Alkaline and neutral Comet assay profiles of sperm DNA damage in clinical groups

Ribas-Maynou, Jordi; García-Peiró, Agustín; Abad, Carlos; Amengual, M. J.; Navarro, J.; Benet, Jordi

doi:10.1093/humrep/der461

Cited by 91 publications

(117 citation statements)

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“…Since the OTM values at pH [ 13 were 5.1-15.3 times higher than those at pH 12.1, we deduce that compared with the DNA strand breaks, ALS was the main type of DNA damage produced by monocrotophos exposure. As the neutral comet assay was reported to be more specific for the detection of DSB (Shahidi et al 2007;Cortés-Gutiér-rez et al 2012;Ribas-Maynou et al 2012) and the OTM values at pH 12.1 were 4.9-31.5 times higher than those at pH 8.2-8.5, it was predominantly SSB with much fewer DSB detected by the pH 12.1 comet assay in this study. Consistently, Tice et al (2000) suggested that almost all genotoxic agents induce SSB orders of magnitude more than DSB.…”

Section: Discussionsupporting

confidence: 49%

Induction of DNA base damage and strand breaks in peripheral erythrocytes and the underlying mechanism in goldfish (Carassius auratus) exposed to monocrotophos

Zhao

Wei

Zhang

et al. 2015

Fish Physiol Biochem

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Using goldfish (Carassius auratus) as the model animal, the present study revealed the types of the DNA damage induced by monocrotophos, a highly toxic organophosphorus pesticide, and explored the mechanism underlying the DNA-damaging effect of this pesticide. Results of the alkaline comet assay showed that global DNA damage (including single- and double-strand breaks and alkali-labile sites) in peripheral erythrocytes of goldfish, measured as olive tail moment, was significantly increased by exposure to 0.01, 0.10, and 1.00 mg/L monocrotophos for 24, 48, 96, and 168 h. In particular, alkali-labile sites rather than single- or double-strand breaks, distinguished by the alkaline, pH 12.1, and neutral comet assays, were mainly induced by monocrotophos at 48 h. Oxidative damage in DNA bases and telomeric DNA was investigated by using the alkaline comet assay combined with endonuclease III or formamidopyrimidine DNA glycosylase and with fluorescence in situ hybridization, respectively. Further, glutathione peroxidase activity significantly decreased at 24 h but increased at 96 and 168 h, and malondialdehyde concentrations significantly increased at 48 h but gradually decreased at 96 and 168 h, which indicated an over-production of reactive oxygen species (ROS) at short exposure durations, but effective scavenging at long exposure durations in the peripheral blood tissues. Accordingly, our results suggest that DNA damage induced by monocrotophos in fish blood cells is possibly due to the inhibition of ROS scavenging and resulted accumulation of ROS.

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Section: Discussionsupporting

confidence: 49%

Induction of DNA base damage and strand breaks in peripheral erythrocytes and the underlying mechanism in goldfish (Carassius auratus) exposed to monocrotophos

Zhao

Wei

Zhang

et al. 2015

Fish Physiol Biochem

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“…% Tail DNA represented a threshold value of 15 % for predicting male infertility with an area under the curve 0,837 and Tail Moment represented a threshold value of 15,21 % with an area under the curve 0,796. However, these results are not comparable with previous studies because it is not mentioned which comet measurement were used in these studies [15,18,31]. Higher area under the ROC curve has been shown by % Tail DNA with a high sensitivity and specificity of 0,717 and 0,800 respectively (Table 1, Fig.…”

Section: Discussioncontrasting

confidence: 54%

Fluctuation of “sperm DNA integrity” in accordance with semen parameters, and it’s relationship with infertility

Durmaz

Dikmen

Gündüz

et al. 2014

J Assist Reprod Genet

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Purpose Nowadays sperm analysis is used routinely to assess infertile men, but it's predictive value is still restricted. For this reason, new markers are needed for diagnosis and guide treatment. A DNA damage evaluation method, which is easily applicable for routine analysis is described in this paper to provide an important support in clinical evaluation. Methods In this study, DNA damage evaluation was planned in infertile males with the COMET assay, a method which is simple, cheap, sensitive and reliable. For this purpose, the sperm DNA damage of normospermic, oligospermic, asthenospermic and teratospermic cases were investigated and the relationship between sperm parameters (sperm count, motility, morphology) and DNA damage was assessed. Results and conclusion According to our results we suggest that the factors to blame for sperm DNA damage and semen quality are the same. Also it was found that the infertile males possess substantially more sperm DNA damage than fertile men do. Improving and using the DNA damage evaluation tests seem to improve the results in ART clinics.

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“…According to the same authors, DNA damage in those spermatozoa derives mainly from nuclease activity (35). Of interest, we recently demonstrated that the percentage of dimmer spermatozoa correlates with those of activated caspases and semen apoptotic bodies, also indicating that their damage is due to apoptotic nucleases (26).…”

Section: Figurementioning

confidence: 78%

DNA fragmentation in brighter sperm predicts male fertility independently from age and semen parameters

Muratori

Marchiani

Tamburrino

et al. 2015

Fertility and Sterility

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Objective: To evaluate whether sperm DNA fragmentation (sDF), measured in brighter, dimmer, and total populations, predicts natural conception, and to evaluate the intra-individual variability of sDF. Design: Prospective study. Setting: Outpatient clinic and diagnostic laboratory. Patient(s): A total of 348 unselected patients and 86 proven fertile men. Intervention(s): None. Main Outcome Measure(s): sDF was revealed with the use of terminal deoxynucleotide transferase-mediated dUTP nick-end labeling (TUNEL)/propidium iodide (PI). Receiver operating characteristic (ROC) curves were built before and after matching fertile men to patients for age (76:152) or semen parameters (68:136) or both (49:98). Intra-individual variability of sDF was assessed over 2 years. Result(s): Brighter (area under ROC curve [AUC] 0.718 AE 0.54), dimmer (AUC 0.655 AE 0.63), and total (AUC 0.757 AE 0.54) sDF predict male fertility in unmatched and age-or semen parameters-matched subjects. After matching for both age and semen parameters, only brighter (AUC 0.711 AE 0.83) and total (AUC 0.675 AE 0.92) sDF predict male fertility. At high values of total sDF, brighter predicts natural conception better than total sDF. Intra-individual coefficients of variation of sDF were 9.2 AE 8.6% (n ¼ 25), 12.9 AE 12.7% (n ¼ 53), and 14.0 AE 12.6% (n ¼ 70) over, respectively, 100-day and 1-and 2-year periods, appearing to be the most stable of the evaluated semen parameters. Conclusion(s):The predictive power of total sDF partially depends on age and semen parameters, whereas brighter sDF independently predicts natural conception. Therefore, brighter sDF is a fraction of sDF that adds new information to the routine semen analysis. At high levels of sDF, distinguishing the two sperm populations improves the predictive power of sDF. Overall, our results support the idea that TUNEL/PI can be of clinical usefulness in the male fertility workup. (Fertil Steril Ò 2015;104:582-90. Ó2015 by American Society for Reproductive Medicine.)

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Alkaline and neutral Comet assay profiles of sperm DNA damage in clinical groups

Abstract: Comet assay profiles, applied to different clinical groups, may be useful for determining prognosis in cases of male infertility.

Cited by 91 publications

References 1 publication

Induction of DNA base damage and strand breaks in peripheral erythrocytes and the underlying mechanism in goldfish (Carassius auratus) exposed to monocrotophos

Induction of DNA base damage and strand breaks in peripheral erythrocytes and the underlying mechanism in goldfish (Carassius auratus) exposed to monocrotophos

Fluctuation of “sperm DNA integrity” in accordance with semen parameters, and it’s relationship with infertility

DNA fragmentation in brighter sperm predicts male fertility independently from age and semen parameters

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