The effect of human chorionic gonadotropin-based hormonal therapy on intratesticular testosterone levels and spermatogonial DNA synthesis in men with non-obstructive azoospermia.

The use of hormonal therapy in men with non-obstructive azoospermia (NOA) is controversial because no information is available on how it affects intratesticular testosterone (ITT) levels and spermatogenic cells. The aim of this study was to investigate the ITT level and spermatogonial DNA synthesis, as determined by proliferating cell nuclear antigen (PCNA) expression, before and after human chorionic gonadotropin (hCG)-based hormonal therapy in men with NOA. Twenty patients who failed sperm retrieval procedures using microdissection testicular sperm extraction (micro-TESE) were enrolled in hCG-based hormonal therapy prior to a second micro-TESE. The patients' ITT levels were determined from testicular fluid obtained during the micro-TESE. Spermatogonial PCNA expression was determined by immunohistochemical analysis, and the PCNA labelling index (PCNA-LI) was calculated. During the second micro-TESE, spermatozoa were successfully retrieved from three (15%) of the men who had been treated with hormonal therapy. PCNA-positive cells were predominantly in the spermatogonia and primary spermatocytes, and PCNA-LI was significantly increased after the hormonal therapy. A significant increase in the ITT levels before and after the hormonal therapy (p < 0.0001, 273.6 ± 134.4 and 1348.1 ± 505.4 ng/mL respectively). The sperm-positive group showed a significantly lower basal ITT level compared with the sperm-negative group (p < 0.05). There was a marked increase in the PCNA-LI levels of men treated with both recombinant human follicle-stimulating hormone and hCG. In addition, there was a significant negative association between the increase in PCNA-LI and basal ITT levels at the initial micro-TESE (p < 0.05), but not the stimulated ITT level at the second micro-TESE. HCG-based hormonal therapy significantly raises the ITT level and stimulates spermatogonial DNA synthesis, potentially improving spermatogenesis. ITT optimization plays, at least in part, an important role for stimulating spermatogenesis in men with NOA.

Isolation of antigenic variants from leptospiras grown in vitro and from heart blood of guinea pigs inoculated with a clonized strain of Leptospira.

A clonized culture of Leptospira interrogans serovar copenhageni strain Shibaura (Cl-Shibaura) was inoculated into guinea pigs. The heart blood of the guinea pigs, obtained at the febrile stage and inoculated onto the solid serum medium containing the homologous immune serum, produced large and small colonies. The serological examinations revealed that the large colonies were found mainly to be the antigenic variants, while the small colonies were mostly the parent. The antigenic variants accounted for 16.4% of all the colonies from the blood of the guinea pigs infected with Cl-Shibaura, and for 1.2% of all the colonies from the blood of the guniea pigs infected with Cl-Shibaura, and for 1.2% of all the ll colonies. The serological examinations revealed that the large colonies were found mainly to be the antigenic variants, while the small colonies were mostly the parent. The antigenic variants accounted for 16.4% of all the colonies from the blood of the guniea pigs infected with Cl-Shibaura, and for 1.2% of all the colonies from the culture of Cl-Shibaura in the normal serum medium. Antigenic variants were also isolated in vitro from the culture of 2 other serovars. The fact that the frequency of the antigenic variants of leptospiras was higher in vivo than in vitro is discussed.