Metformin exposure affects human and mouse fetal testicular cells.

BACKGROUND: Metformin is a drug used in the treatment of diabetes and of some disorders related to insulin resistance, such as polycystic ovary syndrome. Gestational diabetes can cause complications for both mother and child, and some studies have shown a beneficial effect of metformin during pregnancy without an increase in perinatal complications. However, the effects on the gonads have not been properly studied. Here we investigated the effect of metformin administered during pregnancy on the development and function of the fetal testis. METHODS: A dual approach in vitro and in vivo using human and mouse models was chosen. Cultures of human and murine organotypic testes were made and in vivo embryonic testes were analysed after oral administration of metformin to pregnant mice. RESULTS: In human and mouse organotypic cultures in vitro, metformin decreased testosterone secretion and mRNA expression of the main factors involved in steroid production. In vitro, the lowest observed effect concentration (LOEC) on testosterone secretion was 50 µM in human, whereas it was 500 µM in mouse testis. Lactate secretion was increased in both human and mouse organotypic cultures with the same LOEC at 500 µM as observed in other cell culture models after metformin stimulation. In vivo administration of metformin to pregnant mice reduced the testicular size of the fetal and neonatal testes exposed to metformin during intrauterine life. Although the number of germ cells was not affected by the metformin treatment, the number of Sertoli cells, the nurse cells of germ cells, was slightly yet significantly reduced in both periods (fetal period: P = 0.007; neonatal period: P = 0.03). The Leydig cell population, which produces androgens, and the testosterone content were diminished only in the fetal period at 16 days post-coitum. CONCLUSIONS: This study showed a potentially harmful effect of metformin treatment on the development of the fetal testis and should encourage future human epidemiological studies.

Effect of mono-(2-ethylhexyl) phthalate on human and mouse fetal testis: In vitro and in vivo approaches.

The present study was conducted to determine whether exposure to the mono-(2-ethylhexyl) phthalate (MEHP) represents a genuine threat to male human reproductive function. To this aim, we investigated the effects on human male fetal germ cells of a 10⁻5 M exposure. This dose is slightly above the mean concentrations found in human fetal cord blood samples by biomonitoring studies. The in vitro experimental approach was further validated for phthalate toxicity assessment by comparing the effects of in vitro and in vivo exposure in mouse testes. Human fetal testes were recovered during the first trimester (7-12 weeks) of gestation and cultured in the presence or not of 10⁻5 M MEHP for three days. Apoptosis was quantified by measuring the percentage of Caspase-3 positive germ cells. The concentration of phthalate reaching the fetal gonads was determined by radioactivity measurements, after incubations with ¹4C-MEHP. A 10⁻5 M exposure significantly increased the rate of apoptosis in human male fetal germ cells. The intratesticular MEHP concentration measured corresponded to the concentration added in vitro to the culture medium. Furthermore, a comparable effect on germ cell apoptosis in mouse fetal testes was induced both in vitro and in vivo. This study suggests that this 10⁻5 M exposure is sufficient to induce changes to the in vivo development of the human fetal male germ cells.

Serum anti-Müllerian hormone levels are negatively related to Follicular Output RaTe (FORT) in normo-cycling women undergoing controlled ovarian hyperstimulation.

BACKGROUND: Since in rodents anti-Müllerian hormone (AMH) has been shown to inhibit antral follicle responsiveness to FSH, we aimed at verifying whether a relationship exists between serum AMH levels and antral follicle responsiveness to exogenous FSH in normo-cycling women. METHODS: Serum AMH, estradiol (E(2)) and FSH levels were prospectively measured on cycle day 3 in patients undergoing controlled ovarian hyperstimulation (COH) with a time-release GnRH agonist and standardized FSH doses. In 162 patients, follicles were counted after pituitary suppression and before FSH administration (baseline; small antral follicles; 3-8 mm), and on the day of hCG (dhCG; pre-ovulatory follicles; 16-22 mm). Antral follicle responsiveness to FSH was estimated by the Follicular Output RaTe (FORT), determined by the ratio pre-ovulatory follicle count on dhCG × 100/small antral follicle count at baseline. RESULTS: Serum AMH levels were positively correlated with the number of small antral follicles at baseline (r = 0.59; P < 0.0001) and pre-ovulatory follicles on dhCG (r = 0.17; P < 0.04). Overall, FORT was 47.5 ± 1.4% and failed to be influenced by the woman's age, BMI or basal E(2) and FSH level. Conversely, multiple regression analysis showed that FORT was negatively correlated with AMH levels (r = -0.30; P < 0.001), irrespective of duration of COH and total FSH dose. CONCLUSIONS: The percentage of follicles that effectively respond to FSH by reaching pre-ovulatory maturation is negatively and independently related to serum AMH levels. Although the mechanisms underlying this finding remain unclear, it is in keeping with the hypothesis that AMH inhibits follicle sensitivity to FSH.

Meiosis initiation in the human ovary requires intrinsic retinoic acid synthesis.

BACKGROUND: The initiation of meiosis is crucial to fertility. While extensive studies in rodents have enhanced our understanding of this process, studies in human fetal ovary are lacking. METHODS: We used RT-PCR and immunohistochemistry to investigate expression of meiotic factors in human fetal ovaries from 6 to 15 weeks post fertilization (wpf) and developed an organ culture model to study the initiation of human meiosis. RESULTS: We observed the first meiotic cells at 11 wpf, when STRA8, SPO11 and DMC1 are first expressed. In culture, meiosis initiation is observed in 10 and 11 wpf ovaries and meiosis is maintained by addition of fetal calf serum. Meiosis is stimulated, compared with control, by retinoic acid (RA) (P < 0.05). No major change occurred in mRNA for CYP26B1, the RA-degrading enzyme proposed to control the timing of meiosis in mice. We did, however, observe increased mRNA levels for ALDH1A1 in human ovary when meiosis began, and evidence for a requirement to synthesize RA and thus sustain meiosis. Indeed, ALDH inhibition by citral prevented the appearance of meiotic cells. Finally, 8 wpf ovaries (and earlier stages) were unable to initiate meiosis whatever the length of culture, even in the presence of RA and serum. However, when human germ cells from 8 wpf ovaries were placed in a mouse ovarian environment, some did initiate meiosis. CONCLUSIONS: Our data indicate that meiosis initiation in the human ovary relies partially on RA, but that the progression and regulation of this process appears to differ in many aspects from that described in mice.

Using Z-scores to compare biometry data obtained during prenatal ultrasound screening by midwives and physicians.

OBJECTIVE: To compare retrospectively the distribution of foetal biometry data as measured by midwives and physicians during second and third trimester screening of an unselected population of pregnant women. METHODS: Standard measurements of biparietal diameter (BPD), head circumference (HC), abdominal circumference (AC), and femur length (FL) were performed by four midwives and ten physicians at 20 to 24 weeks of gestation and at 30 to 34 weeks of gestation as part of routine ultrasound examinations over a 26-month period (Jan. 2005-Mar. 2007). All measurements were converted into Z-scores using different prediction equations. The reference chart best fitting our practice was determined for each fetal parameter (French College of Sonographers for BPD, Chitty et al. for HC and FL, Snidjers and Nicolaides for AC). The means and SDs of the Z-score distributions for data collected by midwives and physicians were compared using Student's t-test for means and the Fisher-Snedecor test for SDs. RESULTS: We retrieved 1566 and 1631 measurements made by midwives and physicians respectively between 20 and 24 weeks of gestation, and 1710 and 1578 measurements made by midwives and physicians respectively between 30 and 34 weeks of gestation. Mean values recorded by midwives were significantly closer to 0 (p < 0.05) for many foetal parameters. SD values were also significantly lower and were below 1. CONCLUSION: In this study, midwives have a greater tendency than physicians to normalize biometry data. Such normalization may hamper the sensitivity of routine ultrasound screening for abnormal foetal growth.

Oocyte in-vitro maturation: BCL2 mRNA content in cumulus cells reflects oocyte competency.

BCL2-associated X protein (BAX) and B-cell leukaemia/lymphoma gene-2 (BCL2), which are, respectively, pro- and anti-apoptotic proteins of the BCL2 gene family, participate in the mitochondria-dependent apoptosis pathway. A correlation between low incidence of apoptosis in cumulus cells and oocyte maturation has previously been suggested in ovarian stimulation. However, little is known in unprimed ovaries. These authors have investigated whether BAX and BCL2 expression in cumulus cells affects the competency of in-vitro matured oocytes. We have studied 100 cumulus-oocyte-complexes (COC) recovered from unprimed ovaries of 13 women diagnosed with polycystic ovary syndrome (PCOS) and undergoing in-vitro maturation (IVM) with their informed consent. COC were matured for 24 h in a specific maturation medium and the cumulus was stripped from the oocyte. BAX and BCL2 mRNA content was measured in each COC using real-time polymerase chain reaction. We found that BCL2 mRNA expression was significantly higher in cumulus cells associated with mature oocytes than those associated with immature oocytes while BAX mRNA concentrations did not vary in cumulus cells. Regarding fertilization, higher BCL2 mRNA content was found in cumulus cells enclosing fertilized oocytes (0.140 versus 0.075; P = 0.03). These results suggest that BCL2 expression is strongly associated with the ability of oocytes to complete nuclear maturation and to be fertilized.

Characterization of human PGD blastocysts with unbalanced chromosomal translocations and human embryonic stem cell line derivation?

Novel embryonic stem cell lines derived from embryos carrying structural chromosomal abnormalities obtained after preimplantation genetic diagnosis (PGD) are of interest to study in terms of the influence of abnormalities on further development. A total of 22 unbalanced blastocysts obtained after PGD were analysed for structural chromosomal defects. Morphological description and chromosomal status of these blastocysts was established and they were used to derive human embryonic stem cell (ESC) lines. An outgrowth of cells was observed for six blastocysts (6/22; 27%). For two blastocysts, the exact morphology was unknown since they were at early stage, and for four blastocysts, the inner cell mass was clearly visible. Fifteen blastocysts carried an unbalanced chromosomal defect linked to a reciprocal translocation, resulting in a positive outgrowth of cells for five blastocysts. One human ESC line was obtained from a blastocyst carrying a partial chromosome-21 monosomy and a partial chromosome-1 trisomy. Six blastocysts carried an unbalanced chromosomal defect linked to a Robertsonian translocation, and one showed a positive outgrowth of cells. One blastocyst carried an unbalanced chromosomal defect linked to an insertion and no outgrowth was observed. The efficiency of deriving human ESC lines with constitutional chromosomal disorders was low and probably depends on the initial morphological aspect of the blastocysts and/or the type of the chromosomal disorders.

A 2-year study on cytomegalovirus infection during pregnancy in a French hospital.

OBJECTIVES: To evaluate the proportion of pregnant women agreeing to cytomegalovirus (CMV) serologic screening. To collect data on CMV infection during pregnancy. DESIGN: Prospective study. SETTING: During two years, all pregnant women were informed on CMV infection. If the patient agreed, serological testing was performed around 12 weeks of gestation (WG) and, if negative, redone around 36 WG. POPULATION: Four thousand two hundred and eighty-seven pregnant women followed from 12 weeks to delivery. METHODS: If the first CMV serologic test was negative, detailed hygiene information was given to the parents. Diagnosis of primary infection was based on the detection of CMV-G, CMV-M and low CMV-G avidity index. When maternal infection was confirmed, diagnosis of CMV congenital infection was done in the newborns by urine culture within the three days following birth. Crude infection-rate data consisted of the number of CMV infection cases and person-time units for both exposed to hygiene CMV information (12 to 36 WG) and unexposed pregnant women (first 12 WG). MAIN OUTCOME MEASURES: Rate of CMV seropositive and seronegative women. Rate of women agreeing for screening. Rate of primary infection. Rate of seroconversion. Number of CMV-infected newborns. RESULTS: Among the 4287 women followed, 3792 were either seronegative or with an unknown immune status. 96.7% out of them agreed for screening. 53.2% were initially CMV-specific IgG negative. Primary infection was detected in nine women between 0 and 12 WG (0.46%) and seroconversion was diagnosed in five women between 12 and 36 WG (0.26%) (mid P = 0.02, 95% CI [1.07-13.6]). CONCLUSIONS: If clear information on CMV infection during pregnancy is given, patients frequently agree to screening. The rate of seroconversion after information, observed in this study, is low after counselling.

Clinical application of fetal urine production rate in unexplained polyhydramnios.

OBJECTIVE: To evaluate the clinical use of hourly fetal urine production rate (HFUPR) in polyhydramnios. METHODS: This was a retrospective review of 33 singleton pregnancies with polyhydramnios, 30 of them unexplained and three due to gastrointestinal atresia. HFUPR was estimated using three-dimensional ultrasound and was compared with recently established nomograms. Abnormal midterm outcome, defined as diagnosis or persistence of pathology after the neonatal period until the age of 2 years, was analyzed according to prenatal HFUPR measurements and other polyhydramnios characteristics. RESULTS: Seventeen of the 30 fetuses with unexplained polyhydramnios had an HFUPR above the 95(th) centile, and five (29.4%) of them developed midterm disorders. None of the 13 with normal HFUPR developed midterm disorders. The HFUPR was 1.9 (SD, 0.7) multiples of the median (MoM) in fetuses with an adverse childhood outcome and 1.4 (SD, 1.2) in fetuses with normal childhood outcome (P = 0.34). In the three fetuses with gastrointestinal atresia, the HFUPR was significantly lower than in those with unexplained polyhydramnios (P = 0.003). CONCLUSION: HFUPR was associated with the mechanism of polyhydramnios but failed to help in the prognosis of unexplained polyhydramnios because of lack of power. Children with prenatal unexplained polyhydramnios and HFUPR above the 95(th) centile should nevertheless receive detailed pediatric follow-up.

[The role of ultrasound examination to evaluate ovarian reserve of infertile patients]. / Place de l'échographie pour le bilan de la réserve ovarienne dans le cadre d'une infertilité

The accurate assessment of ovarian reserve is an essential step before the treatment of infertile couples. Ovarian reserve could either be evaluated through clinical or biological parameters, but ultrasound plays a remarkable role, since it permits the direct visualization and count of ovarian antral follicles. Nevertheless, the available literature data are conflicting about the real sensibility and specificity of this method to predict the exact number of retrieved oocytes or the occurrence of pregnancy after a procedure for medical assisted reproduction. New technologies have been developed, as the 3D-ultrasound, who does not ameliorate ultrasound accuracy, but permits the reduction of the time necessary for patient's examination and virtually eliminates inter- and intra-observer bias. The place of the ovarian artery flow evaluation by Doppler remains unclear and need more studies.

[Is the stability of results in IVF possible?]. / La stabilité des résultats en fécondation in vitro est-elle possible ?

OBJECTIVES: To evaluate quality control impact, in assisted reproductive medecine, in view of the stability of results. MATERIALS AND METHODS: Prospective collection of all IVF cycles at the center of AMP at Antoine-Béclère hospital (Clamart), as well as pregnancy outcomes, from 2002 till 2006. RESULTS: 44.3% in the clinical pregnancy rate and 38.0% in the delivery rate per oocyte retrieval. These rates are stable during the five years studied. The rate of multiple pregnancy is of 30.7%, decreasing, with the decrease in number of transferred embryos. CONCLUSION: Results are stable over five years duration. Quality control seems to be necessary.

[Management of antenatal fetal abdominal tumors. Clues for the diagnosis of a congenital mesoblastic nephroma]. / Conduite à tenir obstétricopédiatrique devant la découverte anténatale d'une masse hyperéchogène rétropéritonéale. Arguments en faveur d'un néphrome mésoblastique congénital.

The prenatal diagnosis of abdominal mass poses the problem of its origin. Renal tumors are rarer than neuroblastoma but they are most often congenital mesoblastic nephroma. The congenital mesoblastic nephroma has a good forecast in spite of a sonographic impressive aspect. MRI can help to locate tumor but cannot tell difference between the different kinds of renal tumor. Prenatal forecast is especially linked with hydramnios and hydrops fetalis. Histolological study of the tumor is important for the prognosis. Two morphological subtypes are currently distinguished: the classic type with a good forecast and the atypical or cellular type. Distant metastases have been related only to the cellular form but especially in infants aged more than 3 months and never in the newborns. The diagnosis of the tumor does not change the mode of delivery except in case of an important volume. Complications are searched during the first days of life: hypertension, hypercalcemia, vomiting, hyperreninemia. Radical nephrectomy is performed after the end of the first week. In case of a classic form, the healing is always obtained. In case of cellular form, distant metastases are searched. In any rate, the follow-up is recommended until the end of the growth.

Stability of the m.8993T->G mtDNA mutation load during human embryofetal development has implications for the feasibility of prenatal diagnosis in NARP syndrome.

BACKGROUND: Mitochondrial DNA (mtDNA) mutations cause a wide range of serious genetic diseases with maternal inheritance. Because of the high transmission risk and the absence of therapy in these disorders, at-risk couples often ask for prenatal diagnosis (PND). However, because heteroplasmy load (coexistence of mutant and wild-type mtDNA) may vary among tissues and with time, the possibility that a single fetal sample may not reflect the whole neonate impedes prenatal diagnosis of mtDNA diseases. METHODS: We performed 13 prenatal diagnoses for the NARP (neurogenic weakness, ataxia, retinitis pigmentosa) m.8993T-->G mtDNA mutation (p.Leu156Arg) in the ATP synthase subunit 6 gene. Analyses were performed on chorionic villous (CVS) and/or amniocyte samples carried out at various stages of pregnancy, using a method enabling quantification of low DNA amounts. RESULTS: Maternal mutant loads ranged from 0 to 75% in blood and had no predictive value for the fetus status, except for women with no detectable mutant DNA, whose fetuses were consistently mutation-free. In 8/13 PND, mutant load was <30%. These children are healthy at 2-7 years of age. In 5/13 PND, mutant load ranged from 65 to 100%, and parents preferred to terminate the pregnancies (15-22 weeks of gestation). Single-cell analysis of 20 trophoblastic cells and 21 amniocytes isolated from two affected fetuses found an average mutant load close to the overall CVS and amniocyte mutant load, despite striking intercellular variation. The m.8993T-->G mutant loads, assessed in 7, 17, 11, and 5 different tissues from 4 terminations, respectively, were identical in all tissues from a given individual (mean (SD) 78 (1.2)%, 91 (0.7)%, 74 (2)%, and 63 (1.6)% for the 4 fetuses, respectively). CONCLUSIONS: Our results indicate that the placental/amniotic mutant loads do reflect the NARP mutant mtDNA load in the whole fetus, even when the sample amount is small, and suggest that heteroplasmy level remains stable during pregnancy, at least after 10 weeks of gestation. Although these data establish the feasibility of PND for this mutation, assessing more precisely the correlation between mutant load and disease severity should further help in interpreting PND results.

Prenatal diagnosis of a patent urachus cyst with the use of 2D, 3D, 4D ultrasound and fetal magnetic resonance imaging.

Patent urachus cyst is a rare umbilical anomaly, which is poorly detected prenatally and frequently confounded with pseudo bladder exstrophy or omphalocele. A 27-year-old woman was referred to our prenatal diagnosis centre at 18 weeks of gestation after diagnosis of a megabladder and 2 umbilical cord cysts. Subsequent 2D, 3D and 4D ultrasound examinations and fetal magnetic resonance imaging (MRI) revealed a typical umbilical cyst and an extra-abdominal cyst, communicating with the vertex of the fetal bladder through a small channel that increased in size when the fetus voided urine. Termination of pregnancy occured at 31 weeks because of associated cerebral septal agenesis, and autopsy confirmed the prenatal diagnosis of urachus cyst. Few cases of urachus cyst diagnosed prenatally are reported in literature, but none were associated with other extra-abdominal disorders and none used 3D, 4D and fetal MRI. Our case illustrated the efficiency in prenatal diagnosis of 3D and 4D ultrasound examinations. This could help pediatrician surgeons to explain to a couple about neonatal surgical repair and plastic reconstruction in the prenatal period.

[Cervical cerclage in 2008]. / Le cerclage du col utérin en 2008.

Cervical cerclage is a common surgical technique that has been used for more than 50 years to prevent preterm deliveries and in the management of a threatened second trimester loss. However, it remains one of the most controversial interventions in obstetrics and this is probably due to difficulties in diagnosing cervical insufficiency, which is based on a history of recurrent second trimester loss or early preterm delivery following painless cervical dilatation in the absence of contractions or bleeding. This article reviews in 2008 the current literature regarding the efficacy of elective cerclage, ultrasound-indicated cerclage, emergency cerclage, and cervico-isthmic cerclage for singletons and multiple pregnancies.

[Development and regulations of testicular functions in the human foetus]. / Ontogenèse et régulations des fonctions testiculaires chez le foetus humain.

Two major functions are assumed by the testis: the production of male gametes (that is, spermatozoa) and the production of steroid hormones. Both two functions are established during fetal life and are essential to the adult fertility and the masculinization of the internal tract and genitalia. For many years, our laboratory has been interested in the ontogeny of those two functions in rodents and, since 2003, in collaboration with gynecology and obstetrics service of professor R. Frydman in Antoine-Béclère hospital, we have studied them in human. The first aim of this work was to improve the global knowledge of the human fetal testis development by using both our experimental data and the literature. Then, we focused on the different defects that can occur during the fetal testis development. Indeed, male reproductive abnormalities have been steadily increasing since the last decades and are thought to be related to the concomitant increase of the concentration of contaminants and particularly of endocrine disruptors in the environment. Thus, we decided to study the effect of endocrine disruptors on human fetal testis and, more particularly, the effect of phthalates, by using an organ culture system developed for human. In contrast to the data obtained in rat, mono (ethylhexyl)-phthalate (MEHP), an active metabolite of the most widespread phthalate in the environment, does not disturb the steroidogenic function. On the other hand, it has a negative effect on the male germ cells number. This study is the first experimental demonstration of a negative effect of phthalates directly on human fetal testis.

[Teaching physician-patient relationship using role-play in obstetrics and gynecology]. / Enseigner la relation médecin-malade en gynécologie obstétrique par des séances de jeu de rôle.

AIM: The aim of this study was to report our experience concerning a role-playing approach for the teaching of physician-patient relationship. METHODS: The role-playing two-day course was designed to be highly interactive for a small group (ten participants). Opinions were gathered by an anonymous structured questionnaire (ten questions) completed by the participants and focused on their view of the play role and the physician-patient relationship. RESULTS: The opinions of the participants were highly positive; all appreciated the courses. The strong emotional involvement was considered beneficial for all of them, sharing emotional aspects of the profession, and usefulness in clarifying opinions on the physician-patient relationship. CONCLUSION: The positive opinions recorded during this experience suggest the benefit of implementing non-conventional educational approaches, such as role-play, to highlight the relative importance of physician-patient relationship in obstetrics and gynecology.

[Amniocentesis practice assessment of the south-west Francilian network during 2003]. / Evaluation de la pratique de l'amniocentèse sur le réseau sud-ouest francilien pendant l'année 2003.

OBJECTIVES: To assess amniocentesis practice of a network during one year. MATERIALS AND METHODS: In a retrospective multicenter study of the south-west Francilian network, we have enrolled 2400 patients who underwent amniocentesis, from 1 January 2003 to 31 December 2003. RESULTS: The rate of amniocentesis was 9.5%. The most frequent indication was a positive maternal serum screening test result (44%). The rate of global fetal losses (spontaneous miscarriage and intrauterine death) was 1.4%. Once the fetuses with aneuploidy and lethal pathology excluded, the rate of global fetal losses potentially related to amniocentesis was 1.21%. The rate of premature rupture of the membranes was 1.12% and prematurity affected 6.5% of the living births. CONCLUSION: Our study has highlighted several practices of amniocentesis within the network. Overall, amniocentesis potentially induces 1.12% of fetal losses. Screening tests are currently used sequentially, which leads to an increase number of amniocentesis and to an increase number of losses of "a priori" healthy fetus. Only the use of a combined screening could lower the frequency of amniocentesis without decreasing the detection rate of chromosome abnormalities.

[Evaluation of the third trimester amniocentesis for fetal karyotyping in women with fear of pregnancy loss]. / Evaluation de l'amniocentèse au troisième trimestre pour le dépistage des anomalies chromosomiques chez les patientes n'acceptant pas le risque de perte foetale.

OBJECTIVES: The aim of this study is to determine the complications of third trimester amniocentesis for fetal karyotyping among women unwilling to accept the fetal loss risks of second trimester amniocentesis. MATERIALS AND METHODS: A retrospective study was carried out from January 1998 to December 2006 of 182 singleton pregnancies that underwent a late amniocentesis (after 32 weeks) for fetal karyotyping. The indications were integrated risk (maternal age, first trimester nuchal translucency, second trimester maternal serum markers) over 1/250 (n=68), isolated maternal age over 38 years (n=51), isolated abnormal second trimester biochemical markers (n=34), history of personal or familial a chromosomal abnormality (n=21) or maternal choice (n=8). Presence of fetal abnormalities at ultrasound or context of viral or parasitologic seroconversion as well as multiple pregnancies were considered as non-inclusion criteria. RESULTS: Median maternal age and gestational age at sampling were 39 years (range 23-48) and 32.4 weeks (29.5-37.6). Median interval between amniocentesis and definitive results of amniocentesis on the one hand, and delivery on the on the hand were 15 days (7-42) and 47 days (8-69), respectively. There were no chromosomal abnormality and non-termination of pregnancy. Nine patients out of 182(5%) had a spontaneous labour followed by premature delivery before 37 weeks and six women (3.3%) among those nine displayed preterm premature rupture of membranes (PPROM). Four patients out of 182 (2%) gave birth before definitive karyotyping result but all of them had a direct fluorescence in situ hybridisation analysis with a normal karyotyping result known well before delivery. CONCLUSIONS: The risk of preterm premature rupture of membrane is 3.3%, with a 5% risk of premature delivery before 37 weeks. This late procedure provides a safe reassurance to women who are unwilling to accept the risks of earlier amniocentesis. However, it should only be used in particular situation and in countries were legislation allows late termination of pregnancy.

[Risk of missed diagnosis of 22q11.2 deletion in a fetal cardiac conotruncal malformation when another chromosomal abnormality is detected]. / Risque d'absence de diagnostic prénatal d'une délétion 22q11.2 associée à une autre anomalie chromosomique dans une malformation cardiaque conotroncale.

We present a rare case of prenatal diagnosis of two de novo chromosome structural rearrangements including a translocation (1;3) associated with a 22q11.2 deletion. The amniocentesis was performed because the systematic ultrasound examination revealed: right aortic cross with double aortic arch, with normal size of aorta and pulmonary artery. Our report emphasises that 22q11.2 deletion must be looked for when a fetal cardiac conotruncal malformation is diagnosed, even in the presence of another chromosomal abnormality. In prenatal diagnosis, this can have implication for patient management and genetic counselling.