[Embryo donation in France: practice and difficulties. Strasbourg's experience]. / L'accueil d'embryon en France: pratiques et difficultés. L'expérience de Strasbourg.

OBJECTIVE: The aim of this study was to present the situation of embryo donation in France and around the world, to expound the difficulties of its practice and the results obtained in our centre 3 years after the introduction of this procedure. PATIENTS AND METHODS: Embryo donation in France is controlled by implemented decrees published between 1999 and 2004. The couples, who have stored frozen embryos since at least two years, were contacted for a pluridisciplinary medical consultation. The indication of embryo donation was evaluated for the recipients through a pluridisciplinary approach. RESULTS: Among the interviewed couples, 16.7% have chosen embryo donation but only half of them have completed the procedure (6% of the couples with frozen embryos). The main indications for embryo donation were a double sterility, unexplained genetic disease, ART failures (poor fertilization or bad embryo quality) and oocyte donation when the delay was too long for the couples. The pregnancy rate was 28.6% after the 21 first embryo transfers. DISCUSSION AND CONCLUSION: The results of embryo donation confirm the international experience both considering the poor number of donated embryos, medical indications and results. Embryo donation has its place among ART techniques, but one should not ignore the general debate on ethical questions raised by this procedure.

[Is gonadotropin releasing hormone (GnRH) agonist trigger beneficial or deleterious?]. / Le déclenchement par agonistes de la Gonadotropin releasing hormone (GnRH) est-il bénéfique ou délétère ?

OBJECTIVES: The undeniable asset of the antagonist protocols in in vitro fertilization is the decrease of the risks of ovarian hyperstimulation syndrome, by the use of a release by GnRH agonist. Nevertheless, questioning persist concerning the rates of clinical pregnancies, the oocyte quantity and the empty follicle syndrome. We thus studied these parameters in our center. METHODS: A retrospective study was realized from January 1st, 2013 till July 31st, 2015. The main objective was the evaluation of the rate of clinical pregnancies in antagonist protocol. A first group of 775 cycles have benefited from a release of the ovulation by HCG, while a second group of 204 cycles, by GnRH agonist. The secondary objectives were the oocyte quantity, the rate of ovarian hyperstimulation syndrome, and the rate of empty follicle syndrome. RESULTS: No statistically significant difference was found between both groups concerning the rates of clinical pregnancies, oocytes quantity, and the rate of empty follicle syndrome, whatever is the type of used release, in fresh embryo transfer. A syndrome of premature ovarian hyperstimulation syndrome was found at 7.9 % of the patients in the group 2 versus 2.3 % in the group 1, with a statistically significant difference (P<0.05). At these patients, a strategy of frozen embryo transfer ("freeze all") was proposed. The accumulated rates by pregnancy in both groups were not statistically different. CONCLUSION: The release by GnRH agonist does not show inferiority in terms of clinical pregnancy, in comparison to HCG.

Structure of the Chlamydomonas reinhardtii cabII-1 gene encoding a chlorophyll-a/b-binding protein.

Gene cabII-1 is a light regulated gene that encodes the precursor of a major chlorophyll-a/b-binding protein in Chlamydomonas reinhardtii. It is a member of a small gene family composed of about 3-7 members. Nucleotide sequencing data and S1 mapping reveal that the cabII-1 gene is interrupted by three introns. Except for the transit peptide and the N-terminus, the cabII-1 gene product is similar to cabII proteins in higher plants. The cabII-1 gene in C. reinhardtii appears to be an intermediate between type-I and type-II cabII genes described in higher plants.

[Future perspectives: oocyte cryopreservation and in vitro maturation]. / Perspectives d'avenir: la congélation et la maturation in vitro des ovocytes.

After treatment for neoplasia, a young patient can nowadays hope to be pregnant. The aim of this paper is to establish a review of the literature about cryopreservation of oocytes and ovarian tissue, and in vitro maturation of oocytes. These studies also concern patients for which ovarian stimulation is difficult, such as PCOS. These techniques could also facilitate the creation of an oocyte bank as the one already existing for sperm. So far the main studies published, in spite of very timid first results, allow us to look to a great future for cryopreservation and in vitro maturation of oocytes.

[Towards a single embryo transfer: predictive value of uterine and embryological parameters in assisted reproduction outcome]. / Vers un transfert mono-embryonnaire : prise en compte conjointe des paramètres embryonnaires et utérins en assistance médicale à la procréation.

OBJECTIVE: The transfer of a single embryo would avoid obstetrical and neonatal complications due to multiple pregnancies. We studied the clinical value of both uterine and embryological parameters to define precise conditions allowing a single embryo transfer without decreasing pregnancy rates. PATIENTS AND METHODS: Endometrial parameters expressed by a uterine score together with biological criteria of 131 in vitro fertilization or intracytoplasmic sperm injection attempts were retrospectively analysed. RESULTS: Two hundred and sixty-two day-3 embryos were replaced through 131 transfers. Fifty-seven pregnancies were induced and 16 twins were obtained. The clinical pregnancy rate was 35.9% and the embryonic implantation rate 24.0%. After the transfer of two embryos, successful implantation was determined by the occurrence of top-quality embryos and simultaneously by a receptive endometrium. The uterine pulsatility index was significantly decreased for twin compared to singleton pregnancies. DISCUSSION AND CONCLUSION: This study confirms that a uterine score constitutes a powerful tool for evaluating the uterine receptivity. This parameter has to be taken into account as well as the embryonic quality, in order to optimise the success rate. Young patients with at least two top-quality embryos available, a high uterine score and a low pulsatility index, have a high risk of multiple births and are suitable for a single-embryo transfer.

[Abnormal bacterial colonisation of the vagina and implantation during assisted reproduction]. / Colonisation bactérienne vaginale anormale et implantation en assistance médicale à la procréation.

OBJECTIVE: To evaluate the efficiency of our treatment of vaginal infection for couples included in an IVF program. PATIENTS AND METHODS: Microbiologic screening of vaginal flora and semen has been performed one month prior to in vitro fertilization for 951 couples in 2000. Antibiotic treatment was prescribed in case of positive culture. RESULTS: Positive microbial growths were observed from endocervical and vaginal cultures in 218 women (22.9%). The clinical pregnancy rate was 30.29% in the group of patients without growth and 30.27% in the group with positive microbial growth. The implantation rate was significantly diminished in case of bacterial growth: 14.6 compared to 19.3% (P <0.02) for sterile endocervical culture. Five main bacterial species were found at the cervical level: Candida albicans (69 cases), Ureaplasma urealyticum (49 cases), Gardnerella vaginalis (43 cases), Streptococcus B or D (24 cases) and Escherichia coli (22 cases). Positive cultures from both vagina and semen were observed for 77 couples whose clinical pregnancy rate was 19.5 vs 36.2% in case of vaginal infection alone (P <0.01) with a spontaneous miscarriage rate of 46.7 compared to 17.6% (P <0.01). DISCUSSION AND CONCLUSION: Endocervical microorganisms, even treated with adapted antibiotics, may affect embryonic implantation. Positive culture from both female and male partner may enhance this negative effect. In this case, the best strategy would be to cancel the IVF treatment.

[Twin birth after preimplantation diagnosis for cystic fibrosis]. / Naissance gémellaire après diagnostic préimplantatoire pour mucoviscidose.

OBJECTIVE: Cystic fibrosis is a common autosomal recessive disease most often caused by a deletion (delta F508) in the CFTR gene. It is the most common indication for preimplantaion genetic diagnosis which allows genetic analysis of embryos obtained after in vitro fertilization and transfer of unaffected embryos into the patient's uterus. PATIENTS AND METHODS: We report the first preimplantation genetic diagnosis performed in Strasbourg for a couple at risk of having a child affected by severe cystic fibrosis due to a homozygous delta F508 mutation. Three days after fertilisation, embryos obtained after intra-cytoplasmic testiculare sperm injection were biopsied and analysed. PCR amplification of the genomic fragment containing the delta F508 locus allowed detection of the delta F508 mutation and transfer only of the unaffected embryos. RESULTS: Three embryos were transferred after this preimplantation genetic diagnosis. A twin pregnancy was obtained and the babies born from this cycle are both exempt from the mutation. CONCLUSIONS: Preimplantation genetic diagnosis for the cystic fibrosis delta F508 mutation is now available in our centre. In this report, we could resolve both the problem of infertility and the risk of transmission of a severe form of cystic fibrosis. Preimplantation genetic diagnosis is also available for other mutations involved in cystic fibrosis and also for other genetic diseases.

[Evolution of homocysteine during ovarian stimulation for IVF or ICSI]. / Evolution de l'homocystéine lors d'un cycle d'induction ovarienne pour FIV ou ICSI.

OBJECTIVE: To determine the homocysteine evolution during ovarian stimulation in IVF or ICSI protocols and in, a second time, to evaluate the role of hyperhomocysteine as thrombotic risk factor for the treated patients. MATERIAL AND METHODS: Plasma homocysteine was determined three times for each of 31 women included in an IVF/ICSI program. Dosages were realised before stimulation, after gonadotrophin-releasing hormone agonist treatment (GnRH) and on the day of hCG injection. Vitamin B12 and folates were determined before stimulation. In case of hyperhomocysteinemia, a research of APCR (Activated Protein C Resistance) was realised. RESULTS: Five hyperhomocysteinemia cases were discovered (16.12% of studied population). APCR was found in a patient with hyperhomocysteinemia (14 mumol/L, before stimulation). Molecular biology has confirmed an heterozygous mutation of factor V Leiden. During the ovarian stimulation the evolution of homocysteine was independent of the 17 beta oestradiol evolution. CONCLUSION: The prevalence of hyperhomocysteinemia was not significative according to the limited size of the studied population. The increase of oestradiol during induction protocols is unrelated to the homocysteine level. This work must be continued with largest population to have better knowledge of the prevalence of hyperhomocysteinemia among women included in ovarian stimulation protocols.

[Pregnancy by micromanipulation in a case of excretory azoospermia associated with a natural spermatocele]. / Grossesse obtenue par micromanipulation dans un cas d'azoospermie excrétoire associée à la présence d'une spermatocèle naturelle.

Microinjection of capacitated sperm into the perivitelline space of oocytes has been performed in a case of male infertility attributed to excretory azoospermia. In this particular case, a spermatocele detected five years ago was observed, which provide motile spermatozoa. These sperm collected by direct puncture into the cyst, were capacitated using discontinuous Percoll gradient. After ovarian stimulation, seventeen oocytes were collected. Nine of them were used for classical IVF whereas the other eight were submitted to microinjection. Only one oocyte of the second group fertilized and cleaved 48 hours later. A clinical pregnancy was achieved after embryo transfer and a normal, healthy boy delivered at term.

Optimal management of extreme oligozoospermia by an appropriate cryopreservation programme.

BACKGROUND: Severe oligozoospermia is characterized by sperm count fluctuations that may result in insufficient quantities of motile sperm for ICSI on the day of oocyte retrieval, thus necessitating testicular biopsy. To avoid this, we proposed that patients, with transient azoospermia or repeatedly low sperm counts, make a safety pool of frozen spermatozoa before ICSI attempts. METHODS: Seventy cryptozoospermic (<10(3) spermatozoa/ml) and 46 oligozoospermic patients (10(3)-10(5)/ml) were included. Although all oligozoospermic patients succeeded in sperm banking, only 44 of 70 cryptozoospermic patients were successful. Others underwent testicular extraction of spermatozoa. The ICSI results for frozen sperm from cryptozoospermic patients were compared with those obtained with fresh sperm from a group of normal patients (>10(5) spermatozoa/ml). RESULTS: In this prospective matched, controlled study, five cryptozoospermic, but no oligozoospermic, patients failed to produce sperm on the ICSI day, and frozen sperm was used instead. Although fertilization and pregnancy rates (per attempt) using fresh (49% and 5/44, respectively) and frozen sperm (54% and one-fifth, respectively) were similar for this cryptozoospermic group, the results for fresh sperm were significantly lower when compared with the control group (66% and 16/43, P < 0.0001, P < 0.001, respectively). In contrast, results for the oligospermic and control groups were similar. CONCLUSIONS: Banking of ejaculated sperm is helpful for cryptozoospermic patients.

Seminal haploid cell detection by flow cytometry in non-obstructive azoospermia: a good predictive parameter for testicular sperm extraction.

BACKGROUND: Testicular sperm extraction (TESE) associated with ICSI gives patients suffering from non-obstructive azoospermia (NOA) the possibility of becoming a father. The success rate of TESE based on sperm recovery is approximately 50%, and the commonly used non-invasive parameters are not predictive enough. Only the invasive testis biopsy has a good prognostic value. The aim of this study was to evaluate the prognostic value of the detection of seminal haploid cells by flow cytometry (FCM) in order to avoid unnecessary testicular biopsy. METHODS: For 37 NOA patients undergoing testicular biopsy, we measured testis size, serum FSH and inhibin B levels and carried out seminal cytology, seminal FCM analysis and histological examination. RESULTS: Sperm were found in 18 biopsies. These results were correlated with cytology, FCM analysis and the histological examination. FCM was more sensitive than cytology (100 versus 59%) but less specific (67 versus 83.5%) whereas the histological observation of complete spermatogenesis appeared to be less sensitive (50%) but more specific (100%). CONCLUSION: Detection of seminal haploid cells by FCM appears to be an interesting non-invasive technique which can predict TESE results and improve the management of NOA patients.

Peritoneal fluid and infertility: a comparison of in-vitro fertilization of oocytes in peritoneal fluid and B2-Menezo medium.

Peritoneal fluid probably has an important and complex role in fertility and infertility. In this study, in-vitro fertilization was attempted in peritoneal fluid and the classical B2-Menezo medium. Clinical criteria concerning male and female fertility and biological factors such as fresh semen quality, sperm survival, sperm-zona pellucida binding, cleavage and pregnancy rates were analysed for each infertile couple. The following results have been obtained. The quality of peritoneal fluid appears to fluctuate greatly from one woman to another. Routine use of peritoneal fluid as a culture medium during in-vitro fertilization should not be encouraged. Peritoneal fluid toxicity may explain some cases of 'unexplained' infertility.

[Follicular fluid: role and importance in the preparation of spermatozoa for fertilization]. / Fluide folliculaire: rôle et importance dans la préparation des spermatozoïdes à la fécondation.

It has been demonstrated that the preovulatory follicular fluid creates a micro-environment which is of importance for normal sperm function. In this paper, we attempt to evaluate the role of human follicular fluid (HFF) on the fertilizing capacity of spermatozoa.

Prognostic value of objective semen parameters in an in vitro fertilization program.

PURPOSE: The basic semen parameters seem to have a limited predictive value in male fertility. Could other objective sperm analyses be helpful in the choice of the most adapted assisted procreation technique? METHODS: This study concerns 78 infertile couples with insemination failures. For each semen, 21 objective parameters are analyzed in fresh semen and after sperm selection procedure. The 78 couples are then included in an IVF protocol and classified into two groups: fertile (at least one cleaved embryo is obtained) and infertile. RESULTS: Using multiple variant discriminant factorial analysis, we have found nine nonconventional parameters which induce us to define two classes of semen. These two classes fit with the classification into fertile and infertile groups in 74.4% of the cases. CONCLUSIONS: So these parameters allow us to predict the chance of obtaining embryos during an IVF trial and to choose for each couple the most appropriate technique: IVF or ICSI.

[Therapeutic strategies in azoospermia: management]. / Stratégie thérapeutique dans les azoospermies: mise au point.

ICSI allows use of epididymal or testicular sperm in case of azoospermia. The aim of this study is to propose an algorithm for the management of the different situations of azoospermia according to our personal experience and according to data of literature. The following points are discussed: epididymal or testicular origin of sperm, retrieval technique, contribution of freezing, interest of in vitro culture, side effects and need of a preliminary biopsy. So a therapeutic strategy adapted to both types of azoospermia obstructive and non obstructive can be proposed.

Identification of hyaluronic acid and chondroitin sulfates in human follicular fluid and their effects on human sperm motility and the outcome of in vitro fertilization.

This study measured glycosaminoglycans (GAGs) in human follicular fluid (hFF) obtained from patients undergoing hormonal stimulation with combined GnRH agonist followed by gonadotropin hormone (hMG or FSH) for in vitro fertilization (IVF). The GAGs were partially characterized through the use of various mucopolysaccharidases and then their effects tested on human sperm motility. In hFF, the GAG and protein concentrations were 4.4 +/- 1.3 mg/L and 32.6 +/- 3.2 g/L respectively. Chondroitins (CS) and hyaluronic acid (HA) significantly stimulate sperm motion in comparison to the control. The oocytes inseminated with GAGs-pretreated spermatozoa showed a significantly higher rate of cleavage and pregnancy or delivered number. The present results suggest that hFF is a rich source of sulfated glycosaminoglycan (CS), and non-sulfated glycosaminoglycan (HA). The stimulatory effects of hFF on human sperm motility may well depend on CS and HA. GAGs pretreated sperm may enhance the fertilizing ability of spermatozoa and thus the IVF outcome.

Could in vitro fertilization with a modified sperm preparation technique be an option to micromanipulations?

PURPOSE: This study evaluated the efficiency of in vitro fertilization with a modified sperm preparation technique for male infertility, to avoid systematic recourse to ICSI. METHODS: The sperm function stimulants pentoxifylline, 2-deoxyadenosine, and follicular fluid were used during the sperm preparation of IVF trials for two groups of patients: couples with long-standing infertility and previous IVF failures and couples with male-factor infertility scheduled for their first treatment. RESULTS: Forty-nine attempts were performed in the first group. The overall fertilization rate (percentage of zygotes with two pronuclei 18 hr after sperm insemination) was 30% and the clinical pregnancy rate per transfer was 33%. For the second group, 30 attempts were performed. The fertilization rate was 29.6% and the clinical pregnancy rate was 62.5% per transfer. The total number of recovered motile sperm appears to be a significant parameter: a cutoff value of 0.25 million motile sperm recovered after capacitation treatment has a good prognostic value for choosing between IVF with sperm enhancers and ICSI. CONCLUSIONS: The described modified IVF could constitute an alternative to the systematic use of ICSI.

A case of human pregnancy after microinjection of capacitated sperm into the perivitelline space.

Microinjection of capacitated sperm into the perivitelline space of oocyte was offered to one couple with persistent infertility of mixed origin. The husband's semen was subnormal, whereas his wife had definitive tubal occlusion and polycystic ovaries. Four previous in vitro fertilization (IVF) attempts were performed but no fertilization was obtained. After superovulation, 13 oocytes were collected. Ten were submitted to microinjection and two were damaged during the procedure. One of the remaining eight had two pronuclei 18 hr after microinjection and progressed to a four-cell embryo after 48 hr. After reimplantation, a normal pregnancy was initiated, caryotype (46XX) was checked at 17 weeks. A normal and healthy girl has been delivered at term.

Does body mass index of infertile women have an impact on IVF procedure and outcome?

PURPOSE: The objective was to explore whether body mass index (BMI) of women related to the different parameters of the in vitro fertilization (IVF) procedure and outcome. METHODS: This retrospective study on 398 couples analyzed epidemiological features, characteristics of ovarian stimulation, number and quality of retrieved oocytes, as well as pregnancy outcome according to three groups of BMI values: BMI < 20, 20 < or = BMI < 25 and BMI > or = 25 kg/m2. RESULTS: The prevalence of underweight and overweight women was 21.8% and 22.3%, respectively. The mean ratio follicle-stimulating hormone-luteinizing hormone increased significantly according to BMI. An increase in the mean number of consummated gonadotropin ampoules together with a decrease in the number of collected oocytes was observed in long stimulating protocol cycles when BMI > or = 25 kg/m2. The same observations were made in short stimulation protocol cycles of BMI < 20 and > or = 25 kg/m2. No significant difference could be found in clinical pregnancy and miscarriage rates between underweight, normal weight, and overweight patients. CONCLUSIONS: Both underweight and overweight have negative effects on IVF parameters and outcome leading to decreased chances of pregnancy.