Seminal LDH-C4 isoenzyme and sperm mitochondrial activity: a study in male partners of infertile couples.

Lactate dehydrogenase isoenzyme patterns were obtained in the seman of 93 male partners of infertile couples and 28 proven fertile subjects as a control group. Sperm mitochondrial activity index (SMAI) alongwith the conventional seminal parameters was studied for all the subjects excepting the azoospermic and vasectomised males. Only LDH-C4, a germ cell specific isoenzyme activity varied with the variation in sperm density. LDH-C4 activity varied significantly (p<0.001) within and between different groups. Lactate dehydrogenase-C4 activity was absent in 17 azoospermic samples, confirming its germinal epithelial origin, as well as in 8 samples of vasectomised males. In one azoospermic sample, there were many immature germ cells along with surprisingly high LDH-C4 activity suggesting more activity of germinal epithelium associated with high LDH-C4 activity. LDH-C4 activity was reduced significantly in oligozoospermic samples in proportion to sperm density, thus confirming strong correlation (p<0.001) between LDH-C4 and sperm density. There was statistically significant correlation between LDH-C4 and percentage sperm motility as well as between LDH-C4 and Sperm Mitochondrial Activity Index (SMAI) (probability varying from p<0.05 to p<0.01 in different groups), but no such correlation was found between LDH-C4 and sperm morphology. The data confirms LDH-C4 as a germinal epithelial marker. Its relationship with percentage sperm motility is suggestive of definite role of LDH-C4 in evaluation of the spermatozoal quality, similarly its relationship with Sperm Mitochondrial Activity Index (SMAI score) suggest the role of LDH-C4 in metabolism of the spermatocytes and sperms, though further studies are required for clear and detailed understanding of its metabolic role in semen.

The role of hypo-osmotic swelling test in recurrent abortions.

The role of male factors in abortions has not been studied extensively. We undertook this study to determine if there was any relationship between hypo-osmotic swelling test score and recurrent abortions. This was a cross-sectional case control study conducted at the semen analysis laboratory at a tertiary level referral centre. The male partners of 25 patients who had three or more first trimester abortions of unknown etiology were chosen as cases. Controls were 26 unmarried volunteers with unknown fertility potential to represent the entire population. The conventional seminal parameters were studied according to WHO guidelines. The HOS test score were obtained by the method advocated by Jayendran et al. There were no statistically significant differences in mean sperm count, percentage motility and morphology among the cases and controls. We found a low hypo-osmotic swelling test score among the cases [55.7 +/- 1.197 (SEM)] than the controls [69.3 +/- 1.143 (SEM)] which was statistically significant [P < 0.001]. The normal spermatozoal membrane is the prerequisite for the specialized cell-to-cell communications and cell-to-cell binding. In spite of apparently normal semen analysis, subtle membrane defects in the spermatozoa, which could be the cause of defective membrane functions in the embryo effecting miscarriages, can be elucidated by the hypo-osmotic swelling test.

Acrosome intactness and seminal hyaluronidase activity: relationship with conventional seminal parameters.

Seminal hyaluronidase activity was estimated after liquefaction in semen samples of 100 male partners of infertile couples including 16 azoospermic (no spermatozoon) men and 48 fertility proven men by a method based on measurement of the area of digestion of substrate (hyaluronic acid) in agar plate. Semen samples were also evaluated for Acrosomal Intactness (AI) test except the azoospermics of the studied samples. Seminal hyaluronidase activity was completely absent in azoospermic specimens confirming its cellular origin. Seminal hyaluronidase activity was found to be significantly correlated, statistically, with sperm density (r = 0.708, p < 0.001), % motility (r = 0.6478, p < 0.001) and % normal sperm morphology (r = 0.5724, p < 0.001). Acrosomal Intactness (AI) test scores were also well correlated with sperm density (r = 0.6477, p < 0.001), % motility (r = 0.5965, p < 0.001) and % normal morphology (r = 0.6237, p < 0.001). Both values were higher in semen samples with normal routine parameters (proven fertility and normozoospermic infertile groups) than those compared with abnormal routine parameters (oligozoospermic). We also found very highly significant correlation (r = 0.8442) between seminal hyaluronidase activity and Acrosomal Intactness scores, statistically (p < 0.001). This could be because; normal germinal semineferous epithelium generates abundant number of sperms with normal motility and morphology that are also having intact acrosome. Intact acrosome prevents loss of acrosomal enzymatic activity (e.g. hyaluronidase) until released after liquefaction during seminal analysis and during acrosomal reaction in female genital tract prior to fertilization. Seminal hyaluronidase activity, thus determined, is primarily dependent upon the intact status of acrosome. As each sperm contributes to the seminal hyaluronidase activity, it is directly correlated with sperm density; but at the same time it exhibits goods correlation with % motility and % normal morphology. Therefore AI score and seminal hyaluronidase activity can be considered as good indicators of sperm function.