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- W2188374992 abstract "Summary Introduction: Infertility affects approximately 15% of all couples trying to conceive, male factor is the sole or contributing factor in roughly half of cases infertility. Men with normal semen analysis may also have a high degree of DNA fragmentation, which can be a major cause of undiagnosed/unexplained infertility. In this work, we estimate the level of damage DNA, using a modified protocol of the Sperm Chromatin Dispersion (SCD) test. Materials and Methods: We performed a DNA controlled denaturation to generate restricted single-stranded DNA (ssDNA) motifs from DNA breaks. Later, nuclear proteins were removed to form nucleoids with a central core and a peripheral halo of dispersed DNA loops. Such DNA halos are absent or extremely small in nucleoids with nonfragmented DNA. To demonstrate the sensitivity of the SCD test, aliquots of sperm samples, from two different subjects were exposed to increasing concentrations of the nitric oxide donor sodium nitroprusside (SNP), for 1 hour at room temperature, to induce DNA damage. To estimate interobserver variability in sperm cells scoring, three technicians analyzed 500 sperm cells from each sample, three times a day. Means of DNA fragmentation level obtained by the observers of each sample were compared. Results and Conclusions: The modified protocol of SCD test was sensible to detect DNA fragmentation from 60 µM of SNP. Analysed samples showed a dose-response effect by SNP. There wasn't significant differences (Pearson Test χ 2 , P>0.05) in the mean of sperm cells with fragmented DNA observed, among the observers. SCD is a simple, fast, cheap and reproducible technique does not require complex or expensive equipments. Sperm DNA damage can be determined with accuracy using this assay, in laboratories with a basic instrumentation. The analysis can be carried out with conventional bright-field microscopes." @default.
- W2188374992 created "2016-06-24" @default.
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- W2188374992 date "2007-01-01" @default.
- W2188374992 modified "2023-09-24" @default.
- W2188374992 title "DNA DAMAGE MEASUREMENT USING A MODIFIED PROTOCOL OF SPERM CHROMATIN DISPERSION TEST" @default.
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