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Objective: To study the resistance of human spermatozoa to cryoinjury in repeated cycles of thaw-refreezing by using the fast liquid nitrogen vapor method. Materials and Methods: Semen specimens were obtained from sixteen normal and oligozoospermic individuals who required disposal at the sperm bank. Five of them had testicular cancer. Specimens were thawed and an aliquot was removed for analysis. The remaining specimens were refrozen without removing the cryomedia. Repeated freeze-thaw cycles were performed until no motile sperm were observed. Sperm motility, number of motile spermatozoa and viability were determined after thawing. Resistance to cryoinjury was compared between groups and also after each refreezing cycle within groups. Results: Motile spermatozoa were recovered after five and two refreeze-thawing cycles in normozoospermic and oligozoospermic specimens, respectively. There were no significant differences in the recovery of motile spermatozoa between thaws within each group of normal and oligozoospermic specimens, but percentage motility and total number of motile spermatozoa were significantly lower in the oligozoospermic one. Specimens from men with cancer were exposed to six refreeze-thawing cycles. Although recovery of motile spermatozoa was significantly impaired after each thawing, there were no significant differences in the recovery of motile sperm between thaws in cancer and non-cancer groups. Conclusions: Human spermatozoa resist repeated cryopreservation using the fast liquid nitrogen vapor method. Normozoospermic specimens withstand refreezing for an average two cycles longer than oligozoospermic ones. Specimens from cancer patients seem to resist repeated cryoinjury similarly to non-cancer counterparts. Resistance to repeated cryoinjury was related to the initial semen quality.

OBJECTIVE:We evaluated the effectiveness of sperm washing using the discontinuous two-layer density gradient centrifugation method to remove antisperm antibodies attached to the sperm surface. METHOD: We prospectively enrolled sixty-six men with unexplained infertility who were seeking evaluation. Each patient delivered one semen specimen for the study. We determined antisperm antibody levels using the direct immunobeads test. Specimens were classified into two groups according to the pre-washing levels of antibody-bound spermatozoa: group 1 (low antisperm antibodies levels, immunobeads test , 20%; n ¼ 54), and group 2 (high antisperm antibodies levels, immunobeads test $ 20%; n ¼ 12). Sperm washing was carried out using the discontinuous two-layer colloidal density gradient centrifugation method. Pre-and post-wash levels of antisperm antibodies were compared in the groups. RESULTS: The pre-and post-wash percentage of spermatozoa with antisperm antibodies attached to their surface was 11% and 6.5% in group 1 (mean difference ¼ 40%; p , 0.01), and 30% and 19.5% in group 2 (mean difference ¼ 36.8%, p ¼ 0.02), respectively. The effectiveness of density gradient centrifugation in removing antisperm antibodies was not different between the groups, but individual variations from 252.3% to 23.9% were observed. CONCLUSIONS: Sperm washing by density gradient centrifugation is an overall effective method to remove antibodies bound to sperm membranes, regardless of the levels of antisperm antibodies in the neat semen. Due to an inter-individual variation in the effectiveness of the method, we recommend that each patient be tested before applying sperm washing by density gradient centrifugation in intrauterine insemination.

positive result was considered to urethritis if the mean count of >10.000/mL in the first-voided urine was counted.RESULTS: The mean age of the patients was 34.5;3.8 years. The incidence of leukocytospermia was 22.3% (1115/5000). Patients were classified in two groups: infertile leukocytospermic (n¼ 1115) and non-leukocytospermic group (n¼3885). The concentration of leukocytes was not significantly different between these two groups. Semen volume and sperm concentration were not significantly affect by the presence of leukocytes. Sperm motility was sigificantly lower in leukocytospermic infertile group (47.9; 9.3%) compared to infertile non-leukocytospermic group(68.5; 10.5%)(p<0,0035). The percentage of sperm morphologically normal was lower in infertile leukocytospermic group(7.3;1.9%) compared to infertile non-leukocytospermic group(9.4;1.8%). From the patients with leukocytospermia, 555 (49.8%) were diagnosed with asymptomatic urethritis. CONCLUSIONS: Our study demonstrated a significant correlation between increased seminal leukocytes concentration and the prevalence of abnormal semen parameters. We also concluded a high number of infertile patients present with asymptomatic urethritis confirmed by urethral smear and first-voided urine.OBJECTIVE: There is considerable variability in the literature regarding the incidence of sertoli-cell only (SCO)/germ cell aplasia in nonobstructive azoospermia. Our objective was to define the incidence of SCO histopathology in a modern series of patients.

OBJECTIVE:We evaluated the effectiveness of sperm washing using the discontinuous two-layer density gradient centrifugation method to remove antisperm antibodies attached to the sperm surface. METHOD: We prospectively enrolled sixty-six men with unexplained infertility who were seeking evaluation. Each patient delivered one semen specimen for the study. We determined antisperm antibody levels using the direct immunobeads test. Specimens were classified into two groups according to the pre-washing levels of antibody-bound spermatozoa: group 1 (low antisperm antibodies levels, immunobeads test , 20%; n ¼ 54), and group 2 (high antisperm antibodies levels, immunobeads test $ 20%; n ¼ 12). Sperm washing was carried out using the discontinuous two-layer colloidal density gradient centrifugation method. Pre-and post-wash levels of antisperm antibodies were compared in the groups. RESULTS: The pre-and post-wash percentage of spermatozoa with antisperm antibodies attached to their surface was 11% and 6.5% in group 1 (mean difference ¼ 40%; p , 0.01), and 30% and 19.5% in group 2 (mean difference ¼ 36.8%, p ¼ 0.02), respectively. The effectiveness of density gradient centrifugation in removing antisperm antibodies was not different between the groups, but individual variations from 252.3% to 23.9% were observed. CONCLUSIONS: Sperm washing by density gradient centrifugation is an overall effective method to remove antibodies bound to sperm membranes, regardless of the levels of antisperm antibodies in the neat semen. Due to an inter-individual variation in the effectiveness of the method, we recommend that each patient be tested before applying sperm washing by density gradient centrifugation in intrauterine insemination.