Evaluation of the Effect of Vitamin E on Reproductive Parameters in Morphine-Treated Male Mice.

Background: Morphine is a narcotic pain reliever that is prescribed to reduce postoperative pain and can produce reactive oxygen species (ROS). Therefore, it can have negative effects on spermatogenesis and male fertility. Vitamin E is an effective antioxidant which plays an important role in membrane lipid peroxidation due to increased ROS. The present study aimed to evaluate the effects of vitamin E and morphine on sperm parameters, level of malondialdehyde (MDA), and diameter of seminiferous tubules in morphine-treated mice. Methods: In this experimental study, 80 mice were divided into ten groups (n=8) including control, normal saline, vehicle, morphine, various doses of vitamin E (100, 200, 300 mg/kg), and morphine plus vitamin E (100, 200, 300 mg/kg) groups. The groups were followed up for 30 consecutive days. Sperm parameters, testis weight, the diameter of seminiferous tubules, and the level of MDA were analyzed and compared. Findings: Data analysis showed seminal parameters decreased significantly (excluding sperm count) and there was an increase in the level of MDA in morphine-treated mice compared with the normal saline group (P<0.05). Administration of E100 to morphinetreated mice did not show a significant difference in the evaluated parameters compared with the morphine group. However, E200 and E300 significantly reduced MDA and improved sperm parameters (P≤0.05). Conclusion: The results showed co-administration of vitamin E in high doses (200 & 300) could prevent the deleterious effects of morphine on some reproductive parameters and decrease the level of MDA in morphine-treated mice.

Male factor testing in recurrent pregnancy loss cases: A narrative review.

Recurrent pregnancy loss is a distinct disorder defined as the loss of at least 2 pregnancies before the 20 th wk of gestation. With half of the genome of the embryo belonging to the father, the integrity of the sperm genome is crucial for a successful pregnancy. Semen analysis is recommended for men in such cases to evaluate sperm concentration, morphology, vitality and motility. However, other important sperm parameters such as sperm epigenetics, aneuploidy, Y chromosome microdeletion and chromatin integrity also correlate with successful pregnancy and delivery rate. This article examines the use of different sperm tests and their importance in male partners of women suffering from recurrent pregnancy loss.

Effect of Selenium and Vitamin E on the Level of Sperm HSPA2+, Intracellular Superoxide Anion and Chromatin Integrity in Idiopathic Asthenoteratozoospermia: A Double-Blind, Randomized, Placebo- Controlled Trial.

PURPOSE: Male infertility accounts for about half of all infertility cases. Asthenoteratozoospermia is a severe form of male infertility. Free radicals play an important role in infertility. In a study we found that asthenoteratozoospermic men had a lower mean percentage of sperm HSPA2+ and higher intracellular anion superoxide than normozoospermia. Antioxidants are thought to be able to counteract the negative effects of free radicals. We explored the efficacy of vitamin E in combination with Se on the level of sperm HSPA2+, intracellular anion superoxide and chromatin integrity in these patients. MATERIALS AND METHODS: 60 patients entered the study. They were randomized to treatment group of oral Se (200 µg) in combination with vitamin E (400 units) for 3 months (n= 30) or placebo (n= 30). Semen samples were obtained and assessed for sperm parameters, intracellular O2-, protamine deficiency, sperm HSPA2+ and apoptotic spermatozoa at baseline and after treatment phase. RESULTS: There were no significant differences in baseline semen parameters, intracellular O2- protamine deficiency, sperm HSPA2+ and apoptotic spermatozoa between the treatment and placebo groups. There was a statistically significant decrease in sperm apoptosis and the level of anion superoxide (P=.001) and an increase in sperm motility and viability (P=.001) in the treated group, but no significant difference was found in the percentage of sperm HSPA2+ and sperm protamine deficiency compared with baseline. Moreover, no significant change was found in these parameters in placebo group after 3 months. CONCLUSION: Our results showed that administration of vitamin E and selenium for three months may improve sperm motility and viability by decreasing intracellular anion superoxide and sperm apoptosis in asthenoteratozoospermic infertile men. We suggest that consuming these supplements before assisted reproductive technology (ART) may improve outcomes in these patients.

Comparison between sperm parameters and chromatin in recurrent pregnancy loss couples after antioxidant therapy.

BACKGROUND AND AIMS: Recurrent Pregnancy loss (RPL) is a heterogeneous disease. The role of maternal factor is clear but the relationship between the paternal factors remains uncertain. It has been shown that increase the level of Reactive Oxygen Species (ROS) and decrease the antioxidant levels in men can lead to RPL. New researches show treatment with antioxidant can improved sperm parameter. The aim of the present study was to assess the effect of vitamin E plus Zinc therapy on sperm parameters and chromatin quality in couples with RPL. METHODS: In this clinical trial study, 60 RPL patients were selected from RPL clinic. Standard sperm parameters were analyzed and then male partners were intake vitamin E and Zinc in antioxidant therapy plan for 90 days. After that, sperm parameters were analyzed again. Sperm chromatin assay was reviewed before and after vitamin E and Zinc therapy by cytochemical assay including aniline blue (AB), chromomycin A3 (CMA3), toluidine blue (TB). To assess DNA fragmentation index, TUNEL test was used. Data were analyzed and compared before and after treatment. RESULTS: data analysis showed all sperm parameters significantly improved after treatment (P < 0.001). The number of AB+ and TB+ sperms were decreased significantly after vitamin E and Zinc therapy (P = 0.0001). Decrease in DNA fragmentation in post treatment group in comparison to pre-treatment was statistically significant. CONCLUSION: Supplemental of vitamin E plus Zinc may improve sperm parameters chromatin quality and decrease sperm DNA fragmentation in RPL patients.

Cytochemical tests to investigate sperm DNA damage: Assessment and review.

Male factor infertility has been diagnosed as the cause of infertility in about 20% of infertile couples. Sperm analysis is the most common method for diagnosing infertility in a laboratory. However, approximately 15% of infertile men have a normal sperm analysis. Therefore, the result of a routine sperm analysis often cannot be a definitive diagnosis for male factor infertility. Also, approximately 8% of infertile men with normal sperm parameters have high levels of abnormal sperm DNA. This indicates the role of the integrity of sperm DNA in male infertility. Here, we review the current tests available to evaluate the sperm DNA integrity along with their benefits and limitations.

Evaluation of Sperm Chromatin Integrity Using Aniline Blue and Toluidine Blue Staining in Infertile and Normozoospermic Men.

BACKGROUND: Male infertility is defined as a man lost his ability to fertilize a fertile female naturally. Diagnosis of male infertility cannot be made just according to basic semen analysis. It is necessity to have specific tests for evaluation of chromatin integrity. In this study, an attempt was made to evaluate the sperm chromatin quality in fertile men and infertile subgroup. METHODS: Among 1386 couples, 342 men were categorized into normospermia and 1044 were infertile and they were referred to Yazd Research and Clinical Center for infertility treatment. Standard semen analysis and sperm nuclear maturity tests including aniline blue (AB) and toluidine blue (TB) staining were done. Data were analyzed by SPSS software. The p≤0.05 was considered statistically significant. RESULTS: The mean value of TB staining was significantly higher in infertile group compared to normospermic group (p=0.005). Mean of sperm normal morphology was lower in idiopathic infertile men in comparison with normozoospermic men (p= 0.001). The highest negative correlation was obtained between sperm count and AB staining. Progressive motility was negatively correlated with AB and TB staining in both groups but there was no significant difference between AB staining and progressive motility in men normospermia group. CONCLUSION: Sperm chromatin staining using AB and TB showed a negative association between sperm chromatin condensation with sperm count, normal morphology and progressive motility. It seems that the AB and TB test may be useful for the assessment of male fertility potential.

The etiologies of DNA abnormalities in male infertility: An assessment and review.

The sperm DNA damage may occur in testis, genital ducts, and also after ejaculation. Mechanisms altering chromatin remodeling are abortive apoptosis and oxidative stress resulting from reactive oxygen species. Three classifications of intratesticular, post-testicular, and external factors have been correlated with increased levels of human sperm DNA damage which can affect the potential of fertility. Lifestyle, environment, medical, and iatrogenic factors might be considered to cause dysmetabolism to make distracting interactions and endocrine disrupting compounds. As a result, these may induce chromatin/DNA alteration in germ cells, which may be transmitted across generations with phenotypic consequences. Alcohol consumption may not increase the rate of sperm residual histones and protamine deficiency; however, it causes an increase in the percentage of spermatozoa with DNA fragmentation and apoptosis. In a medical problem as spinal cord injury, poor semen parameters and sperm DNA damage were reported. Infection induces reactive oxygen species production, decreases the total antioxidant capacity and sperm DNA fragmentation or antigen production that lead to sperm dysfunctions and DNA fragmentation. While reactive oxygen species generation increases with age, oxidative stress may be responsible for the age-dependent sperm DNA damage. The exposing of reproductive organs in older men to oxidative stress for a long time may produce more DNA-damaged spermatozoa than youngers. Examining the sperm chromatin quality in testicular cancer and Hodgkin's lymphoma patients prior to chemotherapy demonstrated the high incidence of DNA damage and low compaction in spermatozoa at the time of the diagnosis. In chemotherapy cycle with genotoxic agents in cancer patients, an increase in sperm DNA damage was shown after treatment. In overall, those factors occurring during the prenatal or the adult life alter the distribution of proteins associated with sperm chromatin induce changes in germ cells which can be detected in infertile patients.

Evaluation of intracellular anion superoxide level, heat shock protein A2 and protamine positive spermatozoa percentages in teratoasthenozoospermia.

BACKGROUND: Teratoasthenozoospermia (TA) is a severe form of male infertility with no clear etiology. OBJECTIVE: To compare the level of intracellular anion superoxide (O2-), heat shock protein A2 (HSPA2) and protamine deficiencies in ejaculated spermatozoa between teratoasthenozoospermic and normozoospermic men. MATERIALS AND METHODS: In this case- control study, semen samples of 20 infertile men, with TA (with normal morphology lower than 4%_ and total motility lower than 40% ) as the case group and 20 normozoospermic fertile men as the control group were evaluated for intracellular O2- and HSPA2 by flow cytometry and protamine deficiency by Chromomycin A3 (CMA3) test. RESULTS: The rate of CMA3+ spermatozoa in the case group was higher than controls (p=0.001). The percentages of HSPA2+ spermatozoa in the cases were significantly lower than controls (p=0.001). Also, intracellular O2- levels in the case group were significantly higher than controls (p=0.001) and had positive correlations with sperm apoptosis (r=0.79, p=0.01) and CMA3 positive sperm (r=0.76, p=0.01), but negative correlations with normal morphology (r=-0.81, p=0.01) and motility (r=-0.81, p=0.01). There was no significant correlation between intracellular O2- and HSPA2 in the case group (r=0.041, p=0.79). CONCLUSION: We suggest that the increase in intracellular O2-, decrease in spermatozoa HSPA2+, and high percentages of spermatozoa with immature chromatin might be considered as etiologies of infertility in TA patients.

Etiologies of sperm oxidative stress.

Sperm is particularly susceptible to reactive oxygen species (ROS) during critical phases of spermiogenesis. However, the level of seminal ROS is restricted by seminal antioxidants which have beneficial effects on sperm parameters and developmental potentials. Mitochondria and sperm plasma membrane are two major sites of ROS generation in sperm cells. Besides, leukocytes including polymer phonuclear (PMN) leukocytes and macrophages produce broad category of molecules including oxygen free radicals, non-radical species and reactive nitrogen species. Physiological role of ROS increase the intracellular cAMP which then activate protein kinase in male reproductive system. This indicates that spermatozoa need small amounts of ROS to acquire the ability of nuclear maturation regulation and condensation to fertilize the oocyte. There is a long list of intrinsic and extrinsic factors which can induce oxidative stress to interact with lipids, proteins and DNA molecules. As a result, we have lipid peroxidation, DNA fragmentation, axonemal damage, denaturation of the enzymes, over generation of superoxide in the mitochondria, lower antioxidant activity and finally abnormal spermatogenesis. If oxidative stress is considered as one of the main cause of DNA damage in the germ cells, then there should be good reason for antioxidant therapy in these conditions.

Mycobacterium tuberculosis infection in women with unexplained infertility.

BACKGROUND: Genital tuberculosis (GTB) is an important cause of female infertility, especially in developing countries. The positive results of polymerase chain reaction (PCR) in endometrial GTB in the absence of tubal damage raise the possibility of the detection of sub-clinical or latent disease, with doubtful benefits of treatment. OBJECTIVE: To evaluate the mycobacterium tuberculosis infection in endometrial biopsy samples collected from unexplained infertile women attending Yazd Research and Clinical Center for Infertility by using PCR techniques. MATERIALS AND METHODS: In this cross sectional study, 144 infertile women with unexplained infertility aged 20-35 years old and normal Histro-saplango graphy findings were enrolled. Endometrial biopsy samples from each participant were tested for mycobacterium tuberculosis detecting by PCR. In 93 patients, peritoneal fluid was also taken for culture and PCR. RESULTS: The PCR results of endometrial specimens were negative in all cases, demonstrating that there was no GTB infection among our patients. CONCLUSION: Our results showed that GTB could not be considered as a major problem in women with unexplained infertility. Although, studies have indicated that PCR is a useful method in diagnosing early GTB disease in infertile women with no demonstrable evidence of tubal or endometrial involvement.