Relationship between sperm protamine deficiency and apoptosis in couples with unexplained repeated spontaneous abortions

Background: Etiology of more than half of Recurrent Spontaneous Abortion. The etiology of more than 50 percent of Recurrent Spontaneous Abortions [RSA] cases has been remained unexplained. It is supposed that RSA may have "paternal effect" due to supply 50% of embryonic genomic content by male gamete

Objective: The aim of present study was to evaluate the role of sperm apoptosis and protamine deficiency at same time in RSA cases

Materials and Methods: Forty fertile [control] and 40 unfertile men with RSA [case] were enrolled in this case-control study. Semen analysis was performed in accordance with WHO criteria and sperm apoptosis and protamine deficiency were evaluated by cell apoptosis detection kit and chromomycin A3, respectively

Results: Results showed significant different between normal morphology and total motility in two groups. Case group had higher percentage of spermatozoa with protamine deficiency and apoptosis compared to controls significantly

Conclusion: Our results showed that in cases of RSA, in addition to abnormal sperm parameters, we have a high percentage of spermatozoa with protamine deficiency and apoptosis and these two anomalies may consider as important causes of idiopathic recurrent abortions. It should be advised that sperm chromatin and DNA examinations are useful tools in the process of RSA treatments

Sperm parameters, protamine deficiency, and apoptosis in total globozoospermia

Globozoospermia is a severe form of teratozoospermia [incidence < 0.1%] in infertile men that is characterized by round headed sperm and acrosomeless in semen. To compare the semen parameters, protamine deficiency, and apoptosis in ejaculated spermatozoa between globozoospermic and normozoospermic men. Thirty six semen samples were divided into two groups including 15 infertile men with total globozoospermic [> 90% round-headed sperm] and 21 healthy donors with normal spermograms as controls. Semen analysis was performed according to World Health Organization criteria [2010]. Sperm protamine deficiency was assessed using Chromomycin A3 [CMA3] staining and the rate of apoptotic spermatozoa was evaluated with TUNEL assay. Sperm concentration, motility, and normal morphology in globozoospermic men were significantly decreased compared with controls [p<0.05]. The rate of CMA3-reacted spermatozoa [CMA3+] in globozoospermic men was higher than controls [65.93 +/- 11.77 vs. 21.24 +/- 7.37, respectively, p<0.0001]. The rate of apoptotic spermatozoa [TUNEL positive] were significantly increased in globozoospermic cases with respect to the controls [17.60 +/- 10.72 and 5.95 +/- 3.02, respectively, p<0.0001]. There was no significant correlation between sperm protamine deficiency and apoptosis in globozoospermic men. Globozoospermic samples contain a higher proportion of spermatozoa with abnormal chromatin packaging and DNA fragmentation than normozoospermic samples. Therefore, in addition to absence of acrosome in the spermatozoa of globozoospermic patients, the high percentage of spermatozoa with immature chromatin and apoptotic marker may be considered as the other etiologies of infertility in these patients

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

Seminal bacterial contaminations: probable factor in unexplained recurrent pregnancy loss

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 This study consisted of 30 fertile men [group A] and 30 infertile [group B] men with unknown RPL. 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. 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. 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

frequency of Yq microdeletion in azoospermic and oligospermic Iranian infertile men

About 15% of couples have infertility problems which 40% of them are related to the male factors. Genetic factors are candidate for about 10% of male infertility conditions. Among these, AZFa, AZFb, AZFc and AZFd regions on the Yq are considered most important for spermatogenesis. Microdeletions of these regions are thought to be involved in some cases of azoospermic or oligospermic infertile men. We studied the prevalence of AZF microdeletions among Iranian infertile men with non-obstructive azoospermia and oligospermia. A total of 50 Iranian azoospermic and oligospermic infertile men were selected for case group and 50 men with normal spermogram as control group. The molecular study of Y chromosome microdeletions was done by multiplex polymerase chain reaction [M-PCR] method by using of 13 sequence tagged site [STS] markers from AZF region. Four [8%] patients showed Y chromosome microdeletions among case group, deletion in AZFc region was the most frequent [80%] followed by AZFb [20%], in AZFa and AZFd region we did not detect any deletions. No deletion was detected in control group; the ratio of Y chromosome microdeletion in azoospermic men was higher than this ratio in oligospermic men [19% [3/16] among azoospermic men and 3% [1/34] among oligospermics]. Serum FSH level in men with microdeletions was higher than this level in men with no deletions [p=0.034]. Because of relatively high prevalence of microdeletions on the long arm of Y chromosome among Iranian azoospermic and oligospermic patients, screening of this microdeletion may be advised to infertile men particularly azoospermic and oligospermic men before using assisted reproductive treatments