maryam arbabian

Sperm motility is a fundamental factor for sperm penetration into the oocyte and fertilization. According to the World Health Organization (WHO) 2021, less than 42 percent of motility is called asthenozoospermia, which is one of the most common causes of male infertility. This retrospective cohort study aims to investigate sperm DNA damage with TUNEL and SCSA methods and its relationship with sperm parameters, age, and body mass index (BMI) in severe asthenospermia (<2% sperm motility) and normozoospermia.

The study parameters between 111 subjects with severe asthenozoospermia and 113 subjects with normozoospermia were investigated. The 2010 World Health Organization guidelines were used to evaluate sperm parameters, and TUNEL and SCSA methods were used to assess sperm DNA damage. The statistical analysis was done using the t-test of two independent samples. Pearson's correlation coefficient was used to investigate the relationship between sperm DNA damage and sperm parameters, age, and BMI P<0.05 was considered significant.

The mean seminal volume, sperm concentration and count, and the percentage of sperm total motility and progressive motility were significantly lower in severe asthenospermic subjects compared to normozoospermic subjects (P<0.001). In addition, the mean percentage of sperm DNA damage in severe asthenozoospermic individuals was significantly higher than in normozoospermic individuals (P<0.001). There was also a positive and significant relationship between sperm DNA damage and the age of severely asthenozoospermic men.

Sperm motility, sperm DNA damage, and the father's age are essential in the successful conception, development, and health of the embryo. Therefore, the evaluation of sperm DNA damage is recommended in examining male fertility and selecting the appropriate treatment approach for men with severe asthenozoospermia.

Practical Implications:

Considering that the sperm genome constitutes 50 percent of the genetic material of the next generation, evaluating sperm DNA health to determine the appropriate treatment approach in infertile men can be influential in the success of assisted reproductive techniques and in maintaining the next generation’s health.

Antioxidant apigenin (AP) is a natural, non-mutagenic, and less toxic flavonoid with pharmacological anti-cancer, anti-viral, anti-bacterial, anti-apoptotic, and anti-inflammatory activities. This antioxidant is easily received by the cell, binds to sperm DNA, and forms a DNA-AP complex, thereby protecting sperm DNA. The present study was conducted to determine the antioxidant effect of AP on human sperm quality after freezing-thawing.

In this descriptive-analytical study, 10 normozoospermic samples underwent freezing-thawing conditions, and sperm functional tests were investigated in different AP concentrations, including 0.4 mM, 0.2 mM, 0.1 mM, and 0.05 mM.

The quality of total sperm parameters and functional tests decreased after freezing compared to before freezing. Among the AP concentrations, only in the 0.2 mM AP concentration, the improvement of the additional histone percentage, protamine deficiency, and sperm DNA health were observed compared to the control; this finding was not statistically significant.

The use of AP with a concentration of 0.2 mM during freezing-thawing culminates in improving sperm functional tests.

Implementation of sperm preparation techniques based on cellular and molecular characteristics can improve the clinical outcomes of couples with male factor infertility. These methods attempt to select better sperm compared to classical methods of preparation such as swim-up and density gradient centrifugation (DGC). In this view, the aim of this study was the comparison of clinical outcomes of magnetic-activated cell separation (MACS) followed by DGC or DGC alone in infertile men undergoing intracytoplasmic sperm injection (ICSI). For this prospective single parallel blind clinical trial study, 206 infertile couples with male factor infertility and having abnormal sperm morphology higher than 96% were included. 106 and 100 couples were considered for the study (MACS-DGC) and control group (DGC), respectively. Clinical outcomes of ICSI; fertilization, embryo quality, and implantation, pregnancy rates were compared between two groups. Mean of fertilization (80.19 ± 1.88 vs. 75.63 ± 2.06, P=0.1), top embryo quality on the day 3 (30.22 ± 3.59 vs. 17.96 ± 2.9, P=0.009), clinical pregnancy (30.76% vs. 22.22%, P=0.19), and implantation rate (18.12% vs. 10.42%, P=0.04) were higher in the study group compared to the control group. Sperm preparation by MACS followed by DGC in teratozoospermic men could improve the clinical outcomes after ICSI (Registration number: IRCT201610317223N8).

Density gradient centrifugation (DGC) and Zeta procedures are used for separation of normal sperm for treatment of infertility. Previous studies showed that Zeta method was more efficient in separation of normal sperm with intact chromatin compared to DGC. In this study, we aimed to compare postacrosomal sheath WW domain-binding protein (PAWP) protein as one of the factors involved to oocyte activation between two methods. Semen samples were collected from 23 men with normal sperm parameters (concentration, motility and morphology) according to World Health Organization criteria. Semen samples were washed with VitaSperm media and each sample were divided into three groups; control, DGC and Zeta. The percentage and intensity of PAWP were assessed by flow cytometry for each group. In addition, localization of PAWP in spermatozoa was detected by immunostaining procedure. Percentage of PAWP-positive spermatozoa was significantly higher in separated spermatozoa from DGC procedure in comparison with control and Zeta groups (P<0.001), while intensity of PAWP was significantly (P<0.001) higher in separated spermatozoa from Zeta method in comparison with control and DGC procedures. PAWP appear to be localized mainly in equatorial segment and post acrosome sheath of sperm head in all the groups. The Zeta method could separate low percentage of PAWP-positive spermatozoa compared to DGC method but such sperm appear higher intensity of PAWP than DGC. Therefore, separated spermatozoa by Zeta method possibly has better chance in activation of oocyte. However, further studies are needed to confirm this result.

Sperm-specific phospholipase C zeta (PLCζ) protein is expressed during spermatogenesis and plays a main role during oocyte activation. In this study, the expression of sperm PLCζ protein and DNA damage in fertile and infertile men was assessed.
In this case-control study, semen samples were collected from 15 fertile and 10 infertile men candidated for intra-cytoplasmic sperm injection (ICSI). Sperm parameters, the expression of PLCζ protein, and DNA damage were assessed by the World Health Organization (2010) protocol, Western Blot, and TUNEL assay, respectively.
The quality of sperm parameters were significantly lower in the infertile men compared with the fertile men. In addition, the expression of PLCζ protein was significantly lower, and percentage of sperm DNA damage were significantly higher in infertile men than fertile men.
Our results clearly showed that low or absence expression of PLCζ and sperm DNA damage could be considered as factors involved in failed fertilization in these infertile men. Therefore, the evaluation of some tests such as PLCζ protein and chromatin tests to assess fertilization potential of a semen sample for infertile men are recommended.

Varicocele is characterized by dilation and abnormal tortuosity of the spermatic veins in testes and it has long been implicated as a major cause of treatable in fertile men. Testicular function in individuals with varicocele progressively damaged but the exact mechanisms involved in initiation and development of dysfunction of testis in these individuals are not fully known. Therefore, the aim of this review article was to study known pathophysiological mechanisms in the incidence of varicocele.
Literature search in different electronic databases such as Science Direct, Pubmed and Scopus, as well as other data sources were related to ISI between 1989-2015.
Varicocele is often associated with abnormal spermatogenesis, elevated testicular temperature and oxidative stress. This condition is lead to reduction of semen quality and fertility.
The microsurgical varicocelectomy method is now the preferred approach between most urologists due to low recurrence rates and complications compared to other treatment suggested by urologists.

Flow cytometry (FCM) has been extensively used to study mammalian sperm in the areas of clinical andrology and reproductive toxicology. FCM provides a powerful advantage over microscopy technique in terms of rapid, accurate and reproducible technology for the quantification of various cell characteristics, including chromatin status. During spermiogenesis, histones are replaced by protamines resulting in a very condensed structure of sperm chromatin. Infertile men have an increased sperm histone: protamine ratio than fertile counterparts. Chromomycin A3 (CMA3) staining represents a useful tool for assessing the packaging quality of sperm chromatin and allows indirect visualization of protamine deficiency. Routinely, fluorescence microscope is used for evaluation of protamine deficiency by CMA3. Considering the advantages of FCM and increasing use of CMA3 in assessment of protamine deficiency in the literature and its possible use as a diagnostic test, the aim of this study is to standardize this procedure for routine laboratory analysis. Semen samples were collected from 85 infertile men who referred to Isfahan Fertility and Infertility Center. A portion of semen sample was used for routine semen analysis according to WHO criteria and the remainder were evaluated to standardize CMA3 staining procedure for fixation, the number of sperm and duration of exposure to CMA3. The results were compared with standard fluorescent microscopic procedure. Percentage CMA3 positive sperm were compared between flow cytometry and standard fluorescent microscopic procedure. Our results show that fixation, the number of sperm and duration of exposure to CMA3 can affect on FCM outcomes. In addition we show that the samples can be fixed, stained with CMA3, stores and then assessed for FCM. The optimal conditions for FCM assessment of CMA3 are: fixation, concentration of 0.25 mg/ml, sperm density of 2 million/ml and exposure for 60 minutes.