Sperm chromatin dispersion test before sperm preparation is predictive of clinical pregnancy in cases of unexplained infertility treated with intrauterine insemination and induction with clomiphene citrate

Background/aims: A large proportion of men with normal sperm results as analyzed using conventional techniques have fragmented DNA in their spermatozoa. We performed a prospective study to examine the incidence of DNA fragmentation in sperm in cases of couples with previously unexplained infertility and treated with intrauterine insemination. We evaluated whether there was any predictive value of DNA fragmentation for pregnancy outcome in such couples. Methods: The percentage of DNA fragmentation and all classical variables to evaluate sperm before and after sperm treatment were determined. We studied the probable association between these results and pregnancy outcome in terms of clinical and ongoing pregnancy rate per started first cycle. We also assessed the optimal threshold level to diagnose DNA fragmentation in our center. Results: When using threshold levels of 20, 25, and 30%, the occurrence of DNA fragmentation was 42.9, 33.3, and 28.6%, respectively. Receiver operating characteristic (ROC) analysis of all cases revealed an area under the curve of 80% to predict the clinical pregnancy rate per cycle from testing the sperm motility (a + b) before treatment. We failed to generate an ROC curve to estimate pregnancy outcome from the amount of DNA fragmentation before treatment. However, when selecting only those men with a pretreatment DNA fragmentation of at least 20%, the pretreatment result was statistically different between couples who achieved a clinical pregnancy and those who did not. Conclusion: DNA fragmentation is often diagnosed in couples with unexplained infertility. Each center should evaluate the type of test it uses to detect DNA fragmentation in sperm and determine its own threshold values

High-throughput on-chip DNA fragmentation

free microfluidic deoxyribonucleic acid (DNA) fragmentation chip that is based on hydrodynamic shearing. Genomic DNA has been reproducibly fragmented with 2-10 kbp fragment lengths by applying hydraulic pressure ΔP across micromachined constrictions in the microfluidic channels. The utilization of a series of constrictions reduces the variance of the fragmented DNA length distribution; and parallel microfluidic channels design eliminates the device clogging

Effects of chronic buproprion and nicotine administration on cell genesis and DNA fragmentation in adult rat dentate gyrus

Previous experiments have shown that chronic subcutaneous administration of nicotine dose-dependently inhibits the acquisition and retention of a spatial task in the Morris water maze and reduces cell genesis in the dentate gyrus (DG) of adult rats.1 In the present study, the effects of nicotine and buproprion, an atypical antidepressant used in smoking cessation, on dentate gyrus cell genesis and DNA fragmentation were investigated. The results show that nicotine, chronically infused for 21 days, suppressed cell genesis and enhanced DNA fragmentation in the DG, an effect not attenuated by co-administration of buproprion.peer-reviewe

Apoptosis in human unfertilized oocytes after Intracytoplasmic Sperm Injection

Objective To investigate the presence of programmed cell death in unfertilized oocytes after intracytoplasmic sperm injection (ICSI), assuming that previous apoptotic events could be correlated with the fertilization failure. Design Comparison of the rate of DNA fragmentation in human oocytes at different stages of maturation soon after pick-up (control) and in unfertilized oocytes after ICSI treatment. Setting In vitro fertilization (IVF) laboratory with extensive ICSI experience. Patient(s) Sixty-three patients undergoing assisted fertilization by ICSI. Intervention(s) Terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) assay and anticaspase-3 cleaved immunoassay to detect apoptosis in control and ICSI-treated oocytes. Main Outcome Measure(s) Differences in the percentage of oocytes demonstrating DNA fragmentation between control oocytes and unfertilized ICSI treated oocytes at different stages of maturation. Result(s) The DNA fragmentation, by TUNEL assay, appeared in all the immature control oocytes, but only 37% of mature oocytes showed DNA fragmentation. This DNA fragmentation was observed in 88.8% of the oocytes unfertilized after ICSI; furthermore, DNA fragmentation appeared as well in the sperm injected into the cytoplasm. Conclusion(s) The study has shown DNA fragmentation in human oocytes unfertilized after ICSI. The evidence is confirmed as well in control oocytes, free from in vitro culture or manipulation stress. Caspase-3 immunoassay suggests the presence of apoptosis. The high percentage of oocytes demonstrating DNA fragmentation in the unfertilized oocytes could be correlated with fertilization failure

Correlation of Paternal Homocysteine Level and Sperm DNA Fragmentation in Couple with Idiopathic Recurrent Early Pregnancy Loss

Background: Recurrent pregnancy loss is the traumatic event for couples in the effort to obtain offsprings. Various female etiologies have been extensively studied, but more than half of remain unknown. As a result of unification of sperm and oocyte, an embryo combined with any defect during spermatogenesis will also affect the quality of embryo consequently determining the pregnancy outcomes. Routine semen analysis failed to support the evidence of influence of defective sperm in recurrent pregnancy loss. Currently, examination of sperm DNA fragmentation has been added to evaluate the quality of sperm beside the routine semen analysis. We hypothesized that high sperm DNA fragmentation plays a role in the incidence of idiopathic recurrent early pregnancy loss. Beside that, the cause of sperm DNA fragmentation are numerous and remain controversial. This study was conducted to determine the impact of paternal hyperhomocysteinemia on high sperm DNA fragmentation and incidence rates of idiophatic recurrent early pregnancy loss. Material and methods: Fourty partners of idiopathic recurrent early pregnancy loss caes and 40 cases of control from normal male fertile population were included in this study. Blood and semen samples were collected for routine semen analysis, sperm DNA fragmentation, serum and seminal homocysteine. The results were then analyzed to determine the association between sperm and DNA fragmentation, serum homocysteine, seminal homocysteine and incidence rates of idiopathic recurrent early pregnancy loss. Results: incidence rates of idiopathic recurrent early pregnancy loss was significantly associated with sperm DNA fragmentation (p<0.05) and serum homocysteine (p<0.05). Idiopathic recurrent pregnancy loss partners had significantly higher DFI (p<0.05). Hyperhomocysteinemia appears to be associated with significantly increase of sperm DNA fragmentation (p<0.05). Conclusion: Paternal serum homcysteine and high sperm DNA fragmentation was significantly correlated with the incidence rates of idiopathic recurrent early pregnancy loss. Key words : hyperhomocysteinemia, DFI, idiopathic recurrent early pregnancy los

Separate metabolic pathways leading to DNA fragmentation and apoptotic nuclear chromatin condensation

Apoptosis is the predominant form of cell death observed in a variety of physiological and pathological conditions such as cancer involution, insect metamorphosis, the development of the immune and nervous systems, and embryogenesis. The typical nuclear changes taking place in apoptotic cells include extensive condensation of chromatin and internucleosomal DNA fragmentation into units of 200 base pairs. However, the mechanisms responsible for both chromatin condensation and DNA fragmentation have yet to be elucidated. In this study, micrococcal nuclease and the divalent cations, Ca2+ and Mg2+, were applied to isolated nuclei in an attempt to reconstitute in vitro the digestion of genomic DNA associated with apoptosis. Micrococcal nuclease was found to induce a typical pattern of DNA fragmentation, but did not give rise to chromatin condensation, whereas Ca2+/Mg2+ induced both chromatin condensation and DNA fragmentation in isolated mouse liver nuclei. When the endonuclease inhibitor ZnCl2 was used, the DNA fragmentation induced by Ca2+/Mg2+ in nuclei could be completely inhibited, but chromatin condensation still occurred. For comparison, intact liver cells were treated with valinomycin, a potassium ionophore, which gave rise to an atypical cell death, with chromatin condensation appearing without DNA fragmentation. Our results suggest that endonuclease activation in apoptosis is neither necessary nor sufficient to induce chromatin condensation, and that DNA fragmentation and chromatin condensation may be triggered through separate pathways during apoptosis

FRAGMENTASI DNA SPERMATOZOA PADA PEKERJA BENGKEL PENGECATAN MOBIL DI MALANG

Objective: The aim of this study was to prove that exposure to benzene, toluene, and xylene in automobile painters are risk factors for the increase in sperm DNA fragmentation. Method: This research was observational descriptive cross-sectional study design. Research conducted at the Laboratory of Biochemistry-Biomolecular, Faculty of Medicine, Brawijaya University. The research sample are the semen of 15 married automobile painters and 12 control workers. The sample group is distinguished through air samples which were analyzed to detect benzene, toluene, and xylene and these chemicals was detected in automobile painters’ working environment. All of the ejaculate were tested by TUNEL Assay in combination with Propidium Iodide (PI) and analyzed by flowcytometry. TUNEL assay results are then compared statistically to assess the difference, and analyzed the risk of an increase in sperm DNA fragmentation in automobile painters. Result: The results showed the control workers have an average of 14.9% of sperm DNA fragmentation and automobile painters have an average value of 47.66% of sperm DNA fragmentation. Comparison test showed a statistically significant difference, p = 0.000 (p <0.05, CI 95%). For risk factors, an analysis with chi-square using the two groups of workers and two groups of TUNEL assay results which is categorized into high risk and low risk of DNA fragmentation with the cut-off of 30%. Statistical analysis showed an increased risk of sperm DNA fragmentation in automobile painters is significant, p = 0.001 (p <0.05, CI 95%). Conclusion: It is concluded that there are differences in sperm DNA fragmentation in automobile painters compared to control workers and there is a risk of an increase in sperm DNA fragmentation in automobile painters. Keywords: fragmentation, DNA, spermatozoa, TUNEL, propidium iodide, flowcytometr

Lower sperm DNA fragmentation after r-FSH administration in functional hypogonadotropic hypogonadism

Abstract PURPOSE: An observational clinical and molecular study was designed to evaluate the effects of the administration of recombinant human FSH on sperm DNA fragmentation in men with a non-classical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. METHODS: In the study were included 53 men with a non-classical form of hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia. In all patients, sperm DNA fragmentation index (DFI), assessed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) in situ DNA nick end-labelling (TUNEL) assay, was evaluated before starting the treatment with 150 IU of recombinant human FSH, given three times a week for at least 3 months. Patients' semen analysis and DNA fragmentation index were re-evaluated after the 3-month treatment period. RESULTS: After recombinant human FSH therapy, we did not find any differences in terms of sperm count, motility and morphology. The average DNA fragmentation index was significantly reduced (21.15 vs 15.2, p 15 %), while no significant variation occurred in the patients with DFI values ≤15 %. CONCLUSIONS: Recombinant human FSH administration improves sperm DNA integrity in hypogonadotropic hypogonadism and idiopathic oligoasthenoteratozoospermia men with DNA fragmentation index value >15 %

Sperm DNA fragmentation, recurrent implantation failure and recurrent miscarriage

Evidence is increasing that the integrity of sperm DNA may also be related to implantation failure and recurrent miscarriage (RM). To investigate this, the sperm DNA fragmentation in partners of 35 women with recurrent implantation failure (RIF) following in vitro fertilization, 16 women diagnosed with RM and seven recent fathers (control) were examined. Sperm were examined pre- and post-density centrifugation by the sperm chromatin dispersion (SCD) test and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. There were no significant differences in the age of either partner or sperm concentration, motility or morphology between three groups. Moreover, there were no obvious differences in sperm DNA fragmentation measured by either test. However, whilst on average sperm DNA fragmentation in all groups was statistically lower in prepared sperm when measured by the SCD test, this was not seen with the results from the TUNEL assay. These results do not support the hypothesis that sperm DNA fragmentation is an important cause of RIF or RM, or that sperm DNA integrity testing has value in such patients. It also highlights significant differences between test methodologies and sperm preparation methods in interpreting the data from sperm DNA fragmentation tests