[Prospective evaluation of the impact of sperm characteristics on the outcome of intra-uterine insemination]. / Evaluation prospective de l'impact des paramètres spermatiques sur le succès des inséminations intra-utérines.

OBJECTIVES: Thresholds of sperm parameters associated with intrauterine insemination (IUI) success are controversial. This entails heterogeneous practices and leads to low success rate (approximately 8% per attempt). The aim of this study was to evaluate the influence of sperm parameters on the outcome of IUI. PATIENTS AND METHODS: This study included 248 cycles of IUI. Parameters of the raw and selected sperm, as well as the main female characteristics, have been prospectively tabulated and compared, according to the presence or not of a clinical pregnancy. RESULTS: In all, 28 clinical pregnancies were obtained (11.3% per cycle). Thresholds of 10% for the forward motility "a" in the whole sperm and of 20% for the percentage of typical spermatozoa in the selected sperm were related to the success of IUI. The number of motile inseminated spermatozoa, concentration and overall motility of sperm, determined before or after selection, did not have any significant influence on the outcome of IUI. DISCUSSION AND CONCLUSION: Thresholds determined in our study should help to optimize the use of IUI.

[Is there an interest in the addition of hyaluronan to human embryo culture in IVF/ICSI attempts?]. / Y a-t-il un intérêt à utiliser un milieu de transfert embryonnaire enrichi en acide hyaluronique lors de tentatives de FIV/ICSI ?

OBJECTIVE: Several studies have been performed up to date, in order to assess whether the addition of hyaluronan (HA) in human embryo culture could improve the probability of pregnancy, leading, however, to controversies. The aim of the present study was to analyze whether the use of a HA-enriched transfer medium increases clinical pregnancy (PR) and implantation rates (IR), compared with the use of a conventional transfer medium. PATIENTS AND METHODS: All IVF/ICSI attempts from unselected women were included from January 2009 to July 2009 (study group). All embryo transfers (ET) were performed using a HA-enriched media. IVF outcomes of this series were compared to those of a retrospective series, including continuous IVF/ICSI attempts from January 2008 to July 2008 where ET were classically performed (control group). Primary endpoints were PR and IR, compared firstly in all patients of the study and control groups and then regarding woman's age and presence of previous implantation failures. RESULTS: In all, 292 (study group) and 296 (control group) IVF/ICSI attempts were included. PR (32.9%) and IR (22.2%) were significantly increased in the study group compared with those of the control group (PR=25.0%; IR=15.4%; p<0.05). We observed that PR and IR were statistically increased only in subgroups of women >or=35 years or in case of at least one previous implantation failure (p<0.05). DISCUSSION AND CONCLUSION: The use of a HA-enriched embryo transfer medium is beneficial since associated with increased PR and IR. This type of media should be used in infertile women >or=35 years or needing at least a second IVF/ICSI attempt.

[A prospective auto-controlled study to evaluate the interest of selected sperm morphology to inseminate oocytes during IVF]. / Evaluation d'une procédure d'insémination des ovocytes en FIV classique tenant compte de la morphologie des spermatozoïdes sélectionnés.

OBJECTIVES: Since we observed in our laboratory a significant decrease of the fertilization rate in the classical IVF procedure compared to ICSI, we wanted to evaluate a new insemination procedure, using sperm morphology of selected semen as a criterion taken into account to calculate the number of mobile sperm to inseminate. PATIENTS AND METHODS: Therefore, we designed a prospective autocontrolled study which took place from September 2006 to May 2007. All IVF attempts resulting from infertile couples were included in the study if a previous analysis (lesser than one year) of sperm parameters performed in our laboratory was assessed as normal. The retrieved oocytes were separated into two groups (sibling ovocytes), inseminated randomly either with the usual procedure (control group) or with the tested protocol (tested group). Fertilization rate was the primary end point to evaluate this assay. Polyspermy rates were also compared. Subsequently, embryonic quality and development were assessed to eliminate an oxidative stress impact. Paired Student t-test was applied for statistical analysis. RESULTS: In all, 130 couples were included. The diploid fertilization rate was significantly increased in the tested group compared to the control group (66.9% vs 61.3%; p=0.017). No statistically significant difference was showed between the polyspermy rate and numbers of (i) good quality embryos and (ii) blastomeres per embryo into the two groups. DISCUSSION AND CONCLUSION: Compared with the conventional insemination procedure, the fertilization rate benefited from the tested insemination method in our laboratory without damaging embryonic growth. In the limit of our procedure, selected sperm morphology should be used to inseminate an appropriate number of spermatozoa within oocytes, even in case of normal semen values.

[Prospective evaluation of elective single-embryo transfer versus double-embryo transfer following in vitro fertilization: a two-year French hospital experience]. / Evaluation prospective du transfert sélectif d'un embryon: deux années d'expérience.

OBJECTIVE: Multiple embryo transfer is responsible for a high rate of multiple pregnancies (ICSI), with subsequent risks of premature birth and perinatal death. This prospective non randomized study aimed to assess the ability of an elective single-embryo transfer (eSET) policy to reduce the twin pregnancy rate, compared to a double embryo transfer (DET) approach. PATIENTS AND METHODS: Between March 2005 and May 2006, 180 eligible women were proposed to benefit from an eSET transfer rather than a DET. Inclusion criteria were (i) age less than 37 years old; (ii) at least two good quality embryos available (three to five cells at day 2 or six to nine cells at day 3; less than 20% fragmentation and the absence of multinucleates blastomeres), after IVF or ICSI and (iii) no more than one previous failed treatment cycle. Outcome analysis included cycles with frozen-thawed embryo transfer (FET). RESULTS: According to patients' decision, 107 and 73 women had an eSET (59.4%) and a DET (40.6%) respectively. No differences were found between eSET and DET groups regarding demographics and biologicals parameters. The clinical pregnancy rate (PR) per transfer was 43.9% in eSET group and 57.5% in DET group (p=0.07). The twin pregnancy rates were 0 and 14.3%, in eSET and DET groups, respectively (p=0.007). The cumulative PR per patient, including the outcome of performed FET cycles, was 63.6% in eSET group and 61.6% in DET group. In this case, the cumulative twin pregnancy rates were 2.9 and 15.6% in eSET and DET groups, respectively (p=0.02). DISCUSSION AND CONCLUSION: Our data show that in a selected population of women, transferring one fresh embryo and then, if required, one or two frozen-thawed embryos significantly reduces the twin pregnancy rate without decreasing the overall pregnancy rate. This study supports the policy of eSET in this subgroup of patients.

Interest of growth hormone-releasing hormone administration for improvement of ovarian responsiveness to gonadotropins in poor responder women.

OBJECTIVE: We have investigated the beneficial effect of a somatotroph axis stimulation on ovarian response to gonadotropin. DESIGN: Growth hormone-releasing hormone (GH-RH) was administered in a prospective study in women undergoing an in vitro fertilization protocol. PATIENTS: Twelve patients were selected for their poor ovarian response to previous stimulations using gonadotropin-releasing hormone analog (GnRH-a) and human menopausal gonadotropins (hMG). INTERVENTIONS: Five hundred micrograms of GH-RH1-29 were administered two times daily concomitantly with GnRH-a and hMG from day 2 of the cycle to the time of ovulation. MAIN OUTCOME MEASURES: Stimulation of somatotroph axis was appreciated by measuring over-night urinary growth hormone (GH) output, plasma GH, and insulin-like growth factor I (IGF-I) and follicular fluid (FF) IGF-I. The effects of GH-RH administration on ovarian function were determined by plasma estradiol levels and follicular data. RESULTS: Administration of GH-RH was associated with a significant improvement of urinary (P less than 0.025) and plasma (P less than 0.001) GH concentrations and of the hormonal response to hMG (P less than 0.01). Levels of IGF-I followed a biphasic plasma variation, and a slight increase in recruited follicles, retrieved oocytes, and FF IGF-I content was also observed. CONCLUSIONS: Activation of the somatotroph axis by GH-RH enhances the hormonal ovarian response to hMG and may be an adjunctive therapy to improve follicular maturation.

[Spontaneous pregnancies and premature menopause]. / Grossesses spontanées et ménopause précoce.

Hypoestrogenic hypergonadotrophic amenorrhea occurring before the age of 40 years is generally considered irreversible. The term "premature menopause" used to define this condition seems a misnomer, and "premature ovarian failure" (POF) is preferred on both a psychological and a physiological basis. POF is not uncommon considering the incidence rate of 1-2% of women during their theoretically reproductive life. These circumstances are particularly painful when a child is desired. However, since a long time the notion of permanent cessation of ovarian function is challenged and intermittent and transient return of ovarian function has been described with 5-10% chance of pregnancy often more than 12 months after the beginning of the amenorrheic status. A multicentric retrospective study conducted by the GEDO reports 27 pregnancies out of 518 patients waiting for an in vitro fertilization with oocyte donation because of POF or occult ovarian insufficiency. Considering this work and a literature review, we try to propose part of understanding of the basic POF physiopathology, and to examine the factors which could act on the prognosis and the clinical management.

[Oocytes in vitro maturation: results and future in humans]. / Maturation in vitro des ovocytes : bilan et perspectives dans l'espèce humaine.

Mature oocytes are rare and highly specialized cells. In vitro maturation of human oocytes is an emerging assisted reproductive technology allowing to produce more mature oocytes without ovarian stimulation. Whereas in vitro maturation is technically more demanding than conventional in vitro fertilization for the laboratory, it carries many potential advantages, for example, in terms of lower treatment heaviness and removal of risk of severe ovarian hyperstimulation syndrome for the patients. Although the technology is still experimental, oocytes in vitro maturation has been successfully used and pregnancies and live births have been reported. Despite these successes, the overall efficiency of in vitro maturation remains low and this procedure must still be improved. The different steps of in vitro maturation process are shown and discussed as well as results in terms of pregnancy and live birth rates.

Short gamete co-incubation during in vitro fertilization decreases the fertilization rate and does not improve embryo quality: a prospective auto controlled study.

PURPOSE: Evaluate the effect of short gamete incubation on fertilization rate and embryo quality. METHODS: A prospective study has been performed. Two thousand five hundred and forty seven sibling oocytes from 240 couples undergoing IVF attempts were allocated to a short (1 h) or a standard (18 h) insemination procedure. Diploid fertilization rate (two pronuclei, 2PN), polyspermy (>2PN) and embryo quality were compared. RESULTS: The fertilization rate was statistically lower in the short insemination group compared to the standard insemination one (64.9% and 70.1%; P = 0.039), with a similar polyspermy rate observed between the two groups. A slight, but non significant, increase was observed concerning good embryo quality rate in the short insemination group when compared to the standard insemination, both at day 2 (60.1 vs. 58.1%; P = 0.06) and day 3 (53.2 vs. 48.5%; P = 0.22). CONCLUSION: This new study highlights that a 1 h gamete exposure decreases the fertilization rate and does not improve embryo quality compared with a standard 18 h insemination procedure.

A prospective randomized study to assess the benefit of partial zona pellucida digestion before frozen-thawed embryo transfers.

BACKGROUND: Assisted hatching (AH) in fresh embryo transfer (ET) could be associated with increased implantation rates. However, very few prospective randomized studies have specifically addressed the issue of AH during frozen-thawed embryo transfers (FET) cycles, those that have reported controversial results. The aim of this study was to evaluate the benefit of an enzymatic zona pellucida treatment of frozen-thawed embryos before transfer. METHODS: This was a prospective study including 125 non-donor FET cycles from 125 infertile couples. FETs were randomly allocated into AH group (n = 61, embryos pretreated with pronase 5 IU/ml for 1 min at 37 degrees C) or control group (n = 64, untreated embryos). Zona pellucida thickness was measured for each transferred embryo. The main outcome parameters were clinical pregnancy and implantation rates. RESULTS: The two groups were comparable regarding mean women's age, duration and indications of infertility, IVF outcome after fresh ETs, numbers and quality of fresh and frozen embryos, frozen-thawed embryo survival rates and blastomeres survival indexes. Despite a statistically significant decrease of zona pellucida thickness after pronase treatment [(mean +/- SD) 18.5 +/- 2.25 versus 14.5 +/- 2.75 microm; P < 0.0001], implantation (9.6 versus 9.2%) and clinical pregnancy rates (18.0 versus 17.2%) were not statistically different after FETs, with a similar mean number of embryos transferred between AH and control groups, respectively. CONCLUSION: Within the constraints of our protocol, partial enzymatic digestion of zona pellucida by pronase was not related with any benefit of the FET outcome especially concerning the implantation ability of frozen-thawed embryos.

Chromosome segregation in an infertile man carrying a unique pericentric inversion, inv(21)(p12q22.3), analysed using fluorescence in situ hybridization on sperm nuclei: significance for clinical genetics. A case report.

We report the case of a 40-year-old patient referred to our centre after 3 years of infertility. Karyotyping with the aid of fluorescence in situ hybridization (FISH) analysis showed a unique pericentric inversion of chromosome 21:46,XY,inv(21)(p12q22.3). This type of intrachromosomal structural rearrangement can lead to chromosome imbalance in offspring by producing unbalanced gametes if an odd number of crossover events occur within the inverted segment. Therefore, partial trisomy/monosomy with clinical consequences can be observed in the progeny of carriers. Semen samples from the inversion carrier were analysed by FISH using a combination of probes [a subtelomeric 21q probe and a locus-specific Down's syndrome critical region (DSCR) probe] to evaluate the proportion of recombinant chromosomes. Sperm-FISH analysis of 3400 spermatozoa revealed a 67.4% rate of balanced chromosomes (normal or inverted). The frequencies of recombinant chromosomes with duplication of the long arm and deletion of the short arm, and vice versa, were 11.2 and 21.4%, respectively. The risk for the couple of conceiving a child with an unbalanced chromosome 21 is estimated to be around 32%. This case study shows the utility of sperm-FISH analysis in the genetic counselling of a pericentric inversion in a male carrier to assess the frequency of recombinant chromosomes and therefore evaluate the probability of having a normal conception.

World Health Organization grade 'a' motility and zona-binding test accurately predict IVF outcome for mild male factor and unexplained infertilities.

BACKGROUND: The aim of this study was to determine the pronostic value of a sperm-zona pellucida (ZP) binding assay, combined with World Health Organization (WHO) grade 'a' sperm motility on the day of the IVF attempt, to predict sperm fertilizing ability in unexplained and moderate male factor infertilities. METHODS: In total, 84 couples (64 unexplained infertility; 20 male factor) underwent both a sperm-ZP binding assay and an IVF attempt, irrespective of the test's result. The test was negative when grade 'a' motility was #5% and/or the ZP binding index was <0.7. Fertilization and pregnancy rates were related to the test's results. RESULTS: Thirty-one patients had a negative test (group N) and 53 a positive test (group P). A difference was observed concerning the fertilization rate [median (range): 0 (0-75%) versus 50 (0-100%); P = 0.0001] and the number of cycles with fertilization rate <20% (65 versus 23%; P = 0.0002) between groups N and P respectively. In the group of unexplained and male factor infertilities, the test showed a sensitivity of 83 and 60%, specificity of 50 and 90%, positive predictive value of 76 and 86%, and negative predictive value of 61 and 69% respectively. CONCLUSION: Sperm-ZP binding test, combined with WHO grade 'a' motility assessment, is relevant to prevent IVF fertilization failures in unexplained infertility and, most particularly, in moderate male factor infertility.

[Quantitative cytologic and histologic study of the maternal hepatocyte and lobule on the 18th day of gestation in the Wistar rat]. / Etude cytologique et histologique quantitative de quelques aspects de l'hépatocyte maternel et du lobule au 18e jour de la gestation chez le rat "Wistar".

With the object of studying the part taken by liver cells and lobules in mother's liver weight increase (+42%), planimetric cells areas were measured. The main results are: same diversity of cells size as in reference samples, perilobular cells remaining the larger. In the same way, one can note a small increase of cells size for pregnant animals: perilobular cells (+10%), nucleus (+26%), nucleolus (+19%), cytoplasms (+12%). These facts demonstrate the metabolic part played by liver during pregnancy but do not explain the liver weight increase. But planimetrical investigations of overall lobular size lead to an increase of 40%. Finally the increase of liver weight might be explained by an increase of cells number.

[Study of the weight of the pregnant rat and the maternal liver: their relation to biometric parameters of the developing embryo and placenta]. / Etude pondérale de la ratte gravide et du foie maternel: ses relations avec quelques paramètres biométriques de l'embryon et du placenta en développement.

Observations carried out on a large sample of pregnant rats, their fetuses and placentas, lead to the following findings: continuous increase of liver weight along pregnancy from 2nd to 20th days, mainly within the 2nd week (organs making), to reach limit value on the 18th day. This liver weight increases roughly along with embryos and placentas weights specially from 14th to 20th days. At the 18th day the mother's liver weight depends on embryos and placentas weights. This weight of the mother's liver increased with embryos and placentas number but does not depend on resorption number. The overall mother's weight is proportional to liver weight and embryos number. All these facts underline metabolic relations between mother and fetus during pregnancy.

[Morphometric study of ultrastructural modifications in the maternal hepatocyte during normal gestation in the Wistar rat]. / Etude morphométrique des modifications ultrastructurales de l'hépatocyte maternel au cours de la gestation normale chez la rate Wistar.

Quantitative changes in hepatocyte ultrastructures during normal gestation were studied in Wistar rats with morphometric methods. On the 18th day of the gestation, variations in nuclearcytoplasmic ratio, size of R.E.R., mitochondria, lysosomes and microbodies were observed, with variations according to the localisation of hepatocyte inside the lobule. We report the most obvious effects as follows: An increase of R.E.R. in the central and perilobular zones. Mitochondria are larger and the rounded forms are more numerous inside the two lobular zones. The number of lysosomes and microbodies are only elevated in the perilobular cells. In conclusion, it is suggested that the hepatocyte organelles are significantly increased during gestation. This is probably due to the metabolic activation. These cellular modifications are only one aspect of the liver increase.

Optic atrophy, microcephaly, mental retardation and mosaic variegated aneuploidy: a human mitotic mutation.

A 12-year-old girl presents with optic atrophy, pale papilla, amblyopia and microcephaly (-3 s.d.) with mild mental retardation and facial dysmorphism. She had mitral insufficiency with mitral prolapse and moderate short stature (-2.5 d.s.). She had normal flash visual evoked potentials, normal electroretinograms and electrooculograms and normal cranial CT scan as well as other lab tests to rule out malformations, tumors or multiple sclerosis. Her lymphocyte karyotype showed a variegated mosaicism with: 2 cells with 49, XX, +mar,+mar,+mar; 21 cells with 48, XX, +mar,+mar; 57 cells, with 47, XX,+mar; 20 cells with 46,XX; while parental karyotypes were normal. This syndrome therefore associates optic atrophy, mental retardation and microcephaly and short stature with chromosomal instability in the form of variegated mosaicism.

Factors influencing human sperm kinematic measurements by the Celltrak computer-assisted sperm analysis system.

This study examines the effect of varying several factors, both extrinsic and technology-dependent, on the reconstruction of human sperm trajectories and the derived kinematic measurements using videotapes and the Motion Analysis Celltrak/S instrument. In semen samples from normal healthy men, curvilinear (VCL) and straight line velocities (VSL) were found to increase 1.5-fold, and linearity (LIN) of trajectories and amplitude of lateral head displacement (ALH) increased 1.25-fold when the temperature of analysis was raised from 24 to 37 degrees C. Only VCL and VSL were found to increase significantly between 24 and 37 degrees C for sperm samples selected by Percoll gradient and incubated in a capacitating medium. An analysis chamber of 20 microm depth was found to be adequate for seminal sperm samples while for Percoll-selected sperm samples the analysis in a 50 microm depth provided the highest proportions of spermatozoa with the highest VCL and the largest ALH. The grey level detection threshold required careful adjustment: using a threshold lower than the optimal threshold produced spurious sperm trajectories for seminal sperm samples and rejected some trajectories for Percoll-selected sperm samples. Definition of the appropriate frame rate and maximum burst speed was critical for valid trajectory reconstruction and therefore adequate derived kinematic measurements. Optimal values of these parameters were found to be 30 Hz and 400 microm/s for seminal spermatozoa and 60 Hz and 700 microm/s for selected spermatozoa. The optimal values of 'ALH path-smoothing factor' used to calculate average path and ALH were 5-10 points for seminal spermatozoa analysed at 30 Hz and 15-20 points for selected spermatozoa analysed at 60 Hz. We propose a set of standard conditions for reliable kinematic analysis of human spermatozoa using the Celltrak/S system.

De novo interstitial deletion of the long arm of chromosome 7:46,XY,del(7)(q23;q32).

A de novo interstitial deletion of the long arm of chromosome 7 is reported in a newborn boy. Our observation is compared with seven others deletions of the same bands. Clinical features showed the following: hypotonia, microcephalia, difficulty in swallowing, low-set dysplastic ears, an abnormal cry, upslanting and small palpebral fissures, and abnormalities of the hands and feet. Delayed mental and physical development is the general rule, and visceral malformations are uncommon. Our patient had genital abnormalities and a cardiac malformation.

[Results of oocyte donation in France (Study Group on Oocyte Donation)]. / Bilan du don d'ovocytes en France (GEDO).

The French study group on oocyte donation, named GEDO, reports the results concerning the oocyte donation activity in France during 1998, including information from all the centers in effective operation except one. These data describe the recipients' and donors' situation and give the analysis of the results obtained.

[Results of oocyte donation in France (GEDO)]. / Bilan du don d'ovocytes en France (GEDO).

The French study group on oocyte donation, named GEDO, reports the results concerning the oocyte donation activity in France from 1994 to 1997, with the informations of all the centers, in effective operation. These data describe the recipients' and donors' situation and give the analysis of the results obtained.

Cytogenetic study of human oocytes uncleaved after in-vitro fertilization.

Chromosome analysis of oocytes uncleaved after IVF allows the cause of the failure of cleavage to be determined and shows the incidence of chromosome disorders among human oocytes. A total of 198 uncleaved oocytes fixed 40 h after insemination were successfully analysed according to Tarkowski's air-drying method: 78.3% were unfertilized and arrested in metaphase II. Among them, 70% were normal (23,X) and 30% aneuploid (16% were hypohaploid, 14% were hyperhaploid). The incidence of chromosome breaks was 18%. In 12.1% of the oocytes, sperm chromosome condensation appeared premature usually in the G1 phase. This was especially observed in idiopathic infertility (7% of fertilized oocytes versus 2% in tubal infertility cases). In 8.1% of the cases, chromosome analysis showed diploidy which may be interpreted by either an absence of extrusion or a reintrusion of the polar body or by first cleavage failure during mitosis. In 1% of the cases triploidy was observed. Our results show that the main reason for failure of cleavage is related to failure of fertilization (78.3%). However, premature condensation of sperm chromosomes at the G1 phase appears to be quite frequent. This may be involved in the aetiology of some cases of idiopathic infertility. Finally, the high rate of chromosomal disorders (30%) in human oocytes may explain the high rate of chromosomal abnormalities in preimplantation embryos.