A series of studies aiming at introducing an effective treatment for idiopathic oligozoospermia was conducted in a step-wise fashion spanning over a 20-year period. The concept was that co-administration of an accessory gland-stimulating androgen, testosterone undecanoate (40 mg t.i.d.) and the FSH raising anti-oestrogen tamoxifen citrate (10 mg b.i.d.) may improve sperm parameters. A prerequisite for such an effect was the demonstration that testosterone undecanoate had no suppressing action on pituitary-testicular axis. In this context, initial studies demonstrated no change in basal or stimulated gonadotrophin and testosterone secretion in short- or long-term protocols. Two subsequent trials with this combination showed a marked improvement of sperm parameters and pregnancy incidence, with a seasonal variation noted in response to treatment, this being higher during the cold seasons of autumn and winter. Regarding the mechanism of testosterone undecanoate's action, a recent study from our unit showed that its administration resulted in a marked rise of serum DHT levels. Because this steroid is an epididymal function promoter, it appears that its contribution in the combination is mediated mainly through its DHT raising effect. By and large, this empiric approach for the treatment of idiopathic oligozoospermia was satisfactorily documented after a 20-year investigative saga.

The effect of testosterone (T) on germinal epithelium development and maturation was studied in prepubertal male rabbits. For this purpose, implants of free T (50 mg) or placebo (PL) were placed intratesticularly in 55-60-day-old animals. Each group was divided into sub-groups additionally treated with 0.9% NaCl, FSH/LH (3.0 IU) or cyproterone acetate (CA, 2.5 mg) over a 45-day period. Initial and final evaluations included measurements of testicular volume, testicular biopsy score count (TBSC) and plasma T, FSH and LH concentrations. At the end of the experiment, increment values (final-initial evaluation) of the parameters examined showed the following differences: (1) mean TBSC in T-implanted, irrespective of additional treatment, was higher than that of PL-implanted rabbits (P less than 0.01); (2) mean plasma T, FSH and LH values were significantly different in T- and PL-implanted rabbits, with higher T (P less than 0.05) but lower FSH and LH increments (P less than 0.01 for both) in the former group; (3) marked differences amongst types of additional treatment, irrespective of the implant used, were found for TBSC, T, FSH and LH (P less than 0.01 for all); animals treated with CA had markedly lower increments than other treatment groups. From these findings, it may be concluded, that in the prepubertal rabbit T plays an important role in development and maturation of the testis, but this effect probably requires the synergistic action of endogenous gonadotrophins.

Background Assessment of testosterone undecanoate’s (TU) presence in the functional compartments of the male reproductive tract has never been performed despite the evidence that its documented beneficial effect in male infertility might be mediated through an epididymal action and this study was set to examine this possibility. Materials and methods In 18 normozoospermic volunteers TU has been administered (40 mg t.i.d.) for 6 days with serum measurements of TU, total testosterone (T), DHT, E2, SHBG, FSH, LH, and PRL before and at the end of medication. Steroid hormones (T, E2, and TU) were also assayed in seminal plasma. In a selected group of 7 men with previously diagnosed non-obstructive azoospermia TU, T, and E2 were assayed in the extracts of testicular biopsy material taken before ICSI and at the end of the same medication. Results A marked rise of serum DHT (average 148%, P < 0.001) has been found after treatment, whereas T, E2, FSH, LH, SHBG, and PRL did not significantly change. Measurable amounts of TU were found in the serum of all men but only in 6 cases in seminal plasma (11.1 ± 8.0 ng/mL) and all of them in semen delivered 7–8 h after the last TU capsule was taken. In dilution fluid from testicular tissue extracts, no detectable amounts of TU were found whereas mean values of 92.5 ± 54.3 pg/mL and 43.8 ± 16.3 ng/mL for E2 and T were observed. Positive correlations among TU and E2, T or DHT concentrations were found in serum samples ( P < 0.01, 0.02, and 0.002) as well as between E2 and T ( P < 0.01), E2 and DHT ( P < 0.001), or T and DHT ( P < 0.001). Conclusion It is concluded that TU was identified and measured for the first time in seminal plasma of a fair percentage (33%) of men on this medication and was associated in all men with a marked rise of DHT concentration, a known epididymal function promoter, in the absence of an effect on pituitary and gonadal activity. On this evidence, it appears that a beneficial effect of TU on epididymal function may be a distinct possibility.

Sertoli cell functional reserve was assessed in normozoospermic men and oligozoospermic patients and its prognostic potential was evaluated for patient selection and treatment. For the first objective, three groups of normo-follicle-stimulating hormone (FSH)/normozoospermic fertile men (n:12), normo-FSH/oligozoospermic (n:21) and hyper-FSH/oligozoospermic subfertile men participated in the study whereas for the second objective 24 normo-FSH oligozoospermic patients volunteered for a pilot therapeutic trial. For the first part, high purity (hp) FSH (225 i.u., i.m.), human chorionic gonadotropin (hCG) (1500 i.u., i.m.) or their combination was given separately at weekly intervals, with samplings at 0, 3, 24 and 48 h. For the pilot trial, rec-FSH (150 i.u./48 h, i.m.) or placebo were prescribed for 6 months. The main outcome measures for the study were inhibin-B (inh-B), insulin-like growth factor (IGF)-I, testosterone and oestradiol concentrations and the main sperm parameters. Bolus administration of hp-FSH or hp-FSH/hCG combination in normozoospermic men resulted in a significant rise of inh-B in normozoospermic men (mean +/- SD, basal: 183.8 +/- 24.2 pg/mL in hp-FSH and 175.2 +/- 23.5 in hp-FSH/hCG treatment; 48 h: 256.1 +/- 34.2 and 246.3 +/- 19.0, respectively, p < 0.001 for both). In oligozoospermic groups basal inh-B concentration was lower than in normozoospermic men (normo-FSH: 117.4 +/- 16.5, hyper-FSH: 81.2 +/- 19.8, p < 0.001 for both) with a post-stimulation increase noted only in normo-FSH patients (hp-FSH 24-h: 132.8 +/- 19.7, p < 0.01; hp-FSH/hCG 0 min: 105.7 +/- 20.1, 24-h: 119.5 +/- 20.6, p < 0.05). Total sperm number and progressive motility showed significant improvements (p < 0.05 for both) after 6 months of rec-FSH treatment in the group of patients with a satisfactory response to hp-FSH stimulation. In conclusion, the basal and reserve activity of Sertoli cells, as judged by inh-B secretion, was higher in normozoospermic than in dyspermic men, with a better therapeutic outcome noted in those patients with an adequate response to hp-FSH stimulation.

Evaluation of sex steroids in cervical mucus was performed at different phases of spontaneous or clomiphene-citrate-induced ovulatory cycles. To this end, 11 women with normal ovulatory cycles and 9 subjects with polycystic ovary syndrome of comparable age and body mass index were investigated. Serum and cervical mucus samplings were assessed for 17β-estradiol (E2), progesterone, testosterone, and sex hormone binding globulin levels at the pre-, peri-ovulatory, and mid-luteal phases of the cycle. The cervical mucus maturation index also was estimated in all women. Measurable amounts of E2 were found in most mucus samples with a cyclic variation in all cases. The highest E2 and mucus maturation index values coincided, but both lagged by 24 h behind the serum mid-cycle peak of this steroid. Detectable amounts of progesterone were found in the luteal phase, testosterone was present at low levels throughout the cycle, but sex hormone binding globulin was undetectable in all cervical mucus samples. Differences between spontaneous or drug-induced ovulatory cycles were not found. It is concluded that sex steroids are present in human cervical mucus, showing variations similar to those in peripheral blood. The significance of these findings is not clear at present, but it is probably related to the cyclic changes of cervical epithelium and gland secretion. An important implication of the absence of measurable sex hormone binding globulin amounts in cervical mucus is that the free fraction of sex steroids present in that fluid are presumably higher, and therefore, expected to exert greater biologic activity than in peripheral blood.

Objective: To evaluate the effects of T undecanoate given as a supplementary treatment with tamoxifen citrate (TAM) or hMG on pituitary and Leydig cell function in men with idiopathic oligozoospermia.Design: A total of 48 normogonadotropic men with idiopathic oligozoospermia were allocated in to six groups (n = 8 per group) treated with placebo, 40 mg T undecanoate three times per day, 10 mg TAM two times per day, T undecanoate and TAM, 75 IU/d hMG, and T undecanoate and hMG. All groups were evaluated with standard GnRH, thyrotropin-releasing hormone, and hCG tests before and on the final day of 3 months on treatment with measurements of FSH, LH, thyroid-stimulating hormone (TSH), PRL, T, E(2), 17-hydroxyprogesterone, sex hormone-binding globulin, and seminal analyses (at least twice each time).Results: Basal and stimulated concentrations and incremental FSH and LK values showed no differences among TAM or hMG and TAM + T undecanoate or hMG + T undecanoate treated groups. Basal, stimulated, and incremental values for TSH and PRL were elevated markedly during treatment in most groups in comparison to placebo. Basal, stimulated, and incremental T and E(2) values were similar in active treatment groups except that higher T concentration was found in TAM + T undecanoate as compared with T undecanoate only treated men. Finally, significant improvements were noted in important seminal parameters and particularly in the functional sperm fraction of the TAM + T undecanoate group as compared with single treatment with TAM.Conclusion: These results indicate that T undecanoate in combination with TAM or hMG not only had no adverse effects on pituitary and Leydig cell activity but also seemed to improve important seminal parameters and signify that androgens may be tried as a supplementary treatment to conventional regimes in idiopathic oligozoospermia.

Objective: To evaluate Leydig and Sertoli cell response to prolonged pulsatile stimulation with hMG after pituitary desensitization with the GnRH agonist (GnRH-a) triptoreline in normogonadotropic men with abnormal semen analyses.Design: A group of four oligozoospermic men were investigated in the following manner: [1] basal and GnRH-hCG stimulated activity were assessed in all volunteers; [2] a long-acting form of the GnRH-a triptoreline (3.75 mg every month for 3 months) was given, and its effectiveness was evaluated on day 20; and [3] on that day hMG pulsatile administration was introduced (150 IU per 24 hours in 90-minute pulses) with serial hourly sampling (6 to 7 hours) for measurement of FSH, LH, T, E(2), and inhibin on days 20, 41, and 90 from the first GnRH-a injection.Results: Initial evaluation showed normal basal, GnRH, and hCG-stimulated hormone concentrations. Pituitary and gonadal activity were effectively suppressed by GnRH-a when tested on day 20. Pulsatile hMG had no immediate stimulatory effect on gonadal activity (day 20). However, on middle and final evaluations (days 41 and 90), basal T, E(2), and inhibin had risen to pre-GnRH-a levels, and, moreover, distinct secretory pulses were seen for these hormones.Conclusion: These findings indicate that suppression of pituitary gonadotropin activity with triptoreline combined with pulsatile hMG stimulation offers a new, useful tool for investigation of the male reproductive system in oligozoospermic men.

The effect of induced peripheral anosmia on gonadal development and maturation was investigated in sexually immature male rabbits. Furthermore, the effect of agents promoting gonadal maturation (LHRH, hMG or methyl testosterone) on testicular development was examined in anosmic rabbits. Peripheral anosmia was induced by spraying the olfactory mucosa with 5% ZnSO4 solution; its effects were evaluated after a 45-day period, corresponding to the duration of spermatogenesis. Evaluation was based on measurement of body-weight, testicular size, testicular biopsy score count (TBSC) and a standard LHRH test (0 and 30 min) involving measurement of the blood levels of FSH, LH and testosterone before and at the end of the test. Markedly lower final and incremental values were noted in anosmic, compared to intact, animals for body-weight (P less than 0.001), TBSC (P less than 0.001), FSH (P less than 0.01) and LH (P less than 0.05). On the other hand, treatment of the anosmic rabbits with 0.9% saline, resulted in lower FSH, TBSC and testicular size increments than in rabbits treated with LHRH, hMG or testosterone, while testosterone levels and body-weight increments were similar in all groups. These findings indicate that induced peripheral anosmia is probably responsible for the inadequate gonadal maturation in prepubertal anosmic male rabbits. This relationship was confirmed by the observed stimulatory effect of administration of agents activating pituitary gonadotrophin secretion or gonadal function in anosmic animals.

The incidence of thyroid disease was examined prospectively in 97 consecutive patients with breast cancer (Group I) and was compared with that of 61 women with cystic breast disease (Group II) and that of 60 control women (Group III) with no breast problems. Thyroid enlargement was found in 47%, 49%, and 22% of those in Groups I, II, and III, respectively, and treatment with thyroid hormones was taken by 9.3% (I), 8.2% (II), and 5.0% (III) of the cases. The mean thyroid-stimulating hormone (TSH) concentration in those in Group I (5.4 +/- 2.2 microU/ml) was significantly higher than in Groups II (3.9 +/- 1.9, P less than 0.01) and III (4.0 +/- 1.8, P less than 0.001), whereas thyroid microsomal antibodies were detected in 13.4% (I), 9.1% (II), and 1.7% (III); mean triiodothyronine (T3) and thyroxin (T4) concentrations were similar in the three groups. When both TSH and T3 concentrations were taken into account, 24% and 17% of the patients, respectively, from Group I were no longer in the Group III range; the corresponding figures for Group II were 13% and 23%, respectively. These results indicate that breast cancer and thyroid disease are probably related, but not in a specific way, since benign mastopathy also seems to be associated with thyroid disturbances.

The endocrine milieu on which spermatogenesis and sperm maturation mainly depend was evaluated quantitatively with simultaneous measurements of FSH, LH, PRL, testosterone, estrone, estradiol (E2), and sex hormone-binding globulin concentrations in spermatic venous plasma, antecubital plasma, seminal fluid, and wash fluid from vas deferens in 16 normospermic men and 24 oligospermic patients. Anesthesia and surgical stress caused a rise of only PRL and E2 (P less than 0.001-0.01). Mean FSH, LH, and PRL levels were comparable in antecubital and spermatic venous plasma, and antecubital values were higher in oligospermic patients for FSH and LH (P less than 0.05-0.001). Mean (+/- SD) T levels were similar for normospermic and oligospermic men in spermatic venous plasma (473 +/- 75 and 439 +/- 270 ng/ml), in antecubital plasma (6.5 +/- 1.3 and 6.6 +/- 1.8), and in seminal fluid (0.3 +/- 0.1 for both). Minute quantities of testosterone were detected in pooled wash fluid (0.08 ng). For E2, similar concentration gradients from high to low levels were found in normospermic and oligospermic men (spermatic venous plasma = 926 +/- 205 pg/ml and 1090 +/- 262; antecubital plasma = 31.0 +/- 12.0 and 28.4 +/- 1.9; seminal fluid = 14.3 +/- 2.3 and 12.0 + 2.8). Estrone was also high in spermatic venous and low in antecubital plasma but higher in seminal fluid than in antecubital plasma. Sex hormone-binding globulin levels were slightly though not significantly lower in spermatic venous (23 +/- 10 nmol/liter) than in antecubital plasma (28 +/- 6), but not measurable in seminal fluid. These results define important aspects of the endocrine milieu prevailing in the male reproductive tract and demonstrate a change of the relative activity of androgens and estrogens from the testis to the seminal fluid.

Prolactin (PRL) concentration was estimated in milk and blood of women with various types of galactorrhea (n = 53) and was compared with lactating (n = 17) and normally menstruating women (n= 36). Mean milk (± SD) PRL concentration in lactating women (160±66 μg/l) was similar to that of galactorrhea due to pituitary adenoma (149±87 μg/l, n = 24), thyroid dysfunction (193±72 μg/l, n= 8), functional causes (192±91 μg/l, n = 10) or recent pregnancy (198±44 μg/l, n = 2), but higher than that of sex steroid-induced galactorrhea (85±53 μg/l, n = 9). Plasma PRL concentration in lactating women (208±102 μgl) was higher than in normal control women (14±5 μg/l) and patients with galactorrhea due to thyroid dysfunction (36±15 μg/l), functional causes (16±8 μg/l), drugs (20±4 μg/l), recent pregnancy (22±3 μg/l) or pituitary adenoma (145±53 μg/l). The milk to plasma PRL concentration ratio was 0.8 ± 0.4 in lactating women and 1.1 ± 0.7 in patients with adenoma but significantly higher (p = 99% two-sided) in galactorrhea due to thyroid dysfunction (4.4±2.1), drugs (3.4±1.1) or functional causes (12.3±4.3). Bromocriptine administration reduced PRL in both fluids. It is concluded that in women with galactorrhea milk PRL concentration is similar to that of nursing mothers, but plasma levels of this hormone are significantly lower than that of the latter group in all but the pituitary adenoma related galactorrheas.

Prolactin (Prl) exerts important biological actions on the mammary gland being responsible, with other hormones, for milk production. Its presence and concentration in milk of nursing mothers (Healy et al. 1980) and in breast duct – fluid of nonlactating women (Wynder & Hill, 1977) has been determined in recent years. It has been suggested that milk Prl estimations in association with pre – and post – suckling serum determinations of this hormone may become a potentially satisfactory procedure for establishing Prl requirements for efficient milk production (Healy et al. 1980). Furthermore, from a physiopathological point of view, Prl in breast fluid has been listed amongst the factors associated with the induction and development of experimental breast cancer, thus providing a possible link between high Prl concentration in breast duct – fluid and the incidence of breast cancer in Western women. In view of these associations, and the absence of any

The male reproductive system has the distinction that being itself hormonally active, through its endocrine secretions, at the same time is a target system receiving not only the biological action of hormones from the anterior pituitary gland but also from products of its own endocrine activity. Furthermore, the reproductive system operates in a very sophisticated fashion, with a number of interrelated functional compartments which accommodate the successive steps of a very delicate reproductive activity. In this context, as functional compartments could be defined the testicular tissue (TT) reflecting the endocrine and tubular activity of the testis, the blood of spermatic vein (SVB), reflecting indirectly the endocrine function of the gland, the peripheral blood (cubital blood–CB), expressing the availability of hormones at tissue level, the hormone content of the vas deferens fluid, expressing the endocrine environment in a part of the tract critical for sperm maturation and, finally, the seminal plasma

Pituitary function was assessed by measuring serum prolactin thyroid stimulating hormone (TSH), follicle stimulating hormone (FSH) and luteinizing hormone (LH) before and after LH releasing hormone (LH-RH) (100 mg intravenously) and sulpiride (100 mg intramuscularly) administration in 13 young women in a reformatory institution, and in six employees of the same establishment with regular cycles, serving as control group. Ovarian endocrine activity was assessed with basal oestradiol estimations. Seven out of the 13 restricted women had secondary amenorrhea that began shortly after their imprisonment and six had regular cycles. Basal oestradiol concentration was similar in the three groups, but prolactin response to sulpiride was significantly higher in the control group than that in either group of inmates. Basal FSH, LH and TSH concentrations as well as FSH and TSH responses were not different in the three groups, but the LH response to LH-RH was significantly higher in the group of inmates with amenorrhea than that in the two remaining groups.

The effect of clomiphene citrate (CG) on sex hormone binding globulin (SHBG) was studied in 10 oligozoospermic patients with varicocele and 6 normospermic men. Plasma SHBG, testosterone (T), oestradiol (E2), FSH, LH, Prolactin (Prl), thyroxine (T4) and 17-OH-progesterone (17-OH-P) were determined before and during medication. SHBG concentration rose from 38.1 +/- 18.3 to 54.3 +/- 16.0 nmol/l (P less than 0.01), while T and E2 showed significant increases from 31.2 +/- 10.8 nmol/l and 24.6 +/- 5.4 pg/ml to 52.0 +/- 3.6 and 43.3 +/- 14.9, respectively in the oligozoospermic patients, with similar rises noted in the normospermic men. FSH, LH and 17-OH-P were markedly elevated while on CC, but Prl and T4 remained unchanged. The findings of this study indicated the CC causes an increase of SHBG concentration, which is probably related to the rise of E2 concentration. This SHBG change, combined with the intrinsic oestrogenic activity of CC might be one of the factors responsible, through a decrease of free T and a T to E2 imbalance, for the lack of significant effect on parameters of seminal quality in so treated patients.