DNA flow cytometry of left and right testes in normospermic patients affected by left varicocele.

To assess whether changes in spermatogenesis are present in men suffering from varicocele who are still normospermic, a comparative flow cytometric analysis of the left and right testicular DNA content was performed on 26 young normospermic males (sperm density > 20 x 10(6)/ml), with varying degrees of asthenospermia. Cell samples were obtained by fine needle aspiration biopsy. Flow cytometric analysis revealed four peaks in the nuclear DNA content: (i) two peaks for haploid cells (1-A), the first composed of highly condensed nuclear cells (1Ac), essentially spermatozoa, and the second of less condensed cells, essentially spermatids (1-Anc): (ii) a third peak of diploid cells (2-D): somatic cells, G1-stage spermatogonia, primary and secondary spermatocytes and (iii) a fourth peak of tetraploid cells, essentially postleptotene primary spermatocytes and G2-M-stage spermatogonia (4-T). Flow cytometry showed the left testis to have a lower percentage of haploid cells than the right (mean 48.4 +/- 17.9 versus 57 +/- 15.4%, P < 0.05). Significantly fewer condensed cells were found on the left side than on the right (respectively 19.7 +/- 11.2 versus 31.5 +/- 13.5%, P < 0.004). The diploid cell percentage was significantly higher in the left testis than in the right (37.0 +/- 18.5 versus 25.5 +/- 9.6, P < 0.003). No statistically significant differences were found in respect of percentages of either non-condensed and tetraploid cells (respectively 26.6 +/- 14.8 and 11.3 +/- 5.6 on the left and 25.9 +/- 10.3 and 12.4 +/- 6.2 on the right). Flow cytometric analysis of cadaver biopsy tissue showed no statistically significant difference between left and right testicles in respect of the percentages of haploid, diploid and tetraploid cells. The reduced percentage of haploid cells and the increase in diploid cells observed in the left testis of our subjects indicate that the testicular function is impaired to a greater extent in the testis ipsilateral to varicocele than in the contralateral testicle.

Clinical and hormonal aspects of male hypogonadism in myotonic dystrophy.

In order to study male hypergonadotropic hypogonadism as completely as possible, and to evaluate its possible effects on muscle atrophy and sexuality, RIA or IRMA methods were used to measure the levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin, total (T) and free (FT) testosterone, estradiol (E), dihydrotestosterone (DHT), sex hormone binding globulin (SHBG), androstenedione (A) and 17-OH-progesterone (17-OH-P) in 29 patients with myotonic dystrophy (MD). The mean hormonal levels +/-SD were: LH 8.0 +/- 4.4 mIU/ml, FSH 17.4 +/- 11.5 mIU/ml, A 200 +/- 130 ng/dl (all higher than in controls); T 406 +/- 290 ng/dl, FT 22.7 +/- 7.0 pg/ml, DHT 55.5 +/- 29.7 ng/ml (all lower than in controls). The low FT and DHT levels (never previously studied in MD) confirm the androgenic deficiency. The high androstenedione levels and low testosterone concentrations suggest defective enzyme 17-dehydrogenase. The duration of the disease correlated with both testosterone (r = -0.56) and FT levels (r = -0.59), showing that hypogonadism tends to worsen progressively. When the patients were divided into three groups on the basis of the severity of muscle involvement (A, B and C), LH and FSH levels were higher in group C (more severe disease) than in group A, respectively 9.3 +/- 4.7 and 20.6 +/- 12.3 mIU/ml versus 4.8 +/- 0.9 and 8.4 +/- 3.8, p < 0.03; T levels were lower in group C than in group A, 337.3 +/- 263.4 ng/dl versus 649.7 +/- 320.3 (p < 0.03); however, there was no significant difference in the FT levels of the three groups, which may imply that hypogonadism is unlikely to have a direct effect on muscle atrophy. About 25% of our patients were impotent; these subjects had higher LH and FSH (p < 0.001) and lower FT levels than the patients who were not impotent (p < 0.03). However, hypogonadism may not be the only cause of impotence as all of the impotent patients belonged to group C and had a very high (CTG)n triplet expansion. We hypothesise that hypogonadism and sexual impairment could be partially due to a muscle cell alteration: i.e. a dysfunction of both the testicular peritubular myoid cells and of the corpus cavernosum smooth muscle.

Postzygotic instability of the myotonic dystrophy p[AGC] in repeat supported by larger expansions in muscle and reduced amplifications in sperm.

We have analysed the [AGC] expansion in leucocytes, muscle and sperm from 17 individuals affected by myotonic dystrophy (DM). Skeletal muscle showed a larger repeat number than leucocytes in the same patient. A similar degree of expansion was detected in differently affected muscles of a single patient. The germline mutation (< or = 350 repeats) was expanded in somatic cells of the progeny in all patients examined. Our results provide evidence of an early postzygotic instability of the [AGC] repeat in DM.

Bone mass and mineral metabolism in Klinefelter's syndrome.

A reduced bone mineral density (BMD) is frequently observed in hypogonadal males; however, very little is known on bone and mineral metabolism in Klinefelter's syndrome (KS). In this study 32 XXY KS patients and 24 healthy age-matched male controls were examined. Serum total and free testosterone (TT and FT) were significantly lower in patients than in controls (TT in KS, 15.1 +/- 7.8 nmol/l; controls, 30.4 +/- 9.1; p < 0.001. FT in KS, 81.8 +/- 24.9 pmol/l; controls, 135.7 +/- 16.4; p < 0.001). 17 beta-Estradiol was slightly higher in KS patients (KS, 49.0 +/- 27.1 pg/ml; controls, 39.3 +/- 16.4 pg/ml), but the difference was not significant. BMD, measured at the spine (L2-4) and at the proximal epiphysis of the left femur, was similar in patients and in the control group (spine: KS, 1.016 +/- 0.142; controls, 1.085 +/- 0.144 g/cm2; p = not significant. Femoral neck: KS, 0.926 +/- 0.149; controls, 0.926 +/- 0.122 g/cm2; p = not significant). Bone GLA protein (BGP) was significantly higher in the KS group (12.7 +/- 4.8 vs 8.9 +/- 5.2 ng/ml; p < 0.02), while serum calcium, serum phosphate, calciotrophic hormones and the fasting urinary hydroxyproline/creatinine ratio (OHP/Creat) were similar in the two groups. A positive relationship between FT and both spine and femoral BMD was found in KS patients. Furthermore, OHP/Creat ratio was inversely related to BMD at the femur, and positively related to BGP in KS patients, but not in normal subjects. These findings suggest that (1) KS patients have normal bone mass, most probably because the hypogonadism is moderate; and (2) patients with lower bone mass appear to have higher bone turnover.

Male hypogonadism in myotonic dystrophy is related to (CTG)n triplet mutation.

The Authors considered the relationship between hypogonadism in myotonic dystrophy (MD) and MT-PK gene mutation. Twenty-seven subjects were studied, and the (CTG)n amplification varied from 70 to 1520 (mean 661 +/- 463). Hypergonadotropic-hypogonadism with LH levels of 6.94 +/- 3.87 and FSH 14.54 +/- 9.58 IU/L was present; testosterone still showed normal values (505.7 +/- 376.2 ng/dl), but 44.4% of patients had abnormal serum level less than 250 ng/dl. We found a significant correlation (p < 0.001) between CTG repeat size and levels of both LH and FSH: these findings suggest that the severity of hypogonadism is related to MT-PK gene mutation.

High intensity physical exercise induced effects on plasma levels of copper and zinc.

Physical exercise leads to many metabolic, cardiovascular, and muscular changes in the body. The trace elements (TE) zinc and copper are directly involved, as enzymatic cofactors, in many of these processes, especially those related to nutrients metabolism, oxygen transport, and formation of usable energy. The effects of high-intensity physical exercise on plasma levels of CU2+ and Zn2+ in 19 subjects are investigated (9 males and 10 females). Plasma copper concentration decreases, and plasma zinc concentration increases, after exercise, in both sexes. After 30 min recovery, both TE concentration values shifts toward rest values in both sexes. These results only partially agree with literature data, probably because we used the treadmill exercise, which makes many muscles work, whereas other authors made their subjects perform a cycloergometer exercise. Physical exercise causes a marked redistribution of TE (copper and zinc) between body stores, bloodstream, and tissues. The condition of high metabolism may lead to a deficiency of TE, requiring supplementation in order to maintain high level performance.

Achievement of spermatogenesis and genital tract maturation in hypogonadotropic hypogonadic subjects during long term treatment with gonadotropins or LHRH.

15 subjects with Hypogonadotropic Hypogonadism (HH) were treated with either gonadotropins (13 cases) or pulsatile subcutaneous Luteinizing Hormone Releasing Hormone (LHRH) (2 cases) for up to 42 months, to study the effects of therapy step by step. The following results were obtained: (A) In postpubertal HH (5 cases = Group A), therapy brought about onset of spermatogenesis within 3 months and its normalization within 6 months. In HH of prepubertal onset (10 cases = Group B), spermatogenesis started within 9 to 21 months and became normal in only 3 cases after at least 18 months. The best sperm counts were obtained in Group A in the third month of treatment (41.75 +/- 43.68 mil./ml) and in Group B in the 36th month (14.87 +/- 17.06 mil./ml). Sperm motility was normal in the majority of the cases in Group A from the beginning but did not become normal in Group B. (B) Seminal fructose and zinc were normal from the beginning of therapy in 66% of the cases in both groups. Zinc became normal in 100% within 3 months in Group A, in Group B within 18. Carnitine was normal in 50% of cases in both groups, contemporaneous with sperm appearance. Transferrin was normal in Group A after appearance of spermatozoa, but in Group B never became normal. (C) We hypothesize that the recovery of fertility passes through the following stages: (1) Functional recovery of Leydig cells, followed by seminal vesicles and prostate. (2) Recovery of epididymal function, which probably implies beginning of the tubular function. Recovery of Sertoli cell function occurs with more difficulty.

Gonadotropin decrease induced by prolonged exercise at about 55% of the VO2max in different phases of the menstrual cycle.

In order to evaluate the influence of physical exercise on the hypothalamic-pituitary-ovarian axis, we studied ten women in the early follicular phase (EFP), twelve in the late follicular phase (LEP) and nine in the luteal phase (LP). The test consisted of a 90-minute physical exercise on a motor driven treadmill at 55-60% of VO2max. Blood samples were taken before, during and after the test. Prolactin and cortisol did not increase in any phase. Estradiol showed a significant increase in LFP (from 361.5 +/- 110.6 to 472.7 +/- 138.9 pmol/L) and in LP (from 457.2 +/- 94.6 to 555.3 +/- 96.9 pmol/L) but not in EFP. Progesterone levels increased significantly only in LP (from 28.2 +/- 6.7 to 33.5 +/- 6.7 mmol/L): Luteinizing hormone (LH) levels decreased significantly in all phases: from 13.7 +/- 2.0 to 10.5 +/- 1.1 IU/L in EFP; from 14.6 +/- 2.1 to 11.5 +/- 1.9 IU/L in LFP and from 7.5 +/- 1.3 to 5.7 +/- 1.0 IU/L in LP, while follicle stimulating hormone (FSH) levels decreased only in LFP (from 8.1 +/- 0.5 to 6.7 +/- 0.6 IU/L). Our exercise protocol (prolonged, continuous and moderate) was able to cause a decrease in gonadotropins levels, and this phenomenon is not due to changes in the other tested hormones.

[Pulsatile treatment with gonadotropin-releasing hormone (GnRH)]. / La terapia pulsatile con Gonadotropin-releasing hormone (GnRH).

The different ways of administration condition the frequency of the pulsatile GnRH, because of the different rate of depot. High frequencies (60-90/min) can be reached only with the intravenous route which is suitable for ovulation induction: ovulation is reached in 73-92% of women affected by hypothalamic amenorrhoea and in 41-51% of women affected by polycystic ovarian syndrome (previously suppressed with buserelin); overstimulation risk is lesser than during therapy with gonadotropins. Subcutaneous route is suitable for puberty induction which needs long-term treatment (18-24 months); the results are generally good, except in hypopituitaric patients. Intranasally route needs frequencies greater than 150-180 min: low frequencies administration (3 times/day) is sufficient to treat cryptorchidism and to reach good results (30-70%); moreover intranasally route can be useful to maintain the results reached with long term therapy with LHRH or gonadotropins.

[Treatment of male hypogonadotropic hypogonadism]. / La terapia dell'ipogonadismo ipogonadotropo maschile.

Males affected by hypogonadotropic hypogonadism can be treated with androgen replacement therapy, if they do not wish fertility. In order to limit or avoid androgen toxicity on the liver, it is possible to use testosterone undecanoate (which is absorbed in the gut by lymphatic system) at the dose of 160-240 mg/die or testosterone esters administered intramuscolarly at the dose of 250 mg/month. Estradiol and DHT derived from testosterone catabolism can be in excess therefore they can be provoke toxic phenomena, even if slight, such as gynecomastia or prostatic diseases. If patients wish fertility, they must be treated with gonadotropins or pulsatile LHRH. Therapeutic effects are very different depending on the different origin of the hypogonadism. In postpubertal onset hypogonadotropic hypogonadism, the response is constant and rapid; sperm count normalization can be reached within 6 months with the only hCG. Prepubertal onset hypogonadotropic hypogonadal men need hu-FSH too and longer treatment (18-24 months); sperm count normalization can be reached in less than half case. Nevertheless fertility can be reached even in oligozoospermic stage. Negative prognostic factors are: pan-hypopituitarism, cryptorchidism, how old are the patients at the beginning of the treatment and small testis volume. It is not yet clear if pulsatile LHRH therapy is profitable in terms of therapeutic results.

Sex hormones and sex hormone binding globulin in males with compensated and decompensated cirrhosis of the liver.

The present work investigates the sex hormone profiles in 50 male patients with liver cirrhosis of different etiology according to the degree of liver dysfunction. The only hormonal impairment in well-compensated cirrhotics (group A) was an increase in mean serum concentrations of estrone, androstenedione, and sex hormone binding globulin. In decompensated cirrhotic patients with ascites (group B), low mean levels of total and free testosterone were found along with normal gonadotropins mean levels. Estrone and androstenedione levels were still elevated, whereas sex hormone binding globulin levels were not different from controls. In decompensated cirrhotics patients with encephalopathy (group C), total and free testosterone mean levels were lower than in group B, and LH mean levels were elevated; estrone levels were markedly high, but androstenedione levels were subnormal; sex hormone binding globulin concentrations were again not different from controls. The few patients with high prolactin levels belonged primarily to this group. Estradiol mean levels were not significantly elevated in any of the groups. It is concluded that the various hormonal patterns of gonadal failure and of the impairment of steroid metabolism and transport, observed in cirrhosis, can be attributed to the degree of liver dysfunction.

Childhood's enuresis in the history of women with hyperprolactinemic amenorrhea.

Hyperprolactinemia, or elevated levels of prolactin in blood, is a normal physiologic post-partum response in lactating women. Non-lactating women with hyperprolactinemia often present during the reproductive years since they may have amenorrhea, galactorrhea, or both. Hypersecretion of prolactin is most commonly due to pituitary adenomas. Women with hyperprolactinemic amenorrhea are often quite anxious, depressed and hostile. It has been hypothesized that these psychological symptoms might antecede the onset of hyperprolactinemia and that hyperprolactinemia may be associated with early developmental problems and may be psychogenic in origin. Twenty patients with hyperprolactinemic amenorrhea and twenty-one normoprolactinemic patients with amenorrhea had an interview covering psychiatric history in order to establish whether they had ever met DSM-III criteria for functional nocturnal enuresis at one time during their childhood. While seven out of twenty (35%) patients with hyperprolactinemic amenorrhea were found to have had functional enuresis during their childhood, only two out of twenty-one (9.5%) normoprolactinemic amenorrheic women reported having had functional enuresis. The difference between the two groups was statistically significant (chi-squared: 3.88; p less than 0.05). We postulate that early stress and developmental problems may present in children as psychological distress and functional enuresis and in women as psychological symptoms (e.g., anxiety and depression) and hyperprolactinemic amenorrhea.

Testicular function and prolactin responsiveness to TRH and cimetidine after renal transplantation.

The hypothalamic-pituitary-testicular axis and the regulation of prolactin secretion were investigated in eleven male renal transplant recipients. Mean serum levels of testosterone and estrone were normal, whereas those of androstenedione and estradiol were low. Mean basal luteinizing hormone (LH) levels were slightly elevated, but the peak responses to 50 micrograms i.v. gonadotropin-releasing hormone (GnRH) were not dissimilar from controls. Both basal and GnRH-stimulated follicle-stimulating hormone (FSH) levels were elevated (p less than 0.02-0.05) and also positively correlated with the time spent on hemodialysis (p less than 0.005-0.002). Basal prolactin (PRL) levels were normal, in all subjects. Nine out of 11 patients had a normal PRL response to Thyrotropin-releasing Hormone (TRH). However only six out of 11 had a normal response to 200 mg i.v. Cimetidine (Cim). Three subjects normally responding to TRH failed to respond to Cim. Uremic primary hypogonadism is not fully reversed by renal transplantation: a slight defect in the pituitary LH release may persist and the impairment of the tubular testicular function is left unchanged. While uremic hyperprolactinemia is corrected, the responsiveness to PRL-stimulating agents, particularly Cim, is not restored to normal, reflecting a derangement at the pituitary as well as the hypothalamic level.

Hyperprolactinemia and psychological distress in women undergoing chronic hemodialysis.

In order to investigate the possible relationship of hyperprolactinemia to psychological distress in patients undergoing chronic hemodialysis, 19 uremic women were evaluated by a semistructured interview and administered the Kellner Symptom Questionnaire. Group A (10 uremic women with hyperprolactinemia) did not differ significantly in anxiety, depression, somatization and hostility from group B (9 normoprolactinemic uremic women). Both groups rated themselves more depressed and hostile than a normal control group of 10 women, and hyperprolactinemic patients were also significantly more anxious than the normal controls.

Male hypogonadism of uremic patients on hemodialysis.

Primary hypogonadism occurring among uremic men on hemodialysis has been widely investigated, yet few data are available concerning the general pattern of steroidogenesis. In 161 hemodialysis patients and in 83 healthy subjects, serum levels of gonadotropins (LH and FSH), prolactin (PRL), testosterone (T), androstenedione (A), estrone (E1), estradiol (E2), and dehydroepiandrosterone-sulphate (DHEA-S) were assessed through RIA methods. Mean +/- SD hormone levels were: LH 45.6 +/- 41.1 mIU/ml, FSH 16.3 +/- 16 mIU/ml, PRL 42.4 +/- 69.1 ng/ml, A 0.83 +/- 0.27 ng/ml, E1 64.3 +/- 31.7 pg/ml, all higher than controls; T 289 +/- 125 ng/100 ml, E2 11.8 +/- 3 pg/ml, and DHEA-S 1.4 +/- 1.4 micrograms/ml, all lower than controls. The A/T and E1/E2 ratios were also higher than controls and showed a good positive linear correlation (r = 0.40; p less than 0.001) between each other. The uremic damage acts at the testis level, impairing the activity of the enzyme 17-beta-hydroxysteroid-dehydrogenase (17-OHSD), even if a derangement of the peripheral interconversion between steroids cannot be excluded.

Osteoporosis and decline of gonadal function in the elderly male.

The bone mineral content was evaluated in 30 male subjects aged between 60 and 90 years using the relief of the percent cortical area (PCA) at the level of the second phalanx of the left-hand index finger, by Garn's method. This was to evaluate the rate of bone loss with increasing age. Testosterone, androstenedione, estrone, 17 beta-estradiol plasma levels were determined in all subjects by the RIA method. 60% of our patients showed increased bone resorption (PCA less than 55%); in these subjects testosterone and androstenedione plasma levels were significantly lower than in subjects not affected by osteoporosis. A positive linear correlation is evident between PCA and testosterone, androstenedione and estrone plasma levels. Thus, like in women, decline of gonadic function determines an increased bone resorption in men too.

Autoantibodies against Leydig cells in patients after spermatic cord torsion.

This study is aimed at searching for the presence of circulating antibodies against frozen sections of human testis, ovary and trophoblast in patients that had spermatic cord torsion. Sixty-eight sera samples were studied. Nine patients (13.2%) were positive for organ specific anti-testis autoantibodies. Six patients were positive for antibodies against Leydig cells: five were positive only with the indirect immunofluorescence technique of complement fixing (ITT/CF), the sixth patient was positive only with the indirect immunofluorescence technique (ITT). The other three patients were positive for antibodies against germ line cells: two patients were positive with both techniques, the third was positive only with indirect immunofluorescence technique. Eight of these patients were negative for antibodies against adrenal cortex while only one case was positive with indirect immunofluorescence technique both on adrenal cortex and Leydig cells. Human lyophilized testis absorbed the reactive antibodies against Leydig cells and germ line cells, while adrenal cortex and lyophilized testosterone were ineffective. This study shows the identification of a specific antibody against Leydig cells and germ line cells in patients after spermatic cord torsion.

Prolactin, aggression and hostility: a discussion of recent studies.

Several studies are summarized in which the relationship of high prolactin levels and self-rated anger-hostility was examined. The Symptom Questionnaire, a state measure which contains an anger-hostility scale, was included in all studies. Women with hyperprolactinemic amenorrhea were found to have higher hostility scores than amenorrheic women with normal prolactin levels. In another study, hyperprolactinemic women were found to have higher hostility scores than female family practice patients, random employees and there was a nonsignificant trend for higher hostility scores than in female nonpsychotic psychiatric outpatients. In both studies, depression and anxiety were also significantly higher. When bromocriptine, a prolactin lowering drug, was administered to hyperprolactinemic women in a double blind crossover study, there was a significant and progressive decrease of hostility, depression and anxiety while on bromocriptine, parallel with the decrease in prolactin and no change on placebo. Post-partum women who had high prolactin levels were significantly more hostile than a control group of employees and as hostile as hyperprolactinemic women. Hyperprolactinemic males were no more hostile than controls. The relationship of prolactin to post-partum aggression in mammals is briefly reviewed. The findings are inconclusive; in the three species studied, postpartum aggression is perhaps enhanced, but does not depend on high prolactin levels. There are no studies on the relationship of prolactin levels and violence in women. Hostility associated with high prolactin levels in postpartum women is perhaps a phylogenetic remnant which may have had the evolutionary advantage of protecting the young.

Hyperprolactinemia and sexual disturbances among uremic women on hemodialysis.

The investigation of a sample of 99 women on maintenance hemodialysis has shown the presence of sexual disturbances to a great extent: the rate of sexual intercourse and the ability to reach orgasm were significantly lower than in age-matched control women. 80% declared a reduction in their sexual desire and the frequency of intercourse was also lower as compared to the period prior to dialysis. Ageing decreased the sexual activity in both the ill and healthy population, but in uremic patients the sexual activity ended at an earlier age. The patients with hyperprolactinemia reported lower frequencies of intercourse and lower percentages of orgasm than normoprolactinemic ones. The incidence of sexual dysfunction and the role of hyperprolactinemia in this respect were similar to those which are found among male patients on hemodialysis.