2009 Vienna - European Society of Human Genetics

Reproductive genetics
P04.23
Results of prenatal cytogenetic and molecular genetic study of
the high risk chromosomal or monogenic diseases patients
N. Huleyuk, H. Makuh, M. Tyrkus, O. Nechay, J. Korinets, O. Malanchuk, L.
Melenchuk;
Institute of Hereditary Pathology of AMS Ukraine, Lviv, Ukraine.
Results of 162 prenatal diagnostics have been analysed. 144 prenatal
cytogenetic studies have been performed. In 20 amniocyte cultures
numeral (8 cases) and structural (12 cases) changes of the karyotype
were detected. In 10 cases chromosomal rearrangements arise due to
translocations in parents. Balanced chromosomal translocations were
detected in 8 amniocyte cultures. Robertsonian translocations were
found in 5 cases: 14;21 _ 3 cases, 13;15 and 13;22 _ one case each.
In 11 of cases, imbalanced karyotype was diagnosed: trisomy of 21-st
and 18-th chromosome; simple and mosaic forms of gonosomal monosomy;
additional marker chromosomes; mosaicism 46,XX/92,XXXX
(30:20). Non-balanced derivative chromosomes were detected in
two foetuses _ 46,XY,der(4),t(4;7)(q35;q31.1)mat and 46,XY,der(10),
t(10;7)(q26,13; p21.2)pat.
Prenatal molecular genetic diagnostics of families with heterozygous
mutations carriers, whose mutations lead to cystic fibrosis and Nijmegen
syndrome, was performed. DNA was extracted from cells of amniotic
fluids or chorion.
13 prenatal diagnostics of Cystic Fibrosis in families of heterozygous
carriers of CFTR gene mutations have been performed. As a
result, following foetus genotypes were identified: F508del/F508del
- 3, F508del/1717-1G>A - 1, F508del/wt - 6, N1303K/wt - 1, wt/wt
- 2. 5 prenatal molecular genetic diagnostics of Nijmegen breakage
syndrome in families of heterozygous carriers of 657del5 mutation of
NBN gene were performed. 2 foetuses were homozygous for 657del5
mutation of the NBN gene (genotype 657del5/657del5) and 3 cases
-heterozygous for this mutation (genotype 657del5/ wt). The prenatal
conformation of monogenic or chromosomal diseases mostly caused
termination of pregnancy by decision of family.
P04.24
immunogenetic markers of secondary infertility
D. Zastavna, O. Terpyljak, J. Zahanjach, N. Helner;
Institute of Hereditary Pathology Academy of Medical Sciences of Ukraine, Lviv,
Lviv, Ukraine.
In marital couples with secondary infertility and multiple first trimester
miscarriages, HLA-antigen distribution in A-, B-loci and a single nucleotide
polymorphism (SNP 1082 G->A) of the promoter region of IL-10
were studied. 157 individuals with secondary infertility with 2 or more
involuntary first trimester miscarriages and 64 individuals with primary
infertility were examined. The control group included 227 healthy individuals
with healthy children. It was determined that immunogenetic
markers of secondary infertility were HLA-antigens A10, B41 and B38,
with primary infertility associated with B7 andA19. The results also
show that more than 50% investigated couples had homologous HLA
genotypes. We studied the SNP1082 G->A of the IL-10 promoter region
in 50 couples with secondary infertility and recurrent loss of first
trimester pregnancies. We determined an allele A (low expressing allele)
frequency of 38,2%, and a G allele (high expressing) frequency of
61,8%. We observed a statistically significant, compared to the control
group, increase of high expression GG-genotype frequency, and a statistically
significant decrease of normal expression IL-10 gene (AGgenotype)
frequency. This suggests that IL-10 is active in the pathogenesis
of recurrent miscarriages. It was confirmed that the presence
of common HLA-antigens in marital couples and an increase of high
expression associated GG-genotypes at SNP -1082 G->A pIL-10 were
prognostically unfavourable for completion of pregnancy.
P04.25
couples with recurrent spontaneous abortions in moldova
(genetic characteristics).
V. C. Sacara, L. P. Rusu, V. V. Egorov, A. N. Misina;
Centre of reproductive health and medical genetics, Chisinau, Republic of
Moldova.
Infertility and spontaneous abortions are a major medical-social problem
in the conditions of demographic crisis existing in the Moldova
during the last decade. It’s claimed that all couples with reproductive
problems needs complex investigations witch include medico-genetics
analysis.
Materials and methods. In this study, 49 couples with miscarriage
passed medico-genetic counseling with cytogenetic and molecular genetic
analyses. Cytogenetic features of peripheral blood lymphocytes
cultivated according to standard techniques and DNA analyses to estimate
the differential risk associated with DQ genotypes.
Results. Analyses of family history revealed what in 67.39% of couples
were cases of recurrent spontaneous abortions (RSA), in 30.43% of
couples were cases of RSA with congenital anomalies, and in 2.17%
RSA and single-gene disorders. The analysis of TORCH-infection revealed
that test for serum IgG were positive in 45.13% women with
RSA, in 42.86% women with RSA and congenital anomalies. Cytogenetic
analysis showed different chromosomal aberrations involved
1, 9, 15, 17, and 22 chromosomes in 8% of cases. Were investigated
32 persons (total 64 chromosomes) by molecular analysis of HLA
haplotypes DQA1 and DQB1, discovered prevailed haplotypes of
DQA1*0101/0102 (35,9%) as well as DQA1*0501 (43,6%) in group
of patients and in control group. Haplotype DQB1 *0201 (26.6%) was
more frequent in patients as well as in control group (25.0%).
Conclusions. These data indicated that medico-genetic counseling
with cytogenetic and molecular genetic investigations may be indicative
in diagnosis of causes of RSA. Cytogenetic analysis could be valuable
for these couples when clinical data fail to clarify the cause.
P04.26
Role of mitochondria in repeated pregnancy loss
S. M. Seyedhassani1,2 , M. Houshmand2 , S. M. Kalantar1 , G. Modabber2 , R.
Mirfakhrai2 , A. Ebrahimi2 , A. Rasti1 , A. Aflatoonian1 ;
1Research and clinical center for infertility, Yazd, Islamic Republic of Iran,
2National institute of genetic engineering and biotechnology, Tehran, Islamic
Republic of Iran.
Introduction: Mitochondria are small structures in cells that generate
energy for the cell to use. All mitochondria are inherited from the mother’s
ovum. About 1 in 300 couples involve with Repeated Pregnancy
Loss (RPL) and the main part of them remains unknown. The aberrant
expression of apoptotic related genes is seen in RPL. It seems internal
apoptotic pathway and mitochondria have important role in fertilization
and proliferation of the cells.
Methods: In total 96 females who were suffered from idiopathic RPL.
Four multiplex PCR are done on each sample for detection of deletions.
D-loop part is analyzed by PCR-sequencing method. Bax and
Bcl2 genes is evaluated by PCR-sequencing method for promoter regions
and PCR-SSCP for exones.
Results: No deletions were found in 96 DNA samples. Mononucleotide
repeat (poly C) from 303 to 315 nucleotide positions (D310) exhibited a
polymorphic length variation (among 89 cases; 7C in 43, 8C in 34, 9C
in 8, and ≥10C in 4 females. Many sequence alterations identified in
D-loop region of cases, that will be described as mtDNA haplogroups
or novel nucleotide variants. Nucleotide change in Bax gene was seen
in promoter region at -55 A>G.
Discussion: Some of these nucleotide alterations might be involved
in RPL and could be included in a panel of molecular biomarkers for
susceptibility in pregnancy loss and even failure of in-vitro fertilization.
We believe that mutation in Bax and Bcl2 genes will lead to early apoptosis.
The results can be used in assessment of RPL.
P04.27
Blood pressure in 8 and 10- year-old singleton icsi children
F. Belva 1 , R. Painter 2 , J. De Schepper 3 , T. Roseboom 4 , I. Liebaers 1 , M. Bonduelle
1 ;
1 Medical Genetics, UZ Brussel, Brussels, Belgium, 2 Obstetrics and Gynaecology,
AMC, Amsterdam, The Netherlands, 3 Pediatric Endocrinology, UZ Brussel,
Brussels, Belgium, 4 Clinical Epidemiology and Biostatistics, AMC, Amsterdam,
The Netherlands.
Introduction: To evaluate if the in-vitro procedure in humans has longterm
consequences on the cardiovascular functioning, longitudinal
blood pressure measurements were compared between children born
after intra-cytoplasmic sperm injection (ICSI) and after spontaneous
conception (SC).
Material and Methods: Longitudinal questionnaire data and parameters
of physical examination of 8-year-old ICSI children were compared
with results of peers born after SC. At the age of 10 years, 108
of the initial recruited150 ICSI children were re-examined and 93 out of
0