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The present study was performed to investigate the relation of Chlamydia trachomatis infection to sperm chromatin/DNA integrity in a population of infertile men (male partner of infertile couples) from Iran. Blood, semen and first-void urine samples were obtained from 250 infertile men. Data were analysed with regard to the results of (i) serological analysis for specific antibodies to C. trachomatis in serum; (ii) the presence of C. trachomatis and DNA in first-void urine; and (iii) in the semen sample of the male partner, in addition to sperm analysis, four different tests (aniline blue, chromomycin A3, acridine orange and TUNEL) were used to detect sperm chromatin and DNA abnormalities. The main conclusions of the results were: (i) no evidence of C. trachomatis infection in semen samples was found; (ii) sperm DNA fragmentation and chromatin studies were not correlated with C. trachomatis diagnosis; (iii) the percentage of DNA fragmentation is positively correlated with the percentage of immotile sperm but negatively with semen volume, normal morphology; and (iv) in sperm chromatin evaluations, only the percentage of chromatin protamination was related to male age., (© 2017 Blackwell Verlag GmbH.)

Globozoospermia is a severe form of teratozoospermia with low incidence in infertile patients, considered as one of the important causes of male infertility. The objective was to investigate the chromatin/DNA integrity as well as apoptosis in ejaculated spermatozoa of cases with partial or total globozoospermia. Fifty-seven semen samples were divided into three groups of partial globozoospermia (n = 17), total globozoospermia (n = 10) and normozoospermia (control; n = 30). Sperm chromatin condensation, DNA integrity and apoptosis were assessed using cytochemical assays. The results showed significant differences in sperm parameters of count and motility between two case groups versus controls. The percentages of spermatozoa with abnormal chromatin packaging and protamine deficiency were significantly higher in total and partial globozoospermic men compared to normozoospermic samples. Also, the rates of TUNEL-positive spermatozoa were significantly increased in both globozoospermic cases with respect to the control (18.3 ± 10.1 and 12.3 ± 9.2 versus 5.9 ± 3 respectively). However, no significant differences were noticed between two subgroups of patients with regard to sperm DNA denaturation, DNA fragmentation and apoptosis. Abnormal chromatin packaging, DNA damage and apoptosis were significantly higher in cases than controls. The sperm chromatin/DNA anomalies may be considered as one of the main aetiology of ART failure in globozoospermic patients., (© 2017 Blackwell Verlag GmbH.)

Objectives: To evaluate the effect of cord injury on (1) sperm parameters and (2) DNA chromatin status., Design: Case-control study., Setting: Data were collected from men referred to Research and Clinical Center for Infertility, Yazd, Iran., Participants: Thirty infertile men with the presence of any level of spinal cord injury (SCI) were compared with 30 healthy donors with definite fertility and normal sperm parameters., Interventions: Not applicable., Outcome Measures: Sperm chromatin integrity was assessed using aniline blue (AB), chromomycin A3 (CMA3), toluidine blue (TB), and acridine orange (AO) assays. The rate of apoptotic spermatozoa was evaluated with terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) staining., Results: Sperm concentration, motility, and morphology in men with SCI were significantly decreased compared with control group (P < 0.05). In addition, with regard to cytochemical staining and TUNEL test, the rate of reacted spermatozoa was increased significantly in SCI group when compared with the controls (P < 0.05). The majority of AB, TB, AO, and CMA3-reacted spermatozoa were higher than the "cut-off" value in men with SCI, as were the number of apoptotic spermatozoa stained with TUNEL., Conclusion: Results showed that SCI disturbs sperm parameters, nuclear maturity, and DNA integrity of spermatozoa. Therefore, the production of spermatozoa with less condensed chromatin and more apoptotic rate increases after cord injury and this may be one possible cause of infertility following SCI.

The aim of this study was to examine the possible relationship between sperm DNA integrity and chromatin packaging evaluated by cytochemical assays, traditional sperm parameters and recurrent spontaneous abortion (RSA) of unknown origin. In this cohort study, 40 couples with a history of RSA and 40 couples with proven fertility were considered as case and control groups respectively. The semen samples of all husbands were analysed for sperm parameters and also sperm chromatin and DNA integrity assessed using cytochemical tests including aniline blue (AB), chromomycin A3 (CMA3), toluidine blue (TB), acridine orange (AOT) and nuclear chromatin stability assay. Among different sperm parameters, only slow motility was significantly different between the two groups. In sperm chromatin evaluations, there were significant differences between the two groups in all of the tests. In addition, the majority of semen samples in RSA patients exhibited upper percentages of abnormal spermatozoa than the cut-off values regarding different cytochemical assays. Our study showed that in the cases of RSA, slow motility had a significant reduction in comparison with controls and also spermatozoa of men from RSA group had less chromatin condensation and poorer DNA integrity than spermatozoa that obtained from fertile men with no history of RSA., (© 2011 Blackwell Verlag GmbH.)

Varicocele occurs in approximately 15% to 20% of the general male population and it is the most common cause of poor semen production and decreased semen quality. It has been demonstrated that patients with varicocele have a significantly higher DNA fragmentation index (DFI) and spermatozoa with nuclear anomalies than healthy fertile men. Therefore, the aim of this study was to evaluate sperm chromatin integrity in these patients. Sixty men referring to the andrology laboratory were categorised into three different groups: 20 infertile men with varicocele, 20 infertile men with abnormal semen parameters and 20 fertile men who had normal spermatogram were considered as control group. Semen analysis was performed according to WHO criteria. To evaluate sperm chromatin quality and DNA integrity, after fixation of sperm smears, aniline blue, toluidine blue, chromomycin A(3) and acridine orange staining were applied in three groups. The slides were analysed by light and fluorescent microscopy and to determine the percentage of mature or immature spermatozoa, 200 spermatozoa were counted in each slide. The results showed that the rates of aniline blue-reacted spermatozoa were significantly higher in infertile and varicocele patients than in the normal group (P < 0.001). In addition, with regard to chromomycin A(3), acridine orange and toluidine blue staining, there was a significant difference between the three groups (P < 0.001). The results showed that the varicocele samples contain a higher proportion of spermatozoa with abnormal DNA and immature chromatin than those from fertile men as well as infertile men without varicocele. Therefore, varicocele results in the production of spermatozoa with less condensed chromatin and this is one of the possible causes of infertility due to varicocele.

Summary: The present study was performed to investigate the relation of Chlamydia trachomatis infection to sperm chromatin/DNA integrity in a population of infertile men (male partner of infertile couples) from Iran. Blood, semen and first‐void urine samples were obtained from 250 infertile men. Data were analysed with regard to the results of (i) serological analysis for specific antibodies to C. trachomatis in serum; (ii) the presence of C. trachomatis and DNA in first‐void urine; and (iii) in the semen sample of the male partner, in addition to sperm analysis, four different tests (aniline blue, chromomycin A3, acridine orange and TUNEL) were used to detect sperm chromatin and DNA abnormalities. The main conclusions of the results were: (i) no evidence of C. trachomatis infection in semen samples was found; (ii) sperm DNA fragmentation and chromatin studies were not correlated with C. trachomatis diagnosis; (iii) the percentage of DNA fragmentation is positively correlated with the percentage of immotile sperm but negatively with semen volume, normal morphology; and (iv) in sperm chromatin evaluations, only the percentage of chromatin protamination was related to male age. [ABSTRACT FROM AUTHOR]

Introduction: Sperm chromatin condensation is a complex process. During this process 85% of histones are replaced by protamines. Protamine deficiency has a negative impact on male fertility, fertilization and early embryonic development. The main goal was to evaluate the rate of spermatozoa with residual histones in a large community of infertile patients. Materials and Methods: A total of 1394 men referred to andrology laboratory during 5 years (2008-2013), were divided into two groups. 1044 men who had abnormal sperm parameters were considered as case and 350 normozoospermic men as normal groups. Semen analysis was performed according to WHO guidelines. Sperm chromatin integrity was assessed using Acidic Aniline Blue (AAB) assay. Results: The rate of AB-reacted spermatozoa were significantly higher in case group compared to the normal group (54.29±19.12 vs. 49.78± 18.32, respecfively, p<0.0001). There was a negative correlation between progressive motility, normal morphology and AB-reacted spermatozoa in infertile patients (Pearson correlation= -0.24, p<0.0001 and Pearson correlation= -0.22, p<0.0001, respectively). Conclusion: The infertile men with abnormal semen parameters showed a higher proportion of sperm cells with residual histones and immature chromatin than those men with normal semen indices. Therefore, it seems evaluation of sperm DNA/chromatin integrity could be regarded as a part of male infertility work-up. [ABSTRACT FROM AUTHOR]

Introduction: Globozoospermia is a severe form of teratozoospermia with very low incidence in infertile patients that is characterized by round sperm head and lack of acrosome. It's considered as one of the important causes of male infertility which the success rate in assisted reproductive technology (ART) cycles is also low. The goal was to compare the semen parameters and chromatin/DNA integrity as well as apoptosis in ejaculated spermatozoa between globozoospermic and normozoospermic men. Materials and Methods: In total 57 men were divided into two groups including globozoospermic (n=27) and normozoospermic men as controls (n=30). Semen analysis was performed according to WHO criteria (2010). Sperm chromatin condensation and DNA integrity were assessed using cytochemical tests including: Aniline blue (AB), Toluidine blue (TB), Chromomycin A3 (CMA3) and Sodium dodecyl sulfate (SDS) for chromatin compaction and Acridine orange (AO), Sperm chromatin dispersion (SCD) and TUNEL assays for DNA structure and apoptosis detection. Results: There were significant differences regarding sperm count, motility and normal morphology between two groups. The percentage of abnormal chromatin packaging/ DNA integrity (using AB, TB, AO and SCD tests) was significantly higher in globozoospermic men compared to normozoospermic samples. The rate of spermatozoa with protamine deficiency (CMA3+) showed an increase in golobozoospermic patients when comparison with controls (63.59±13.29 vs. 24.17±9.5, respectively, pO.OOOl). it should be noted that in SDS test, we didn't any significant difference between groups. But, the rate of TUNEL positive spermatozoa were significantly increased in globozoospermic cases respect to the controls (14.81±9.91 vs. 5.95± 3.02, respectively, p<0.0001). There was no significant correlation between sperm DNA denaturation, DNA fragmentation and apoptosis in globozoospermic men. Our data showed significant correlation between single strand DNA (AO+) and progressive motility (p=0.036). Conclusion: The rate of spermatozoa with abnormal chromatin packaging, DNA damage and apoptosis were significantly higher in globozoospermic samples than normal fertile men. Elowever, the sperm chromatin/ DNA anomalies may be considered as one of the main etiologies of ART failure in these patients. [ABSTRACT FROM AUTHOR]

Introduction: It is estimated that about 50% of causes of recurrent pregnancy loss (RPL) cases remain unknown. Sperm factors are suggested to have probable role in cases with RPL. The goal was to determine the possible relationship between semen bacterial contaminations with unexplained RPL. Also, the correlation between number of bacterial colony and sperm chromatin condensation was examined. Materials and Methods: This study consisted of 30 fertile men (group A) and 30 infertile men (group B) with unknown RPL in their wives. Semen collection and analysis were done according to WHO manuals. Sperm count and motility were evaluated by Makler chamber. Eosin-Nigrosin and Papanicolaou staining methods were applied for viability and morphology assessment, respectively. The semen samples from both groups were cultured for aerobic bacteria. Aniline blue (AB) and toluidine blue (TB) staining methods were applied for evaluating sperm chromatin condensation. Results: The numbers of colonies were significantly higher in group B when compared to group A. Also, S. aureus and E. coli contaminations showed significant differences between two groups. Both AB+ and TB+ sperm cells showed significant increase in group B compared to group A. There was a significant negative correlation between colony number and progressive motility (p=0.01), and sperm viability (p=0.007). In addition, positive correlations were found between colony number and AB+ (p=0.001) and TB+ (p=0.004) as well. Conclusion: Bacterial contaminations in semen of men from RPL couples had significantly higher levels when compared to fertile controls. Presence of microorganisms in semen may be correlated with irregular sperm parameters and quality. [ABSTRACT FROM AUTHOR]