2009 Vienna - European Society of Human Genetics

Cytogenetics
quality as compared to conventional semen analysis. The aim of this
study was to examine the role of apoptosis and reactive oxygen species
(ROS) in inducing DNA damage in ejaculated spermatozoa. METH-
ODS: We examined spermatozoa for apotosis by flow cytometry, DNA
damage by comet assay and raw semen for Reactive Oxygen Species
(ROS) by chemiluminescence in 43 idiopathic infertile patients having
normal semen parameters, 67 infertile men with abnormal semen
parameters and in 29 fertile controls. RESULTS: Apoptosis in infertile
men with normal semen parameters was significantly higher (p=0.009)
as compared to controls but non significantly different from infertile
men with abnormal sperm parameters (p=0.317). ROS values were
significantly correlated with apoptosis levels in patients with normal
and abnormal semen parameters. (p=0.023; p=0.0121 respectively)
but not in the fertile controls (p=0.067). Non significant difference in
Olive Tail Moment (OTM) was obtained in patients with normal and
abnormal semen parameters. However significant difference for OTM
was observed in the fertile controls and patients with normal and abnormal
semen parameters (p=0.014; p=0.029). CONCLUSION: DNA
damage analysis provides additional dimension to semen analysis and
has the advantage of rapidly performed and interpreted. Thus DNA
damage evaluation should be included in the diagnostic workup of infertile
men.
P03.198
Analysis of 5t allele of the CFTR gene intron 8 in men with
azoospermia and oligozoospermia from Ukraine
O. A. Fesai, S. A. Kravchenko;
Institute of Molecular Biology and Genetics,, Kiev, Ukraine.
Men with azoospermia or oligozoospermia due to germ cell failure
were suggested to have an increased risk of carrying CFTR gene mutations.
5T is one of the alleles found at the polymorphic Tn locus in
intron 8 of the CFTR gene. A stretch of 5, 7 or 9 thymidine residues
is found at this locus. Less efficient splicing will occur when a lower
number of thymidines are found, resulting in CFTR transcripts that lack
exon 9 sequences.
In the present study we evaluated the incidence of the 5T allele in men
with azoospermia and oligozoospermia from Ukraine.
Isolated DNA from blood samples of 153 infertile men (60 - azoospermia
and 93 - oligozoospermia) and of 102 fertile men (control group)
has been amplified by polymerase chain reaction targeting the 5T allele
of the CFTR gene intron 8. For the 5T allele discrimination fragment
analysis of Cy5-labeled PCR products on an automated DNA
analyzer “A.L.F.-express” were used.
The percentage of patients with infertility who had 5T allele was significantly
higher (p0.05).Obtained
data suggests that 5T allele could be involved in the process of spermatogenesis
or sperm maturation in azoospermic and oligozoospermic
males.
P03.199
the frequency of b2/b4, b2/b3 and gr/gr deletions in Russian
infertile men
T. M. Sorokina, V. B. Chernykh, L. V. Shileiko, L. F. Kurilo, O. P. Ryzhkova, A.
V. Polyakov;
Research Centre for Medical Genetics, Russian Academy of Medical Sciences,
Moscow, Russian Federation.
Introduction: Y chromosome microdeletions are commonest genetic
cause of male infertility. In contrast to complete AZF deletions partial
AZFc region deletions is still under evaluated. The aim of our study
was to analyze of frequency of partial AZFc deletions in Russian infertile
men.
Materials and Methods: We investigated a cohort of 1510 Russian
men from infertile couples. Complete AZF deletions were detected according
to Laboratory guidelines for molecular diagnosis of Y-chromosomal
microdeletions (EAA/EMQN, 1999), with some modifications.
Partial deletions in AZFc region were tested by multiplex PCR of following
STS loci: sY142, sY1197, sY1192, sY1291, sY1206, sY1054
and sY1125.
Results: In total AZF deletions were detected in 268 (17.8%) examined
individuals. Complete AZFc (b2/b4) deletions were found in 54 (3.6%)
infertile men. Other complete (classic) AZF deletions (AZFa, AZFb and
AZFb+c) were revealed in 19 (1.3%) patients. Partial AZFc deletions
were detected in 195 (12.9%) in examined individuals. Most common
types of partial AZFc deletions were b2/b3 and gr/gr deletions. Their
frequencies were 8.5% and 3.6%, respectively. Deletion b1/b3 was revealed
in one patient. Other types of deletions partially covering AZFc
region were detected in 11 (0.7%) examined men. Various degrees of
spermatogenesis defects (from asthenozoospermia to azoospermia)
have been found in patients with partial AZFc deletions.
Conclusion: Obtained data demonstrated high prevalence of partial
AZFc deletions in Russian infertile men. Partial deletion b2/b3 is commonest
the Y chromosome microdeletion among Russian infertile
men.
P03.200
A case of 46,XX male syndrome
S. Gunes 1 , G. Okten 1 , M. Mercimek 2 , A. Tukun 3 , T. Ozcelik 4 , R. Asci 2 , H. Bagci 5 ;
1 Ondokuz Mayis University, Medical Biology Department, Medical Genetics
Section, Samsun, Turkey, 2 Ondokuz Mayis University, Department of Urology,
Samsun, Turkey, 3 Ankara University, Medical Genetics Department, Ankara,
Turkey, 4 Bilkent University, Molecular Biology and Genetics Department, Ankara,
Turkey, 5 Ondokuz Mayis University, Medical Biology Department, Samsun,
Turkey.
46,XX male syndrome is a rare sex chromosome disorder occurring
about 1 in 25 000 males. XX male syndrome mostly results from
unequal crossing over between X and Y chromosome during male
meiosis. Point mutations, deletions or translocations of sex determining
region Y gene (SRY) the most common causes of sex reversal.
Approximately 80% of 46,XX males have translocation of SRY region
onto an X chromosome. SRY positive chromosomal aberrations arise
due to unequal recombination between Xp and Yp terminal regions
during paternal meiosis. In this report, we present the clinical, cytogenetical,
molecular cytogenetical, molecular data and X chromosome
inactivation pattern of a 16-year-old patient referred to urological clinic
for evaluation of small testes and small penis. Chromosomal analysis
revealed 46,XX karyotype. SRY amplification was positive and is confirmed
by fluorescence in situ hybridization (FISH). FISH test showed
the presence of SRY region translocated to the short arm of the X chromosome.
Analysis of these cases illustrated that conventional cytogenetic,
FISH and SRY amplification techniques are useful for accurate
diagnosis and genetic counseling.
P03.201
Ambiguous Genitalia: Diagnosis and management with special
reference to parental consanguinity and age at diagnosis
S. M. Tayel 1 , H. S. Kassem 2 , I. Marzouk 3 , N. A. Abukarsh 4 , H. N. Sallam 5 ;
1 Faculty of Medicine & Suzanne Mubarak Regional Centre for Women’s Health
& Development, Alexandria, Egypt, 2 Clinical Genomic Centre, Faculty of
Medicine, Alexandria, Egypt, 3 Genetics Unit, Pediatrics Department, Faculty of
Medicine, Alexandria, Egypt, 4 Histology Department, Al-Fateh Faculty of Medicine,
Tripoli, Libyan Arab Jamahiriya, 5 Faculty of Medicine & Suzanne Mubarak
Regional Centre for Women’s health & Development, Alexandria, Egypt.
Diagnosis of ambiguous genitalia (AG) represents an enormous challenge.
One hundred six cases were ascertained over the past 10 years
(1999-2008) in three Arab countries (Egypt, Libya, and Saudi Arabia)
by clinical evaluation, blood karyotyping, hormones and biochemistry,
pelvic imaging, and gonadal biopsy and SRY detection when indicated.
Results revealed MSHP in 66 cases, FSHP in 29 cases, 5 cases with
genetic syndromes, 5 cases with gonadal dysgenesis (GD) and one
true hermaphrodite (THF) with the 46,XX karyotype. Age of diagnosis
ranged between newborn and 36 years (mean is 5.6 years). Parental
consanguinity was observed in 82 cases (74 of them showed autosomal
recessive disorders as a cause for their genital ambiguity). Six
families had more than one sib affected (2-6). Proper gender reassignment
was performed in 15 cases where patients with complete form
of TFS were assigned the female sex while partial TFS, GD and THF
were assigned either gender according to the feasibility of surgical
reconstruction, the functioning gonads, pattern of expected hormonal
pubertal changes and the patient and family desire (gender identity).
Four cases rejected gender reassignment due to late diagnosis (12-23
years), religious and psychosocial reasons. This study highlights the
high frequency of AG due to parental consanguinity and the late onset
at diagnosis in Arab countries which might correlate with religious