[A possible therapeutic improvement of the antibacterial effect by monitoring the production of leukocyte free oxygen radicals during antibiotic treatment of infertile patients with chronic bacterial prostatitis]. / Possibile ottimizzazione terapeutica dell'effetto antibatterico attraverso il monitoraggio della produzione leucocitaria di RLO durante antibiotico-terapia in pazienti infertili con prostatite cronica batterica.

In the male chronic accessory gland bacterial infections (AGBI), antibiotic treatment (AT) efficacy usually evaluates the antimicrobial outcome through one or more spermiocolture (SC) become negative. Recently, bacterial olipeptide fMPL has been used to detect a specific Radical Oxygen Species production by leukocytes (L-RLO), even when they are present at low concentrations in sperm fractions specimens. In the male AGBI could be present endogenous fMLP at sufficient levels to produce L chemiotaxis in the semen, and secondly active their specific L-RLO production, till when bacteriospermia remained positive. In the Percoll 50% (Pc50%) fraction, at higher L concentrations, from 22 infertile patients affected by chronic bacterial prostatitis (BP) and enrolled to a randomly AT treatment with a Quinolone (n = 12) or a Macrolide (n = 10), through two secondary milestones (possible elevated basal L-RLO in the pre-treatment, T0; significant changes in the basal and fMLP-stimulated L-RLO production within each treatment group and between groups), we would verify if a normalized L-RLO production could be taken as break-point for the AT withdrawal before checking SC for control. Indeed, in T0 all patients had positive SC and exhibited both basal and fMLP-stimulated L-RLO levels higher than those observed in 2 control groups (group cA = 10 fertile men, without chronic BP; group cB = 10 patients affected by chronic abacterial prostatitis (AP). On the 3rd AT cycle for 14 days, together in 20/22 AT-treated patients, basal and fMLP-stimulated L-RLO levels become low or normal, as well as their SCs become negative (CFU/ml = 0), whilst in a third control group (group cC) of 10 not-AT-treated BP patients, through matched-follow-up observations, these L-RLO values were always elevated and their SC remained positive (CFU/ml > or = 10(5)). In chronic BP patients, AT seems to demonstrate both antimicrobial effectiveness and reduction of L-RLO production with values similar to those of control group cA. The monitored L-RLO values during AT could be useful in order to ottimize antimicrobial effect: this tool being able to previse SC outcome, could be assumed to define clearly AT break-point and/or cycle numbers.

Effectiveness of long-acting gonadotrophin-releasing hormone agonist treatment in combination with conventional therapy on testicular outcome in human orchitis/epididymo-orchitis.

A group of 24 patients suffering from acute post-pubertal orchitis/epididymo-orchitis were prospectively randomized to treatment over a total period of 3 months with a combined conventional plus long-acting gonadotrophin-releasing hormone analogue (GnRHa) (leuprolide, 3.75 mg every 27 days) therapy (n = 13, group A) or conventional therapy alone (n = 11, group B) respectively, in order to verify any differences in clinical and sperm outcome during treatment and follow-up. Seven patients affected by uncomplicated mumps (group C), receiving GnRHa for 3 months, served as an additional control group. Combined leuprolide plus conventional therapy provided better outcomes than conventional therapy alone in terms of scrotal ultrasound scans and serum antisperm antibodies (ASA) at time of presentation and monthly thereafter throughout the study: the most solid data relate to testicular atrophy in three out of 11 patients from group B and their sAb positivity persisting, as opposed to zero out of 13 in group A. Moreover, semen analyses performed at 3-6 months after therapy withdrawal revealed in groups A and C mean total sperm counts and percentages of oval forms significantly higher than those observed in group B. It is concluded that GnRHa treatment is a promising pharmacological tool with no side-effects, effective in reducing testicular vascular leakages in male orchitis.

[Abnormal production of oxygen free radicals by low leukocytospermia in purified fractions of semen from infertile patients]. / Abnorme produzione di radicali liberi di ossigeno originati da bassa leucocitospermia in frazioni purificate del seme di pazienti infertili.

Seminal leucocytes (WBC) play an important role, whereas this is still unknown. Quantitative normal threshold-value for leucocytospermia (Ls) (WBC < or = 1 million/ml) seems not to be a safety predictive marker of male accessory gland infection (MAGI) and/or MAGI-related adverse effects on semen quality and/or fertility because several drawbacks: a) inadequate staining techniques are responsible of an underestimated WBC concentration in spermatic fractions (100%) obtained from high-density Percoli gradient (Pc 100%); b) Ls could be correlated far better to the clinical findings and/or outcomes if some biochemical data on L metabolism are also studied (Radical Oxygen Species-ROS-generation, phagocytosis, leucotoxins). In order to point out importance about an joint morphological/biochemical analysis about Ls, we recruited 76 selected infertile (from 2-7 yrs) patients (aged 32-45 yrs) who, in their 50% Percoli fractions, had previously produced WBC-ROS > 95th percentile values registered from our control group (n = 28, fertile men). In all infertile population we have analysed through Pc100 fraction either the percentage of WBC-negative patient's specimens (group A) and within WBC-positive ones if a significant difference was present in the maximal WBC-ROS production among different subgroups at WBC concentrations (group B:0.01-0.09, group C:1-10, group D: > 10 x 10(4) WBC/10(7) spermatozoa).(ABSTRACT TRUNCATED AT 250 WORDS)

Involvement of corticotropin-releasing hormone and endogenous opioid peptides in prolactin-suppressed gonadotropin-releasing hormone release in vitro.

Corticotropin-releasing hormone (CRH) has been shown capable of inhibiting hypothalamic gonadotropin-releasing hormone (GnRH) release through activation of an endogenous opioid peptide (EOP)-dependent mechanism. Recently, we have shown that prolactin (PRL) stimulates CRH release and inhibits GnRH release by hypothalami explanted from male rats. Thus, the present study was undertaken to investigate whether the inhibitory effect of PRL on GnRH release in vitro is mediated by CRH and/or EOP. To this aim, the release of GnRH in response to PRL was evaluated in presence of CRH9-41 alpha-helical (CRH-9-41), a CRH receptor antagonist, and/or naloxone (NAL), a nonselective opioid receptor antagonist, using a static hypothalamic organ culture system which enabled us to evaluate immunoreactive GnRH (iGnRH) release from individually incubated longitudinally halved hypothalamic. As previously shown, PRL at the concentration of 100 nmol/l inhibited basal iGnRH release by about 35%. CRH9-41 or NAL overcame the inhibitory effect of PRL on iGnRH release in a concentration-dependent fashion. The simultaneous co-incubation with both antagonists was not more effective than each single antagonist. CRH9-41 did not have any effect on basal iGnRH release whereas NAL, as previously reported, increased it. In addition, PRL at the concentration of 100 nmol/l stimulated basal hypothalamic beta-endorphin (beta-EP) release. In conclusion, these data show that antagonism to CRH receptors counteracts the suppressive effects of PRL upon GnRH release and that PRL is able to stimulate hypothalamic beta-EP release in vitro.

Effect of acetylcarnitine treatment in oligoasthenospermic patients.

Acetylcarnitine (AC), present in human spermatozoa and seminal fluid, plays an important role in sperm metabolism. To further investigate the effect of AC on sperm quality, AC (4 g/day) was given to 20 patients with idiopathic oliogasthenospermia for 60 days. AC had no effects on sperm density and total motility, but it did significantly increase progressive sperm motility (mean +/- SEM: 21.7 +/- 3.2% vs 38.2 +/- 4.7). The increment in sperm motility was sustained ( > or = 40%) in 12 patients (mean increment 2.7 fold). This parameter returned to basal value 4 months after therapy discontinuation. Five pregnancies occurred during treatment and only 2 during the 4 months follow-up ensuing therapy discontinuation.

Therapy with human chorionic gonadotrophin alone induces spermatogenesis in men with isolated hypogonadotrophic hypogonadism--long-term follow-up.

The effects of long-term (14-120 months) hCG-treatment of 17 male patients affected by isolated hypogonadotrophic hypogonadism (IHH) on testicular volume, plasma testosterone levels, and sperm concentration were assessed. Mean testicular volume increased from 3.8 +/- 0.2 (Mean +/- SEM) ml to a maximal of 14.9 +/- 1.1 ml after 22.2 +/- 2.3 months of hCG treatment. Maximal testicular volume correlated positively with the volume recorded before the patients had undergone any previous treatment. Testicular growth was also analysed by sorting the patients into two sub-groups according to whether their initial testicular volume was less than 4 ml (small testis subset, STS) or greater than or equal to 4 ml (large testis subset, LTS), supposedly indicating complete or partial gonadotrophin deficiency, respectively. Testicular volumes in the LTS group were always greater than those of the STS. Plasma testosterone levels reached adulthood values during hCG treatment and no statistically significant difference was detected between LTS and STS patients with IHH. Thirteen patients (70%) became sperm-positive during treatment with hCG alone; five out of eight (60%) were STS patients and eight out of nine (90%) were LTS. In addition, LTS patients always had a greater sperm output than did STS patients. Sperm concentration correlated positively with maximal testicular volume, but not with patient age, length of treatment, or initial testicular volume. The administration of hMG to eight of these patients caused an increase in testicular volume in two patients but the mean volume was not statistically different from that recorded at the end of treatment with hCG alone. Similarly, sperm concentration improved in three patients but again it did not differ significantly from that achieved in the course of hCG treatment. It is noteworthy that one patient became sperm-positive after the addition of hMG to his therapeutic regimen. Among sperm-positive patients attempting conception, seven out of 10 succeeded, two of whom were from the STS group. In summary, this study indicates that hCG alone is an effective treatment to induce complete spermiogenesis in IHH patients regardless of their initial testicular volume. However, a number of IHH patients may benefit from the addition of hMG in terms of testicular volume, sperm output, and pregnancy outcome.

[Antibiotic therapy in infertile subjects with chronic gonococcal infections: measurement of sperm output]. / Antibioticoterapia in soggetti infertili con infezione cronica da gonococco: mediazione dell'output spermatico.

Several antibiotics of choice for the treatment of Gonorrhoea have proved to be effective in the acute forms of this disease. Men infected with gonococci may not develop or may not notice any symptoms at all: thus, complications of gonococcal infection can cause semen inadequacy and infertility. We are not aware of any previous paper on the treatment of chronic asymptomatic gonorrhoea (IUGC) and the cytotoxic effects in sperm maturation (so called "stress pattern tubular syndrome") and/or motility possibly mediated by some antigonococcal agents after prolonged period of administration. In IUGC the aims of therapy should be both the bactericidal activity and the improvement in the seminal quality. One-hundred eleven out of 785 male partners of infertile marriages were studied. They had negative post coital test and culture positive for N. Gonorrhoea in their semen. They were subdivided in six random groups and were treated with Ceftazidime (CFZ = 20 cases), Cefonicid (CFN = 20 cases) Spectinomycin (SPM = 15 cases), Aztreonam (AZT = 20 cases), Piperacillin (PP = 18 cases) and Doxycycline (DXC = 18 cases) respectively. After 7 days of treatment (C1) seminal cultures became negative in 25 cases (22.5%) and no seminal parameter showed significant variation among groups. When the treatment was repeated other twice (C3. = 1 week every month for 3 months) the antigonococcal efficacy was complete (100%), but various effects on seminal quality (Density, D; Motility, M, Oval forms, O; Sperm precursors, Sp; nemaspermic penetration depth in bovine cervical mucus PN; rate of pregnancies) were observed.

Generation of reactive oxygen species in subgroups of infertile men.

The capacity to generate reactive oxygen species (ROS), both basally and after stimulation with the calcium ionophore A23187, was examined in the motile fraction of sperm isolated after swim-up from the semen of 10 naturally fertile men and three groups of infertile patients. The latter included: (1) men with a non-bacterial inflammation of the genital tract (n = 10); (2) men unable to impregnate their partners during an intra-uterine insemination programme (IUI) (n = 8) and their matched controls (n = 6); and (3) men with hypogonadotrophic hypogonadism (HH) who remained infertile after induction of spermatogenesis with gonadotrophin or gonadotrophin-releasing hormone therapy (n = 3) and their matched controls (n = 3). The levels of ROS production were elevated in the sperm of some infertile men with inflammation of the genital tract compared to those found in 10 naturally fertile men. In addition, sperm from those patients who remained infertile after an IUI programme produced higher amounts of ROS compared to their control group who became fertile. Similarly, the production of ROS by sperm from three patients with HH who remained infertile was significantly higher than those of the three men who became fertile. These data suggest that an excessive production of ROS by sperm may explain some cases of idiopathic male infertility.

lntra- and Inter-Individual Variability in Growth Hormone Responses to Growth Hormone-Releasing Hormone*.

Abstract Normal subjects show a wide range of growth hormone (GH) responses to growth hormone-releasing hormone (GHRH) stimulation, but it is uncertain whether this variability reflects differences among individuals or whether it would also be observed on repeated tests of the same subject. To clarify this, we tested nine normal men repeatedly with iv bolus doses of 1 mug/kg GHRH(1-44)NH(2). Most subjects showed wide variations in their GH responses on repeated testing, and the intra-individual variability was nearly as great as the inter-individual variability in responses, accounting for about two-thirds of the overall variance. A minority of subjects had lower and less variable responses. Ultradian fluctuations in hypothalamic somatostatin secretion may account for this marked intra-individual variability.

Pituitary and adrenal response to ovine corticotropin-releasing hormone in women with polycystic ovarian syndrome.

It has been hypothesized that there is an adrenal abnormality in the polycystic ovary syndrome (PCO). This study was undertaken to examine this hypothesis in a more physiological way, by enhancing the ACTH secretion in response to ovine corticotropin releasing hormone (oCRH) injection so that adrenal androgen and glucocorticoid responsiveness to endogenous stimulation could be examined. Plasma ACTH and the ACTH and cortisol (F) response to oCRH were normal. The plasma T and dehydroepiandrosterone (DHEA) responses were also normal. The androstenedione (A) response, however, was exaggerated. This study supports the hypothesis that the adrenal gland in patients with PCO produces increased amounts of androstenedione in response to ACTH stimulation.

Dynamics of plasma gonadotropin and sex steroid release in polycystic ovarian disease after pituitary-ovarian inhibition with an analog of gonadotropin-releasing hormone.

To assess the dynamics of the suppression and recovery of plasma gonadotropins and sex steroids during and after inhibition of pituitary-ovarian function by a long-acting agonist GnRH-analog (GnRH-A), eight patients with polycystic ovarian disease were treated with 12 micrograms/kg X day GnRH-A for 56 consecutive days. In response to GnRH-A, these patients had a sharp and pronounced decline of their initially elevated immunoreactive LH and bioactive LH (bioLH) levels. Plasma immunoreactive FSH levels declined more rapidly than did bioLH, but the FSH decline was less sustained. Plasma testosterone, androstenedione, and estrone (E1) levels also declined during GnRH-A administration. The pattern of plasma androgen decrease resembled that of bioLH. There was a positive correlation between bioLH and the two androgens (r = 0.85; P less than 0.05, by Spearman's rank correlation, for both hormones). Cessation of GnRH-A administration was followed by prompt progressive increases in gonadotropin and androgen concentrations to pretreatment values. FSH recovered faster than bioLH. BioLH plasma concentrations reached pretreatment values by day 28. The recovery of plasma androstenedione and testosterone levels correlated positively with that of bioLH. Although plasma E1 levels were higher during the recovery period than during treatment, they never reached the concentrations found during the basal period, whereas estradiol concentrations were slightly but not significantly higher than those in the basal period. As a consequence, the E1 to estradiol ratio, very high in the basal period, approximated unity during recovery. These data indicate that hyperandrogenism in polycystic ovarian disease is gonadotropin dependent and accompanied by a relative abundance of LH bioactivity basally and during GnRH-A administration. Thus, the relative increase in bioLH secretion appears to be independent of the rate of gonadotropin secretion and the circulating sex steroid concentrations.

Failure of GnRH analogue to inhibit serum concentrations of testosterone and 17 alpha-hydroxyprogesterone in hCG-substituted hypogonadotropic hypogonadism.

Gonadotropin-releasing hormone analogues (GnRH-A) induce inhibition of testicular function and reduction of serum testosterone (T) in man, but the mechanism involved is still debatable. To elucidate it we studied six patients with hypogonadotropic hypogonadism (HH) in chronic substitution with hCG for correction of androgen deficiency symptoms, and evaluated the effect of addition of GnRH-A to the hCG therapy on plasma levels of T and 17 alpha-hydroxyprogesterone (17 OHP). All patients were treated with 1000 U of hCG in every 3rd day for 24 weeks. After 8 weeks of this regimen, GnRH-A, Buserelin (D-Ser-TBU-EA-LHRH), 200 micrograms per day sc, was added and given for 8 weeks. After cessation of analogue administration patients were followed for 8 further weeks. The levels of the two steroids did not differ markedly in the pre- and post-GnRH-A period. GnRH-A given for two months did not lower T or 17 OHP levels as in eugonadal men after similar treatment. The median T concentrations during GnRH-A tended to be increased, with plasma values higher (P less than 0.025) than the peak values observed during hCG alone. Since administration of Buserelin did not inhibit hCG-sustained steroid levels in these HH patients, it is conceivable that GnRH-A may have lacked a direct inhibitory gonadal effect in such experimental conditions.

Effect of gonadotrophin-releasing hormone analogue (GnRH-A) administration on serum gonadotrophin and steroid levels in patients with polycystic ovarian disease.

A gonadotrophin-releasing hormone (GnRH) analogue, D-Ser[TBU]LRH-EA10, (GnRH-A), at a dose of 200 micrograms was given daily for 2 months to 6 women with polycystic ovarian disease (PCO). Prior to therapy the patients presented elevated LH, testosterone (T), oestrone (E1) and dihydrotestosterone (DHT) in the circulation. In response to GnRH-A, these subjects exhibited a marked decrease in circulating T, DHT and androstenedione (A) levels as measured 24 h after GnRH-A injection, by 4 weeks and onwards (P less than 0.05). After 2 weeks of daily administration, the serum LH profile, evaluated by sampling at 2, 4, 7 and 24 h after injection of GnRH-A, was not different from baseline, whereas after 4, 6 and 8 weeks the levels were significantly lower (*P less than 0.01). The profile of serum T levels was unmodified at the second week, but significantly decreased thereafter (*P less than 0.01). At the end of treatment, the E1 concentrations, elevated in pre-injection condition, were markedly decreased. These data demonstrate that in PCO subjects, GnRH-A significantly lowered the elevated levels of androgens commonly found in these patients. The close correlation observed between reduced serum LH and androgen concentrations suggests that pituitary desensitization could be responsible for the reduction in androgen levels, and may be evidence for a gonadotrophin dependence of the elevated concentrations of T in these patients.

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.

The prolactin-secreting system in relation to aging.

It is known that an increase in serum PRL can be responsible in men of impotence. The frequent sexual disturbances that are present in aging have suggested that in this age the PRL levels may be increased. Some authors have found elevated levels of PRL and an alteration in PRL response to pharmacological stimuli in elderly subjects. Other authors suggest that this increase of serum PRL in aging may be develop for determinating prostatic disease. In the present study we not have found differences in serum PRL between adult men, elderly subjects and prostatic subjects, while the responses to pharmacological stimuli observed in prostatic patients was higher than that of old age subjects. This finding is suggestive of an alteration in PRL-secreting system operating in aging.

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.

Effect of CRF on the release of anterior pituitary hormones in normal subjects and patients with Cushing's disease.

Patterning of plasma ACTH, beta-EP/beta-LPH and cortisol in response to ovine CRF (1 microgram/kg b.w.t. injected i.v.), was studied in three normal subjects and in five patients with Cushing's disease, two of whom had undergone total bilateral adrenalectomy. CRF caused in all subjects a prompt and concurrent rise of plasma hormone levels. The hormonal response was of the same magnitude in normal controls and in the three untreated Cushing's patients, but was much greater in the two adrenalectomized subjects. No changes were noted, after CRF, in plasma levels of GH, PRL, TSH, LH and FSH. These findings indicate that CRF specifically promotes the pituitary release of ACTH and ACTH-related peptides both in normal subjects and patients with Cushing's disease. In view of the likely CRF hypersecretion of the adrenalectomized patients, their ACTH and beta-EP/beta-LPH hyperresponsiveness to exogenous CRF would denote that the peptide does not down regulate its own pituitary receptors.

Down-regulation of prolactin secretion in men during continuous thyrotropin-releasing hormone infusion: evidence for induction of pituitary desensitization by continuous TRH administration.

TRH was administered as a 5-h constant rate iv infusion (5 micrograms/min) to seven healthy adult men. Serum samples were collected at regular intervals for measurement of PRL, TSH, and T3. Serum levels of PRL during TRH infusion increased sharply to maximum level by 40 min, and then, despite continued TRH stimulation, PRL levels declined gradually to a plateau value after 100 min. No further rise in serum PRL was observed when a bolus of 200 micrograms TRH was administered to three subjects after 240 min of infusion. Conversely, an iv bolus of sulpiride (25 mg), a dopaminergic antagonist, given to four subjects after 240 min, brought about a marked increase in serum PRL values above the plateau level. These results are consistent with the interpretation that down-regulation in PRL secretion which follows the initial peak of response most likely represents pituitary desensitization to TRH. During the infusion serum TSH increases in two phases. A first phase of secretion was observed by 40 min followed by a plateau, with a second phase of increase occurring between 80-180 min.

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.