OBJECTIVETo identify the effects of overtraining on human sperm DNA.

METHODSMolecular epidemiological investigation of 249 men from different groups (training and non-training) was carried out by using flow cytometer to detect the integrity and damage of in situ DNA of sperm nucleus, and sperm chromatin structure assay was performed.

RESULTSThe average COMPalpha(t) in training group was 11.02% while that in control group was 5.90% (P < 0.01). COMPalpha(t) was significantly correlated with sperm activity (r = 0.41, P < 0.05).

CONCLUSIONOvertraining could induce sperm DNA injury and affect sperm activity, thus to decrease the potentiality of reproduction.

Adult ; Chromatin ; genetics ; metabolism ; DNA Fragmentation ; Exercise ; physiology ; Humans ; Male ; Sperm Motility ; physiology ; Spermatozoa ; cytology ; metabolism

AIMTo investigate whether early apoptotic changes in spermatozoa can be significant markers for sperm quality.

METHODSTwo early apoptotic changes in the semen of 56 men were assessed using Annexin V (AN)/propidium iodide (PI) staining for phosphatidylserine externalization and JC-1 staining for mitochondrial membrane potential (MMP). The results were compared with conventional semen parameters and DNA fragmentation identified using the TUNEL assay.

RESULTSThe different labeling patterns in the bivariate Annexin V/PI analysis identified four distinctive spermatozoa populations. The percentage of AN(-)/PI(-) spermatozoa positively correlated with conventional semen parameters and MMP, but negatively correlated with TUNEL (+) spermatozoa. As for the AN(-)/PI(+) fraction, we found an opposite result in comparison to AN(-)/PI(-) spermatozoa. The level of early apoptotic AN(+)/PI(+) spermatozoa negatively correlated with MMP and sperm motility. The level of late apoptotic AN+/PI+ spermatozoa negatively correlated with conventional semen parameters and MMP, and positively correlated with TUNEL (+) spermatozoa. MMP positively correlated with conventional semen parameters, but negatively correlated with TUNEL (+) spermatozoa.

CONCLUSIONAlthough early apoptotic AN+/PI(-) spermatozoa only negatively correlates with sperm motility, the differences in proportion of each subpopulation of spermatozoa (especially, the percentage of AN(-)/PI(-) spermatozoa), and decreased MMP might be significant markers for diagnosing male infertility. They possibly bring additional information to predict the outcome of in vitro fertilization.

Adult ; Apoptosis ; physiology ; DNA ; physiology ; DNA Fragmentation ; Humans ; Infertility, Male ; diagnosis ; Male ; Membrane Potential, Mitochondrial ; physiology ; Semen ; physiology ; Spermatozoa ; cytology ; physiology

Objective: Sperm DNA fragmentation (SDF) is widely used to predict male infertility and the methods of detecting SDF are varied. This study aimed to compare two methods of SDF detection and investigate the correlation between SDF and sperm quality. METHODS: Using sperm chromatin structure assay (SCSA) and sperm chromatin dispersion test (SCD), we detected SDF in 108 semen samples collected in the Center of Reproduction and Genetics of Suzhou Municipal Hospital. We compared the results of the two methods and analyzed the correlations of SDF routine semen parameters, sperm morphology and the age of the patients. RESULTS: A significant consistency was found in the SDF index (DFI) between the two methods (P<0.01). The DFI was correlated negatively with sperm motility, the percentage of progressively motile sperm, and that of morphologically normal sperm (P <0.01), but positively with the teratozoospermia index (P <0.01 in SCSA and P <0.05 in SCD). The DFI measured by SCSA showed a significantly positive correlation with the patients' age (P <0.01), but not that obtained by SCD. CONCLUSIONS The results of both SCSA and SCD play an important role in predicting sperm quality. As a clinical index, the DFI has a predictive value for male infertility. However, the results of different detecting methods vary widely, which calls for further studies on their standardization.
Chromatin ; genetics ; physiology ; DNA Fragmentation ; Humans ; Infertility, Male ; diagnosis ; Male ; Semen ; physiology ; Semen Analysis ; Sperm Motility ; Spermatozoa ; physiology ; ultrastructure

PURPOSE: Because NELL2 expression is strictly restricted only in neurons in developing and post- differentiated neural tissues, it is thought to be involved in the neuronal differentiation during development and in the maintenance of neuronal physiology in the post-differentiated neurons. In this study, we examined whether NELL2 is involved in the regulation of cell cycle and apoptosis in the hippocampal neuroprogenitor HiB5 cells. METHODS: Effects of NELL2 on the cultured HiB5 cell numbers, DNA fragmentation, and proteins involved in the regulation of the cell cycle were measured. RESULTS: NELL2 induced a decrease in cell numbers and an increase in G1 phase arrest. Moreover, transfection of NELL2 resulted in an increase of DNA fragmentation that shows an evidence of apoptosis. Contents of proteins involved in the regulation of cell cycle were also changed by transfection of NELL2 expression vectors. CONCLUSION: This study suggests that NELL2 plays an important role in the regulation of cell cycle and apoptosis of neurons.
Apoptosis ; Cell Count ; Cell Cycle* ; DNA Fragmentation ; G1 Phase ; Neurons* ; Physiology ; Transfection

The renin angiotensin system (RAS) appears to influence male fertility at multiple levels. In this work, we analyzed the relationship between the RAS and DNA integrity. Fifty male volunteers were divided into two groups (25 each): control (DNA fragmentation ≤20%) and pathological (DNA fragmentation >20%) cases. Activities of five peptidases controlling RAS were measured fluorometrically: prolyl endopeptidase (which converts angiotensin [A] I and A II to A 1-7), neutral endopeptidase (NEP/CD10: A I to A 1-7), aminopeptidase N (APN/CD13: A III to A IV), aminopeptidase A (A II to A III) and aminopeptidase B (A III to A IV). Angiotensin-converting enzyme (A I to A II), APN/CD13 and NEP/CD10 were also assessed by semiquantitative cytometry and quantitative flow cytometry assays, as were the receptors of all RAS components: A II receptor type 1 (AT1R), A II receptor type 2 (AT2R), A IV receptor (AT4R or insulin-regulated aminopeptidase [IRAP]), (pro)renin receptor (PRR) and A 1-7 receptor or Mas receptor (MasR) None of the enzymes that regulate levels of RAS components, except for APN/CD13 (decrease in fragmented cells), showed significant differences between both groups. Micrographs of RAS receptors revealed no significant differences in immunolabeling patterns between normozoospermic and fragmented cells. Labeling of AT1R (94.3% normozoospermic vs 84.1% fragmented), AT4R (96.2% vs 95.3%) and MasR (97.4% vs 87.2%) was similar between the groups. AT2R (87.4% normozoospermic vs 63.1% fragmented) and PRR (96.4% vs 48.2%) were higher in non-fragmented spermatozoa. These findings suggest that fragmented DNA spermatozoa have a lower capacity to respond to bioactive RAS peptides.
Angiotensins ; DNA Fragmentation ; Humans ; Insulin ; Male ; Renin-Angiotensin System/physiology* ; Spermatozoa

Nowadays, the injury in human mitochondrial DNA (mtDNA) is well known to accumulate in various tissues with age. It's significant to further investigate and then apply it to estimation of the age at parenchymas.
Aging/physiology* ; Base Pair Mismatch/genetics* ; DNA Damage/physiology* ; DNA Fragmentation/genetics* ; DNA, Mitochondrial/physiology* ; Gene Deletion ; Humans ; Polymerase Chain Reaction

OBJECTIVETo study the correlation between sperm DNA fragmentation index (DFI), age of male, various parameters of sperm, rates of fertilization, high quality embryo and pregnancy and implantation rates.

METHODSOne hundred and eleven infertile couples were selected randomly, and DFI was tested by flow cytometry for the sperm used for IVF. The patients were divided into different groups according to the DFI scores. The results of each group were analyzed.

RESULTSThe IVF normal fertilization was significantly lower in couples with sperm DFI over 10% (60.5%) than that in couples with DFI below 10% (70.1%) (P<0.05). Significantly positive correlation was found between DFI and the age of male (r=0.624, P<0.05). DFI was also significantly negatively correlated with the percentage of linearly progressive sperm (r=-0.360, P<0.05). There was no significant correlation between the rates of high quality cleaved embryos, pregnancy and implantation rate and sperm DFI.

CONCLUSIONDFI scores are increased with male's age, and it can influence the sperm motility. DFI=10% can be considered as a critical point which can be used to estimate the clinical fertility rate of IVF. But it could not provide relative information about the rates of high quality embryos and pregnancy for infertile couples undergoing IVF procedure.

Adult ; Aging ; genetics ; physiology ; DNA Fragmentation ; Embryo Implantation ; genetics ; Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Regression Analysis ; Spermatozoa ; metabolism ; physiology

Akabane, Aino and Chuzan virus are arthropod-borne (arbo)viruses mainly associated with reproductive failures in cattle. We investigated apoptosis in Vero cells (C-1586) infected with Akabane, Aino and Chuzan virus. The fragmentation of chromosomal DNA was simultaneously detected with the progress of cytopathic effect from 48 hr to 72 hr post infection, depending on viruses. Although the treatment of cycloheximide blocked apoptosis in Vero cells infected with three viruses, actinomycin D did not prevent DNA oligomerization, thus indicating that de novo viral protein synthesis is critical for viral apoptosis. In addition, the activation of caspase-3 was also detected in Vero cells by indirect fluorescent assay. From the present results, it is of future interest whether apoptotic characteristics of these viruses are related to pathogenecity in vivo.
Animals ; Apoptosis/*physiology ; Bunyaviridae/*physiology ; Caspase 3 ; Caspases/metabolism ; Cercopithecus aethiops ; Cytopathogenic Effect, Viral/*physiology ; DNA Fragmentation/physiology ; Dactinomycin ; Enzyme Activation ; Orbivirus/*physiology ; Vero Cells

Objective: To clarify the roles of yam polysaccharide (YPS) in improving sperm viability and protecting sperm DNA integrity in vitro and provide a new approach to the treatment of oligoasthenozoospermia. METHODS: We collected samples by masturbation from 36 normal fertile males aged 27－39 years. Each sample was divided into six groups: blank control or treated with normal saline, vitamin C solution, and YPS solution at low (0.25 mg/ml), medium (1.0 mg/ml) or high concentration (5.0 mg/ml). Using eosin-Y staining, sperm hypotonic swelling (HOS) and sperm chromatin diffusion (SCD) test, we observed the effects of different concentrations of YPS on sperm viability, membrane integrity and nuclear DNA. RESULTS: After 24 and 48 hours of treatment, sperm viability was markedly reduced in the vitamin C (［28.5 ± 3.1］ and ［6.5 ± 1.2］%), low-YPS (［31.3 ± 3.5］ and ［6.5 ± 2.2］%), medium-YPS (［37.1 ± 3.5］ and ［9.5 ± 2.8］%) and high-YPS groups (［38.3 ± 3.3］ and ［9.0 ± 3.2］%) as compared with the blank control (［17.3 ± 2.1］ and ［3.2 ± 1.3］%) (P <0.01) and normal saline groups (［13.4 ± 4.1］ and ［3.1 ± 2.0］%) (P <0.01), and it was significantly higher in the medium- and high-YPS than in the vitamin C group (P <0.05 and P <0.01). The rate of sperm DNA fragmentation was remarkably decreased at 48 hours in the vitamin C (［30.5 ± 3.1］%), low-YPS (［29.4 ± 2.6］%), medium-YPS (［28.5 ± 2.3］%) and high-YPS groups (［27.9 ± 1.9］%) in comparison with the blank control (［41.7 ± 2.2］%) (P <0.01) and normal saline groups (［42.1 ± 3.3］%), markedly lower in the medium- and high-YPS than in the blank control, normal saline and vitamin C groups (P <0.05 or P <0.01), but with no statistically significant difference between the low-YPS and vitamin C groups (P >0.05). CONCLUSIONS Yam polysaccharide can improve sperm viability and protect sperm DNA integrity in vitro.
Adult ; Ascorbic Acid ; pharmacology ; DNA ; drug effects ; DNA Fragmentation ; Dioscorea ; chemistry ; Humans ; Male ; Polysaccharides ; pharmacology ; Semen Analysis ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Vitamins ; pharmacology

AIMTo examine the relationship between sperm DNA damage and sperm nuclear histone (H2B) staining.

METHODSWe evaluated sperm samples from 14 consecutive asthenoteratozoospermic infertile men and six consecutive fertile controls. Sperm nuclear histone (H2B) staining and sperm chromatin integrity (assessed by sperm chromatin structure assay and expressed using the percentage of (i) DNA fragmentation index [% DFI] and (ii) high DNA stainability [% HDS)]) were evaluated.

RESULTSHistone H2B immunocytochemistry demonstrated two nuclear staining patterns: (i) focal punctate staining; and (ii) diffuse staining. Infertile men had a higher mean percentage of spermatozoa exhibiting diffuse H2B staining than did fertile men (7.7% +/- 4.6% vs. 1.6% +/- 1.2%, respectively, P < 0.01). We observed significant relationships between the proportion of spermatozoa with diffuse nuclear histone staining and both sperm % DFI (r = 0.63, P < 0.01) and sperm %HDS (r = 0.63, P < 0.01).

CONCLUSIONThe data demonstrate that infertile men have a higher proportion of spermatozoa with diffuse histone H2B than do fertile men and suggest that sperm DNA damage might, at least in part, be due to abnormally high histone H2B levels.

Adult ; Chromatin ; chemistry ; metabolism ; DNA ; biosynthesis ; DNA Fragmentation ; Histones ; metabolism ; Humans ; Immunohistochemistry ; Infertility, Male ; metabolism ; Male ; Nucleic Acid Denaturation ; Sperm Motility ; physiology ; Spermatozoa ; metabolism