Enhancing detection of gonococcus in ejaculates of adult males using sperm dilution.

Specific cultures were used to detect growth of Neisseria Gonorrhoeae (NG) in 90 ejaculates of partners of childless marriages. Although no gonococcal growth was observed in undiluted semen, 9 out of 68 subjects with silent infection presented growth of NG in seminal plasma after dilution 1:2 with saline. It is concluded that semen dilution increases the chances of detection of NG in semen samples of asymptomatic gonococci carriers.

Kinetics of steroid responsiveness to hCG during chronic inhibition of testicular function by GnRH analogue.

The effect of daily injections of D-Ser-(TBU)6-LRH-EA10 (GnRH analogue (GnRH-A) 100 micrograms sc) on serum testosterone (T), 17 alpha-hydroxyprogesterone (17OHP) and oestradiol-17 beta (E2) was studied in 4 men. During GnRH-A therapy T, 17OHP and E2 were markedly decreased by the end of the second month. Continuous long-term administration of GnRh-A inhibited testicular function. To test whether the biosynthetic pathway was affected by the regimen, a bolus of 2000 U hCG was given to each subject after 10 months of therapy. Evaluation of the kinetics of steroid responsiveness showed a significant release of T in response to the trophic stimulus, with little or no elevation of serum 17OHP and E2. The response seen in these treated men appeared similar to that found in hypogonadotrophic men and prepubertal boys.

hCG-induced maturation of the seminiferous epithelium in hypogonadotropic men.

The effect of long-term hCG administration on sperm output was evaluated in a study in 3 hypogonadal patients with a selective deficiency of gonadotrophins (LH and FSH). The diagnosis of complete hypogonadotropic hypogonadism was based on clinical and hormonal findings as well as testicular histology. Pubertal maturation took place gradually during hCG therapy. 2 out 3 patients, who were azoospermic before treatment, had spermatozoa in their ejaculate after 12 and 24 months of therapy respectively. These effects on spermatogenesis were reversed after hCG withdrawal for 4 months and the patients again became azoospermic. This azoospermia was not reversed by testosterone (T) replacement therapy, or by addition of HMG to T. In vitro, the crude hCG preparation stimulated cAMP accumulation in rat Sertoli cell cultures indicating that this hCG preparation possesses an 'FSH-like' action. The present findings indicate that hCG therapy alone can induce and maintain spermatogenesis in some patients with complete hypogonadotropic hypogonadism.

Evidence for the presence of androgen receptors in purified rat Leydig cells.

Leydig cells, purified on two sequential Percoll gradients to purities of 89 +/- 1%, were used to study the binding of 17-beta-hydroxy-17 alpha-methyl-estra-4,9,11-triene-3-one(3H-R1881). The accumulation of 3H-R1881 in the nuclear fraction of these cells was time- and temperature-dependent. Specific binding was saturable with an apparent Ka of 0.14 nM-1 and a single class of binding sites at a concentration of 721 fmol/mg DNA. A fraction of the bound radioactivity in the nuclear pellet could be extracted with 0.4 M KCl, and a portion of this extracted steroid was associated with macromolecular species. The nuclear accumulation was androgen-specific. These data are consistent with the presence of androgen receptors in rat Leydig cells.

Hydrotestolactone lowers serum oestradiol and PRL levels in normal men: evidence of a role of oestradiol in prl secretion.

The effect on serum PRL levels of lowering serum oestradiol (E2) concentration by short-term administration of an aromatase activity inhibitor, hydrotestolactone (HT), was studied in six healthy male subjects. After HT administration serum E2 levels decreased from 68 +/- 5.8 to 26 +/- 2.5 pmol/l (mean +/- SE, P less than 0.05). These E2 changes were accompanied by a significant decrease in mean 2-h PRL levels from 11.2 +/- 2.1 to 6.5 +/- 1.6 ng/ml mean +/- SE, P less than 0.05). The evaluation of individual percentage change from basal concentrations showed a varying decrease in all subjects. These findings suggest that under physiological conditions E2 may be one of the factors which control blood PRL concentrations in men.

Dynamics of Prl release in galactorrhoeic normoprolactinaemic women.

Prl responsiveness to TRH and sulpiride was evaluated in 14 galactorrhoeic normoprolactinaemic women. Radiological signs of sellar alteration were present in 6 of these patients. Pituitary responses to TRH did not differ from those obtained in a group of 16 healthy women. Serum Prl levels after sulpiride were higher in the patients than in the controls (P less than 0.01). The delta max response was 256 +/- 16 SE vs 158 +/- 10.6 ng/ml in the control group (P less than 0.01). The mean integrated area after sulpiride was also greater 20738 +/- 1263 SE ng/ml/2 h vs 13 188 +/- 408) (P less than 0.01). The markedly enhanced Prl response to sulpiride in our patients with galactorrhoea could be due to a functional disorder of the hypothalamic-pituitary axis or to a lactotrope hyperplasia.

Testicular responsiveness to chronic human chorionic gonadotropin administration in hypogonadotropic hypogonadism.

Steroidogenic responsiveness to long term hCG administration (1500 U three times a week for 23 months) was characterized in 8 males with hypogonadotropic hypogonadism (HH). During hCG treatment, testosterone (T), which was in the prepuberal range under basal conditions, rose considerably to the upper end of the normal range and remained at that level during the 23 months of observation. A 2.5-fold increase was observed in serum levels of 17 beta-estradiol (E2) an increment less than seen with T. The increment in 17 alpha-hydroxyprogesterone was also lower than that in T throughout the study; thus, the 17 alpha-hydroxyprogesterone to T ratio, despite continuous hCG administration, remained low. Serum androstenedione was slightly increased during hCG therapy. No significant changes were observed in serum levels of dehydroepiandrosterone. These data indicate that continuous long term hCG administration stimulated T levels in HH, with a relatively small change in E2. The kinetics of the T and E2 responses to 2000 U hCG, evaluated after 23 months of therapy, indicated that the testicular response was markedly reduced. No increment in T levels was observed at 24 h; the maximal response occurred at 48 h. This pattern of T response supports the idea that partial testicular desensitization occurs in HH patients receiving chronic treatment with hCG.

Dynamic evaluation of prolactin secretion with sulpiride and thyrotrophin releasing hormone in amenorrhoeic and normally menstruating women.

Sulpiride and TRH were used to evaluate the dynamics of prolactin (Prl) release in normoprolactinaemic, amenorrhoeic and normally menstruating women. The two tests were performed in a randomomized order with an interval of 2-3 days. In the normal women the first test was carried out between days 2 and 4 of their cycle. Basal values during the two tests were not statistically different (9.6 +/- 1.2 SE vs. 10.7 +/- 1.4 ng/ml in amenorrhoeic women with sulpiride and 11.3 +/- 1.4 vs. 12.4 +/- 1.7 in amenorrhoeic women during the TRH test). The two tests were performed while the subjects of the two groups had identical serum oestrogen concentrations. The Prl response following TRH administration was similar in both groups (delta max mean 49.5 +/- 12.5 SE and 49.4+/- 8.5 SE in normal and amenorrhoeic women, respectively). In contast, when release by sulpiride administration was studied the normal women had greater Prl release (delta max mean 169.9 +/- 18.2 SE) than amenorrhoeic women (delta max mean 99.8 +/- 9.4). Either the different oestrogen status (lower in amenorrhoeic women than in menstruating women) or the hypothalamic pituitary dysfunction likely to be responsible for the amenorrhoea in our patients, may explain the lower Prl release observed after sulpiride in subjects with amenorrhoea.

Direct evidence in men for a role of endogenous oestrogens on gonadotrophin release.

In six healthy subjects serum oestradiol was selectively decreased by administering an aromatase activity inhibitor, hydrotestolactone (HT). After HT administration serum oestradiol (Oe2) decreased from 18.7 +/- 2.3 (SEM) to 6.7 +/- 0.6 pg/ml whereas testosterone (T) and dihydrotestosterone (DHT) blood levels were not modified. These oestradiol changes were associated with a significant increase in serum LH and FSH concentrations (P less than 0.001). The administration of tamoxifen, an oestrogen antagonist, to 5 subjects caused a sharp increase in LH and FSH levels (P less than 0.001). Oe2 was unchanged after the treatment with tamoxifen, whereas T levels were significantly higher. The sum of these data suggests that oestradiol under physiological conditions plays a specific role in the feedback mechanism of gonadotrophin release.

Some critical considerations on the use of the nomifensine test in the hyperprolactinaemic syndrome.

Nomifensine has recently been proposed as a dynamic test to discriminate tumoral from functional hyperprolactinaemia. In the present study, this test was performed in 9 subjects with radiological signs of sellar alteration and in 6 with no signs of pituitary lesion. Adopting the criterion reported in the previous study, the test was considered positive in 2 subjects of the first group and in 2 subjects of the second group. Therefore, our study indicates that the ability of this test to discriminate subjects with tumorous or non-tumorous hyperprolactinaemia still appears debatable.

Effect of androgens on prolactin (PRL) release in humans.

In order to evaluate whether androgens were able to affect PRL release, testosterone and dihydrotestosterone were injected intramuscularly in male and female subjects. PRL blood levels were not modified by testosterone either in healthy males or in amenorrhoeic women, and PRL release in males proved unaffected by dihydrotestosterone at the dose used.

Effect of cyproterone acetate acutely administered on the pituitary-testicular axis.

The effect of cyproterone acetate (CA) on the pituitary-testicular axis was studied in 6 healthy men. A dose of 300 mg of CA was administered orally in the early morning, after 3 h blood samples were collected using a multiple sample technique. Testosterone (T) levels were decreased by CA in all subjects (p 0.01), LH in all (p less than 0.01) but one whereas 17-hydroxyprogesterone did not show any significant variation. In vitro, using an equilibrium dialysis, CA displaced T from plasma-binding proteins in males at 37 degrees C. The role of testosterone-binding globulin on the effects of CA on the pituitary-testicular axis remains to be clarified.

Decrease of prolactin by methysergide in amenorrheic hyperprolactinemic women.

The effect of methysergide (MES), a serotoninergic antagonist on prolactin (PRL) blood levels was studied in five hyperprolactinemic amenorrheic women. The drug was administered for five days at a daily dosage of 11.2 mg. MES decreased significantly the PRL blood levels in all subjects (p less than 0.01). Since the MES has been shown to have antiserotoninergic effects and since serotonin has been thought to be involved in the control of PRL release, the effects of MES in lowering PRL might be due to a decrease of serotonin tone.

The stimulation of prolactin secretion by sulpiride in "adolescent gynaecomastia".

In order to evaluate the function of the hypothalamic-pituitary-prolactin axis in "adolescent gynaecomastia" (AG), sulpiride was administered to 7 normal boys and 7 boys with AG. The maximum increase in serum prolactin (PRL) above the mean baseline level (deltamax) was used as index of response. The sulpiride induced a greater PRL release in boys with gynaecomastia than in the controls. Our data indicate that boys with gynaecomastia may have a greater pituitary prolactin pool. The results also illustrate the usefulness of specific neurotrophic agents such as sulpiride as important tools for evaluating the function of the hypothalamic-pituitary-PRL axis.

Effect of the alteration of gonadal feed-back on LH and FSH release in men.

To observe the influence of gonadal feed-back on FSH and LH release in men we studied the blood levels of both gonadotropins before and after orchiectomy in eight subjects. In four orchiectomized subjects the LH and FSH release induced by LH-RH was also studied. The LH-RH was also administered in eight patients with primary gonadal diseases. Our findings clearly show that FSH is more increased than LH by orchiectomy. The LH-RH administration in our subjects increased the FSH and LH similarly. These findings suggest that the specificity of pituitary responsiveness to LH-RH is under the influence of gonadal steroids.

Effect of oestradiol on gonadotrophin release induced by LHRH in men.

In order to study the role of the oestradiol on the feed-back mechanism, fourteen men were subjected to LHRH test before and after treatment with oestradiol. The oestrogen was administered i.m., 3 mug/kg/24 h and 150 mug/kg/24 h, in two groups of patients. Doses of 3 and 150 mug respectively produced a reduction of LH release of 74% and 60% below control. FSH response to LHRH was almost abolished by oestrogens. Our findings show that oestrogens in men decrease the pituitary response to LHRH.

Effects of dihydrotestosterone on LH release induced by LH-RH in men.

In order to determine the effect of dihydrotestosterone on the feed back mechanisim of the hypothalamo-pituitary-testis axis, nine healthy subjects were subjected to LH-RH tests before and after treatment with 5alpha-DHT. A double antibody radio-immunoassay technique was used to measure the concentration of LH and FSH in the plasma. Our results clearly show that the pituitary LH release by LH-RH after pre-treatment with 5alpha-DHT is more prononeced. In the light of actual findings from several laboratories it is not possible to interpret clearly the results of our own experiments, and hence we advance only some hypotheses.