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STUDY QUESTION Could whole-exome sequencing (WES) be useful in clinical practice for men with maturation arrest (MA) after a first testicular sperm extraction (TESE)? SUMMARY ANSWER WES in combination with TESE yields substantial additional information and may potentially be added as a test to predict a negative outcome of a recurrent TESE in patients with MA. WHAT IS KNOWN ALREADY At present, the only definitive contraindications for TESE in men with non-obstructive azoospermia (NOA) are a 46,XX karyotype and microdeletions in the azoospermia factor a (AZFa) and/or AZFb regions. After a first negative TESE with MA, no test currently exists to predict a negative outcome of a recurrent TESE. STUDY DESIGN, SIZE, DURATION In a cohort study, we retrospectively included 26 patients with idiopathic NOA caused by complete MA diagnosed after a first TESE. PARTICIPANTS/MATERIALS, SETTING, METHODS Twenty-six men with MA at the spermatocyte stage in all seminiferous tubules, according to a histopathological analysis performed independently by two expert histologists, and a normal karyotype (i.e. no AZF gene microdeletions on the Y chromosome) were included. Single-nucleotide polymorphism comparative genomic hybridization array and WES were carried out. The results were validated with Sanger sequencing. For all the variants thought to influence spermatogenesis, we used immunohistochemical techniques to analyse the level of the altered protein. MAIN RESULTS AND THE ROLE OF CHANCE Deleterious homozygous variants were identified in all seven consanguineous patients and in three of the 19 non-consanguineous patients. Compound heterozygous variants were identified in another 5 of the 19 non-consanguineous patients. No recurrent variants were identified. We found new variants in genes known to be involved in azoospermia or MA [including testis expressed 11 (TEX11), meiotic double-stranded break formation protein 1 (MEI1), proteasome 26s subunit, ATPase 3 interacting protein (PSMC3IP), synaptonemal complex central element protein 1 (SYCE1) and Fanconi anaemia complementation group M (FANCM) and variants in genes not previously linked to human MA (including CCCTC-binding factor like (CTCFL), Mov10 like RISC complex RNA helicase 1 (MOV10L1), chromosome 11 open reading frame 80 (C11ORF80) and exonuclease 1 (EXO1)]. LARGE SCALE DATA Data available on request LIMITATIONS, REASONS FOR CAUTION More data are required before WES screening can be used to avoid recurrent TESE, although screening should be recommended for men with a consanguineous family background. WES is still a complex technology and can generate incidental findings. WIDER IMPLICATIONS OF THE FINDINGS Our results confirmed the genetic aetiology of MA in most patients: the proportion of individuals with at least one pathologic variant was 50% in the overall study population and 100% in the consanguineous patients. With the exception of MEI1 (compound heterozygous variants of which were identified in two cases), each variant corresponded to a specific gene—confirming the high degree of genetic heterogeneity in men with MA. Our results suggest that WES screening could help to avoid recurrent, futile TESE in men with MA in general and in consanguineous individuals in particular, but these results need to be confirmed in future studies before clinical implementation. STUDY FUNDING/COMPETING INTEREST(S) The study was funded by the Fondation Maladies Rares (Paris, France), Merck (Kenilworth, NJ, USA), IRSF (Montigny le Bretonneux, France) and Agence de la Biomédecine (Saint Denis, France). There are no competing interests. TRIAL REGISTRATION NUMBER N/A.

AIM The aim of this study was to investigate whether recurrent pregnancy loss (RPL) is associated with adipokine gene polymorphisms (namely the leptin -2548 (G/A), adiponectin 276 (G/T), and adiponectin 45 (T/G) polymorphisms) and/or adipokine serum levels. METHODS A total of 145 women participated in the study. For the analysis of serum adipokine levels, 19 healthy fertile women (control group) and 60 women suffering from RPL were included. For the polymorphism analysis, 126 women suffering from RPL were included. Serum adipokine levels were determined using a commercial radioimmunoassay kit. Adipokine polymorphisms were analyzed using an allele-specific polymerase chain reaction (PCR). RESULTS Our immunoassays revealed that serum leptin levels were similar in control and RPL groups (17.34 and 20.16 ng/mL, respectively). In contrast, serum adiponectin levels were significantly higher in women with RPL than in controls (9.83 and 6.89 μg/mL, respectively; P < 0.05). Unfortunately, our allele-specific PCR experiments did not reveal any significant differences in allele frequency between women with RPL and NCBI allele frequencies. CONCLUSION This study demonstrates that adiponectinemia is increased in patients suffering from RPL. However, association of adiponectin with adverse pregnancy outcomes remains to be elucidated.

International audience; In recent years, the introduction of novel genome analysis technologies (such as array comparative genomic hybridization) has enabled the prenatal diagnosis of various recurrent copy number variations (CNVs). Some of these CNVs have been linked to a greater susceptibility of developmental and neuropsychiatric disorders; for example, recurrent duplication at the 2q13 locus is associated with developmental delay, dysmorphism and intellectual disability. However, this CNV has low penetrance and variable clinical expressivity. It also can be observed in healthy controls and can be transmitted by unaffected parents, making genetic counseling especially challenging. Here, we report on the inheritance of a 2q13 duplication in an asymptomatic family; the case highlights the role of the family survey in genetic counseling with regard to novel CNVs diagnosed before birth.

Key Clinical Message We report a rare case of recurrent trisomy 21 caused by an isochromosome 21q and what is very likely to be maternal germ-line cell mosaicism. Over 90% of cases of rob(21;21) reported in the literature are due to an isochromosome 21q, with a risk of recurrence of more than 10%.

Microduplication 22q11.2 is primarily characterized by a highly variable clinical phenotype, which ranges from apparently normal or slightly dysmorphic features (in the presence or absence of learning disorders) to severe malformations with profound mental retardation. Hence, genetic counseling is particularly challenging when microduplication 22q11.2 is identified in a prenatal diagnosis. Here, we report on 24 prenatal cases of microduplication 22q11.2. Seventeen of the cases were also reanalyzed by microarray analysis, in order to determine copy number variations (CNVs, which are thought to influence expressivity). We also searched for possible correlations between fetal phenotypes, indications for invasive prenatal diagnosis, inheritance, and pregnancy outcomes. Of the 24 cases, 15 were inherited, six occurred de novo, and three were of unknown origin. Termination of pregnancy occurred in seven cases and was mainly decided on the basis of ultrasound findings. Moreover, additional CNVs were found in some patients and we try to make a genotype-phenotype correlation. We discuss the complexity of genetic counseling for microduplication 22q11.2 and comment on possible explanations for the clinical heterogeneity of this syndrome. In particular, we assessed the co-existence of additional CNVs and their contribution to phenotypic variations in chromosome 22q11.2 microduplication syndrome. © 2014 John Wiley & Sons, Ltd.

Key Clinical Message Translocations between X and acrocentric chromosomes are rare. We report on the inheritance of a familial t(X;15)(p22.3;p10) translocation in a fetus referred for short long bones. Cytogenetic analysis revealed an unbalanced translocation combined with a three-gene nullosomy. After genetic counseling, a prognosis was established and a healthy boy was delivered.

Objective The 22q11.2 deletion (del22q11.2) is one of the most common microdeletions. We performed a collaborative, retrospective analysis in France of prenatal diagnoses and outcomes of fetuses carrying the del22q11.2. Methods A total of 272 fetuses were included. Data on prenatal diagnosis, ultrasound findings, pathological features, outcomes and inheritance were analyzed. Results The mean time of prenatal diagnosis was 25.6 ± 6 weeks of gestation. Most of the diagnoses (86.8%) were prompted by abnormal ultrasound findings [heart defects (HDs), in 83.8% of cases]. On fetal autopsy, HDs were again the most common disease feature, but thymus, kidney abnormalities and facial dysmorphism were also described. The deletion was inherited in 27% of cases. Termination of pregnancy (TOP) occurred in 68.9% of cases and did not appear to depend on the inheritance status. However, early diagnosis was associated with a higher TOP rate. Conclusion This is the largest cohort of prenatal del22q11.2 diagnoses. As in postnatally diagnosed cases, HDs were the most frequently observed abnormalities. However, thymus and kidney abnormalities and polyhydramnios should also be screened for in the prenatal diagnosis of del22q11.2. Only the time of diagnosis appeared to be strongly associated with the pregnancy outcome: the earlier the diagnosis, the higher the TOP rate. © 2014 John Wiley & Sons, Ltd.

Study question Do TNF-308 and -238 polymorphisms impact the embryo implantation rate after in vitro fertilization (IVF) in women without female infertility factor? Summary answer The presence of the TNF-308A allele is associated with high implantation and multiple pregnancy rates in women without known infertility factors after ovarian hyperstimulation with exogenous FSH. What is already known Multiple pregnancies are frequent after the use of Assisted Reproductive Technologies. Single embryo transfer (SET) has been proposed as a simple way to prevent these risks. However, the extension of SET indications to patients not selected based on specific criteria is controversial because of reduced pregnancy rates. To date, the predictive value of the parameters used for SET (age, gynecological history of the patient and uterine characteristics) allows a pregnancy rate of ~30%. Study design, size, duration The potential predictive value of TNF polymorphisms (-308, rs1800629 and -238, rs361525) on implantation rate was evaluated in 424 women requiring IVF due to male fertility factors. This cohort retrospective study was conducted over 4 years in University-affiliated hospitals. Participants, setting, methods The entire patient group included 424 women undergoing intracytoplasmic sperm injection (ICSI) due to male fertility factors without the contribution of any female factor. From among this group, a selected patient group included 120 women with a normal karyotype, age under 38 years, serum follicle-stimulating hormone (Day-3 FSH) levels below 10 IU/l, a long agonist desensitization protocol associated with recombinant FSH treatment and a Caucasian background. Main results and the role of chance The TNF-238 polymorphism was not associated with implantation rate. In contrast, the presence of the TNF-308A allele was associated with increased Day 3-E2 levels as well as higher implantation and multiple pregnancy rates after fresh embryo transfer in women from the entire and selected patient groups. Moreover, in the selected patient group, the presence of the TNF-308A allele was also associated with a decrease in the miscarriage rate. The benefit of the TNF-308A allele in predicting implantation rates was not observed after the use of frozen embryos. Limitations, reasons for caution Future studies are needed to evaluate whether the TNF-308A allele might also be a biomarker in women with infertility factors. Wider implications of the finding The TNF-308A allele may represent a good candidate for a potential predictive, non-invasive biomarker in the SET strategy. However, its impact should be evaluated in prospective studies. Study funding/competing interest This study was conducted with financial support from the French Institute for Health and Medical Research (INSERM), Organon France for a FARO (Fond d'Aide a la Recherche Organon) fellowship (to V.T.) and CHU Nice PHRC (PHRC 09-279).There are no competing interests.

Complex chromosome rearrangements (CCR) with two independent chromosome rearrangements are rare. Although CCRs lead to high unbalanced gamete rates, data on meiotic segregation in this context are scarce. A male patient was referred to our clinic as part of a family screening programme prompted by the observation of a 44,X,der(Y),t(Y;15)(q12;q10)pat,rob(13;14)(q10;q10)mat karyotype in his brother. Karyotyping identified the same CCR. Sperm FISH (with locus-specific probes for the segments involved in the translocations and nine chromosomes not involved in both rearrangements) was used to investigate the rearrangements meiotic segregation products and establish whether or not an inter-chromosomal effect was present. Sperm nuclear DNA fragmentation was also evaluated. For rob(13;14) and der(Y), the proportions of unbalanced products were, respectively, 26.4% and 60.6%. Overall, 70.3% of the meiotic segregation products were unbalanced. No evidence of an inter-chromosomal effect was found, and the sperm nuclear DNA fragmentation rate was similar to our laboratory's normal cut-off value. In view of previously published sperm FISH analyses of Robertsonian translocations (and even though the mechanism is still unknown), we hypothesise that cosegregation of der(Y) and rob(13;14) could modify rob(13;14) meiotic segregation.

Duplication 17p11.2 (Potocki-Lupski syndrome (PTLS) MIM# 610883) is a genomic disorder with an estimated incidence of 1 in 25,000 births. As for other genomic disorders this duplication is typically de novo and is not associated with advanced maternal age or advanced paternal age. Herein we describe a prenatal diagnosis of duplication 17p11.2. This diagnosis was not suspected as the prenatal ultrasound findings were non-specific; however, BACs-on-Beads™ technology and array comparative genomic hybridization (aCGH) confirmed the common ∼3.7 Mb duplication. Evaluation of the foetus following termination of pregnancy revealed mildly dysmorphic features as well as congenital anomalies not previously reported in PTLS, specifically left pulmonary isomerism, an abnormally positioned left coronary orifice and nodular cerebellar heterotopia. This report exemplifies the utility of prenatal testing using new genomic technologies even when there are no multiple anomalies on foetal ultrasound. This report also exemplifies the utility of foetal autopsy in the identification of "occult" congenital anomalies.

Microdeletion and microduplication syndromes are well-known causes of developmental delay and/or malformations of differing severity. It was recently reported that a microdeletion at the 3q13.31 locus is associated with a new syndrome combining developmental delay, postnatal overgrowth and dysmorphic features. However, the reciprocal microduplication has only been described in a few case reports displaying some clinical features of the microdeletion syndrome. Here, we report on a female infant with a 3.34 Mb microduplication of the 3q13.2q13.31 region inherited from her mother. The infant presented with severe intellectual disability, learning difficulties, intrauterine and postnatal growth retardation and skeletal particularities but no dysmorphic traits. This microduplication encompassed the previously described shortest region of overlap, which contains five genes (DRD3, ZNF80, TIGIT, MIR568 and ZBTB20). We reviewed the phenotypes described in the literature on microduplications and in the well-characterized 3q13.31 microdeletion syndrome. In agreement with the literature data, DRD3 and ZBTB20 appear to be strong candidate genes for neurodevelopmental defects and growth retardation. Lastly, we consider the putative mechanism of this rearrangement, which may involve a particular kind of nonallelic homologous recombination of human endogenous retrovirus elements.

Our objective was to identify a marker for oocyte aneuploidy in follicular fluid (FF) in women with an increased risk of oocyte aneuploidy after controlled ovarian hyperstimulation.Three groups of oocytes were constituted for polar body screening by FISH (chromosomes 13, 16, 18, 21 and 22): Group 1, advanced maternal age (n = 156); Group 2, implantation failure (i.e. no pregnancy after the transfer of more than 10 embryos; n = 101) and Group 3, implantation failure and advanced maternal age (n = 56). FSH and other proteins were assayed in the corresponding FF samples.Of the 313 oocytes assessed, 35.78 % were abnormal. We found a significant difference between the follicular FSH levels in normal oocytes and abnormal oocytes (4.85 ± 1.75 IU/L vs. 5.41 ± 2.47 IU/L, respectively; p = 0.021). We found that the greater the number of chromosomal abnormalities per oocyte (between 0 and 3), the higher the follicular FSH level.High FF FSH levels were associated with oocyte aneuploidy in women having undergone controlled ovarian hyperstimulation.

Objective Molecular cytogenetic techniques on uncultured prenatal samples are the sole tests applied in some countries in cases with advanced maternal age (AMA) or increased risk after prenatal screening. Moreover, there is a trend to perform invasive prenatal diagnosis (PD) during the first trimester before ultrasound manifestations, so new rapid and reliable assays are necessary to investigate microdeletions not detectable with the conventional karyotype. We report the validation study of the prenatal bacterial artificial chromosomes-on-Beads™ (BoBs™; CE-IVD), a bead-based multiplex assay detecting chromosomes 13, 18, 21, X/Y aneuploidies and nine microdeletion regions having an overall detection rate of 1/1700. Method We retrospectively studied 408 selected samples and prospectively tested 212 consecutive samples ascertained for conventional karyotyping. Results We did not find false-positive results. Triploidies were not detected. Maternal cell contamination of male samples up to 90% was unmasked inspecting gonosome profiles. Mosaic conditions at 20 to 30% were revealed. Failures were due to low amount of DNA. Conclusion Prenatal BoBs™ is a robust technology for the investigation of fetuses with normal karyotype with or without sonographic abnormalities. Running in parallel with the karyotype analysis, it can be proposed instead of rapid FISH or QF-PCR providing rapid results on common aneuploidies and additional information regarding the microdeletion syndromes. Copyright © 2011 John Wiley & Sons, Ltd.

Array-CGH emergence allowed important diagnosis progress, and a better care of patients in postnatal. So, there is a great temptation to use it also in prenatal diagnosis. The point of view objective is to make a rapid overview of cytogenetic diagnosis evolution during the last 50 years, and to show all questions raised by the use of array-CGH, and problems that could arise in prenatal diagnosis. While array-CGH just comes in genetic laboratories, new diagnosis approaches emerged like whole genome sequencing or non-invasive prenatal diagnosis. The 2nd objective will be to review all these techniques for a probably close future.

Résumé Cette recherche est née de la constatation clinique, à travers les récits de vie d’hommes infertiles, de l’existence de perturbations dans leur filiation. Dans un premier temps, il s’agira de déterminer, à l’aide d’une grille de filiation construite à partir d’une clinique psychanalytique, si, pour certains de ces hommes infertiles, le projet de devenir père pourrait être entravé par leur difficulté à se situer dans leur généalogie et à s’identifier à leur propre père. La comparaison d’un groupe de 30 hommes infertiles avec un groupe de 30 hommes fertiles permet de mettre en évidence l’impact de certains événements dans la filiation sur la capacité à procréer. Le second temps de la recherche sera consacré à une approche psychodynamique qui tentera d’éclairer l’hypothèse d’un lien entre l’impact traumatique de ces événements sur les sujets et sa traduction dans une expression somatique.

Preimplantation genetic diagnosis (PGD) is widely used for women heterozygous for a Robertsonian translocation. Preconceptional diagnosis (PCD), performed before fertilization, may be an alternative to PGD, especially in countries where PGD is restricted or prohibited, as in France. It could also give different information and clarify the influence of reproductive and obstetric history.In our study, translocation was diagnosed before ICSI in five cases (group A), and after newborn or fetal aneuploidy or miscarriage in two cases, (group B).First polar body (PB1) analysis using acrocentric centromeric probes was done for 85 PB1s, and aneuploidy rate was at 42.4%. Oocyte aneuploidy rate differed (p0.0001) between groups A and B (30% vs 84%). Despite the small group sizes, we demonstrate a correlation (p=0.0358) of aneuploidy rate in polar bodies after 2 or more attempts. Three live births were recorded, all in group A.PCD could thus be an alternative to PGD. This pilot study also provides new prognostic information taking into account the women's natural history, but further confirmation is required.

Translocations involving gonosomes are frequent in azoospermic patients and sometimes in oligozoospermic ones, conditions that lead to request for assisted reproduction treatment. This study reports an unexpectedly fertile 49-year-old man bearing a de-novo translocation 46,X,t(Y;10)(q11.2;q15.2) associated with a high chromosomal risk for offspring, and referred for familial investigations after the diagnosis of an unbalanced translocation 46,XX,der(10)t(Y;10)(q11.2;p15.2) in his naturally conceived and mentally retarded daughter. Chromosome molecular investigation confirmed Y long-arm inheritance in the daughter and absence of the Yq deletion in the father. Semen analysis showed a normal sperm count associated with moderate asthenospermia and severe teratospermia. A total of 984 spermatozoa were analysed using fluorescence in-situ hybridization (FISH). Alternate segregation pattern was found in 50.31% of the spermatozoa studied. The frequencies of adjacent I, adjacent II, 3:1 segregation, and diploidy (or 4:0 segregation) were respectively 39.62, 1.63, 7.83, and 0.61%. No interchromosomal effect was observed. This patient is the first fertile man in whom the meiotic segregation pattern of a Y-autosome translocation has been analysed. The imbalance risk was close to those observed for reciprocal translocations, and emphasizes the value of FISH studies in males with a chromosomal translocation in order to provide them a personalized risk evaluation.

Objectives: Etiologic diagnosis of multiple congenital abnormalities (MCAs) is often lacking. Large chromosome abnormalities can be detected by conventional cytogenetic methods, but more subtle chromosome micro-rearrangements and/or de novo abnormalities require multi-FISH analysis, which is hampered by the amount of material available in prenatal testing. Methods: We used the comparative genomic hybridization (CGH) array, Genosensor™ Array 300, to screen for classic microdeletion syndromes and subtelomeric rearrangements in 39 consecutive fetuses with MCAs, after termination of pregnancy, in a prospective study. Thirty-seven of them had a normal karyotype, and two had a de novo unbalanced karyotype that could not be characterized with conventional cytogenetic methods. Results: Two de novo unbalanced karyotypes were characterized by array CGH, and four additional abnormalities were diagnosed: an unbalanced inherited cryptic translocation, a deletion in band 22q11.2, a 1p36 deletion, and a 6p12.1–21.2 duplication. Conclusion: Chromosomal imbalances were therefore detected and/or characterized in 6 of 39 (15.4%) fetuses, indicating the value of routine array CGH in cases of MCAs and in uncharacterized chromosome rearrangements. Extension to all prenatal diagnoses may be warranted when copy number variation is identified and all FISH probes are commercially available.

The aim of this study was to evaluate the stability of the aneuploidy rate of the first polar body. Knowing the stability of the oocyte aneuploidy rate for each patient would allow the first analysis to be used as a prognostic tool for further attempts at intracytoplasmic sperm injection (ICSI). After a first unsuccessful ICSI attempt with preconceptional screening, 24 women underwent a second attempt. First polar body aneuploidy rates were compared in the course of two successive ovarian stimulations. The first polar body was biopsied after laser dissection of the zona pellucida and five chromosomes were analysed using the MultiVysion™ polar body multicolour probe panel. A total of 200 polar bodies were analysed; 91 and 109 in the first and second ICSI attempts, respectively. The total aneuploidy rate was identical in the first and second attempts; 44.0% (40/91) and 44.0% (48/109), respectively. The first evaluation of the aneuploidy rate was statistically (P = 0.0007) correlated with the second, with a correlation coefficient, r = 0.707. The stability of the aneuploidy rate in different cohorts from the same patient, if confirmed in a larger series, makes this parameter a useful tool for counselling couples.

To clarify the mechanisms underlying oocyte abnormalities in meiosis: meiotic nondisjunction of a whole chromosome or premature separation of sister chromatids in two situations of increased chromosomal risk.Preconception diagnosis by first polar-body analysis in two situations of increased chromosomal risk.Departments of reproductive biology, cytogenetics, gynecology, and obstetrics.First polar body analysis was proposed to 76 patients (91 cycles) for advanced age (AMA; n = 30, 36 cycles), recurrent implantation failure (RIF;10 embryos transferred without implantation; n = 32, 36 cycles), or both (AMA + RIF; n = 14, 19 cycles), before their intracytoplasmic sperm injection procedure.First polar-body analysis using fluorescence in situ hybridization.Mechanisms and frequency of aneuploidy.Three hundred eighty-four oocytes were analyzed by fluorescence in situ hybridization, 130 from women38 years of age, 171 from women with RIF, and 83 from women with both indications. The oocyte abnormality rate was similar in the three groups, respectively, 38.5%, 40.4%, and 45.8%. The aneuploidy mechanisms were different for women38 years of age who had no previous implantation failure (AMA) compared with women of whatever age who had implantation failure (P.05 vs. RIF; P.001 vs. AMA+RIF), with, respectively, for the AMA, RIF, and AMA+RIF groups, 72.2%, 56.6%, and 49.2% premature separation of sister chromatids and 27.8%, 43.4%, and 50.8% meiotic nondisjunction. In the two implantation-failure groups, we distinguished a subgroup (22% in the RIF group and 33% in AMA+RIF group) of patients with2/3 abnormal oocytes, suggesting a meiosis alteration.The mechanisms accounting for oocyte aneuploidy differed in the two clinical situations of advanced maternal age and RIF. Advanced maternal-age aneuploidy was linked to a loss of sister chromatid cohesion that led to one single chromatid abnormality, whereas implantation failure is a much more heterogeneous situation.

La cryoconservation de spermatozoides est une technique couramment utilisee en AMP, en particulier dans les cas d’oligospermie severe, dans la crainte d’une aggravation ulterieure. L’objectif de notre etude a ete de rechercher un effet deletere eventuel du processus de migration-congelation-decongelation sur la fragmentation de l’ADN des spermatozoides evaluee par la technique TUNEL. L’etude a porte sur 72 patients repartis en 4 groupes selon leurs caracteristiques spermatiques: groupe 1 [n=20] (parametres «normaux» selon l’OMS), groupe 2 [n=24] (numeration spermatique normale associee a une astheno et/ou une teratospermie), groupe 3 [n=16] (numeration totale comprise entre 5 et 20 millions), groupe 4 [n=12] (numeration totale inferieure a 5 millions). Une evaluation des parametres spermatiques conventionnels et de la fragmentation de l’ADN (technique TUNEL) a ete realisee sur le sperme entier (non migre non congele) et sur le sperme decongele. Une evaluation des taux de fragmentation a egalement ete realisee sur la fraction migree avant congelation pour 20 patients (10 du groupe 1 et 10 du groupe 2); cette analyse n’a pas pu etre effectuee sur les prelevements oligospermiques (groupes 3 et 4). Apres l’ensemble du processus de migration, congelation et decongelation du sperme, le taux de fragmentation diminue dans la population temoin (groupe 1), alors qu’il augmente significativement dans les groupes de patients presentant des parametres spermatiques alteres (groupes 2, 3 et 4). En examinant l’etape intermediaire, la migration diminue les taux de fragmentation dans les 2 groupes etudies par rapport au sperme entier. Donc le taux de fragmentation diminue apres migration, puis s’eleve apres congelation-decongelation a un taux inferieur a celui du sperme entier dans la population temoin, alors qu’il remonte a un taux plus eleve lorsque le sperme est altere. Cependant, cet effet deletere de la congelation sur les spermes alteres reste modere. En conclusion, ces resultats sont plutot en faveur de l’autoconservation de sperme pour les patients oligospermiques severes. Correspondance

BACKGROUND: Maternal ageing is the only aetiological factor unequivocally linked to aneuploidy. Two mechanisms seem to explain these abnormalities in oocytes: non-disjunction and premature unbalanced separation of sister chromatids (PSSC). Previous studies of unfertilized oocytes argue for a major role of PSSC in the aetiology of aneuploidy for women of advanced age, but in vitro ageing of the oocytes could influence the results. METHODS: Owing to the high prevalence of aneuploidy in women of advanced age, chromosomal screening of the first polar body just before ICSI was offered to women (from 38 years of age) included in an assisted reproduction programme. RESULTS: Among 141 oocytes from 29 women (mean age 40 years and 2 months), 43 (30.5%) were abnormal. Sixtyfive abnormalities were found and PSSC was involved in 80% of cases. CONCLUSION: These results are in accordance with previous studies and confirm, in 'fresh' oocytes, the major role of PSSC in the aetiology of aneuploidy in women of advanced age.

RESUME Le taux d’echec du developpement embryonnaire est tres important dans notre espece. On sait depuis longtemps que la cause des fausses couches spontanees est majoritairement chromosomique (66 % des fausses couches). Avec le developpement des techniques d’assistance medicale a la procreation, il est devenu possible de rechercher la part des anomalies chromosomiques beaucoup plus tot, sur les embryons preimplantatoires, ce qui a permis de mettre en evidence un fort taux d’anomalies chromosomiques a ces stades (50 % des embryons concus). Deux mecanismes ont ete identifies comme etant a l’origine des aneu- ploidies ovocytaires, la non-disjonction meiotique (NDM) et la separation prematuree des chromatides sœurs (SPCS). L’etude du premier globule polaire, cellule complementaire de l’ovocyte a la meiose, peut etre realisee par la technique d’hybridation fluorescente in situ (FISH) avant injection intra-cytoplasmique de spermatozoide (ICSI). Ceci a ete realise dans notre equipe dans deux populations distinctes (patientes âgees de plus de 38 ans et patientes en echec d’implantation) et confirme les resultats obtenus sur les ovocytes non fecondes. Ils permettent egalement d’affirmer que la SPCS est le mecanisme majoritairement implique dans les aneuploidies liees a l’âge maternel chez l’humain.

Chez un homme, les translocations impliquant les gonosomes sont le plus souvent responsables d'azoospermie ou parfois d'oligospermie, conduisant les patients a demander une AMP. Nous rapportons ici le cas d'un patient oligosperme, porteur d'une translocation 46,Y,t(X;2)(p21;p25.3) et les modalites specifiques de prise en charge du couple avant ICSI. Apres un conseil genetique, nous avons propose une analyse chromosomique des spermatozoides par cytogenetique moleculaire (FISH) dans le but de connaitre le mode de segregation de la translocation et le risque pour l'embryon et la descendance. Cette etude a montre que 34% des spermatozoides etaient normaux ou equilibres pour les chromosomes etudies. Les 66% des spermatozoides restant presentaient un desequilibre lie a la translocation et/ou a des anomalies de non-disjonction des gonosomes. Devant ce resultat, une nouvelle analyse en FISH, visant a preciser les risques eventuellement accrus de non-disjonction meiotique des chromosomes X et Y, a ete decidee, et sur 1000 spermatozoides analyses 60% seulement etaient normaux pour les gonosomes. Le risque de desequilibre pour la descendance concerne donc non seulement la translocation elle-meme mais egalement les gonosomes (syndrome de Klinefelter ou Turner) par malsegregation des chromosomes sexuels. Suite a ce resultat et en raison de l'âge de la conjointe (42ans), nous avons deconseille l'ICSI au couple au cours d'un nouveau conseil genetique Ce cas souligne l'interet qu'il y a a evaluer par FISH, les consequences d'une translocation dans les spermatozoides d'un patient porteur. L'etude ne doit pas se limiter a l'etude de la translocation elle-meme, mais egalement concerner les anomalies de non-disjonction des gonosomes, frequentes au cours de la spermatogenese et plus particulierement en presence d'une translocation impliquant un gonosome.

Introduction L’insuffisance ovarienne prematuree (IOP) se definit par une amenorrhee et un taux de FSH superieur a 25 UI/L. Dans environ 70 % des cas, aucune etiologie n’est retrouvee. Ces dernieres annees, de nombreux genes candidats ont ete identifies. Cependant, leur imputabilite et leur prevalence restent mal connues. Objectif Evaluer la prevalence de mutations de genes candidats dans une cohorte multicentrique de patientes avec IOP. Materiel et methodes Une analyse par sequencage a haut debit utilisant une puce contenant 18 genes candidats a ete realisee chez 288 patientes. Les anomalies retrouvees ont ete classees en « mutation » ou « variant rare » a l’aide de logiciels de prediction (SIFT, Polyphen-2, mutation taster). Resultats La repartition des patientes etait de 12,5 % de patientes avec une amenorrhee primaire, 87,5 % avec une amenorrhee secondaire. Au moins, une mutation a ete identifiee chez 22 % des patientes et 2 mutations chez 1,4 %. NOBOX etait le gene le plus frequemment mute (9 % des cas). Des variants rares presumes non pathogenes etaient presents chez 33 % des patientes. Parmi les 47 cas ayant une forme familiale, 17 % avaient une mutation (versus 23 % dans les formes non familiales). En cas d’amenorrhee primaire, 28 % des patientes presentaient une mutation. Discussion Cette etude sur une large cohorte a identifie des anomalies genetiques a priori non pathogenes et des mutations chez respectivement 33 et 22 % des patientes. Ces resultats illustrent la precaution necessaire dans l’interpretation des resultats, notamment pour le conseil genetique des patientes avec IOP.

Microdeletion and microduplication syndromes are well-known causes of developmental delay and/or malformations of differing severity. Although homogeneous abnormalities can now be detected relatively easily using microarray technologies, they are more difficult to detect and interpret in cases of mosaicism. Here, we report on a male infant with a mosaic de novo derivative chromosome 9, featuring a 10.2 Mb 5q35 duplication (including the NSD1 gene) and a 687 kb 9q34 deletion (including EHMT1). The infant presented developmental delay, short stature, brachy/plagiocephaly and hyperactivity. The proportion of abnormal cells was 50% in saliva (in a microarray analysis) and 25% in lymphocytes (in a FISH analysis). Despite the low-level mosaicism in lymphocytes, this imbalance appears to be responsible for a distinctive phenotype (suggesting the presence of variable clinical expression and/or major somatic mosaicism).

BACKGROUND A multiple pregnancy is now considered to be the most common adverse outcome associated with in vitro fertilization (IVF). As a consequence, the identification of women with the best chances of embryo implantation is a challenge in IVF program, in which the objective is to offer elective single-embryo transfer (eSET) without decreasing the pregnancy rate. To date, a range of hormonal and clinical parameters have been used to optimize eSET but none have significant predictive value. This variability could be due to genetic predispositions related to single-nucleotide polymorphisms (SNPs). Here, we assessed the individual and combined impacts of thirteen SNPs that reportedly influence the outcome of in vitro fertilisation (IVF) on the embryo implantation rate for patients undergoing intracytoplasmic sperm injection program (ICSI). MATERIALS AND METHODS A 13 gene polymorphisms: FSHR(Asn680Ser), p53(Arg72Pro), AMH(Ile49Ser), ESR2(+1730G>A), ESR1(-397T>C), BMP15(-9C>G), MTHFR1(677C>T), MTHFR2(1298A>C), HLA-G(-725C>G), VEGF(+405G>C), TNFα(-308A>G), AMHR(-482A>G), PAI-1(4G/5G), multiplex PCR assay was designed to genotype women undergoing ICSI program. We analyzed the total patients population (n = 428) and a subgroup with homogeneous characteristics (n = 112). RESULTS Only the VEGF(+405G>C) and TNFα(-308A>G) polymorphisms impacted fertilization, embryo implantation and pregnancy rates. Moreover, the combined VEGF+405.GG and TNFα-308.AG or AA genotype occurred significantly more frequently in women with high implantation potential. In contrast, the VEGF+405.CC and TNFα-308.GG combination was associated with a low implantation rate. CONCLUSION We identified associations between VEGF(+405G>C) and TNFα(-308A>G) polymorphisms (when considered singly or as combinations) and the embryo implantation rate. These associations may be predictive of embryo implantation and could help to define populations in which elective single-embryo transfer should be recommended (or, conversely, ruled out). However, the mechanism underlying the function of these polymorphisms in embryo implantation remains to be determined and the associations observed here must be confirmed in a larger, more heterogeneous cohort.

Genes involved in testicular differentiation, spermatogenesis, proliferation and apoptosis of germ cells have been shown to evolve rapidly and display rapid DNA changes. These genes are therefore good candidates for explaining impairments in spermatogenesis. Initial studies of some of these genes appear to confirm this hypothesis. The RHOXF2 candidate gene belongs to the RHOX family clustered in Xq24 and is specifically expressed in the testis. It contains four exons and codes for a 288 amino acid (aa) transcription factor. It has a high degree of homology (99.9%) with its paralogue RHOXF2B, which is also preferentially expressed in the testis.To sequence RHOXF2 and RHOXF2B in intracytoplasmic sperm injection (ICSI) patients and identify any single-nucleotide polymorphisms (SNPs) associated with impaired spermatogenesis.A cohort of 327 patients in ICSI programmes at Poissy and Bichat hospitals. All patients gave their written, informed consent to participation. One hundred patients had unaffected spermatogenesis and 227 displayed impaired spermatogenesis.The four exons in each of RHOXF2 and RHOXF2B were sequenced in 47 patients with oligospermia or non-obstructive azoospermia. Given that exons 2 and 3 were found to harbour most of the SNPs, only these two exons were sequenced in the remaining 280 subjects.Due to the extremely high degree of sequence identity between RHOXF2 and RHOXF2B, we were not able to distinguish between the sequences of these two genes. Although 9 SNPs were identified, there were no significant frequency differences between ICSI patients with normal vs. impaired spermatogenesis. Two insertions were identified: a 21-nucleotide insertion was retrieved in both groups and a guanine insertion (inducing a premature stop codon) only found in two patients with impaired spermatogenesis.RHOXF2 is a good candidate for rapid evolution by positive selection. Analysis of the polymorphism frequency in exons 2 and 3 did not allow us to correlate the identified SNPs with male infertility. However, a single nucleotide insertion was identified only in men with impaired spermatogenesis. Further work will be needed to establish whether genetic changes in RHOXF2 can give rise to defects in spermatogenesis.Les gènes impliqués dans la différenciation des testicules, la spermatogenèse, la prolifération et l'apoptose des cellules germinales ont été montrés comme ayant une évolution rapide de la séquence d’ADN. Ces gènes sont donc de bons candidats pour expliquer les déficiences de la spermatogenèse. Les premières études semblent confirmer cette hypothèse. Le gèneSéquencerUne cohorte de 327 patients inclus dans un programme d’ICSI. Tous les patients ont donné leur consentement écrit et éclairé à la participation de cette étude. Cent patients n’avaient pas d’altération de la spermatogenèse et 227 avaient une déficience.Les quatre exons deBien que 9 SNPs aient été identifiés, il n’y avait pas de différence de fréquences significatives entre les patients ayant une altération, ou non de la spermatogenèse. Deux insertions ont été identifiées: une insertion de 21 nucléotides retrouvées dans les deux groupes et une insertion d’une guanine (induisant un codon stop prématuré) chez deux patients présentant une altération de la spermatogenèse.

DNA undermethylation is a characteristic feature of ICF syndrome and has been implicated in the formation of the juxtacentromeric chromosomal abnormalities of this rare syndrome. We have previously shown that in female ICF patients the inactive X chromosome (Xi) is also undermethylated. This result was unexpected since female ICF patients are not more severely affected than male patients. Here we show that CpG island methylation is abnormal in some ICF patients but in other ICF patients, the difference in methylation pattern between Xi and Xa (active X) is maintained. The consequences of Xi undermethylation on gene expression were investigated by enzyme assays. They showed that significant gene expression did not correlate with CpG island methylation status. The widespread Xi undermethylation does not affect overall Xi replication timing and does not prevent Barr body formation suggesting that a normal methylation pattern is not required for normal chromatin organization of Xi. Molecular investigation of some X-chromosome intron regions showed that the methylation changes in ICF female patients extend to non CpG islands sequences. Our results suggest that the genetic alteration of DNA methylation in ICF syndrome has little consequence on X chromosome gene expression and chromatin organization.

We report a case of prenatal diagnosis of trisomy 20p resulting from a maternal pericentric inversion. The diagnosis was confirmed on both chorionic villi and amniotic cells. This case underlines the fact that prenatal ultrasound diagnosis of this structural anomaly is difficult. The only early sonographic feature was increased nuchal translucency.

5p deletion syndrome commonly known as cri du chat is well described in affected neonates with catlike cry and hypotonia. Karyotyping will usually show a deletion of the short arm of one chromosome 5 with variable breakpoints. Only a few cases have been reported prenatally, and the fetal form of the syndrome has not been clearly individualised. We report a new case of 5p deletion syndrome diagnosed prenatally in association with Dandy-Walker syndrome and agenesis of the corpus callosum. Other brain anomalies have been reported previously, but this unusual association suggests the use of a specific probe in the investigation of these malformations.

Resume Objectifs Tenter d’analyser le vecu de couples ayant vecu des fausses couches a repetition, en repondant aux questions suivantes : que nous apprennent ces hommes et ces femmes vivant des fausses couches a repetition ? Que pouvons-nous mieux apprehender sur le couple lui-meme ? Patients et methodes Un questionnaire de personnalite approfondi, le MMPI-2, a ete propose a 50 couples ayant vecu des fausses couches repetees. Resultats Grâce a une classification hierarchique, des profils differents apparaissent tant dans le groupe des femmes que dans la groupe des hommes, mettant en evidence une somatisation de la souffrance psychologique ; elle montre aussi l’existence de profils de personnalite particulierement defensifs temoignant d’une restriction des affects chez beaucoup d’hommes de ces couples vivant des echecs repetes de procreation. Cette restriction emotionnelle peut etre facteur de souffrance supplementaire pour leurs compagnes et pour eux-memes. Discussion et conclusion On peut conclure que dans ces conditions, la prise en charge medicale et/ou psychologique devra inclure les deux membres du couple pour favoriser un meilleur ajustement marital et permettre au couple d’aborder la prochaine grossesse, toujours vecue dans l’angoisse d’un nouvel echec, avec une plus grande serenite. Quelques hypotheses etiologiques pourront etre evoquees.

Summary The objective of this study was to assess genome-wide DNA methylation in testicular tissue from azoospermic patients. A total of 94 azoospermic patients were recruited and classified into three groups: 29 patients presented obstructive azoospermia (OA), 26 displayed non-obstructive azoospermia (NOA) and successful retrieval of spermatozoa by testicular sperm extraction (TESE+) and 39 displayed NOA and failure to retrieve spermatozoa by TESE (TESE−). An Illumina Infinium Human Methylation27 BeadChip DNA methylation array was used to establish a testicular DNA methylation pattern for each type of azoospermic patient. The OA and NOA groups were compared in terms of the relative M-value (the log2 ratio between methylated and non-methylated probe intensities) for each CpG site. We observed significantly different DNA methylation profiles for the NOA and OA groups, with differences at over 9000 of the 27 578 CpG sites; 212 CpG sites had a relative M-value >3. The results highlighted 14 testis-specific genes. Patient clustering with respect to these 212 CpG sites corresponded closely to the clinical classification. The DNA methylation patterns showed that in the NOA group, 78 of the 212 CpG sites were hypomethylated and 134 were hypermethylated (relative to the OA group). On the basis of these DNA methylation profiles, azoospermic patients could be classified as OA or NOA by considering the 212 CpG sites with the greatest methylation differences. Furthermore, we identified genes that may provide insight into the mechanism of idiopathic NOA.

Small supernumerary marker chromosomes (sSMCs) are structurally abnormal chromosomes that cannot be characterized by karyotype. In many prenatal cases of de novo sSMC, the outcome of pregnancy is difficult to predict because the euchromatin content is unclear. This study aimed to determine the presence or absence of euchromatin material of 39 de novo prenatally ascertained sSMC by array-comparative genomic hybridization (array-CGH) or single nucleotide polymorphism (SNP) array. Cases were prospectively ascertained from the study of 65,000 prenatal samples [0.060%; 95% confidence interval (CI), 0.042-0.082]. Array-CGH showed that 22 markers were derived from non-acrocentric markers (56.4%) and 7 from acrocentic markers (18%). The 10 additional cases remained unidentified (25.6%), but 7 of 10 could be further identified using fluorescence in situ hybridization; 69% of de novo sSMC contained euchromatin material, 95.4% of which for non-acrocentric markers. Some sSMC containing euchromatin had a normal phenotype (31% for non-acrocentric and 75% for acrocentric markers). Statistical differences between normal and abnormal phenotypes were shown for the size of the euchromatin material (more or less than 1 Mb, p = 0.0006) and number of genes (more or less than 10, p = 0.0009). This study is the largest to date and shows the utility of array-CGH or SNP array in the detection and characterization of de novo sSMC in a prenatal context.

Successful embryo implantation depends on trophoblast proliferation, migration and, lastly, invasion of the endometrium (to anchor the trophoblast to the uterus). This invasion is mediated by locally produced soluble factors. Of these, vascular endothelial growth factor (VEGF) is the best characterized regulator of angiogenesis. Here, we investigate the association between the VEGF + 405 C/G genotype and the recurrence of embryo implantation failure in women undergoing in vitro fertilization (IVF) program with intracytoplasmic sperm injection (ICSI). Forty women with recurrent implantation failure defined by absence of pregnancy after transfer of more than 10 embryos and 131 women control, with at least one live birth after the transfer of fewer than 10 embryos were included. Genomic DNA was analysed with an allele-specific polymerase chain reaction and a Chi-2 test was used to compare the respective VEGF + 405 C/G genotype frequencies in cases and controls. The frequency of the VEGF +405C/C genotype was higher in women with recurrent implantation failure after ICSI-embryo transfer than in controls (17.5 % and 5.3 %, respectively, p = 0.01). The VEGF +405 G/C polymorphism may influence embryo implantation and VEGF + 405 C/C genotype may predispose to recurrent implantation failure after ICSI-ET.

Try to analyse the experience of couples undergoing repeated miscarriages by answering the following questions: what can we learn from these men and women who suffered from repeated miscarriages?A thorough personality questionnaire, the MMPI-2, presented to 50 couples who have had repeated miscarriages.Through a hierarchical classification, different profiles appear in the men's group as well as in the women's group, revealing a somatization of psychological suffering. It is also revealing acute defensive personality profiles showing restricted affects in a lot of these men whose partners have suffered from multiple procreation failures. Such a narrower range of emotions can be a cause of additional pain for their partner and for themselves.We can therefore establish that, in these circumstances, the medical and/or psychological treatment should include both couple members to improve the marital adjustment and ease the couple towards another pregnancy which is always apprehended with the fear of another failure. A few etiological hypothesis may be evoked.

Introduction Obtaining an adequate number of high-quality oocytes is a major challenge in controlled ovarian hyperstimulation (COH). To date, a range of hormonal and clinical parameters have been used to optimize COH but none have significant predictive value. This variability could be due to the genetic predispositions of single-nucleotide polymorphisms (SNPs). Here, we assessed the individual and combined impacts of thirteen SNPs that reportedly influence the outcome of in vitro fertilisation (IVF) on the ovarian response to rFSH stimulation for patients undergoing intracytoplasmic sperm injection program (ICSI). Results Univariate analysis revealed that only FSHR, ESR2 and p53 SNPs influenced the number of mature oocytes. The association was statistically significant for FSHR (p=0.0047) and ESR2 (0.0017) in the overall study population and for FSHR (p=0.0009) and p53 (p=0.0048) in subgroup that was more homogeneous in terms of clinical variables. After Bonferroni correction and a multivariate analysis, only the differences for FSHR and ESR2 polymorphisms were still statistically significant. In a multilocus analysis, only the FSHR and AMH SNP combination significantly influenced oocyte numbers in both population (pA), ESR1(−397T>C), BMP15(−9C>G), MTHFR1(677C>T), MTHFR2(1298A>C), HLA-G(−725C>G), VEGF(+405G>C), TNFα(−308A>G), AMHR(−482 A>G), PAI-1 (4 G/5 G), multiplex PCR assay was designed to genotype women undergoing ICSI program. We analyzed the overall study population (n=427) and a subgroup with homogeneous characteristics (n=112).

Using prenatal BACs-on-Beads technology, the first prenatal case of Williams-Beuren syndrome (WBS) was diagnosed. In light of this result, an ultrasound scan confirmed the presence of well-characterized features of WBS. This case report emphasizes the fact that new genomic technologies will generate prenatal information and provide helpful additional information.

While the incidence of predisposition to aneuploidy in the oocyte increases with age, there is also evidence of increased incidence in young women with recurrent miscarriage, recurrent aneuploidy, or recurrent implantation failure after in vitro fertilization. There is evidence from mouse models and from observations in humans that follicle-stimulating hormone (FSH) probably has a direct or indirect effect on the occurrence of oocyte aneuploidy. It seems that increased endogenous or exogenous FSH could induce meiotic disruption. Although the effect of FSH may explain the age-related increase in aneuploidy rate, many questions remain regarding young women, even if their FSH level is sometimes increased. Disruption of meiotic gene expression caused by exposure to environmental contaminants or by gene defects could also predispose to oocyte aneuploidy. Such abnormalities could impact on the oocyte pool, recombination and synapsis during fetal life, or oocyte growth.

Le diagnostic preconceptionnel (DPC), decrit pour la premiere fois en 1990 (1), est considere, par beaucoup, comme une alternative au diagnostic preimplantatoire (DPI) et au screening preimplantatoire des aneuploidies (SPI) (2). Il s’agit non pas du diagnostic d’une anomalie embryonnaire, mais du diagnostic indirect, sur globule polaire (GP), de la constitution genetique de l’ovocyte. A l’oppose du DPI, fait sur blastomere (3), le diagnostic preconceptionnel s’eff ectue sur le premier GP avant la fecondation, voire sur les deux globules polaires apres la fecondation.

L’infertilite peut etre consideree comme une maladie multifactorielle qui touche aujourd’hui plus de 15% des couples. Dans un tiers des cas, une cause purement feminine est retrouvee, dans 21% des cas une cause uniquement masculine, et une cause a la fois feminine et masculine est objectivee dans 39% des cas. Enfin, dans 7% des cas, aucune cause n’est trouvee (infertilite inexpliquee dite idiopathique) (1).

Molecular cytogenetic techniques on uncultured prenatal samples are the sole tests applied in some countries in cases with advanced maternal age (AMA) or increased risk after prenatal screening. Moreover, there is a trend to perform invasive prenatal diagnosis (PD) during the first trimester before ultrasound manifestations, so new rapid and reliable assays are necessary to investigate microdeletions not detectable with the conventional karyotype. We report the validation study of the prenatal bacterial artificial chromosomes-on-Beads™ (BoBs™ ; CE-IVD), a bead-based multiplex assay detecting chromosomes 13, 18, 21, X/Y aneuploidies and nine microdeletion regions having an overall detection rate of 1/1700.We retrospectively studied 408 selected samples and prospectively tested 212 consecutive samples ascertained for conventional karyotyping.We did not find false-positive results. Triploidies were not detected. Maternal cell contamination of male samples up to 90% was unmasked inspecting gonosome profiles. Mosaic conditions at 20 to 30% were revealed. Failures were due to low amount of DNA.Prenatal BoBs™ is a robust technology for the investigation of fetuses with normal karyotype with or without sonographic abnormalities. Running in parallel with the karyotype analysis, it can be proposed instead of rapid FISH or QF-PCR providing rapid results on common aneuploidies and additional information regarding the microdeletion syndromes.

Array-CGH emergence allowed important diagnosis progress, and a better care of patients in postnatal. So, there is a great temptation to use it also in prenatal diagnosis. The point of view objective is to make a rapid overview of cytogenetic diagnosis evolution during the last 50 years, and to show all questions raised by the use of array-CGH, and problems that could arise in prenatal diagnosis. While array-CGH just comes in genetic laboratories, new diagnosis approaches emerged like whole genome sequencing or non-invasive prenatal diagnosis. The 2nd objective will be to review all these techniques for a probably close future.

Preconception diagnosis requires first polar body biopsy. When the hole in the zona pellucida is made with a laser beam, heat propagation could, like the biopsy itself, be deleterious. Our aim was to evaluate the effect of this technique on human in vitro matured oocyte and embryo development.One hunded fifty five retrieved immature oocytes from 75 women, matured in vitro, were distributed in 3 groups: 50 oocytes in a control group, without laser drilling and first polar body biopsy, 52 oocytes in a group with only laser drilling, and 53 oocytes in a group with both laser drilling and first polar body biopsy. Safety was evaluated using four criteria: [1] oocyte lysis rate, [2] oocyte activation rate, [3] oocyte development after calcium ionophore treatment, [4] and embryo chromosome breakage incidence after Tarkowski preparation.No difference in the four criteria was observed between the 3 oocyte groups.We did not find evidence of deleterious effect of laser drilling and first polar body biopsy on in vitro matured oocytes, according to our criteria.