Congenital bilateral absence of the vas deferens: the impact of spermatogenesis quality on intracytoplasmic sperm injection outcomes in 108 men.

In azoospermic men with congenital bilateral absence of the vas deferens (CBAVD), it is not known whether the outcomes of intracytoplasmic sperm injection (ICSI) depend on the quality of testicular spermatogenesis (as determined histopathologically). We retrospectively studied the impact of spermatogenesis quality on ICSI outcomes in 108 azoospermic men with CBAVD consulting in a university hospital's department of andrology and reproductive biology. As part of an ICSI program, sperm samples were obtained from the epididymis [by microsurgical epididymal sperm aspiration (MESA); n = 47] or the testis [by testicular sperm extraction (TESE); n = 14] or both (MESA + TESE, n = 47). In the TESE group (i.e., TESE-only and MESA + TESE), spermatogenesis was normal in 21 of the 108 men (19.4%) and hypospermatogenesis occurred in 33 (30.5%). The fertilization rate was significantly lower in the hypospermatogenic group than in the normospermatogenesis group (65.6 and 72.9%, respectively; p = 0.02); this was also true for the embryo cleavage rate (88.6 and 92.1%, respectively; p = 0.007), and the proportion of embryos with fewer than 30% of enucleate fragments (79.5 and 86.9%, respectively; p = 0.02). Our study results showed that impaired spermatogenesis had a negative impact on certain early-stage biological outcomes of ICSI. In CBAVD, male factors are likely to exert a harmful effect on the early stages of embryo development.

A predictive score for testicular sperm extraction quality and surgical ICSI outcome in non-obstructive azoospermia: a retrospective study.

BACKGROUND: In non-obstructive azoospermia (NOA), testicular sperm extraction (TESE) is successful in ≈ 50% of cases. A parameter for predicting TESE quality and pregnancy rates after ICSI of testicular spermatozoa is still lacking. METHODS: We retrospectively evaluated the total testicular volume (TTV), hormone levels and TESE quality in 280 patients with NOA. After successful TESE, the characteristics of the ICSI cycles and the take-home baby rates were evaluated. RESULTS: TESE was successful in 149 patients (53.2%). In a multivariate logistic regression analysis, only TTV, FSH and inhibin B were correlated with the TESE outcome. A score including these three parameters was the best predictor of successful TESE (positive likelihood ratio: +3.01). When the score was <18.5, TESE was successful in 77.4% of cases and 'sperm rich' (i.e. yielding >100 spermatozoa) in 91.1% of cases; 42.8% of couples took a baby home. The take-home baby rate did not depend on whether the score was <18.5 or between 18.5 and 3700. If the score was >3700, TESE was successful in 37.8% of cases and 'sperm rich' in 14.3% of cases; only one couple took a baby home (a rate of 7.7 versus 42.8% when the score was <18.5; P< 0.001). Owing to low sperm retrieval on the day of oocyte retrieval, fewer oocytes were injected when the score was >3700 than when <3700. CONCLUSIONS: Thanks to better knowledge of TESE quality, the present score could help to improve care and pre-ICSI counseling for patients with NOA.

[Cryptorchidism: from physiopathology to infertility]. / Cryptorchidie: de la physiopathologie à l'infertilité.

Cryptorchidism is a defect of testicular descent. Hormonal, genetic and environmental factors might also contribute to the aetiology of cryptorchidism and its increased incidence in recent years in industrialised countries. Cryptorchidism itself might be considered a complex disease. This testicular pathology represents the best-characterized risk factor for reduced fertility and testicular cancer. The purpose of this review is to better understand physiopathology and mechanisms of infertility in men with history of cryptorchidism.

[Immunocytochemical analysis of human prolactin receptors using anti-idiotypic antibodies in human breast cancer]. / Analyse immunocytochimique des récepteurs prolactiniques (R-PRL) humains à l'aide d'anticorps antiidiotypes dans les cancers du sein humain.

In this study, we have prepared antiidiotypic antibodies specific of human prolactin receptors (PRL-R) in order to localize these receptors in breast cancer. Antiidiotypic antibodies were prepared using anti-human prolactin (anti-hPRL) sera obtained from New Zealand rabbits. 25 breast cancer were PRL-R positive using radio receptor assay. For immunohistochemistry, the tissues fixed with Nakane and treated by NH4Cl and Saponin to improve staining specificity, were incubated with antiidiotypic antibodies (1/100e). Cytoplasm and membrane of human breast cancer cells were PRL-R positive in 20/25 breast cancer with antiidiotypic antibodies. There was no staining with preimmune serum and immunohistochemical staining elimination by prior absorption with anti-hPRL antibodies.

[Analysis of the results obtained in fertilization in vitro according to the degree and type of teratospermia]. / Analyse des résultats obtenus en fécondation in vitro selon le degré et le type de tératospermie.

The aim of this study was to evaluate the consequences of sperm morphology abnormalities on fertilization capacity. We selected 115 couples who consulted for in vitro fertilization at the Salengro Maternity Ward (Lille, France). Teratospermia was > 40% in all. The mean rate of cleavage was 23% and was significantly lower than in a population which had had in vitro fertilization for a tube indication (63%, p < or = 0.001). In addition, this level was significantly correlated with teratospermia (p < or = 0.05). The ROC method demonstrated that above the threshold of 74% teratospermia, fertilization decreases significantly. We distinguished several types of abnormal morphology and demonstrated a weak correlation between head abnormalities and the rate of cleavage, a stronger correlation between abnormalities of the intermediary piece and rate of cleavage and finally the absence of any correlation between flagella abnormalities and the rate of fertilization. Teratospermia is therefore an important factor in evaluating fertilization capacity. A better selection of normal spermatoids could improve the chance of success in cases of male sterility.

Circannual rhythms of sperm parameters of fertile men.

Mean monthly values for ejaculate volume, sperm density, and percentage of motile sperm were calculated from semen analyses performed on volunteered donors to the laboratory from 1979 to 1986. The data of 4,196 ejaculates were computerized by the single Cosinor method for searching on circannual rhythms of the parameters. The study does not detect circannual rhythms, neither of the ejaculate volume nor of the percentage of motile sperm. A statistically significant high-amplitude seasonal variation was detected in sperm court, the highest values being recorded in late winter and early spring, the lowest values being recorded in late summer.

Immunohistochemical detection of prolactin and its receptors in human testis.

Prolactin (PRL) and PRL receptors were localized by light microscopic immunohistochemistry in the human testes on interstitial Leydig-like cells. Similar stainings were obtained with both monoclonal antihuman PRL antibodies and polyclonal antibodies raised against rabbit mammary gland PRL receptors. Moreover, the detection of PRL binding sites by radio-receptor assay on the same samples gave a confirmation of these results. This suggests that PRL might act directly on Leydig cells in the man, in the same way as in the rodent.

Modulation of prolactin receptors (PRL-R) by lactogenic and steroid hormones in human breast cancer cells in long-term tissue culture (T-47D).

In order to improve the knowledge of prolactin receptors (PRL-R) in human breast tumors, we studied PRL-R modulation by lactogenic and steroid hormones in the PRL-R rich human breast cancer cell-line, T-47 D. The PRL-R were assayed on a preparation of cell total membranes. We demonstrated an abnormal homologous in vitro regulation of PRL-R. Concentrations of human growth hormone (hGH) greater than 500 ng/ml were required to cause a decrease in PRL-R, with a maximal down-regulation at 2000 ng/ml and for 48 hours. Human placental lactogen (hPL) induced a decrease in PRL-R at concentrations greater than 500 ng/ml but later than hGH; ovine prolactin (oPRL) had no effect on PRL-R. Moreover, we also demonstrated that progestins specifically modulated the expression of PRL-R in T-47D cells: Org 2058, a synthetic progestin induced a statistically significant increase in PRL-R after a twenty-four hour incubation period: this effect was already observed at 10(-9) M and was maximal for 10(-6) and 10(-5) M (186% +/- 3.5% (+/- SEM) for total PRL-R). At 10(-6) M, the stimulation occurred early at three hours and was maximal at twenty-four hours. Conversely estradiol (10(-9) to 10(-6) M), cortisol (10(-9) to 10(-6) M), dexamethasone (10(-9) to 10(-5) M) and RU 486 (10(-9) to 10(-5) M), a progestin and glucocorticoid antagonist, had no effect on PRL-R levels. The Org 2058 PRL-R stimulation was abolished in the presence of RU 486. The abnormal PRL-R down-regulation in the human breast cancer cell-line, T-47D, may contribute a growth advantage to these malignant cells over normal tissues. The progestin PRL-R dependence suggests that high levels of PRL-R may reflect a functional progesterone receptor (Pg-R) and a highly hormone-dependent-phenotype of the tumor. These results support a potential role of PRL in the etiology of breast tumors and may have important implications in the management of human breast cancer.

[Correlation of the penetration test of zona-free hamster ovocytes and other sperm parameters. Value of this test in evaluating male infertility]. / Corrélation entre le test de pénétration des ovocytes de hamster dépellucidés et les autres paramètres du sperme. Valeur de ce test dans l'évaluation de la fertilité masculine.

On investigating infertility in 162 couples the zona-free hamster egg penetration test was carried out. As a control we tested the ejaculates of 34 fertile men. There is a significant correlation between the percentage of oocytes that were penetrated and the count (p less than 0.001) in motility after one hour (p less than 0.001) and the count after migration (p less than 0.001). The outcome in 129 infertile couples made it possible for us to confirm that the percentage of oocytes penetrated, the initial motility and the number of motile sperms after migration are the best parameters for assessing sperm function that are known (p much less than 0.001) for investigating male fertility. Finally a study of the test in fertile sperm donors made it possible for us to raise statistically significantly the numbers of pregnancies per artificial insemination cycle when the percentage of oocytes penetrated is higher than 50% (p much less than 0.001). Although the egg penetration test did not prove male sterility it can be considered an extra test to be carried out over and above the sperm count and post-coital test in evaluating the fertilisation capacity of sperm.

Characterization of insulin-like growth factor 1 receptors (IGF1-R) in human breast cancer cell lines.

The binding characteristics of IGF1 on membranes prepared from 5 human breast cancer cell lines were investigated in detail. The presence of one class of high affinity IGF1 binding sites was demonstrated (BT-20: n = 230 fmol/mg protein, Ka = 0.7 nM-1; MCF-7: n = 124 fmol/mg protein, Ka = 1 nM-1; T-47D: n = 61 fmol/mg protein, Ka = 1.1 nM-1; HBL-100: n = 18 fmol/mg protein, Ka = 3.2 nM-1; MDA-MB-231: n = 7 fmol/mg protein, Ka = 2.8 nM-1). Chemical cross-linking of 125I IGF1 to breast cancer cell membranes then sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed one major band of relative molecular weight 130000. The specificity of these receptors was studied: native or recombinant IGF1 had the same potency to inhibit 125I IGF1 binding; IGF2 was able to compete for binding, whereas insulin competed with a potency lower than 1/100 that of IGF1. These characteristics of IGF1 binding sites in breast cancer cell membranes correspond to the previously described binding unit of type I IGF receptors (IGF1-R). Finally we determined that for a routinely used standard assay, it was necessary to incubate for 5 h at 4 degrees C a high amount of membrane protein (400 micrograms) and 200,000 cpm of tracer. Considering the known effect of IGF1 on breast cancer cell multiplication, it is tempting to suggest that this factor might play a major role in the growth of breast cancer: the measurement of IGF1-R, using this standardized method, will give an assessment of these tumors IGF1 sensitivity; it can be performed on the membrane fraction obtained when preparing cytosol for steroid receptor assay.

[Peripheral effects of prolactin in reproductive function. II. Female reproductive function]. / Effets périphériques de la prolactine dans la fonction de la reproduction. II. Fonction de reproduction femelle.

Prolactin (PRL) has an important peripheral role to play in female reproductive function. This hormone, in physiological concentrations, is necessary so that the follicle can mature properly. It also helps the maturation of oocytes and is essential for the physiological action of the corpus luteum so that progesterone production can be stimulated. It inhibits its catabolism and is responsible for keeping up the numbers of LH and oestradiol receptors. It works through the intermediary of prolactin receptors which are localised on the granulosa cells. The mechanism is through immuno-cytochemistry and it can be distinguished biochemically (Ka = 0.5 10(10) M). Probably, when levels are high it upsets follicular maturation by inhibiting the biosynthesis of the oestrogens and also by stimulating the secretion in the granulosa cells of a substance that inhibits oocyte maturation: and prolactin exercises a luteolytic action by stimulating the catabolism of the hormone and lessening the numbers of LH receptors. On the other and, there are high concentrations of PRL in amniotic fluid. As endometrial cells undergo the phenomenon of decidualization they produce PRL which is identical biochemically with the pituitary hormone. Certain steroids seem to regulate this synthesis of PRL. This hormone must play a role in fetal osmoregulation through the intermediary of prolactin receptors.

[Peripheral effects of prolactin in reproductive function. I. Male reproductive function]. / Effets périphériques de la prolactine dans la fonction de reproduction. I. Fonction de reproduction mâle.

Prolactin plays a peripheral role in male reproductive function just as it does in female function. Prolactin, through the medium of immunocytochemistry acts on testicular steroidogenesis through prolactin receptors which are sited on the Leydig cells. It alters the number of LH receptors and therefore the sensitivity of the testis to central stimulation and equally interferes with androgen synthesis. In the areas of physiological concentration the principal effect seems to be to stimulate secretion of testosterone by keeping up the number of LH receptors; but when there is acute hyperprolactinaemia, testosterone secretion is lowered or perhaps not changed at all because of a "post-receptor effect", in spite of there being larger numbers of LH receptors. The direct effect of prolactin on spermatogenesis has not yet been worked out and the results that have been obtained are controversial. Prolactin exerts a direct stimulating effect on the growth of prostatic cells working synergistically with testosterone through specific prolactin receptors. It could play a role in certain cancers of the prostate.

Presence and characterization of insulin-like growth factor 1 receptors in human benign breast disease.

Insulin-like growth factor 1 binding sites were characterized in human benign breast disease. We demonstrated the presence of one high affinity binding site. Chemical cross-linking of [125I]IGF1 to benign breast disease membranes in reducing condition and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed one band with an apparent Mr of 130,000. The specificity of the binding was studied: IGF 2 was a good competitor whereas insulin competed for binding with a potency lower than 1/100 that of IGF1. This IGF1 binding corresponded to the previously described type I IGF receptor (IGF1-R). IGF1-R was assayed in 35 cases of benign breast disease and two samples of normal breast tissue. Forty-three per cent of the lesions were IGF1-R positive. The mean geometric level of specific binding was 1.98% in the whole population, it was significantly lower in adenofibromas (1.55%) than in epithelial hyperplasia (2.5%); it was 2% in dystrophic disease. IGF1-R was undetectable in normal tissue. Considering our previous results showing that almost all the breast cancers contained IGF1-R, these data suggest that the increase in IGF1-R could be a marker of malignant tumor development.

[Copper - a major contraceptive agent?]. / Le cuivre, agent contraceptif majeur?

PIP: The effect of copper ions on the motility of human sperm was studied in vitro to determine whether copper is toxic to sperm. Sperm samples from 30 men of proven fertility who had high quality sperm were collected, separated from the seminal fluid, and resuspended in Menezzo, a chemical medium designed to resemble the secretions of the Fallopian tubes. Menezzo contains all the known constituents which aid sperm capacitation, including mineral salts, organic compounds, free amino acids, and enzymes. Its pH is between 7.4 and 7.6. After 1 hour of incubation the dead and weak sperm were separated out and only mobile sperm were tested. Crystallized copper sulfate was added to 4 of the 5 sperm-Menezzo mixtures. The final concentrations were of 1 millimolar, .1 millimolar, .01 millimolar, and .001 millimolar. Sperm motility showed a statistically significant decrease in the concentration of 1 millimolar but only after a latency period of at least 5 hours. At lesser concentrations and in shorter time periods no significant difference was observed compared to the control. The copper released from a copper IUD into the uterine fluid does not exceed a concentration of .1 millimolar. The lower concentration and the long latency period before the toxic effect manifests itself, seem to indicate that copper has no direct contraceptive effect. Copper is however known to promote inflammatory reactions, and it may work in vivo through the intermediacy of the leukocytes in the uterine cavity, either by directly stimulating phagocytosis or by release of substances toxic to sperm.^ieng

Immunohistochemical localization of enkephalins in adult rat testis: evidence for a gonadotrophin control.

The recent detection of immunoreactive POMC derived peptides in the male reproductive tract raised the possibility that these hormones might regulate reproductive function. In addition, a variety of studies suggested that POMC derived peptides have paracrine effects in the testis. In this present study, after HCG addition to the organ culture medium, Sertoli cells cytoplasma became immunoreactive with the enkephalins antisera.