Apoptosis in preimplantation mammalian embryo and genetics.

The presence of dead cells in the preimplantation mammalian embryo has been well described. Since Kerr et al. (1972), it has become apparent that these cells die by apoptosis, a form of programmed cell death.

Cyclodextrin enhances spermicidal effects of magainin-2-amide.

Magainins are antimicrobial peptides with known spermicidal activity. Their activity is inhibited by cholesterol present in eukaryotic cell membranes. Pretreatment of spermatozoa with methyl-beta-cyclodextrin, which extracts cholesterol from cell membranes and induces capacitation, sensitizes them to magainin-2-amide as shown by a decrease in human sperm motility determined by computer-assisted sperm analysis and a concomitant decrease in sperm viability, as measured by MitoTracker(R) Red CMXRos labeling.

Proteasomes in human spermatozoa.

In the present study we describe the localization of proteasomes in human spermatozoa by means of immunolabelling with different monoclonal and polyclonal antibodies detected by confocal microscopy. Western blotting confirmed the specificity of the antibodies and has shown that proteasomes are present in spermatozoa and in seminal fluid. In spermatozoa proteasomes are concentrated in the neck region where the centrioles are located.

Localization of proteasomes in human oocytes and preimplantation embryos.

In the present study we describe the localization of proteasomes in human oocytes, apoptotic preimplantation embryos, and triploid preimplantation embryos by means of immunolabelling with the MCP21 monoclonal antibody detected by confocal microscopy. While in the oocytes proteasomes are scattered throughout the cytoplasm, in the pronuclear zygote they appear to concentrate at the periphery of the cytoplasm and do not enter the pronuclei. During early cleavage stages, proteasome immunolabelling is concentrated in the nuclei, while the examination of triploid blastocysts showed that proteasomes had a similar cellular distribution to somatic cell lines, i.

A randomized prospective study comparing pregnancy rates after clomiphene citrate and human menopausal gonadotropin before intrauterine insemination.

Objective: To determine whether hMG offers an advantage over clomiphene citrate (CC) in achieving pregnancy after IUI with husband's sperm.

Design: Randomized prospective trial.

Setting: Infertility patients in a university teaching hospital.

The mean of sperm parameters in semen donations from the same donor. An important prognostic factor in insemination.

We analysed 12,100 consecutive cycles of artificial insemination by donor spermatozoa in 1901 infertile couples. In our analysis, particular attention was given to finding an appropriate way of taking into account the respective effects of female and male factors on the pregnancy success rate and the level at which these factors act (cycle vs. woman and donation vs.

Applications of laser scanning confocal microscopy to the observation of human oocytes and embryos.

Laser scanning confocal microscopy (LSCM) is a powerful and useful tool in developmental biology. The diverse applications of LSCM in biomedical field has led to advances in the microscopes themselves and the synthesis of novel specific probes for the observation of biological structures and the hypothesis of physiological process. LSCM was used to visualize the cellular actin cortex together with the chromatin in human arrested preimplantation embryos and in unfertilized oocytes obtained by in vitro fertilization.

Morphological and cytogenetic observations of unfertilized human oocytes and abnormal embryos obtained after ovarian stimulation with pure follicle stimulating hormone following pituitary desensitization.

Morphological and cytological observations of 189 unfertilized oocytes and 40 abnormal embryos obtained from 32 patients in a routine in-vitro fertilization programme were performed. Both the oocytes and the embryos were mounted whole to preserve the original topology of all the structural elements. With the applied protocol of ovarian stimulation associating pituitary desensitization and follicle stimulating hormone stimulation, a high degree of immaturity of the unfertilized eggs was observed in comparison with previous reports.

Six year follow-up of cryopreserved human embryos.

In 1987, we became aware of the importance of remaining in contact with couples whose embryos had been cryopreserved for > 1 year. As a result, a questionnaire was designed to follow the fate of these embryos. Of 407 couples with cryopreserved embryos, 262 couples opted to use them within 1 year with the intention of fulfilling a parental plan.

Cumulative conception rate following intrauterine artificial insemination with husband's spermatozoa: influence of husband's age.

This paper presents the analysis of 901 cycles of intrauterine artificial insemination with the husband's spermatozoa (AIHIU) in 274 couples who obtained 80 pregnancies. The cumulative pregnancy rate after three cycles of AIHIU was 22% and reached 39% after six cycles. Univariate analysis disclosed two factors of poor prognosis: duration of infertility > 3 years (P = 0.

[Clomiphene citrate or hMg: which ovarian stimulation to chose before intra-uterine inseminations? A meta-analysis].

This meta-analysis compares different ovulation inductions before intrauterine insemination. Under hMG, the results seem to be better, but this has to be taken with caution as studies are seldom randomized.

[Study of different components in spermatic function after failure of in vitro fertilization].

The fertilizing potential of sperm depends on numerous properties which influence its functional competence. In order to understand the causes of IVF failure, we studied some of these properties-movement characteristics, hyperactivated motility, acrosome reaction-for infertile sperm (0% in vitro fertilization) and compared to those of fertile sperm (> or = 50% fertilization, control group). Movement characteristics, assessed by video-micrographic analysis, are significantly different for the 2 groups: infertile sperm register the lowest values for motility characteristics and hyperactivation.

Kinetic activity of spermatozoa from fertile subjects and asthenozoospermic infertile patients.

The motility characteristics of spermatozoa from asthenozoospermic semen were investigated and compared to the same parameters in fertile semen. The motility characteristics assessed by the CellSoft semen analyser (CRYO Resources Ltd, NY) were the following: curvilinear velocity (VCL), straight line velocity (VSL), amplitude of lateral head displacement (ALH), linearity (%LIN), and beat cross frequency (BCF). Analysis of the data indicated a decreased kinetic activity in the spermatozoa from the asthenozoospermic group which is expressed as a highly significant decrease (P < 0.

[Psychological approach of the infertile male].

Male infertility is usually anxiogenic, and it is often responsible for sexual dysfunctions due to the cultural indissociability of the virility and fertility concepts on the one hand, and to the timing of sexual intercourse in the ovulation period on the other hand. The intrusion of medicine in the life of the infertile couple frequently amplifies the existing psychological problems and discloses other problems by questioning the representation systems of identity, procreation and filiation.

Spontaneous and ionophore induced acrosome reaction in asthenozoospermic infertile semen.

Spontaneous and ionophore-induced ability of spermatozoa to acrosome-react was examined in asthenozoospermic infertile patients and fertile donors. Spermatozoa were washed free of seminal plasma and capacitated in B2 medium for 2 h at 37 degrees C. Subsequently 5, 10, 20 and 30 microM A23187 (final concentrations) were added to equal aliquots of these samples and incubated for an additional 30 min.

Hyperactivated motility of sperm from fertile donors and asthenozoospermic patients before and after treatment with ionophore.

Hyperactivated motility of capacitated sperm was studied before and after contact for 30 min with the Ca2+ ionophore A23187. Sperm from fertile donors and asthenozoospermic infertile patients were incubated in the presence of increasing concentrations of A23187 in the medium. At 5-10 microM, the ionophore induced a significant increase (P less than 0.

[The value of the level of alpha-1,4-glucosidase and seminal l-carnitine in patients with oligoasthenospermia].

Samples of sperm have been obtained from 95 who consulted us for infertility. In each case seminal plasma was examined for levels of alpha-1,4-glucosidase and L-carnitine. Our results have led us to fix the threshold value of 42.

Improvement of human early embryo development in vitro by coculture on monolayers of Vero cells.

Human "spare" embryos, judged unsuitable for freezing because of their poor quality, were cocultured for 5 days on a "Vero" cell layer. These epithelial cells were selected because kidney and genital tract have a common embryologic origin and "Vero" cells are a safe and highly controlled cellular support used for vaccine production. In the control group, the embryos were cultured in culture medium alone (B2 + 15% serum).

HLA antigens and infertility with sperm autoantibodies in man.

Vasectomized men with sperm auto-antibodies (S.A.A.

Stimulation of motility and energy metabolism of spermatozoa from asthenozoospermic patients by 17 beta-estradiol.

Spermatozoa from nine asthenozoospermic patients were incubated for 3 h in Krebs Phosphate Ringer (KRP), supplemented or not with 17 beta-estradiol. 17 beta-estradiol increased the mean velocity and maintained the percentage of motility during the first 2 h of incubation. Oxydative metabolism and intracellular ATP concentrations were enhanced, too, whereas glycolysis remained unchanged.

Evaluation of sperm motility by optical diffractometry of spermatograms.

Evaluation of sperm motility based on analysis of spermatograms by optical diffractometry was performed. Spermatograms are defined as images of sperm tracks obtained on microphotographs by dark field illumination with long exposure time. Single track analysis proved that the details concerning single tracks are "seen" and recognized by the technique of optical diffractometry.

Effects of estradiol and its antagonist tamoxifen on motility and metabolism of human spermatozoa.

An investigation was conducted to evaluate the motility and metabolism of human spermatozoa incubated at 37 degrees Celsius for 3 hours in a krebs phosphate Ringer solution containing different concentrations of 17beta-estradiol and tamoxifen. 17beta-estradiol (17beta E2) stimulated significantly the motility of human spermatozoa. The oxidative metabolism and the intracellular ATP concentrations are enhanced with 10-20 mcg/ml of 17beta E2; the lactate production remains unchanged however.