Role of male factor in recurrent abortion and in vitro fertilization failure has not been fully defined yet and there is much controversy about evaluating male patients with normal semen analysis. One of the factors that might help establish the male role is DNA fragmentation index. However, strong correlation between this factor and quality of semen, has caused many clinicians to believe that it does not help in abortion and implantation failure. We aim to assess this factor in our patients. In a prospective observational study, we assessed age, duration of infertility, undesired fertility related events (assisted reproductive techniques attempts and abortions), semen parameters and DNA fragmentation index in patients with multiple abortions or in vitro fertilization failures and analysed the results by statistical software SPSS version 24. DNA fragmentation index was remarkably correlated with age, duration of infertility and semen parameters. Among all groups in our study, patients with abnormal semen analysis had statistically significant higher level of DNA fragmentation. Ten percent of patients with normal or slightly abnormal semen analysis had abnormally high SDFI (sperm DNA fragmentation index). Checking DNA fragmentation index is recommended in all couples with fertilization problems even in the presence of normal semen analysis. It might be more reasonable to assess it in aged men, long duration of infertility or candidates with remarkable semen abnormality.
- Keywords
- Abortion, DNA fragmentation index, In vitro fertilization, Intracytoplasmic sperm injection, Male, Sperm,
- MeSH
- Semen Analysis MeSH
- Fertilization in Vitro methods MeSH
- DNA Fragmentation MeSH
- Abortion, Habitual * MeSH
- Humans MeSH
- Infertility, Male * diagnosis genetics MeSH
- Aged MeSH
- Semen MeSH
- Spermatozoa MeSH
- Pregnancy MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Aged MeSH
- Pregnancy MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Observational Study MeSH
Obesity represents a growing problem due to its impacts on human health and reproduction. In this study, we analysed semen quality, sperm DNA integrity and gene-specific CpG methylation in 116 healthy men from normal population. The men were divided into three groups according to their body mass index (BMI), and their ejaculates were analysed using standard methods, sperm chromatin structure assay (SCSA), methylation next generation sequencing (NGS) and amplicon sequencing. The sperm methylation NGS revealed six significantly differentially methylated regions (DMRs). Using subsequent targeted amplicon sequencing in 116 men, two of the DMRs were proved as differentially methylated in sperm of men with normal BMI vs. BMI ≥ 25. The DMRs were located in the EPHA8 and ANKRD11 gene. Also, we detected a significant decline in the EPHA8, ANKRD11 and CFAP46 gene methylation in association with increasing BMI values. The genes EPHA8 and ANKRD11 are involved in the nervous system and brain development; the CFAP46 gene plays a role in a flagellar assembly and is associated with sperm motility. Significantly lower rates of motile and progressive motile sperm were observed in men with BMI ≥ 30. Our results show that excess body weight can modify CpG methylation of specific genes, affect sperm motility, and compromise sperm chromatin integrity. These factors can stand behind the observed reduced fertility in men with obesity. The methylation changes might be transmitted to their offspring through sperm, and become a basis for possible developmental and reproductive issues in the next generation.
- Keywords
- BMI, Body mass index, DNA fragmentation, DNA methylation, Obesity, Semen, Sperm,
- MeSH
- Semen Analysis * MeSH
- Chromatin * metabolism MeSH
- CpG Islands MeSH
- Adult MeSH
- Body Mass Index * MeSH
- Humans MeSH
- DNA Methylation * MeSH
- Young Adult MeSH
- Sperm Motility genetics MeSH
- Obesity genetics MeSH
- Spermatozoa * metabolism MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Young Adult MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Chromatin * MeSH
The epidemiological relationship between tuberculosis cases in a prison and between cases within five families was investigated. Therefore, the isolated Mycobacterium tuberculosis strains were subjected to restriction fragment length polymorphism (RFLP) analysis using insertion sequence IS6110 as a probe. In case of 11 patients, the expected links of transmission were confirmed by RFLP typing. In contrast, in case of 4 patients the conclusion of classical contact tracing were not in agreement with the DNA fingerprinting results. These findings reinforce the usefulness of this recently developed technique as an additional tool in contact tracing. The IS6110DNA fingerprints of all strains investigated consisted of 7 to 13 bands and showed a high degree of polymorphism. Comparison of these fingerprints with those recorded in the Czech Republic previously, revealed the presence of a predominant DNA fingerprint type, without a known connection between the cases. Furthermore, other patterns found in the present study showed a high degree of similarity with the previously obtained fingerprints. Part of the patients were sampled twice. All of these double isolates showed identical fingerprints, confirming the previously described stability of IS6110DNA fingerprints. In contrast, from one couple of strains, isolated from husband and wife both suffering from tuberculosis, a slight change in one of the two patterns was observed. The patterns shared 10 band positions, confirming the expected relationship between these cases, but one of the patterns contained one additional band.
- MeSH
- DNA Fingerprinting * MeSH
- Humans MeSH
- Mycobacterium tuberculosis genetics MeSH
- Polymorphism, Restriction Fragment Length MeSH
- Contact Tracing methods MeSH
- Tuberculosis transmission MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
A detailed survey of mitochondrial and chloroplast diversity in eight populations of Silene vulgaris from Central Europe was conducted for comparison with previously published data on diversity from S. vulgaris populations in the introduced range. Mitochondrial DNA (mtDNA) variation around the coxI gene was assessed with Southern blotting/restriction fragment length polymorphism methods. Chloroplast variation was assessed by sequencing the intergenic spacer separating the trnH and psbA genes. Thirty mtDNA haplotypes and 24 chloroplast DNA (cpDNA) haplotypes were found within 86 individuals. The overall genetic diversity h (0.941 for mitochondrial, and 0.893 for chloroplast markers) and within-population diversity were higher than reported in previous population studies of S. vulgaris in the USA and Europe. The frequency of private alleles was surprisingly high - more than 90% for both kinds of markers. Most of our populations were large and located in relatively undisturbed meadows, whereas surveys in Virginia consisted of smaller roadside populations. The slow rate of population turnover in European populations is discussed as a factor responsible for the relatively high diversity of S. vulgaris in undisturbed areas of its native range. Association between mtDNA and cpDNA haplotypes was also demonstrated. Finally, gender and mtDNA haplotype were associated in the Alps populations, where females were very rare.
- MeSH
- DNA, Chloroplast genetics MeSH
- Genetic Variation * MeSH
- Haplotypes genetics MeSH
- Crosses, Genetic MeSH
- DNA, Mitochondrial genetics MeSH
- Molecular Sequence Data MeSH
- Polymorphism, Restriction Fragment Length MeSH
- Reproduction genetics MeSH
- Base Sequence MeSH
- Sequence Analysis, DNA MeSH
- Silene genetics MeSH
- Blotting, Southern MeSH
- Geography MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Comparative Study MeSH
- Geographicals
- Europe MeSH
- Names of Substances
- DNA, Chloroplast MeSH
- DNA, Mitochondrial MeSH
Markers of apoptosis were followed in batch hybridoma cultures carried out in protein-free medium. Samples were collected on day 0, representing early exponential phase (viability 91%), and on day 8, corresponding to late stationary phase (viability 8%). The apoptotic index reflecting the relative number of bodies insoluble in 6 M guanidinium hydrochloride in the culture of day 8 (30%) exceeded markedly the index in the culture of day 0 (2.5%). A gel chromatography on Sepharose 2B was developed for quantitative evaluation of fragmented cellular DNA. This analysis, including a correction for nonspecific fragmentation, showed that on day 8 more than 30% of cellular DNA was fragmented, whereas on day 0 it was less than 5%. Control necrotic cells prepared by rapid killing in 1% sodium azide displayed a low apoptotic index (2.4%) and low DNA fragmentation. Electrophoretic patterns in agarose gel showed a typical "ladder" of fragments in the DNA sample of day 8. The demonstration of fragmented cellular DNA and of the high incidence of apoptotic bodies at late stationary phase adds substantial weight to the view that in hybridoma cultures apoptosis represents the prevalent mode of cell death.
The purpose of the present study was to investigate the impact of carcinogenic polycyclic aromatic hydrocarbons and volatile organic compounds on sperm quality in a group of city policemen in Prague during a period of increased concentrations of ambient air-pollutants (winter season) compared to a period of low exposure (spring). Polymorphisms in metabolic genes (CYP1A1, EPHX1, GSTM1, GSTP1, GSTT1), folic acid metabolism genes (MTR, MTHFR) and DNA repair genes (XRCC1, XPD6, XPD23, hOGG1) were evaluated in these men as potential modifiers of associations between air pollution exposure and changes in sperm quality. The study population was a group of 47 policemen working in the center of the city. Seasonal differences in exposure were verified by ambient and personal monitoring. Markers of sperm injury included semen volume, sperm concentration, sperm morphology, sperm motility, and sperm DNA damage measured with the sperm chromatin structure assay The sperm chromatin structure assay (SCSA) includes a measure of DNA damage called DNA Fragmentation Index (DFI). The % of cells with detectable DFI (detDFI) by this assay includes sperm with either medium or high DNA damage; the term hDFI is used to define the % of sperm with only high DNA damage. The assay also detects immature sperm defined by high density staining (HDS). No significant differences were found in any of the standard semen parameters between the sampling periods except for vitality of sperms. Both DFI and HDS were significantly higher in winter than in spring samples for all men and for non-smokers. At the bivariate level, significant associations between hDFI or detDFI and polymorphisms of the repair genes XRCC1, XPD6 and XPD23 were observed. In multivariate models, polymorphisms of the genes XPD6, XPD23 and CYP1A1MspI were associated with hDFI and HDS. Moreover, HDS was significantly associated with polymorphisms in GSTM1 gene.
- MeSH
- Chromatin genetics MeSH
- Cytochrome P-450 CYP1A1 genetics metabolism MeSH
- Adult MeSH
- DNA Repair Enzymes genetics metabolism MeSH
- DNA Fragmentation drug effects MeSH
- Genotype MeSH
- Glutathione Transferase genetics metabolism MeSH
- Cotinine urine MeSH
- Smoking MeSH
- Folic Acid metabolism MeSH
- Air Pollutants, Occupational adverse effects MeSH
- Humans MeSH
- Police MeSH
- Polymerase Chain Reaction MeSH
- Polymorphism, Restriction Fragment Length MeSH
- Polymorphism, Genetic * MeSH
- DNA Damage genetics MeSH
- Spermatozoa drug effects MeSH
- Xeroderma Pigmentosum Group D Protein genetics metabolism MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Chromatin MeSH
- Cytochrome P-450 CYP1A1 MeSH
- DNA Repair Enzymes MeSH
- ERCC2 protein, human MeSH Browser
- glutathione S-transferase M1 MeSH Browser
- Glutathione Transferase MeSH
- Cotinine MeSH
- Folic Acid MeSH
- Air Pollutants, Occupational MeSH
- Xeroderma Pigmentosum Group D Protein MeSH
Previous studies have provided evidence for an association between exposure to high levels of air pollution and increased DNA damage in human sperm. In these studies DNA damage was measured using the sperm chromatin structure assay (SCSA) wherein the percentage of sperm with abnormal chromatin/fragmented DNA is determined and expressed as % DNA fragmentation index (%DFI). Here we extend these observations to address the following hypothesis: men who are homozygous null for glutathione-S-transferase M1 (GSTM1-) are less able to detoxify reactive metabolites of carcinogenic polycyclic aromatic hydrocarbons (c-PAHs) found in air pollution. Consequently they are more susceptible to the effects of air pollution on sperm chromatin. Using a longitudinal study design in which men provided semen samples during periods of both low (baseline) and episodically high air pollution, this study revealed a statistically significant association between GSTM1 null genotype and increased SCSA-defined %DFI (beta=0.309; 95% CI: 0.129, 0.489). Furthermore, GSTM1 null men also showed higher %DFI in response to exposure to intermittent air pollution (beta=0.487; 95% CI: 0.243, 0.731). This study thus provides novel evidence for a gene-environment interaction between GSTM1 and air pollution (presumably c-PAHs). The significance of the findings in this study with respect to fertility status is unknown. However, it is biologically plausible that increases in %DFI induced by such exposures could impact the risk of male sub/infertility, especially in men who naturally exhibit high levels of %DFI.
- MeSH
- Gene Deletion MeSH
- Adult MeSH
- DNA Fragmentation MeSH
- Genotype MeSH
- Glutathione Transferase genetics MeSH
- Air Pollutants toxicity MeSH
- Humans MeSH
- DNA Damage genetics MeSH
- Spermatozoa drug effects metabolism MeSH
- Air Pollution adverse effects analysis MeSH
- Check Tag
- Adult MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- Geographicals
- Czech Republic MeSH
- Names of Substances
- glutathione S-transferase M1 MeSH Browser
- Glutathione Transferase MeSH
- Air Pollutants MeSH
The effects of air pollution on men's reproductive health can be monitored by evaluating semen quality and sperm DNA damage. We used real-time PCR to analyse the effects of air pollution on sperm mitochondrial DNA copy number (mtDNAcn) and deletion (mtDNAdel) rates in semen samples collected from 54 men in two seasons with different levels of industrial and traffic air pollution. MtDNAdel rates were significantly higher following the high exposure period and were positively correlated with mtDNAcn. However, we did not find any difference in mtDNAcn between the two seasons. MtDNAcn was positively correlated with the DNA fragmentation index and the rates of sperm with chromatin condensation defects, previously assessed by sperm chromatin structure assay, and negatively correlated with sperm concentration, progressive motility, viability, and normal morphology. This indicates that mtDNAcn is more closely associated with male fertility than mtDNAdel rates. In contrast, mtDNAdel might be a more sensitive biomarker of air pollution exposure in urban industrial environments.
- MeSH
- Semen Analysis * MeSH
- Chromatin MeSH
- Humans MeSH
- DNA, Mitochondrial genetics MeSH
- Sperm Motility MeSH
- Spermatozoa MeSH
- DNA Copy Number Variations MeSH
- Air Pollution * adverse effects MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Chromatin MeSH
- DNA, Mitochondrial MeSH
Obesity can adversely affect human health, including fertility. While obesity can disturb the hormonal profile of the female organism and is associated with fertility loss, little is known about what effect male obesity has on fertility. The present study analysed sperm samples of 153 donors. The men were selected from couples attending an infertility clinic, who had tried for 12 months or more to achieve pregnancy without success. The age of the men under investigation was recorded, and their body mass index (BMI) was calculated. All semen samples were assessed for volume, concentration, motility and morphology. Sperm chromatin integrity was measured by sperm chromatin structure assay. Quality of sperm chromatin condensation was assessed by toluidine blue, aniline blue and chromomycin A3 staining. We can conclude that the impact of elevated BMI on the parameters investigated (basic semen parameters, chromatin integrity and chromatin condensation) was not proven in this study. On the other hand, ejaculate quality appeared to be affected by ageing. The impact was reflected by chromatin integrity, which is a factor that can substantially affect fertility in men, rather than by basic sperm parameters.
- MeSH
- Semen Analysis * MeSH
- Chromatin genetics MeSH
- Adult MeSH
- DNA Fragmentation MeSH
- Body Mass Index MeSH
- Middle Aged MeSH
- Humans MeSH
- Sperm Motility MeSH
- Infertility, Male etiology MeSH
- Obesity complications MeSH
- Sperm Count MeSH
- Semen MeSH
- Spermatozoa cytology MeSH
- Aging * MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Chromatin MeSH
Circulating extracellular DNA (ecDNA) is known to worsen the outcome of many diseases. ecDNA released from neutrophils during infection or inflammation is present in the form of neutrophil extracellular traps (NETs). It has been shown that higher ecDNA concentration occurs in a number of inflammatory diseases including inflammatory bowel disease (IBD). Enzymes such as peptidyl arginine deiminases (PADs) are crucial for NET formation. We sought to describe the dynamics of ecDNA concentrations and fragmentation, along with NETosis during a mouse model of chemically induced colitis. Plasma ecDNA concentration was highest on day seven of dextran sulfate sodium (DSS) intake and the increase was time-dependent. This increase correlated with the percentage of cells undergoing NETosis and other markers of disease activity. Relative proportion of nuclear ecDNA increased towards more severe colitis; however, absolute amount decreased. In colon explant medium, the highest concentration of ecDNA was on day three of DSS consumption. Early administration of PAD4 inhibitors did not alleviate disease activity, but lowered the ecDNA concentration. These results uncover the biological characteristics of ecDNA in IBD and support the role of ecDNA in intestinal inflammation. The therapeutic intervention aimed at NETs and/or nuclear ecDNA has yet to be fully investigated.
- Keywords
- PAD4, cell-free DNA, deoxyribonuclease activity, neutrophil extracellular traps, ulcerative colitis,
- MeSH
- Biomarkers metabolism MeSH
- Deoxyribonucleases metabolism MeSH
- DNA blood metabolism MeSH
- Endoscopy MeSH
- Extracellular Traps drug effects metabolism MeSH
- Extracellular Space metabolism MeSH
- Colitis blood chemically induced pathology MeSH
- DNA, Mitochondrial blood MeSH
- Mice, Inbred C57BL MeSH
- Mice MeSH
- Ornithine analogs & derivatives pharmacology MeSH
- Protein-Arginine Deiminase Type 4 metabolism MeSH
- Dextran Sulfate MeSH
- Streptonigrin pharmacology MeSH
- Intestines drug effects pathology MeSH
- Intestinal Mucosa drug effects pathology MeSH
- Severity of Illness Index MeSH
- Inflammation blood pathology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Biomarkers MeSH
- Deoxyribonucleases MeSH
- DNA MeSH
- DNA, Mitochondrial MeSH
- N-alpha-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine amide MeSH Browser
- Ornithine MeSH
- Protein-Arginine Deiminase Type 4 MeSH
- peptidylarginine deiminase 4, mouse MeSH Browser
- Dextran Sulfate MeSH
- Streptonigrin MeSH
