
Honoraria
Widra, E. A.
Counsyl, Paid consultant
Willey, R. G.
Ferring International Pharmascience
Center US, Inc., Full-time
company employee
Williams, L. A.
AbbVie,Inc, Full-time company
employee
Winston, N.
Pfizer, Direct stockholder; Merck,
Direct stockholder
Wolff, E. F.
OvaScience, Grant recipient
Woodrum, D. A.
BioSentry, Paid consultant
Wright, K.
Alcon, Full-time company employee
Wu, H.
Celmatix Inc., Full-time company
employee
Wyatt, M.
Genesis Genetics, Full-time company
employee
Yang, Z.
ZytoGen, Company officer
Yankov, V. Ferring Pharmaceuticals, Full-time
company employee
Yarnall, S. Recombine, Full-time company
employee; Recombine, Stock
options
Yurttas Beim, P.
Celmatix Inc, Company officer
Zajic, S.
Merck & Co., Inc., Full-time
company employee; Merck & Co.,
Inc., Direct stockholder
Zgodic, A.
Celmatix, Full-time company
employee
Zhang, H.
BMS, Full-time company employee
Zimmerman, R.
Foundation for Embryonic
Competence, Full-time company
employee; BioReference
Laboratories, Full-time company
employee
FERTILITY & STERILITY Ò
e387