Standard Sperm Parameters Research Articles

Correlation between standard sperm parameters and sperm DNA fragmentation from 11,339 samples

Conventional semen parameters have long been considered fundamental in male fertility analyses. However, doubts have been raised regarding the clinical utility of the assessment of spermatozoa (sperm) DNA damage. In this retrospective study, we investigated the potential correlation between conventional semen parameters and semen DNA fragmentation (SDF) assessed as sperm DNA damage, in 11,339 semen samples collected between January 2019 and June 2022. We observed significant negative correlations between the DNA fragmentation index (DFI) and sperm viability (correlation coefficient [r] = −0.514) as well as progressive sperm motility (r = −0.512, p < 0.05). Samples were categorized into three groups according to DFI levels (Groups A, B, and C: ≤15%, 15 < DFI ≤30%, and >30%, respectively). Furthermore, the percentage of semen samples with normal sperm conventional parameters in Groups A, B, and C was 76.7% (4369/5697), 61.4% (2351/3827), and 39.7% (721/1815), respectively. Moreover, according to the reference values of conventional sperm parameters, the samples were divided into Groups F, G, and H with all normal, only one abnormal, and > two abnormal parameters, respectively. In addition, the proportions of samples with abnormal DFI values (>30) in Groups F, G, and H were 9.7% (721/7441), 23.1% (618/2676), and 39.0% (476/1222), respectively. Multivariate logistic regression models demonstrated that sperm vitality, progressive sperm motility, normal sperm form, total sperm count, semen volume, age, and some sperm kinematics collectively improved the area under the receiver operating characteristic curve (AUROC) to 0.861, surpassing the predictive value of a single predictor of pathologically damaged sperm DNA. Our study suggests that samples with abnormal sperm parameters may have a higher likelihood of high DNA fragmentation. Furthermore, certain semen parameters could be potential indicators of sperm DNA fragmentation, aiding sperm selection in assisted reproductive procedures.

Effects of male hepatitis B virus infection and serostatus on sperm quality, pregnancy outcomes, and neonatal outcomes following intrauterine insemination.

To evaluate the impact of male hepatitis B virus (HBV) infection and serostatus on sperm quality, pregnancy outcomes, and neonatal outcomes following intrauterine insemination for infertility. We retrospectively analyzed data from 962 infertile couples undergoing intrauterine insemination treatment at a single center. The case group comprised 212 infertile couples with male HBV infection, and the control group comprised 750 noninfected infertile couples. The couples were further divided into subgroups according to their hepatitis B e antigen (HBeAg)/anti-HBe status: hepatitis B surface antigen (HBsAg)+HBeAg- (group A), HBsAg+HBeAg+ (group B), and HBsAg-HBeAg- (control group). The main outcome parameters, including the seminal parameters, clinical pregnancy rate, miscarriage rate, live birth rate, preterm delivery rate, multiple pregnancy rate, delivery type, birth weight, and sex ratio, were compared. A lower sperm acrosin activity, higher cesarean rate, and newborn sex ratio were observed in the HBV-infected group and group A in comparison with the control group (P < 0.05). However, the standard sperm parameters, clinical pregnancy rate, miscarriage rate, live birth rate, preterm delivery, and birth weight showed no statistically significant differences among the groups. Male HBV infection does not adversely impact standard sperm parameters or pregnancy outcomes but can influence sperm acrosin activity and some neonatal outcomes. Moreover, the effect may vary among different HBV serostatuses.

α/β-Hydrolase D16B Truncation Results in Premature Sperm Capacitation in Cattle.

Recently it was shown that a specific form of male infertility in Holstein cattle was caused by a nonsense variant in the α/β-hydrolase domain-containing 16B (ABHD16B) gene resulting in a protein truncation at amino acid position 218 (p.218Q*) and loss of function. Lipidomics showed that the absence of ABHD16B influenced the content of phosphatidylcholine (PC), ceramide (Cer), diacylglycerol (DAG), and sphingomyelin (SM) in variant carrier sperm membranes. However, the exact cause of infertility in affected sires has remained unclear until now. To elucidate the cause of infertility, we analyzed (i) standard sperm parameters (i.e., total sperm number, morphological intact sperm, total sperm motility), (ii) in vitro fertilizability and effects on early embryonic development, and (iii) sperm survival rates (i.e., capacitation time). The affected spermatozoa showed no changes in the usual sperm parameters and were also capable of fertilization in vitro. Furthermore, the absence of ABHD16B did not affect early embryonic development. Based on these results, it was concluded that the affected spermatozoa appeared to be fertilizable per se. Consequently, the actual cause of the inability to fertilize could only be due to a time- and/or place-dependent process after artificial insemination and before fertilization. A process fundamental to the ability to fertilize after insemination is capacitation. Capacitation is a biochemical maturation process that spermatozoa undergo in the female genital tract and is inevitable for the successful fertilization of the oocyte. It is known that the presence and concentration of certain sperm membrane lipids are essential for the correct course of capacitation. However, precisely these lipids are absent in the membrane of spermatozoa affected by the ABHD16B truncation. Since all other causes of fertilization inability were excluded in the previous experiments, consequently, the only remaining hypothesis was that the loss of function of ABHD16B leads to a capacitation disruption. We were able to show that heterozygous and homozygous affected spermatozoa exhibit premature capacitation and therefore decay before fertilization. This effect of the loss of function of ABHD16B has not been described before and our studies now revealed why sires harboring the variant in the ABHD16B gene are infertile.

STUDY OF SPERM DNA FRAGMENTATION IN MALE PARTNERS OF INFERTILE COUPLES

Background: Infertility affects 15% couples of reproductive age in those 35% with male factor. Semen analysis is cornerstone to evaluate male factor infertility. Sperm chromatine dispersion test is light weight and fast test Normal sperm creates sperm halo. If sperm DNA fragments exceeds 30% sperm quality is significantly reduced. Aim: To study the Role of DNA fragmentation in male partner of infertile couple. Results: The mean age of cases was 31.71 years and that of control was 32.63 years with majority of cases (43.3%) were in age group 31-35 years and controls were (56.7%) in age group 25-30 years. The age difference was statistically non-significant as p-value is greater than 0.05. The mean BMI in cases was 22.9 kg/m2 and that of control was 24.5 kg/m2 with majority of cases (73.33%) and controls (53.3%) had BMI between 18.5-24.9 kg/m2. The difference was statistically non-significant. Here in cases 56.7% were tobacco chewer followed by 30% were smoker, 26.7% were alcoholic, 6.7% had other habits and 1% had no personal history. In control 33.33% were tobacco chewer followed by 23.3% were smoker, 6.7% were alcoholic and 36.7% had no personal history. In cases 83.3% had DFI greater than equal to 30 and 16.7% had DFI less than 30. In control 20% had DFI greater than equal to 30 and 80% had DFI less than 30. The mean DFI in cases was 37.26 and that of control was 23.64. The difference was statistically significant Conclusion: Sperm DNA integrity measurement is more reproducible and more objective than conventional parameters. Our study indicates that the levels of sperm DNA fragmentation in men with subnormal semen parameters were significantly higher compared to the levels in men with normal semen parameters. SDI may reveal a hidden abnormality of sperm nuclear DNA in infertile men classified as idiopathic based on apparently normal standard sperm parameters. This test has an important diagnostic and prognostic value in the evaluation of male infertility, particularly in relation to assisted reproductive technologies. Keywords: DNA fragmentation, DFI

The Negative Impact of Varicocele on Basic Semen Parameters, Sperm Nuclear DNA Dispersion and Oxidation-Reduction Potential in Semen.

Since varicocele is so common in infertile men, this study intends to analyse the relationships between varicocele and conventional semen characteristics, sperm nuclear DNA dispersion and oxidation-reduction potential (ORP) in semen. Varicocele-positive and varicocele-negative infertile men (study groups) showed significantly lower standard sperm parameters and higher sperm DNA fragmentation (SDF) and ORP in semen than healthy volunteers and subjects with proven fertility (control groups). A lower proportion of low SDF levels (0–15% SDF) and higher incidence of high SDF levels (>30% SDF), as well as a higher prevalence of high ORP values (>1.37 mV/106 sperm/mL), were found in the study groups vs. the control groups. Moreover, infertile men had significantly lower odds ratios (ORs) for low SDF levels and significantly higher ORs for high SDF levels and high ORP. SDF and ORP were negatively correlated with sperm number, morphology, motility and vitality. Furthermore, a significant positive correlation was found between SDF and ORP. The obtained results suggest that disorders of spermatogenesis may occur in varicocele-related infertility. These abnormalities are manifested not only by reduced standard semen parameters but also by decreased sperm DNA integrity and simultaneously increased oxidative stress in semen.

Can leukocytospermia predict prostate cancer via its effects on mitochondrial DNA?

Leukocytospermia was previously reported to affect sperm quality by the production of reactive oxygen species (ROS) leading to oxidative stress (OS). In turn, OS decreases sperm functional integrity, increases sperm DNA damage and ultimately alters fertility status. To elucidate the impact of leukocytospermia on sperm nuclear DNA integrity and mitochondrial DNA (mtDNA) structure, we conducted a study including 67 samples from infertile patients with low level of leucocytes (Group 1: n=20) and with leukocytospermia (Group 2: n=47). In addition to standard sperm parameters' assessment, we measured the levels of inflammation biomarkers [interleukin-6 (IL-6) and interleukin-8 (IL-8)] and evaluated the oxidative status [malondialdehyde (MDA) and enzymatic and non-enzymatic antioxidants]. In addition, we evaluated the level of sperm nuclear DNA fragmentation and analysed mitochondrial DNA (mtDNA) of sperm cells by sequencing of 5 genes [cytochrome oxidase I (COXI), cytochrome oxidase II (COXII), cytochrome oxidase III (COXIII), adenosine triphosphate synthase 6 (ATPase 6) and adenosine triphosphate synthase 8 (ATPase 8)]. As expected, patients with leukocytospermia had significantly higher MDA levels (32.56±24.30 nmole/ml) than patients without leukocytospermia (17.59±9.60nmole/ml) (p<.018). Also, sperm DNA fragmentation index (DFI) was significantly higher in Group 2 (33.05±18.14%) as compared to Group 1 (14.19±9.50%) (p<.001). The sequencing of mtDNA revealed a high number of substitutions in Group 2 (n=102) compared to Group 1 (n=5). These substitutions were observed mainly in COXI. Among COXI substitutions found in Group 2, twelve changes were previously described in patients with prostate cancer and six of them were shown associated with this pathology. These findings suggest that leukocytospermia may predispose to the manifestation of prostate cancer through modification of mitochondrial DNA and this may be promoted by OS.

Relationship between nuclear DNA fragmentation, mitochondrial DNA damage and standard sperm parameters in spermatozoa of infertile patients with leukocytospermia

The association of leukocytospermia with male fertility is still under debate. Our objective was to evaluate the association of leukocytospermia with sperm parameters, mitochondrial DNA (mtDNA) variations, and seminal concentration of several oxidative stress and inflammatory cytokines in Tunisian infertile men. The studied patients were divided into two groups: patients without leukocytospermia (Group 1) and patients with leukocytospermia (Group 2). DNA fragmentation significantly increased in group 2 (31.41 %) compared to group 1 (14.68 %) ; (p < 0.001). A total of 115 nucleotide substitutions in mitochondrial DNA were depicted, among which 113 were previously identified. The number of substitutions was more elevated in group 2. Leukocytospermic group had significantly higher MDA (nmole/mL) levels than patients without leukocytospermia (34±24.43 vs 18.94±15.96 ; p=0.001), GSH (μg/mL) levels were also higher compared to the control group (126.53±22.87 vs 79.4±19.38 ; p < 0.001), SOD (U/mg of protein) levels were higher but without reaching the statistical significance (89.74±74.85 vs 67.56±37.11 ; p = 0.25) ; whereas seminal CAT (μmole H2O2/min/mg of protein) levels were lower in this group (10.66±14.32 vs 27.35±25.28 ; p = 0.012). No statistically significant differences between the two groups of patients were found in the levels of inflammatory cytokines. However, IL-8 level was positively correlated with DNA fragmentation and negatively correlated with vitality. These findings confirm the association between leukocytospermia and sperm DNA damage.

Positive effect of nutraceuticals on sperm DNA damage in selected infertile patients with idiopathic high sperm DNA fragmentation.

The use of nutraceuticals to improve sperm parameters and male fertility is debatable, even if evidence suggests that selected infertile patients might benefit from their use. In particular, oxidative stress might play a role in idiopathic male infertility, leading to sperm membrane damage and high sperm DNA fragmentation (SDF). The aim of this study was to evaluate, in selected idiopathic infertile men with high SDF, the effect on sperm DNA damage and on standard semen parameters of a nutraceutical formulation containing myoinositol, alpha lipoic acid, coenzyme Q10, selenium, zinc and B vitamins. The study included 60 idiopathic infertile men with DNA fragmentation index (DFI) >20%. Semen analysis and DFI determination were assessed at baseline and after three months of nutraceutical treatment. Primary outcome was change in DFI. Semen volume, sperm concentration, total sperm count, sperm motility and sperm morphology did not change after treatment. Instead, sperm vitality significantly increased (65.9±11.8% pre-treatment vs. 69.4±9.4% post-treatment, P<0.05) and DFI significantly decreased (33.5±10.1% pre-treatment vs. 26.8±8.7% post-treatment, P=0.0001) after treatment. The percentage of men with normal standard sperm parameters significantly increased (15% vs. 30%, P<0.05). The mean decrease in DFI was -6.7±1.4% and the percentage of men with DFI ≤30% after treatment was 75.0% compared to 48.3% pre-treatment (P<0.005). Higher pre-treatment DFI (and no other parameters) correlated with greater DFI reduction after treatment. Nutraceuticals might be effective in idiopathic infertile men with high DFI to reduce SDF, increase sperm vitality and globally improve semen parameters.

Semen bacterial contamination: Antibiotics susceptibility pattern and standard sperm parameters in men of Birjand, 2017

Background and Aim: Urinary tract infections can be one of the most important causes of infertility in men. Identification of semen bacterial contamination and using of appropriate antibiotic treatment can cause to improvement sperm parameters. The aim of this study was to evaluate the frequency of bacterial contamination of semen and its effect on standard sperm parameters. Materials and Methods: In this descriptive-analytical study, the semen analysis of 40 men referring to Birjand laboratories was performed by using the CASA model spermogram device. The Eosin-Nigrosin staining was used to evaluate of sperm viability. After semen samples were cultured on microbial culture media, bacterial contamination was assessed by using conventional microbiological methods. The determination of antibiotic susceptibility of bacterial isolates was performed by the disk diffusion method. Results: In the present study, 30% of semen 12 samples were detected with bacterial contamination, the most frequent of which was related to Escherichia coli (41.7%). Statistical analysis showed that there was a significant relationship between bacterial contamination of semen and history of urinary tract infection (UTI) (P=0.001). There was a significant decrease in the motility, viability and sperm count in subjects with bacterial semen contamination (P&lt;0.05). Nitrofurantoin, Gentamicin and third-generation of Cephalosporins were reported as the most effective treatment options for reducing bacterial contamination in semen. Conclusion: The bacterial contamination of semen can significantly reduce the motility, viability and sperm count. Due to the high prevalence of bacterial contamination in semen and its significant association with sperm factors, the microbial screening of infertile couples without clinical symptoms is essential. Key Words: Antibiotic Resistance; Bacterial Contamination; Infertility; Semen fluid; Sperm

Semen Bacterial contamination: Antibiotics susceptibility pattern and standard sperm parameters in men of Birjand,2017

Background and Aim: Urinary tract infections can be one of the most important causes of infertility in men. Identification of semen bacterial contamination and using of appropriate antibiotic treatment can cause to improvement sperm parameters. The aim of this study was to evaluate the frequency of bacterial contamination of semen and its effect on standard sperm parameters. Materials and Methods: In this descriptive-analytical study, the semen analysis of 40 men referring to Birjand laboratories was performed by using the CASA model spermogram device. The Eosin-Nigrosin staining was used to evaluate of sperm viability. After semen samples were cultured on microbial culture media, bacterial contamination was assessed by using conventional microbiological methods. The determination of antibiotic susceptibility of bacterial isolates was performed by the disk diffusion method. Results: In the present study, 30% of semen 12 sampleswere detected with bacterial contamination, the most frequent of which was related to Escherichia coli (41.7%). Statistical analysis showed that there was a significant relationship between bacterial contamination of semen and history of UTI(Urinary ??? infection) (P = 0.001). There was a significant decrease in the motility, viability and sperm count in subjects with bacterial semen contamination (P &lt;0.05). Nitrofurantoin, Gentamicin and third-generation of Cephalosporins were reported as the most effective treatment options for reducing bacterial contamination in semen. Conclusion: The bacterial contamination of semen can significantly reduce the motility, viability and sperm count. Due to the high prevalence of bacterial contamination in semen and its significant association with sperm factors, the microbial screening of infertile couples without clinical symptoms is essential. Key Words: Antibiotic Resistance; Bacterial Contamination; Infertility; Semen fluid; Sperm

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.

The higher risk for sperm DNA damage in infertile men.

Supplementary assays are needed for determination of relationships between sperm biomarkers and fertility potential. Therefore, our research was designed to determine the extent of sperm DNA fragmentation (SDF) and establish a discriminating threshold of SDF for fertility potential. Semen characteristics were evaluated according to World Health Organization recommendations, and SDF was assessed by sperm chromatin dispersion test on ejaculated spermatozoa from infertile and healthy normozoospermic men. A higher proportion of SDF was noted in infertile men (median 23.00%) than normozoospermic (median 14.00%). Significantly less subjects (17.03%) with low SDF level (≤ 15%) and more (35.17%) with high SDF level ( > 30%) were found for the infertile group vs the normooospermic (57.90% and 5.26%, respectively). Infertile group had significantly lower odds ratio (OR) for having a low SDF level (OR: 0.1493) and higher OR for having a high SDF level (OR: 9.7627). Receiver operating characteristic analysis [area under curve (AUC) = 0.785] revealed that 20% SDF is predictive value for discriminating between infertile and normozoospermic subjects. SDF was negatively correlated with the sperm number, morphology, progressive motility and vitality but positively with the teratozoospermia index. Our study demonstrates: (1) a significant difference in the extent of SDF and in the risk for having damaged sperm DNA between infertile and normozoospermic men, (2) > 20% SDF has negative predictive value for fertility potential, (3) coexistence of abnormal standard sperm parameters with sperm chromatin damages. Therefore, SDF should be considered as a highly valuable indicator of male fertility potential.

The association of seminal leucocytes, interleukin-6 and interleukin-8 with sperm DNA fragmentation: A prospective study.

The effect of seminal leucocytes on sperm DNA integrity has been discussed controversially in literatures. Moreover, the studies investigating the in vivo effect of pro-inflammatory cytokines interleukin-6 and interleukin-8 on sperm DNA fragmentation are scarce and inconsistent. The association of standard sperm parameters with sperm DNA fragmentation is also a matter of ongoing discussion. Hence, the aims of this study were, first, to evaluate the effect of seminal leucocytes, interleukin-6 and interleukin-8 on sperm DNA integrity and, second, to examine whether standard semen parameters are associated with sperm DNA fragmentation. Seminal leucocytes, interleukin-6, interleukin-8 and standard semen parameters, including total sperm number, sperm concentration, progressive motility, nonprogressive motility, immotility and normal morphology, were determined in 134 consecutive men. The concentrations of seminal leucocytes, interleukin-6 and interleukin-8, did not correlate with sperm DNA fragmentation. In contrast, total sperm number, sperm concentration, progressive motility, nonprogressive motility and normal morphology exhibited significant inverse correlations with sperm DNA fragmentation. Immotile spermatozoa were directly correlated with sperm DNA fragmentation. In conclusion, seminal leucocytes, interleukin-6 and interleukin-8, are not associated with sperm DNA fragmentation. Poor standard semen parameters are significantly related to the high levels of sperm DNA fragmentation.

PO-01-075 Effects of a nutraceutical compound combined with Avanafil on standard sperm parameters, percentage of mature spermatozoa, and sperm capacity to undergo hyperactivation

The main objective of this randomized-controlled trial was to evaluate the effects of a nutraceutical compound (NC) and the effects of avanafil on sperm functional assays administered either alone or in combination. Men with oligoasthenospermia (n=217) were treated daily for 90 days with either an NC (45men, Group-A), L-carnitine (44men, Group-B), NC plus avanafil (43men, Group-C) or avanafil (43men, Group-D); another group of 42 oligoasthenospermic men (Group-E) received no treatment. Sperm parameters were performed before and after the end of treatment in each Group A,B,C and D respectively. The same sperm parameters were evaluated in each participant of Group-E before and at the end of the experimental period. Wilcoxon test for paired observations and Chi-square test (Yates’ correction) were employed for statistical analysis to evaluate, within each group, differences in the mean values (prior to and at the end of the experimental period) for each quantitative and qualitative parameter respectively. A probability P<0.05 was considered to be statistically significant. All assays were conducted in a blind fashion.

Mind-Body Interventions Significantly Decrease Oxidative DNA Damage in Sperm Genome: Clinical Implications

Mind-body interventions (MBIs) have been broadly categorized into a group of interventions which facilitate mind’s capacity to affect the functions of the body. There is a growing body of evidence suggesting the adoption of MBIs like yoga, meditation, Tai Chi, and Qigong as an adjunct in the management of various complex lifestyle-related disorders. This review summarizes the importance of simple yoga- and meditation-based lifestyle intervention as a critical component of male infertility therapy. Defective chromatin integrity is one of the hallmarks of male factor infertility. Regular practice of yoga and meditation affects and targets the whole body, decreases free radical levels, and causes collateral increase in levels of antioxidants, not only resulting in improvement in standard sperm parameters, but also becoming ideal in treating oxidative stress and oxidative DNA damage and modulating levels of sperm transcripts through affecting the sperm methylome. This may aid in reversing testicular aging and improving the overall health and quality of life of male infertility patients and of the next generation.

IMPACT OF SINGLE AND MULTIPLE SPERM ABNORMALITIES AND LOW-LEVEL LEUKOCYTOSPERMIA ON SPERM DNA

&#x0D; Background and objective&#x0D; The aim of the present study was to identify the impact of defective standard sperm parameters individually and in combination on DNA damage in a large cohort of infertile men.&#x0D; Material and methods&#x0D; Retrospective analysis of semen characteristics was conducted on 436 patients. DNA fragmentation analysis was performed by using the terminal deoxynucleotidyl transferase (TdT)-mediated fluoresce-in-dUTP nick end labeling (TUNEL) assay. Sperm parameters were arranged into different categories such as normospermia asthenospermic, teratospermic, asthenoteratospermic, and oligoasthenoteratospermic. GraphPad Prism version 7 software was used for data analysis.&#x0D; Results&#x0D; Our results suggest that the mean percentage of DNA damage was proportionally higher than the semen abnormalities. Sperm with 3 abnormal parameters showed significantly higher DNA damage, suggesting that sperm having more than 2 abnormalities are more likely to have higher DNA damage.&#x0D; Conclusion&#x0D; Sperm motility had significant correlation and is supposed to be a predictor for these tests, while morphology was the second standard sperm parameter inversely correlated with sperm DNA damage. Patients demonstrating low levels of leukocytospermia should be advised sperm DNA testing before assisted reproductive technology treatment. However, there is a clear need for more research studies to further address these issues.&#x0D; &#x0D; &#x0D;

DNA methylation level of spermatozoa from subfertile and proven fertile and its relation to standard sperm parameters.

The epigenetic mechanism plays an important role in spermatogenesis such as DNA methylation where this episode is represented by either switching genes on or off. Twenty-eight samples (14 case and 14 controls) were subjected to Infinium 450K BeadChip arrays to identify genomic regions that differ in sperm DNA methylation patterns in the subfertile compared to the proven fertile group. Then two CpGs were validated by deep bisulphite sequencing on 82 sperm samples. The results screening study revealed eight CpGs were significantly different in their sperm DNA methylation levels between cases and control group. The results of the validation study for the two CpGs (cg19779893 and cg19406113) showed that a significant variation in the methylation level at 2 CpGs of 3 CpGs related to cg19779893 site amplicon in cases compared to the controls. Moreover, six CpGs related to the cg19406113 site amplicon showed significant differences in sperm DNA methylation between the cases and the control group. Furthermore, there was a significant decrease in the sperm parameters in the cases compared to the control group. This study found two CpGs altered in their sperm DNA methylation levels. In addition, a strong association was found between changes in the sperm DNA methylation levels in these CpGs sites and sperm parameters.

Relationship between nuclear DNA fragmentation, mitochondrial DNA damage and standard sperm parameters in spermatozoa of fertile and sub-fertile men before and after freeze-thawing procedure.

The aim of this study was to assess the stability of nuclear and mitochondrial DNA (n-DNA and mt-DNA) of spermatozoa under freeze-thawing and to find out the correlation between them and their association with standard sperm parameters. Forty-three semen samples were collected from fertile (G.1; n=29) and sub-fertile (G.2; n=14). N-DNA fragmentation was determined by TUNEL assay and mt-DNA using caspase 3 staining. Each semen sample was frozen at -196°C by the programmed freezer. Freeze-thawing decrease vitality, total motility and membrane integrity from (43.02±22.74%; 31.63±18.15%; 51.5±24.82%) to (22.71±17.3%; 9.21±6.61%; 34.64±19.92% respectively [p<.001]). G.1 native spermatozoa stained positive with TUNEL and caspase 3 were (14.85±17.6% and 5.8±11.59%) and increased after freeze-thawing to 27.54±19.74% (p=.004) and 7.3±6.13% (p=.01) respectively. In G.2, TUNEL and caspase 3 were (19.84±17.52% and 7.53±8.56%) and increased to (29.48±16.97% [p=.03] and 10.21±11.73%). In conclusion, freeze-thawing process affects not only semen parameters but also n-DNA and mt-DNA. Therefore, n-DNA and mt-DNA could be used as sensitive parameters for assessment of the cryodamage of human spermatozoa.

Assessment of male factor involved in recurrent pregnancy loss: A preliminary study

Abstract This study was conducted to assess the male factor as standard sperm parameters, age and chromosome abnormalities from men whose partners had a history of recurrent pregnancy loss (RPL). A 2 years retrospective study was carried out in 149 couples with 2 or more miscarriages. They referred to as for genetic counseling in the laboratory of histology housed in a Faculty of Medicine of Sfax, with genetic laboratories. Chromosomal analysis was performed using RHG banding in peripheral blood and semen samples were analyzed from men according to World Health Organization guidelines. 67 men were selected of a total of 149 couples whose partners were healthy and with normal karyotype. About them, the mean age was 33 years [from 22 to 53], and altered semen parameters were detected in more than 25%. Abnormal karyotype was detected in 4 men with an overall incidence of 5.97%. The commonest abnormalities appear to be reciprocal translocation. So, this data emphasizes the importance of assessment of male factor in addition to standard female factors for evaluating the risk for RPL.

Impact of cigarette-smoking on sperm DNA methylation and its effect on sperm parameters.

DNA methylation is an epigenetic modification of the genome. The purpose of this study was to determine the influence of cigarette-smoking on sperm DNA methylation from a genomewide survey of sperm samples and to ascertain its effect on sperm parameters. Twenty-eight sperm DNA samples (from 14 fertile smokers as a case study and 14 proven fertile nonsmokers as controls) were subjected to Infinium 450K BeadChip arrays to identify the changes in the DNA methylation level between the two groups. Then, deep bisulphite sequencing was used to validate five CpGs on 78 samples. The results from the Infinium 450K found that only 11 CpGs showed a significant difference in DNA methylation between the case and the control groups. Five CpGs of the eleven (cg00648582, cg0932376, cg19169023, cg23841288 and cg27391564) underwent deep bisulphite sequencing where cg00648582, related to the PGAM5 gene, and the cg23841288 CpGs, related to the PTPRN2 gene amplicons, showed a significant increase in their DNA methylation level in more than one CpG in the case group. In contrast, a significant decrease was found at cg19169023 and at its various neighbouring CpGs in the TYRO3 gene-related amplicons. Furthermore, this study demonstrated a significant correlation between the variation in sperm DNA methylation level and standard sperm parameters in the case group.