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Among infertile couples, approximately half have to face with male infertility. In men with non-obstructive azoospermia, surgical retrieval testicular sperm extraction (TESE) of spermatozoa can be attempted, but with low success rates. A specific biomarker that could predict residual spermatogenesis would obviously be of interest before performing TESE. Thus, our aim was to identify biomarkers of residual spermatogenesis in seminal plasma of patients with non-obstructive azoospermia (NOA) using an isotope-coded protein label (ICPL)-based proteomic strategy coupled with conventional protein assay. This retrospective study was conducted in 40 men with NOA at the University Hospital of Nantes and Proteomics Core facility Biogenouest - Inserm U1085 - IRSET, Rennes, France. Comparative ICPL proteomic screening of frozen seminal plasma and correlation with TESE outcome allowed the identification of some differentially expressed proteins. Among them, lectin galactoside-binding, soluble 3 binding protein (LGALS3BP) expression was further confirmed using conventional protein assay, and its interest as a predictor of TESE outcome was then evaluated and compared with conventional clinical and hormonal markers of residual spermatogenesis. Among the 12 differentially expressed proteins identified with comparative ICPL proteomic strategy, seminal LGALS3BP expression was found to be significantly higher in men with successful TESE. Finally, comparative ICPL proteomic screening of seminal plasma appears to be a promising approach for the identification of biomarkers of residual spermatogenesis. LGALS3BP, associated with clinical and hormonal markers, could potentially be used as a predictive marker of successful TESE outcome in patients with NOA., (© 2013 American Society of Andrology and European Academy of Andrology.)

Background: As cancer therapy can be harmful to spermatogenesis, men are generally advised to cryopreserve sperm before gonadotoxic treatment. Here, we compared fresh and frozen-thawed sperm quality in patients according to cancer type, and reported use rate in subsequent assisted reproductive technology (ART) cycles., Methods: This retrospective analysis of a sperm cryopreservation program in all men suffering from cancer was conducted at the University Hospital of Nantes between 1997 and 2007., Results: A total of 1042 men were referred for sperm cryopreservation during this period (438 testicular cancer, 184 other solid cancers, 278 lymphoma and 142 other haematological malignancies). A total of 2577 ejaculates from 1009 men were studied. Sperm characteristics and sperm sensitivity to the freezing process varied according to the cancer type. Frozen sperm was used in 82 patients, with 112 in vitro fertilization and 66 intrauterine insemination cycles performed, leading to a clinical pregnancy rate per cycle of 23.2 and 12%, respectively., Conclusion: Sperm quality and motility alteration during the freezing process can differ according to the type of cancer, but sperm frozen before gonadotoxic treatment in cancer patients can be successfully used in ART.

Sperm parameter values have been shown to decline with age, according to conventional sperm analysis. However, the effect of age on sperm kinematic parameters has been rarely studied, especially in young fertile men. Here, we studied Computer-Assisted Sperm Analysis (CASA) parameters in a large cohort of men with proven fertility, in order to determine if there is a decline with age in this young fertile population. This retrospective analysis of CASA parameters was conducted on all donors included in the sperm donor programme in the Assisted Reproductive Techniques (ART) Centre of the University Hospital of Nantes between 2006 and 2009. Sperm concentration, motility, and kinetic parameters were recorded by a HTM-Ceros system and compared in 3 groups of sperm donors according to their age: <35 years, 36-40 years, and 41-44 years. A total of 362 ejaculates from 138 donors were analyzed. Values for ALH, VCL, LIN, and STR significantly decreased with age. Sperm concentration, motile sperm proportion, and other kinetic parameters did not differ significantly among the groups. The use of CASA allowed the identification of ALH, VCL, LIN, and STR age-related decrease in young men with proven fertility.

The objective of this study was to determine if characteristics of sperm motion determined by computer-aided semen analysis (CASA) after thawing and preparation on discontinuous gradient could predict pregnancy outcome after intrauterine insemination (IUI) from frozen donor sperm. A retrospective analysis of 100 non-selected women undergoing 171 consecutive donor insemination cycles was conducted between January 2006 and April 2007. Semen samples from all donors were analysed after thawing and density gradient preparation. Women who became pregnant and those who did not were comparable in terms of age, ovarian stimulation regimen and indication of IUI with donor semen. Pregnancy rate per cycle was 21.8%, and pregnancy occurred after 2.5 IUI cycles on average. Motility parameters of sperm measured by CASA (VAP, VCL, VSL, LIN, STR, and ALH) and total spermatozoa concentration after preparation on discontinuous gradient showed no difference in both groups. Progressive and total motile spermatozoa concentration, as well as progressive and total motile percentages was significantly higher in pregnancy group. The receiver operating characteristic (ROC) curve analysis showed that total motile percentage >17% and motile concentration >0.9 x 10(6)/mL best predicted pregnancy. In a multivariate analysis, only total motility percentage was able to predict pregnancy. Sperm motility parameters of frozen-thawed prepared donor sperm obtained by CASA do not seem to predict pregnancy in IUI cycles. Total motile and progressive percentages and concentrations remain the best prognostic elements for pregnancy in IUI with donor semen.

The involvement of proteins extracted from spermatozoa of fertile semen in sperm-zona binding was examined under hemizona assay conditions. One droplet of a suspension of spermatozoa was exposed to sperm proteins and then tested for zona binding, while a parallel semen suspension droplet incubated with culture medium served as a control. The reliability of the test was increased by relating the number of spermatozoa bound to each inseminated hemizona to the surface area of the hemizona and expressed as the binding index. For spermatozoa incubated with extracted proteins, the binding index was greater than (P = 0.001) that of controls (125.2 +/- 45.1 versus 63.6 +/- 29.2, respectively). As a first control, two other protein sources (fetal calf serum and human follicular fluid) were tested in the hemizona assay. No significant differences were found in zona binding for other protein-exposed spermatozoa compared with controls. As a second and reverse control, exposure of one hemizona to sperm proteins before insemination with untreated spermatozoa induced a marked decrease (P = 0.0003) in sperm binding, compared with that of the matched hemizona not exposed to sperm proteins (control) (3.4 +/- 1.4 versus 74.5 +/- 6.8, respectively). Taken together, these findings confirm the involvement of extracted sperm proteins in sperm-zona interactions. Therefore, in the cases in which fertilization in vitro fails because of a lack of sperm-zona binding, incubation of deficient spermatozoa with proteins extracted from spermatozoa of fertile ejaculates should restore their ability to interact with the oocyte and, thus, should enhance the prognosis for in vitro fertilization.

Objective: To evaluate the utility of zona drilling for human severe sperm alterations in in vitro fertilization (IVF)., Design: A prospective randomized study was conducted according to random assignment of collected oocytes to a control or zona drilling procedure., Setting: Reproductive Biology Laboratory, University Hospital of Nantes, France., Patients: Twenty infertile couples with severe semen defects and previous IVF failure., Interventions: Ultrasound-guided transvaginal follicular aspiration., Results: Fertilization rates were compared to assess the advantages of zona drilling: 93% (80/86) drilled oocytes survived and 18.75% (15) were fertilized, whereas only 3% (3/100) control oocytes were fertilized. The polyspermy rate for fertilized drilled oocytes was high (10/15; 66.6%). The normal fertilization rate after zona drilling remained very low (5/86; 5.8%) and was not statistically different from that of control oocytes (3/100; 3%). One implantation occurred after replacement of five drilled embryos in three patients but resulted in early miscarriage. Replacement of the single control embryo led to a progressive pregnancy and normal male birth., Conclusions: This study indicates that zona drilling does not improve IVF results in cases of severe semen alterations.