Humanin (HN) is a peptide showing neuroprotective properties against damage associated with Alzheimer’s disease. In male reproductive system HN is expressed in human and rat testis. This preliminary study aimed to localize HN by immunocytochemistry (ICC) directly in human, mouse and rabbit sperm. Semen samples were processed for the ICC analyses using two different rabbit polyclonal anti-HN antibodies. Western blotting analyses were performed using fresh human semen samples. Normal human, rabbit and mouse sperm, showed HN labeling in sub-acrosomal and mid-piece regions, whereas human abnormally shaped sperm were stained also at acrosomal level and particularly in the flagellum. Western blotting analysis revealed the presence of HN in analyzed human semen samples. HN was localized directly on human, rabbit and mouse spermatozoa. In human samples, we demonstrated a different localization pattern in normally shaped sperm compared to abnormal sperm probably due to a protective effect of HN in pathological sperm.

This retrospective study was aimed at evaluating the effects of cigarette consumption on semen parameters in a group of men with idiopathic infertility. The semen quality of 2 groups of men with idiopathic infertility, smokers (n = 118) and nonsmokers (n = 153), were compared. Conventional semen analysis was performed and sperm morphology was assessed by transmission electron microscopy (TEM). TEM data were elaborated by means of a mathematical formula based on a Bayesian technique able to furnish a fertility index (FI), and the percentages of sperm apoptosis, necrosis, and immaturity. Values of normality recommended by World Health Organization guidelines were used as a control for conventional semen analysis, and values from sperm of 25 men of proven fertility were used for TEM indices. Infertile smoker and nonsmoker patients showed similar sperm parameters, although sperm motility and TEM analysis values in both groups were significantly impaired compared with controls. Smoker patients were then classified as mild (>or=1 and 10 and <20 cigarettes/day), or heavy smokers (>or=20 cigarettes/d). Sperm concentration and FI were significantly (P < .05) different among the 3 considered smoker classes. Comparing the pairs of smoker classes, sperm concentration and FI in heavy smokers were significantly lower (P < .05) than that observed in mild smoker and nonsmoker groups. Although semen quality in males with idiopathic infertility seems not to be dramatically affected by cigarette consumption, heavy smokers show significantly lower sperm concentration and FI: another strong reason to stop smoking.

The present study aims at evaluating the effects of the organic extract of tyre debris (TDOE) from tyre wear on the kinetic and morphologic features of rabbit spermatozoa. Rabbit sperm were incubated for 4 h with 0, 5, 10, 50 and 75 mu g/mL of TDOE. Sperm motility was evaluated by computer-assisted semen analysis. Phosphatidilcholine (PS) externalization (apoptosis) and plasma membrane breakage (necrosis) were assessed using the annexinV/propidium iodide assay. The sperm ultrastructure was observed by scanning (SEM) and transmission electron microscopy (TEM). A relevant decrease in the motility rate, in PS externalization, and in plasma membrane breakage of spermatozoa was observed after incubation with TDOE at concentrations higher than 50 mu g/mL. The most frequent ultrastructural anomalies detected in the analysed specimens were: plasma and/or acrosomal membrane breakage, swollen and disorganized mitochondria, and altered axonemal patterns. Taken together, these results suggest that the organic extract of tyre debris can be toxic to rabbit spermatozoa - affecting their movement and structural integrity - when present in seminal plasma at a concentrations higher than 50 mu g/mL. Although rabbit sperm has been proven to be a suitable model for testing the in vitro effects of many chemical compounds, including TDOE, the obtained results must be considered preliminary and cannot be extrapolated yet to the in vivo outcomes because of scanty data. The results encourage, however, further research in this field.

The role of the male partner in recurrent pregnancy loss (RPL) is not clear. In this study, semen characteristics of 22 men whose partners had experienced RPL were examined by light microscopy. Sperm morphology was analysed by transmission electron microscopy (TEM) and the data were mathematically elaborated to obtain numerical indices expressing the status of an ejaculate: the fertility index and the percentage of apoptosis, necrosis and immaturity. Sperm apoptosis and necrosis were also evaluated by annexin V/propidium iodide assay. To explore the status of meiotic segregation, fluorescence in situ hybridisation (FISH) with probes for chromosomes 18, X and Y, was applied directly on sperm nuclei. Sperm characteristics from a group of men of proven fertility were used as controls. Among the considered sperm characteristics, apoptosis (P < 0.01), 1818YY diploidy (P < 0.05) and 18YY disomy (P < 0.01) scores were significantly higher in men with RPL compared with controls. Our study showed that some patients with normal semen parameters can present a slight increase in aneuploidy compared with controls, indicating a possible involvement of sperm in some cases of RPL. Chromosomal FISH analysis and chromatin tests of sperm could be included in RPL work‐ups when no other cause has been detected.

Little is known about the effect of chronic hepatitis B and hepatitis C on sperm quality. In this study, we analysed sperm quality from selected patients with chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections. Semen samples were examined by light and transmission electron microscopy (TEM). TEM data were elaborated with a mathematical formula able to indicate a fertility index and the presence of the three main sperm pathologies: apoptosis, immaturity and necrosis. Meiotic chromosome segregation was investigated by fluorescence in situ hybridisation carried out on sperm nuclei, using probes for chromosomes 18, X and Y. Despite normal sperm concentration, we observed reduced motility. TEM analysis highlighted that 35.7% of patients showed generally good semen quality. However, significantly higher values of apoptosis and necrosis, compared with controls, were observed, demonstrating spermatogenetic alterations. Regarding meiotic segregation, we found an incidence of disomies similar to that observed in control samples, whereas diploidy resulted higher in HCV patients, without reaching statistical significance. In conclusion, sperm quality in the studied group was not impaired; however, apoptosis and necrosis resulted out of normal range and the fertility index was significantly lower in HCV- and HBV-infected patients versus controls.