Presence of aerobic micro-organisms and their influence on basic semen parameters in infertile men.

Urogenital tract infections in males are one of the significant etiological factors in infertility. In this prospective study, 72 patients with abnormal semen parameters or any other symptoms of urogenital tract infection were examined. Semen analysis according to the WHO 2010 manual was performed together with microbial assessment: aerobic bacteria culture, Chlamydia antigen test, Candida culture, Ureaplasma and Mycoplasma-specific culture. In total, 69.4% of semen samples were positive for at least one micro-organism. Ureaplasma sp. was the most common micro-organism found in 33% of semen samples of infertile patients with suspected male genital tract infection. The 2nd most common micro-organisms were Enterococcus faecalis (12.5%) and Escherichia coli (12.5%), followed by Staphylococcus aureus (7%), Chlamydia trachomatis (7%) and Candida sp. (5.6%). Generally, bacteria were sensitive to at least one of the antibiotics tested. No statistically significant relationship was observed between the presence of aerobic micro-organisms in semen and basic semen parameters: volume, pH, concentration, total count, motility, vitality and morphology.

Testosterone and oestradiol in concert protect seminiferous tubule maturation against inhibition by GnRH-antagonist.

Oestradiol enhances follicle stimulating hormone (FSH) action on seminiferous tubule maturation, but the relative involvement of oestradiol and testosterone remains unclear. This study compares the influences of oestrogen and androgen in FSH and testosterone-deficient rats. Animals were injected daily GnRH-antagonist alone (Ant) or combined with 17ß-oestradiol benzoate (EB), or testosterone propionate (TP), or both from post-natal day (pnd) 5 to 15. Hormone levels, tubule growth, cell numbers, germ cell apoptosis and proliferation, and Sertoli cell maturation were evaluated on pnd 16. Ant decreased serum FSH and testosterone levels to ∼60% and ∼50% of control values, respectively, and decreased tubule growth, Sertoli cell number and maturation. Germ cell number declined by apoptosis. Co-administration of EB stimulated spermatogonia proliferation and maintained FSH levels (86% of control). Tubule growth, Sertoli cell number and spermatocyte apoptosis remained normal after TP co-administration, but Sertoli cell maturation, germ cell number and spermatogonia survival were reduced. Co-administration of EB with TP prevented all inhibitions. In conclusion, administration of oestradiol with testosterone, but neither one alone, protected seminiferous tubule maturation against inhibition caused by Ant-induced disruption. Oestrogen was involved in stimulating germ cell proliferation and the maintenance of Sertoli cell maturation, whereas androgen affected seminiferous tubule growth and spermatocyte survival.

Less advanced testicular dysgenesis is associated by a higher prevalence of germ cell neoplasia.

There is a theory that the more evident clinical signs of testicular dysgenesis, the more frequent the neoplastic lesions are. The aim of this study was to relate the incidence of testicular germ cell neoplastic lesions (overt germ cell tumours--GCT or testicular carcinoma in situ) to the intensity of testicular organogenesis disturbances (dysgenesis). Biopsies were taken from 154 testes of the following patients: 23 patients with GCT in the contralateral gonad (CGCT), 41 patients with undescended testes operated in childhood (UDT), 90 with azoo-/oligozoospermia (A/O) diagnosed because of infertility. Assessment of seminiferous epithelium, number of Leydig cells, areal fraction of intertubular space (IS), morphometric analysis of seminiferous tubules diameter and thickness of tubular wall were performed. Monoclonal antibodies against placental like alkaline phosphatase and cytokeratin 18 were applied. Germ cell neoplastic lesions were detected in 7.1% of testes and were associated with disturbed spermatogenesis. Among testes with disturbed spermatogenesis they were found the most frequently in CGCT (22.2% vs. 11.1% in UDT and 3.8% in A/O), where spermatogenesis had the highest score (5.7 +/- 3.8 points vs. 4.2 +/- 2.7 in UDT and 4.6 +/- 2.9 in A/O). In CGCT, signs of testicular dysgenesis were less advanced: the highest tubular diameter was 164.4 +/- 32.3 microm vs. 163.5 +/- 28.6 in UDT and 161.4 +/- 31.5 in A/O, the lowest thickness of tubular wall was 8.9 +/- 3.2 microm vs. 10.2 +/- 3.6 in UDT and 10.2 +/- 3.2 in A/O, lowest IS was 36.9 +/- 14.9% vs. 47.9 +/- 18.0 in UDT and 46.5 +/- 18.5 in A/O, and the lowest percentage of tubules with immature Sertoli cells was 0.1 +/- 0.4% vs. 4.9 +/- 7.0 in UDT and 5.2 +/- 9.7 in A/O. Results indicate that neoplastic lesions appear only in testes with disturbed spermatogenesis. Worse condition of spermatogenesis is associated by the presence of other dysgenetic features, but neoplastic lesions appear more frequently in testes with the less advanced features of testicular dysgenesis.

Estradiol enhances the stimulatory effect of FSH on testicular maturation and contributes to precocious initiation of spermatogenesis.

Male rats were daily injected with human FSH (hFSH) or estradiol benzoate (EB) or hFSH+EB between day 5 and 15 of life and autopsied on day 16. hFSH accelerated testicular growth, increased number of spermatogonia and serum level of testosterone. hFSH stimulated also spermatogonia differentiation, which resulted in 5-fold increase of the number of spermatocytes. EB given alone induced adverse, inhibitory effects on spermatogenesis and serum testosterone, did not influence serum FSH and LH but increased 14-fold the level of prolactin. Except from testosterone, EB given with hFSH not only overcame inhibitions, but multiplied hFSH stimulatory effects on spermatogenesis up to 30-times of control values. In addition, after FSH+EB premeiotic germ cell ratio reached adult type value precociously. Estradiol may play regulatory roles in testicular maturation (1) inhibitory, direct one or resulting from decrease in testosterone secretion; (2) stimulatory, by enhancement of FSH action, with a possible involvement of prolactin that may act in concert with FSH.

Immunohistochemical diagnosis of preinvasive germ cell cancer of the testis.

The aim of the study was to identify testicular carcinoma in situ (CIS), a precursor of germ cell tumours (GCT), in patients from the high risk groups, using classic and alternative immunohistochemical methods. 70 patients with 46,XY karyotype were examined. Whole gonads or biopsy specimens were fixed in Bouin's fluid. In cases with dysgenetic male pseudohermaphroditism (DMP), gross histopathology revealed sex cord tumour gonadoblastoma in 4 and malignant dysgerminoma in 1 out of 23 patients. In all patients, paraffin sections were treated with antibodies against placental-like alkaline phosphatase (PLAP), a classic immunohistochemical marker of GCT and CIS. CIS was detected immunohistochemically in 10 out of 23 cases with DMP (43.5%), in 1 out of 10 cases with androgen insensitivity syndrome (10%), in 3 out of 18 cases operated previously because of already developed GCT in contralateral testis (16.6%) and in 1 out of 3 patients with cryptorchidism in anamnesis (33.3%). CIS was not found in 16 examined adult infertile men with azoospermia. In addition to PLAP investigation, 12 cases with DMP and 6 cases with GCT were examined using M2A and TRA-1-60 antibodies, the alternative immunohistochemical markers of CIS. While in DMP positive reactions for M2A and TRA-1-60 accompanied PLAP reaction in 1/3 of cases, M2A accompanied PLAP in all cases with GCT. The positive reaction for TRA-1-60 accompanied PLAP and M2A in 1 case with GCT. The results indicate that among different risk groups the highest incidence of CIS occurs in DMP. Screening for CIS is of importance also in cryptorchidism, androgen insensitivity syndrome and in men who underwent gonadectomy because of unilateral GCT. The immunostaining for PLAP seems to be more discriminative procedure. The positive staining of CIS cells with M2A and TRA-1-60 antibodies may be indicative for more advanced neoplastic transformation.