Sperm chromatin and DNA integrity, methyltransferase mRNA levels, and global DNA methylation in oligoasthenoteratozoospermia / 대한생식의학회지

OBJECTIVE: To investigate sperm chromatin/DNA integrity, global DNA methylation, and DNMT mRNA transcription in men with oligoasthenoteratozoospermia (OAT) compared with normozoospermic men. METHODS: Semen samples from 32 OAT patients who comprised the case group and 32 normozoospermic men who comprised the control group were isolated and purified using a standard gradient isolation procedure according to World Health Organization criteria. DNMT1, DNMT3A, and DNMT3B transcripts were then compared between groups using real-time quantitative reverse-transcription polymerase chain reaction. Global DNA methylation in sperm was determined by an enzyme-linked immunosorbent assay. Protamine deficiency and the proportion of apoptotic spermatozoa were evaluated using chromomycin A3 (CMA3), aniline blue (AB), and toluidine blue (TB) staining, as well as the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The p-values < 0.05 were considered to indicate statistical significance. RESULTS: Significantly higher proportions of AB+, TB+, CMA3+, and TUNEL+ spermatozoa, as well as DNMT3A and DNMT3B transcription, were found in the OAT group. Positive correlations were detected between sperm parameters, DNA/chromatin damage, and DNMT3A and DNMT3B transcripts. Global DNA methylation was significantly higher in the OAT patients and had a significant correlation with abnormal results of all sperm chromatin integrity tests, but was not associated with DNMT1, DNMT3A, or DNMT3B expression. CONCLUSION: Oligoasthenoteratozoospermic men showed abnormal sperm parameters, abnormal chromatin/DNA integrity, and a higher global DNA methylation rate, as well as overexpression of DNMT mRNA.

Evaluation of sperm retrieval rate with bilateral testicular sperm extraction in infertile patients with azoospermia

Background: About 10% to 15% of infertile men have azoospermia, which could be obstructive or non-obstructive. Diagnostic biopsy from the testis and recently testicular sperm extraction [TESE] are the most precise investigations in these patients. Testicular biopsy can be done unilaterally or bilaterally. The worth of unilateral or bilateral testicular biopsy in men with azoospermia is controversial

Objective: To evaluate the necessity of bilateral diagnostic biopsy from the testis in new era of diagnosis and treatment of male infertility

Materials and Methods: In this retrospective study, we reviewed the results of testis biopsy in 419 azoospermic men, referred to Yazd Research and Clinical Center for Infertility from 2009-2013. Patients with known obstructive azoospermia were excluded from the study

Results: In totally, 254 infertile men [60.6%] were underwent unilateral TESE, which in 175 patients [88.4%] sperm were extracted from their testes successfully. Bilateral testis biopsy was done in 165 patients [39.4%] which in 37 patients [22.4%], sperm were found in their testes tissues

Conclusion: Due to the low probability of positive bilateral TESE results especially when we can't found sperm in the first side, we recommend that physicians re-evaluate the risk and benefit of this procedure in era of newer and more precise technique of sperm retrieval like micro TESE

Comparison of reactive oxygen species in neat and washed semen of infertile men

Male are involved in near 50% of cases of infertility and reactive oxygen species [ROS] playing an important role in decreasing fertility potential. Accurate measurement of ROS seems to be important in evaluation of infertile male patients. To compare ROS measurement in neat and washed semen samples of infertile men and define the best method for evaluation of ROS in these patients. We measured the level of ROS in semen samples of thirty five non-azoospermic men with infertility. The semen samples were divided into two parts and the semen parameters and ROS levels in neat and washed samples were evaluated. We also evaluated the presence of pyospermia using peroxidase test. The differences regarding sperm count and quick motility were significant in neat and washed semen samples. The mean ROS level was significantly higher in neat samples compared with washed spermatozoa [7.50 RLU vs. 1.20 RLU respectively]. Difference in ROS levels was more significant in patients with pyospermia compared to whom with no pyospermia [378.67 RLU vs. 9.48 RLU respectively]. Our study confirmed that neat or unprocessed samples are better index of normal oxidative status of semen samples. Because we do not artificially add or remove factors that may play an important role in oxidative equilibrium status

effect of oral administration of Pentoxifylline on sperm motility of asthenozoospermic ejaculates from men with or without testicular varicoceles

Pentoxifylline [PX] is a methyxanthin derivative that influences the sperm motion characteristics. In general, PX has been reportedly effective in preserving sperm motility in vitro, also when administered orally to the asthenozoospermic patients. The main objective of this prospective clinical trial study was to rule out the effect of oral administration of PX on sperm progressive motility of asthenozoospermic ejaculates obtained from men with or without mild testicular varicoceles. In addition, the role of patient's age on sperm motility following PX administration was investigated. A total of 68 infertile men with asthenozoospermia were allocated to this study. Following physical examination, 20 cases were found with mild varicocele of testis. A dosage of 400 mg PX/ twice daily for duration of 3 months was administered to each patient. Two semen samples [one before and one after the PX therapy] were evaluated under blind condition. Semen parameters of sperm concentration, total and fast progressive motility [%] and morphology [%] were analyzed for each sample. Also, the sperm motion characteristics of asthenozoospermic patients with testicular varicocele were compared with cases lacking varicocele. The subjects were divided into two age groups of <30 and >/= 30 years old. PX was significantly effective on the fast progressive motility of sperm [p<0.01]. Also, total progressive motility was enhanced from 26.82 +/- 16.8 to 29.60 +/- 22.2 with PX therapy. However, PX did not have any negative effect on other semen parameters. Oral therapy of PX was also effective in improving the fast progressive motility of sperm of samples from cases with or without mild testicular varicocele [p<0.01]. Fast progressive motility was also significantly enhanced in ejaculates of men from both age groups. Our results demonstrate that low dose of oral therapy of PX is significantly useful in enhancing fast progressive motility of sperms from infertile men with asthenozoospermia. Also, testicular varicocele did not interfere with enhancing effect of PX on sperm motility