Are leptin and adiponectin involved in recurrent pregnancy loss?

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.

Testicular Spermatozoa Are of Better Quality Than Epididymal Spermatozoa in Patients With Obstructive Azoospermia.

OBJECTIVE: To assess sperm quality as a function of the sampling site (testis or epididymis) in obstructive azoospermia (OA). MATERIALS AND METHODS: DNA fragmentation rates in spermatozoa sampled from the testis and epididymis (from patients with different etiologies of OA) were assessed in a dUTP nick-end labeling assay. RESULTS: Twenty-one OA patients were included: 5 had congenital bilateral absence of the vas deferens, 8 had genital tract infections, and 8 had idiopathic OA. A total of 8506 spermatozoa sampled from the testis, 18,358 sampled from the caput epididymis, and 18,881 sampled from the corpus/cauda epididymis were assessed. For each patient, spermatozoa from the testis had a lower overall DNA fragmentation rate (6.71% ± 0.75 in average) than epididymal spermatozoa from the caput (14.86% ± 1.89 in average; P = .0007) or the corpus/cauda (32.61% ± 3.11 in average; P < .0001). The DNA fragmentation rates did not differ significantly as a function of the etiology of OA. In this small series, all deliveries were obtained with sperm samples with a low DNA fragmentation rate and delivery rates tended to be higher when testicular sperm (rather than epididymal sperm) was used (35.7% vs 12.1%, respectively; P = .06). CONCLUSION: Our data argue in favor of using testicular sperm (rather than epididymal sperm) for patients with obstructive azoospermia.

Associations between Individual and Combined Polymorphisms of the TNF and VEGF Genes and the Embryo Implantation Rate in Patients Undergoing In Vitro Fertilization (IVF) Programs.

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.

Identity suffering in infertile men.

The suffering caused by infertility in a man can have multiple aspects. It can display a narcissistic dimension, an objectal dimension (object-libido) turned toward others or/and an identity dimension. Two clinical case reports were used here to (i) illustrate all these aspects of infertility suffering, (ii) to evidence the difficulty for infertile men to speak about their infertility and (iii) underlie the importance for professional of medical assisted reproduction to be attentive to this suffering that many men keep silent. An empathetic attention to infertile men may give a way to express this suffering and thus allow the beginning of a psychoanalytic approach which is necessary in infertility and especially for infertile men who do not easily express their suffering.

La souffrance de l'infertilité chez l'homme peut prendre plusieurs aspects. Elle peut avoir une dimension narcissique. Elle peut avoir une dimension objectale, être tournée vers l'autre. Elle peut avoir aussi une dimension identitaire particulièrement douloureuse car elle remet en cause l'identité sexuelle comme l'appartenance à la lignée. Deux cas cliniques vont éclairer cette souffrance que beaucoup d'hommes taisent d'une manière défensive.Cette attitude défensive pourra donner le change et faire croire à une bonne adaptation à la situation. Malgré ou à cause de ce silence les acteurs de l'assistance médicale à la procréation devront rester attentifs à ne pas se laisser abuser par cette apparente absence de difficulté. Une attention empathique pourra donner l'occasion d'une expression de cette souffrance d'infertilité et ouvrir ainsi la possibilité d'une prise en charge psychothérapique jusque là inenvisageable.

The nature of human sperm head vacuoles: a systematic literature review.

Motile sperm organelle morphology examination (MSOME) involves the use of differential interference contrast microscopy (also called Nomarski contrast) at high magnification (at least 6300x) to improve the observation of live human spermatozoa. In fact, this technique evidences sperm head vacuoles that are not necessarily seen at lower magnifications - particularly if the vacuoles are small (i.e. occupying <4% of the sperm head's area). However, a decade after MSOME's introduction, it is still not clear whether sperm head vacuoles are nuclear, acrosomal and/or membrane-related in nature. In an attempt to clarify this debate, we performed a systematic literature review in accordance with the PRISMA guidelines. The PubMed database was searched from 2001 onwards with the terms "MSOME", "human sperm vacuoles", "high-magnification, sperm". Out of 180 search results, 21 relevant English-language publications on the nature of human sperm head vacuoles were finally selected and reviewed. Our review of the literature prompted us to conclude that sperm-head vacuoles are nuclear in nature and are related to chromatin condensation failure and (in some cases) sperm DNA damage.

Le MSOME (motile sperm organelle morphology examination) est une technique d'observation des spermatozoïdes mobiles à fort grossissement (>6300x) à l'aide du contraste interférentiel différentiel de Nomarski. Avec cette technique, des anomalies de la tête spermatique comme les vacuoles peuvent être observées alors qu'elles demeurent souvent invisibles à plus faible grossissement, notamment quand elles sont petites et qu'elles occupent moins de 4% de la surface de la tête. Depuis l'introduction du MSOME dans les années 2000, plusieurs études se sont intéressées à la nature des vacuoles. Sont-elles de nature nucléaire ? de nature acrosomique ? de nature membranaire ? Pour répondre à ces questions, nous avons réalisé une revue de la littérature en suivant les règles PRISMA. Les études publiées sur le sujet entre 2001 et aujourd'hui ont été recherchées dans la base Pubmed en utilisant les mots clés : "MSOME", "human sperm vacuoles" et "high-magnification, sperm". Parmi les 180 études retrouvées, 21 publications écrites en langue Anglaise et traitant de la nature des vacuoles spermatiques ont été sélectionnées et étudiées. Au total, cette revue de la littérature conclut que les vacuoles sont de nature nucléaire, en lien avec une moindre condensation de la chromatine spermatique. Cette moindre condensation chromatinienne représentant un facteur de susceptibilité aux dommages de l'ADN (fragmentation, dénaturation par exemple), les spermatozoïdes vacuolés peuvent aussi présenter plus de dommages de l'ADN que les spermatozoïdes sans vacuole.

Application of a new molecular technique for the genetic evaluation of products of conception.

OBJECTIVES: Karyotyping is a well-established method of investigating the genetic content of product of conceptions (POCs). Because of the high rate of culture failure and maternal cell contamination, failed results or 46,XX findings are often obtained. Different molecular approaches that are not culture dependent have been proposed to circumvent these limits. On the basis of the robust experience previously obtained with bacterial artificial chromosomes (BACs)-on-Beads™ (BoBs™), we evaluated the same technology that we had used for the analysis of prenatal samples on POCs. METHOD: KaryoLite™ BoBs™ includes 91 beads, each of which is conjugated with a composite of multiple neighboring BACs according to the hg19 assembly. It quantifies proximal and terminal regions of each chromosome arm. The study included 376 samples. RESULTS: The failure rate was 2%, and reproducibility >99%; false-positive and false-negative rates were <1% for non-mosaic aneuploidies and imbalances effecting all three BACs in a contig. Detection rate for partial terminal imbalances was 65.5%. The mosaic detection threshold was 50%, and the success rate in macerated samples was 87.8%. The aneuploidy detection rate in samples with cell growth failure was 27.8%, and maternal cell contamination was suspected in 23.1% of 46,XX cultured cells. CONCLUSION: KaryoLite™ BoBs™ as a 'first-tier' test in combination with other approaches showed beneficial, cost-effective and clearly enhanced POC testing.

The vascular endothelial growth factor (VEGF) +405 G/C polymorphism and its relationship with recurrent implantation failure in women in an IVF programme with ICSI.

PURPOSE: 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). METHODS: 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. RESULTS: 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). CONCLUSION: 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.

Genetic polymorphisms influence the ovarian response to rFSH stimulation in patients undergoing in vitro fertilization programs with ICSI.

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 (p<0.01). DISCUSSION: We confirmed the impact of FSHR and ESR2 polymorphisms on the IVF outcome. Furthermore, we showed for the first time that a p53 polymorphism (which is already known to impact embryo implantation) could influence the ovarian response. However, given that this result lost its statistical significance after multivariate analysis, more data are needed to draw firm conclusions. Only the FSHR and AMH polymorphism combination appears to influence mature oocyte numbers but this finding also needs to be confirmed. 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(-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).

Cryopreservation of human spermatozoa decreases the number of motile normal spermatozoa, induces nuclear vacuolization and chromatin decondensation.

Even though cryopreservation of human spermatozoa is known to alter sperm motility and viability, it may also induce nuclear damages. The present study set out to determine whether or not cryopreservation alters motile sperm morphology under high magnification and/or is associated with chromatin decondensation. For 25 infertile men, we used high-magnification microscopy to determine the proportions of various types of motile spermatozoa before and after freezing-thawing: morphometrically normal spermatozoa with no vacuole (grade I), ≤ 2 small vacuoles (grade II), at least 1 large vacuole or >2 small vacuoles (grade III), and morphometrically abnormal spermatozoa (grade IV). The spermatozoa's chromatin condensation and viability were also assessed before and after freezing-thawing. Cryopreservation induced sperm nuclear vacuolization. It decreased the proportion of grade I + II spermatozoa (P < .001). It induced a decrease in the sperm viability rate (P < .001) and increased the proportion of sperm with noncondensed chromatin (P < .001). The latter parameter was strongly correlated with sperm viability (r = 0.71; P < .001). However, even motile sperm presented a failure of chromatin condensation after freezing-thawing, because the proportion of sperm with noncondensed chromatin was correlated with high-magnification morphology (r = -0.49 and 0.49 for the proportions of grade I + II and grades III + IV, respectively; P < .001). Cryopreservation alters the organelle morphology of motile human spermatozoa and induces sperm chromatin decondensation. High-magnification microscopy may be useful for evaluating frozen-thawed spermatozoa before use in assisted reproductive technology procedures (such as intrauterine insemination, in vitro fertilization, and intracytoplasmic sperm injection) and for performing research on cryopreservation methods. If frozen-thawed sperm is to be used for intracytoplasmic sperm injection, morphological selection under high magnification may be of particular value.

Adiponectin and leptin systems in human endometrium during window of implantation.

OBJECTIVE: To measure the expression of adiponectin, leptin, and their respective receptors in the human endometria of fertile women compared with women with unexplained recurrent implantation failure (IF) during the window of implantation. DESIGN: Controlled, prospective, clinical study. SETTING: Teaching hospital and university research laboratory. PATIENT(S): Thirty-one endometrial biopsies from women with IF and 19 fertile controls. INTERVENTION(S): Human endometrial biopsies. MAIN OUTCOME MEASURE(S): Gene and protein expression of endometrial biopsies. RESULT(S): Endometrial leptin expression was significantly lower in the IF group compared with fertile women. In contrast, leptin receptor (Ob-R) expression was higher in endometria of women with IF. Concerning the adiponectin system, adiponectin was expressed to the same extent in both groups. Conversely, the expression of its two receptors, AdipoR1 and AdipoR2, was reduced in endometria of women with IF compared with fertile women. CONCLUSION(S): Although progesterone resistance seems to be a common state of the endometrium in some human reproductive disorders, such as endometriosis or polycystic ovary syndrome, modification in leptin endometrial expression seems to be specific to IF. These results strongly suggest that changes in Ob-R and AdipoR expression profiles [1] should be implicated in the development of uterine receptivity, and [2] may therefore be potential new targets for prediction of IF.

TWEAK appears as a modulator of endometrial IL-18 related cytotoxic activity of uterine natural killers.

BACKGROUND: TWEAK (Tumor necrosis factor like WEAK inducer of apoptosis) is highly expressed by different immune cells and triggers multiple cellular responses, including control of angiogenesis. Our objective was to investigate its role in the human endometrium during the implantation window, using an ex-vivo endometrial microhistoculture model. Indeed, previous results suggested that basic TWEAK expression influences the IL-18 related uNK recruitment and local cytotoxicity. METHODOLOGY/PRINCIPAL FINDINGS: Endometrial biopsies were performed 7 to 9 days after the ovulation surge of women in monitored natural cycles. Biopsies were cut in micro-pieces and cultured on collagen sponge with appropriate medium. Morphology, functionality and cell death were analysed at different time of the culture. We used this ex vivo model to study mRNA expressions of NKp46 (a uNK cytotoxic receptor) and TGF-beta1 (protein which regulates uNK cytokine production) after adjunction of excess of recombinant IL-18 and either recombinant TWEAK or its antibody. NKp46 protein expression was also detailed by immunohistochemistry in selected patients with high basic mRNA level of IL-18 and either low or high mRNA level of TWEAK. The NKp46 immunostaining was stronger in patients with an IL-18 over-expression and a low TWEAK expression, when compared with patients with both IL-18 and TWEAK high expressions. We did not observe any difference for TWEAK expression when recombinant protein IL-18 or its antibody was added, or conversely, for IL-18 expression when TWEAK or its antibody was added in the culture medium. In a pro-inflammatory environment (obtained by an excess of IL-18), inhibition of TWEAK was able to increase significantly NKp46 and TGF-beta1 mRNA expressions. CONCLUSIONS/SIGNIFICANCE: TWEAK doesn't act on IL-18 expression but seems to control IL-18 related cytotoxicity on uNK cells when IL-18 is over-expressed. Thus, TWEAK appears as a crucial physiological modulator to prevent endometrial uNK cytotoxicity in human.

Performance evaluation of microbead and ELISA assays for follicular G-CSF: a non-invasive biomarker of oocyte developmental competence for embryo implantation.

G-CSF in individual follicular fluids correlates with the potential of the corresponding embryo to result in a live birth after transfer in IVF. To evaluate the requirements for routine follicular fluid G-CSF quantification, we compared follicular fluid G-CSF measurements made with two multiplexed microbead assays purchased from Bio-Rad Laboratories and R&D Systems, and a commercial G-CSF ELISA (R&D Systems). Individual follicular fluids (n=139) associated with transferred embryos were analysed to determine cytokine profile and the fate of each transferred embryo was recorded. The effect of multiplexing as well as comparison of the respective performances of the microbead assay with a flow cytometry assay was explored. Multivariable logistic regression analysis was performed and receiver operating characteristic (ROC) analysis was used to determine the performance and sensitivity/specificity of each method for individual follicular fluids. Covariate factors known to influence IVF outcome such as age, serum oestradiol and embryo score were systematically integrated in each analysis. The quantification of follicular fluid G-CSF using microbead assay methodologies, but not ELISA, yielded results showing the utility of follicular fluid G-CSF as a biomarker predictive of a successful delivery (Au(roc): 0.77 [0.68-0.84] (p=0.003) and 0.75 [0.66-0.82] (p=0.004) for Bio-Rad and R&D Systems microbead assays respectively), whereas follicular fluid G-CSF values quantified by ELISA were not predictive (Au(roc):0.61 [0.52-0.70] p=0.84). Microbead assay and flow cytometry appeared similarly efficient for quantifying follicular fluid G-CSF and multiplex versus single-plex assays did not influence the reliability of quantification.

Are zona pellucida laser drilling and polar body biopsy safe for in vitro matured oocytes?

INTRODUCTION: 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. METHODS: 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. RESULTS: No difference in the four criteria was observed between the 3 oocyte groups. CONCLUSIONS: 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.

Sperm chromosome analysis of an infertile patient with a 95% mosaic r(21) karyotype and normal phenotype.

OBJECTIVE: To examine sperm meiotic segregation in a man with mosaic ring chromosome 21. DESIGN: Case report. SETTING: Hospital departments of reproductive biology, cytogenetics, gynecology, and obstetrics. PATIENT(S): One patient referred for cryptozoospermia, heterozygous for a ring chromosome. INTERVENTION(S): Fluorescence in situ hybridization with chromosome 21-specific probes after sperm selection. RESULT(S): A total of 169 spermatozoa were selected; 92.3% carried a normal 21 chromosome, 6.5% the ring chromosome, and 1.2% both. CONCLUSION(S): Ring chromosome frequency in mature sperm cells was low and may be due to preferential meiosis of normal spermatogonia,which could explain the cryptozoospermia and unexpected ratio in this case.

Quality of life of patients undergoing ovarian stimulation with injectable drugs in relation to medical practice in France.

The French study VISION is a multicentric, prospective and retrospective study, designed first to evaluate patients' quality of life during ovarian stimulation for ovulation induction, intrauterine insemination (IUI) or IVF, and secondly to analyse current medical practice in France. Answers were directly entered on a pocket PC. The study was conducted from January to August 2004 and 186 practitioners filled in a total of 1476 questionnaires. The percentages for the different treatments used were analysed according to type of technique [ovulation induction, IUI, IVF/intracytoplasmic sperm injection (ICSI)], type of product used [urinary gonadotrophins, recombinant FSH (rFSH), gonadotrophin-releasing hormone (GnRH) agonists and antagonists] and methods of administration (intramuscularly, subcutaneously, use of Pen-injector, nurses or self-injections). Initial and total amounts of urinary gonadotrophins per cycle were approximately 30% higher compared with rFSH for IUI and IVF. Generally, the impact of these treatments on professional or social life was less than expected. Patients saw self-injection as a significant improvement in their life, especially when using an injection pen, and when other drugs were also self-injected during treatment (human chorionic gonadotrophin, GnRH agonists or antagonists). Clear information is necessary to increase patient's compliance to the treatment. Simplification will make these procedures more patient-friendly and less of a struggle.

Influence of the number of motile spermatozoa inseminated and of their morphology on the success of intrauterine insemination.

BACKGROUND: Although intrauterine insemination (IUI) is one of the most common assisted reproductive technology methods in the world, the relative influence of various semen characteristics on the likelihood of a successful outcome is controversial. The aim of our study was to assess the results of IUI as a function of both the number of motile spermatozoa inseminated (NMSI) and the percentage of morphologically normal spermatozoa after preparation. METHODS: This was a retrospective study of 889 couples who underwent 2564 IUI cycles of ovarian stimulation with HMG or recombinant FSH in our centre between January 1991 and December 2000. RESULTS: A total of 331 clinical pregnancies were obtained, for a pregnancy rate/cycle of 12.91%. When the NMSI was < 1 x 10(6), the pregnancy rate/cycle was significantly lower (3.13%) than in any of the subgroups with NMSI > or = 2 x 10(6). Sperm morphology, assessed before or after preparation, was not in itself a significant factor that affected the likelihood of IUI success. Nonetheless, when the post-migration rate of normal sperm was < 30%, the pregnancy rate/cycle was 5.43% when NMSI was < 5 x 10(6) and 18.42% when NMSI was > or = 5 x 10(6) (P = 0.008). Pregnancy rates did not differ significantly according to NMSI when the percentage of normal sperm after preparation was > or = 30%, or according to percentage of normal sperm when the NMSI was > or = 5 x 10(6). CONCLUSIONS: Our results show that a minimum of 5 x 10(6) motile spermatozoa should be inseminated when the normal morphology of the sperm after preparation is < 30%; the quantity compensates at least in part for the defective quality. If this threshold of NMSI cannot be obtained, IVF should be recommended.

Effects of low concentrations of inhibin B on the outcomes of testicular sperm extraction and intracytoplasmic sperm injection.

OBJECTIVE: To study the effects of undetectable inhibin B concentrations on the outcomes of testicular sperm extraction (TESE) and of intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective study. SETTING: Obstetrics, gynecology, and reproductive biology departments. PATIENT(S): We carried out TESE on 75 men with nonobstructive azoospermia: 42 men had an inhibin B concentration of or = 15 pg/mL (group 2). Twenty-five ICSI cycles were carried out using sperm from men in group 1 (group A1), and 35 ISCI cycles were carried out using sperm from men in group 2 (group A2). The outcomes of ICSI in groups A1 and A2 were compared with those of 81 ICSI cycles performed for obstructive azoospermia (group B). INTERVENTION(S): Testicular sperm extraction, testicular spermatozoa cryopreservation, and ICSI. MAIN OUTCOME MEASURE(S): Testicular sperm extraction outcome, pregnancy, and delivery. RESULT(S): Sperm were significantly less likely to be successfully recovered from men in group 1 than from those in group 2 (21% vs. 48%). The inhibin B concentration was significantly lower in men in whom TESE failed, but the FSH concentration did not differ. The implantation rate per embryo transferred was twofold lower in group A1 (7.4%) than in group B (16%), but this difference is not statistically significant. CONCLUSION(S): Patients with undetectable inhibin B concentration should be informed of the low chances of positive testicular biopsy, and more embryos should be transferred to improve the success rate.

Successful pregnancy and delivery after testicular sperm extraction despite an undetectable concentration of serum inhibin B in a patient with nonobstructive azoospermia.

OBJECTIVE: To describe a successful pregnancy and delivery after testicular sperm extraction (TESE) despite an undetectable concentration of serum inhibin B in a man with nonobstructive azoospermia. DESIGN: Case report. SETTING: Obstetrics and gynecology and reproductive biology departments. PATIENT(S): A 31-year-old woman and a 32-year-old man with nonobstructive azoospermia and an undetectable inhibin B serum level. INTERVENTION(S): TESE, testicular spermatozoa cryopreservation, intracytoplasmic sperm injection (ICSI). MAIN OUTCOME MEASURE(S): Pregnancy and delivery. RESULT(S): Successful pregnancy and delivery of a normal healthy child following a third ICSI cycle with frozen-thawed spermatozoa extracted from the testis. CONCLUSION(S): This case report shows that there is no minimal level of inhibin B below which TESE is always unsuccessful. The delivery of a normal healthy baby is strong evidence to perform TESE in these circumstances.

Biopsied testis cells of four 47,XXY patients: fluorescence in-situ hybridization and ICSI results.

BACKGROUND: A testis biopsy was performed for four non-mosaic 47,XXY azoospermic patients. Spermatozoa were found in three cases and frozen before ICSI. We analysed the various cells found in the four samples by multicolour fluorescence in-situ hybridization (FISH), to evaluate the meiosis and spermatogenesis possibilities of the 47,XXY and 46,XY testis cell lines, and to estimate aneuploidy rate in the resulting spermatids and spermatozoa. METHODS AND RESULTS: Testis diploid cells (either somatic or premeiotic), meiotic, and post-meiotic haploid germ cells were hybridized with probes for chromosomes X, Y and 18. The only patient with no spermatozoa had a homogeneous diploid XXY constitution in the testis; the three other patients presented two cell populations (46,XY and 47,XXY) among their diploid testis cells. All the observed pachytene figures were XY; no XXY pachytene figure was found. The aneuploidy rate among post-meiotic cells for chromosomes X,Y and 18 was 6.75% (5/74). This rate was 1.5% (2/133) for control. Three couples underwent ICSI; four attempts were made, one healthy baby was born. CONCLUSION: FISH results suggest that only 46,XY cells can undergo meiosis.