Application of erythrocyte lysing buffer (ELB) has detrimental effects on human sperm quality parameters, DNA fragmentation and chromatin structure.

Erythrocyte lysing buffer (ELB) facilitates the search for spermatozoa by eliminating erythrocytes in testicular suspension used during the ICSI procedure. This study investigates the effects of ELB on sperm quality parameters, sperm chromatin and sperm DNA fragmentation. Normal ejaculations were used as the model for testicular spermatozoa in this study. After swim-up, the sperm pellets were divided into two parts. Part I, the control (Group A), was diluted with culture media; and Part II, the intervention group (Group B), was diluted with ELB for 10 min. After centrifugation in both groups, the sperm pellets were re-suspended with culture media. The samples were immediately evaluated (A0 and B0) and then evaluated again after 1 hr (A1 and B1). The results indicated ELB decreased the progressive motility (81.60 ± 8.69 vs. 64.69 ± 19.08) and viability (97.62 ± 3.02 vs. 85.91 ± 11.46), in Group A and B, respectively, both immediately and 1 hr after preparation. Also, ELB engendered a significant increase in the DNA fragmentation index both immediately (9.68 ± 3.55 vs. 14.38 ± 6.52) and after 1 hr (10.37 ± 5.03 vs. 19.38 ± 6.39). In conclusion, ELB may damage sperm cells, shown by a decreased motility and viability, and it increased DNA fragmentation. Therefore, the use of ELB in testicular semen handling should be discouraged.

the Short Abstinence May Have Paradoxical Effects On Sperms With Different Level Of DNA Integrity: A Prospective Study.

PURPOSE: To investigate the effect of short abstinence on sperm function tests and semen parameters. MATERIALS AND METHODS: This prospective study included 65 male patients with increased DNA injury in their ejaculated sperm and a history of recurrent pregnancy loss and/or assisted reproductive techniques failures. The effects of antioxidants medical therapy and short abstinence on semen quality were assessed (TUNEL test and CMA3 staining). RESULTS: Antioxidants have statistically significant effects on mean sperm concentration (untreated, 67.51 ± 44.40 million/ml, vs. treated, 56.09 ± 37.85 million/ml; P-value=0.005) and mean TUNEL score (untreated, 24.56% ± 9.49%, vs. treated, 20.64% ± 10.28%; P-value = 0.013). Moreover, a short abstinence period might have positive effects as shown on the TUNEL assay (20.64% ± 10.28 vs. 17.38% ± 8.59 ; P-value = 0.028) and CMA3 staining (47.79% ± 20.78, vs. short 41.92% ± 18.49; P-value = 0.019), when considering all study subjects. However, different results were obtained using more precise analysis based on a TUNEL cutoff score of 20%. The analysis showed that short abstinence might improve sperm DNA integrity in patients with TUNEL score > 20% (mean TUNEL score from 27.85% ± 8.32% to 19.14% ± 8.90% ; P-value =0.001%). However, it might have deleterious effects on sperm DNA integrity in patients with TUNEL score < 20% (mean TUNEL score from 11.89% ± 3.21% to 15.17% ± 7.79%; P-value = 0.045%) Conclusion: Our results showed that short abstinence may not be beneficial in all infertile males, and it should only be used in selected patients with abnormal DNA integrity.

Is there any relationship between human sperm parameters and protamine deficiency in different groups of infertile men?

OBJECTIVE: Abnormality in Histone-Protamine replacements has been indicated to cause sperm DNA damage and infertility. The aim of the present study was to investigate the relationships between sperm parameters in oligospermia, asthenospermia, and teratospermia with protamine deficiency in infertile men. MATERIAL AND METHOD: In this case-control study, we had three experimental groups including oligospermia (n=100), asthenospermia (n=100), and teratospermia (n=100) as well as normospermia (n=100) as controls. Sperm analyses were performed according to the recommendations of the World Health Organization (WHO, 2010) and sperm chromatin quality was assessed using Chromomycin A3 (CMA3) staining for each sample. RESULTS: The comparison of the data between groups indicated that the percentage of spermatozoa with protamine deficiency was significantly different in patients with oligospermia, asthenospermia, and teratospermia when compared with control ones. However, there was no significant correlation between sperm nuclear protamine deficiency and their parameters of the men with teratospermia using CMA3 test. Regarding the oligospermia and asthenospermia semen samples, the findings showed the negative correlations between the sperm nuclear protamine deficiency and progressive motility as well as immobility (p<0.001). CONCLUSION: The higher proportion of spermatozoa with abnormal chromatin packaging was observed in asthenospermic samples than those from other experimental groups as well as controls. It seems that normal morphology cannot have a valuable predictive value for good chromatin quality of spermatozoa, as much as normal motility characteristics, since samples with high mobility rates often have lower protamine deficiencies. The findings may provide a supportable promoting the future wider clinical application of chromatin/DNA integrity testing along with the semen analysis in male infertility.

Iranian temporal changes in semen quality during the past 22 years: A report from an infertility center.

BACKGROUND: Despite numerous reports about temporal changes in semen quality from all over the world, the debates continue. The latest systemic review has shown an overtime decrease in semen quality worldwide. OBJECTIVE: To assess the temporal changes in the semen quality among Iranian population referred to an infertility center. MATERIALS AND METHODS: In this retrospective cross-sectional study, semen parameters including concentration, motility, and morphology were compared between Iranian men reffered to Research and Clinical Center for Infertility, Yazd between 1990 to 1992 (group 1, n = 707) and 2010 to 2012 (group 2, n = 1108). Demographic characteristics and semen analysis were collected from the records. The effect of age on semen parameters was also investigated. RESULTS: Despite the increase in sperm concentration l in group 2, sperm with normal morphology decreased significantly (p < 0.001). Grade-A motility decreased (p < 0.001), grade B motility increased (p < 0.001), and grade C and D motile sperm remained constant (p = 0.303 and p = 0.315, respectively). Also, no significant correlation between the age and semen parameters were observed. CONCLUSION: This study showed inconsistent temporal changes in the participant semen quality. Significant temporal decline were obtained between various semen parameters, sperm morphology and grade A motility. These results should be further evaluated by larger studies in the future.

Evaluation of sperm protamine deficiency and apoptosis in infertile men with idiopathic teratozoospermia.

OBJECTIVE: Sperm morphology plays an important role in infertility, especially in cases of defects in the heads of spermatozoa. Tapered-head or elongated-head spermatozoa are examples of morphological abnormalities. The aim of this study was to compare the semen parameters, levels of protamine deficiency, and frequency of apoptosis between patients with normozoospermia and those with teratozoospermia with tapered-head spermatozoa. METHODS: Fifty-two semen samples (27 patients with tapered-head sperm and 25 fertile men) were collected and semen analysis was performed according to the World Health Organization criteria for each sample. Protamine deficiency and the percentage of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3) staining and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assays, respectively. RESULTS: Sperm concentration, motility, and normal morphology in the tapered-head spermatozoa (cases) were significantly lower than in the normozoospermic samples (controls). CMA3-reactive spermatozoa (CMA3+) in the case group were more common than in the controls. Apoptotic spermatozoa (TUNEL-positive) were significantly more common in the cases than in the controls. CONCLUSION: This analysis showed that tapered-head spermatozoa contained abnormal chromatin packaging and exhibited a high rate of apoptosis, which can be considered to be an important reason for the impaired fertility potential in teratozoospermic patients with tapered-head spermatozoa.