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Study of Low-intensity 2450-MHz Microwave Exposure Enhancing the Genotoxic Effects of Mitomycin C Using Micronucleus Test and comet assay in vitro

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Biomedical and Environmental Sciences 2002年第4期15卷 283-290页

作者： ZHANGMEI－BIAN HEJI－LIANG 等 InstituteofOccupationalandEnivroinmentalHealth MeidcalCoollegeofZhejiangUniveristyHangzhou310031ZhejiangChina InstituteofOccupationalandEnivroinmentalHealth MeidcalCoollegeofZhejiangUniveristyHangzh

Objective To determine the interaction between 2450-MHz microwaves (MW) radiation and mitomycin C (MMC). Methods The synergistic genotoxic effects of low-intensity 2450-MHz microwave and MMC on human lymphocytes were studied using single cell gel electrophoresis (SCGE) assay (comet assay) and cytokinesis-blocked micronucleus (CBMN) test in vitro. The whole blood cells from a male donor and a female donor were either only exposed to 2450-MHz microwaves (5.0 mW/cm2) for 2 h or only exposed to MMC (0.0125 μ/mL, 0.025 μg/mL, 0.05μg/mL and 0.1 μg/mL) for 24 h; and the samples were exposed to MMC for 24 h after exposure to MW for 2 h. Results In the comet assay, the comet lengths ( 29.1 μm and 25.9 μm) of MW were not significantly longer than those (26.3 μrn and 24.1 μm) of controls (P>0.05). The comet lengths (57.4 μm, 68.9 μm, 91.4 μm, 150.6μm and 50.6 μm, 71.7μm, 100.1 μm, 145.1 μm) of 4 MMC groups were significantly longer than those of controls (Pcomet lengths (59.1 μm, 92.3 μm, 124.5 μm, 182.7 μm and 57.4 μm, 85.5 μm, 137.5 μm, 178.3 μm) of 4 MW plus MMC groups were significantly longer than those of controls too (Pcomet lengths of MW plus MMC groups were significantly longer than those of the corresponding MMC doses (P0.05). The MNC rates of 4 MMC groups were 8‰, 9‰, 14‰, 23‰ and 8‰, 8‰, 16‰, 30‰ respectively. When the doses of MMC were 3≥0.05 μg/mL, MNC rates of MMC were higher than those of controls (P<0.05). MNC rates of 4 MW plus MMC groups were 12‰, 13‰, 20‰, 32‰ and 8‰, 9‰, 23‰, 40‰. When the doses of MMC were 5≥0.05 μg/mL, MNC rates of MW plus MMC groups were much higher than those of controls (P<0.01). MNC rates of 4 MW plus MMC groups were not significantly higher than those of the corresponding MMC doses. Conclusion The low-intensi

关键词： Microwaves Mitomycin C comet assay Micornucleus test Synergistic effects

Genotoxicity of Three Avermectins on Polypedates megacephalus Tadpoles Using the comet assay

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Asian Herpetological Research 2016年第4期7卷 251-257页

作者： Baorong GENG Lingling ZHANG Yun JIANG Xiuping HUANG Jinmei DAI College of Life Sciences Fujian Normal University

Avermectins are a new class of macrocyclic lactones derived from mycelia of the soil actinomycete, and are used as effective agricultural pesticides and antiparasitic agents. However, run-off from crops treated with avermectins may contaminate various bodies of water, and accumulated to certain concentrations to impact the development of aquatic animals. Here, we tested the genotoxicity of three avermectins （abamectin, ABM; ivermectin, IVM; and emamectin benzoate, EMB） on Polypedates megacephalus tadpoles by the alkaline single-cell gel electrophoresis assay. Tadpoles were treated for 48 h in the laboratory with different concentrations of these three agents, 0.006, 0.012, 0.018, 0.024, 0.030 mg/L for ABM, 0.003, 0.006, 0.009, 0.012, 0.015 mg/L for IVM and 0.04, 0.06, 0.08, 0.10, 0.12 mg/L for EMB, and then measured their DNA damage by the comet assay tail factor %. The concentrations of resulted in highly significant increases in DNA damage of the tadpoles were found above the concentration threshold of 0.012 mg/ L ABM, 0.003 mg/L IVM and 0.06 mg/L EMB and linear correlations between the intensity of DNA damage and the concentrations of these three avermectins. Our results showed clearly that avermectins caused dose dependent DNA damage on amphibian tadpoles, and there might be a control on the misuse of avermectins.

关键词： Polypedates megacephalus tadpole avermectins abamectin ivermectin emamectin benzoate DNAdamage comet assay

Shortening of alkaline DNA unwinding time does not interfere with detecting DNA damage to mouse and human spermatozoa in the comet assay

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Asian Journal of Andrology 2011年第1期13卷 172-174页

作者： Hirokazu Kusakabe Hiroyuki Tateno Department of Biological Sciences Asahikawa Medical University Asahikawa 078-8510 Japan

The comet assay was performed on mouse and human spermatozoa to examine the effect of alkaline DNA unwinding time. The spermatozoa were treated in vitrowith the DNA-damaging agents, methyl methanesulfonate （MMS） or hydrogen peroxide （Hz02）, and then embedded in agarose gel on glass sl ides. The slides were immersed in alkaline solution （〉pH 13） for 1, 5, 10 and 20 min, and then subjected to the electrophoresis under neutral conditions. In mouse spermatozoa, comet tails seen in solvent controls became brighter and longer as the alkaline DNA unwinding time increased. However, in the MMS-treated mouse spermatozoa, a smaller difference in the damage from that in the solvent control was seen with time within a dose. DNA damage induced by H2O2 could also be detected accurately after alkali treatment for 1-20 min. In human spermatozoa, DNA damage induced by MMS and H2O2 could be detected in a dose-dependent manner after alkali treatment for 1 min. The ability of the comet assay to detect DNA damage was not adversely affected by the short period （1 min） of the alkaline DNA unwinding time.

关键词： alkaline comet assay DNA unwinding human mice spermatozoa

comet assay and cytokinesis-blocked micronucleus test for monitoring the genotoxic effects of X-ray radiation in humans

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Chinese Medical Journal 2000年第10期113卷 47-50页

作者：何继亮陈玮琳金力奋金海燕 Division of Environmental Health School of MedicineZhejiang University

Obejctive To assess the genotoxic effects of X ray radiation on human populations Methods The single cell gel electrophoresis (SCGE) and cytokinesis blocked micronucleus (CBMN) test were applied as biological dosimeters to detect DNA damage and abnormalities in human peripheral lymphocytes of subpopulation exposed to X ray radiation The subjects were divided into four groups: 12 radiation patients; 13 intervention radiation therapy doctors; 32 radiation diagnostians; 28 controls Results The average comet lengths of the four groups were 128 17±4 49?μm, 88 09±5 39?μm, 72 68±2 57?μm and 32 87±0 57?μm, respectively The difference in average comet length between any two groups was highly significant ( P 0 05) Conclusions This study showed that both the comet assay and the CBMN test could be used to monitor populations exposed to X ray radiation, but the comet assay seems to be more sensitive than the CBMN test

关键词： X-ray radiation DNA damage single cell gel electrophoresis assay cytokinesis-blocked micronucleus test comet assay

Forensic application of comet assay:an emerging technique

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Forensic Sciences Research 2017年第4期2卷 180-184页

作者： Ritesh Kumar Shukla Division of Biological and Life Sciences School of Arts and SciencesAhmedabad UniversityAhmedabadIndia

Postmortem interval(PMI)estimation is a recurring problem in the field of forensic *** methods are effective but are insufficient to estimate accurate and precise time of death or *** addition,degradation of biological samples is another major problem in forensic science which affects the investigation process and misleads the *** previous studies reported that DNA fragmentation has strong correlation with *** fragmentation increased with prolonged *** assay is a rapid sensitive,versatile,reliable and cost effective technique that is specifically used for qualitative and quantitative estimation of nuclear DNA *** to this attribute,comet assay can help to estimate accurate and precise time of death for some extent that is for early PMI *** addition,two confounding factors are responsible for DNA fragmentation:(1)micro-organism;(2)environmental ***,comet assay plays a dual role:(1)partially degraded samples get repaired using repair enzyme;(2)accurate time since deposition can be measured without using repair ***,this assay can also help to identify potential exposures of environmental-released chemicals/toxicants and its deleterious effects on human *** this way,comet assay shows its versatile applications that could be useful for forensic ***,with the help of this review,an attempt was made to explore the versatility of comet assay technique for forensic applications and its future perspective.

关键词： Forensic science comet assay PMI single-cell gel electrophoresis DNA fragmentation DNA repair forensic application

The genotoxic effects of benzo[a]pyrene and methamidophos on black porgy evaluated by comet assay

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Chinese Journal of Oceanology and Limnology 2005年第4期23卷 455-460页

作者：刘日先洪华生王新红王克坚王春光 State Key Laboratory for Marine Environmental Science Environmental Science Research Center Xiamen UniversityFujian 361005China

In this study, two common pollutants (benzo[a]pyrene and methamidophos) in marine environment were tested by comet assay for their inducement of in vivo genotoxic effect to the blood cells of black porgy ( Acanthopagrus schlegeli). The fish was exposed to 2 μg/L of benzo[a]pyrene (BaP) and methamidophos, and their mixture. The assay was performed on whole blood at 2 h, 5 h, 24 h and 96 h exposure intervals. A signifi- cant increase in DNA damage was observed in each treatment with the pollutants. Additive effect of BaP and methamidophos was also found in the experiment. However, the decrease ratios of DNA damage for 5 h and 96 h exposure interals compared with 2 h and 24 h exposure ones, respectively, were noticed. This phenomenon may be explained by the function of repairing process via enzyme cytochrome P450 in the animal. Evidence of the genotoxicity of organophosphorus pesticides (OPs) and polynuclear aromatic hydrocarbons (PAHs) on marine fish are discussed in this paper.

关键词： black porgy genotoxic effects benzo[a]pyrene methamidophos DNA damage comet assay

Assessment of Human DNA Repair (NER) Capacity With DNA Repair Rate (DRR) by comet assay

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Biomedical and Environmental Sciences 2005年第2期18卷 117-123页

作者： WEIZHENG JI-LIANGHE LI-PENJIN JIAN-LINLOU BAO-HONGWANG InstituteofOccupationalandEnvironmentalHealth MedicalCollegeofZhejiangUniversityHangzhou310006ZhejiangChina InstiuteofOccupationalandEnvironmentalHealth MedicalCollegeofZhejiangUniversityHangzhou310006ZhejiangChina／／MedicalCollegeofJiaxingUniversityJiaxing314001ZhejiangChina

Objective Alkaline comet assay was used to evaluate DNA repair (nucleotide excision repair, NER) capacity of human fresh lymphocytes from 12 young healthy non-smokers (6 males and 6 females). Methods Lymphocytes were exposed to UV-C (254 nm) at the dose rate of 1.5 J/m2/sec. Novobiocin (NOV) and aphidicolin (APC), DNA repair inhibitors, were utilized to imitate the deficiency of DNA repair capacity at the incision and ligation steps of NER. Lymphocytes from each donor were divided into three grougs: UVC group, UVC plus NOV group, and UVC plus APC group. DNA single strand breaks were detected in UVC irradiated cells incubated for 0, 30, 60, 90, 120, 180, and 240 min after UVC irradiation. DNA repair rate (DRR) served as an indicator of DNA repair capacity. Results The results indicated that the maximum DNA damage (i.e. maximum tail length) in the UVC group mainly appeared at 90 min. The ranges of DRRs in the UVC group were 62.84%-98.71%. Average DRR value was 81.84%. The DRR difference between males and females was not significant (Pcomet assay is a rapid, simple and sensitive screening test to assess individual DNA repair (NER) capacity. It is suggested that the time to detect DNA single strand breaks in comet assay should include 0 (before UV irradiation), 90 and 240 min after exposure to 1.5 J·m-2 UVC at least. The DRR, as an indicator, can represent the individual DNA repair capacity in comet assay.

关键词： DNA repair capacity comet assay UVC Novobiocin Aphidicolin

DNA Damage （comet assay） as Biomarker of Cd Exposure in Marine Seed Scallops Mizuhopecten Yessoensis Age 1 Year

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Journal of Environmental Science and Engineering 2010年第10期4卷 63-69页

作者： V.V. Slobodskova E.E. Solodova V.P. Chelomin V. I.Il＇ichev Pacific Oceanological Institute of the Far Eastern Branch of Russian Academy of Sciences Vladivostok 690041 Russia

Cadmium-induced DNA degradation in gill cells of the scallop Mizuhopectenyessoensis was assessed using the comet assay （single-cell gel electrophoresis）. Accumulation of highly toxic cadmium in the gill cells of bivalve is accompanied by the damage of the cell genome revealed as DNA migration in the comet assay. The main mechanisms of Cd effects on the integrity of the DNA structure are discussed.

关键词： Accumulation cadmium bivalves Mizuhopecten yessoensis genotoxicity comet assay DNA damage

DNA damage levels in systemic lupus erythematosus patients with low disease activity:An evaluation by comet assay

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Advances in Bioscience and Biotechnology 2012年第7期3卷 983-988页

作者： Thais Muniz Montalvao Ana Luisa Miranda-Vilela Mariana Matos Roll Cesar Koppe Grisolia Leopoldo Santos-Neto University Hospital of Brasilia University of BrasiliaBrasiliaBrazil Laboratory of Genetic Toxicology Institute of Biological SciencesUniversity of BrasiliaBrasiliaBrazil Department of Genetics and Morphology Laboratory of GeneticsInstitute of Biological SciencesUniversity of BrasiliaBrasiliaBrazil Faculty of Medicine University of BrasiliaBrasiliaBrazil

Oxidative stress has been implicated in the inflammatory process of Systemic Lupus Erythematosus (SLE), particularly by the formation of anti-DNA autoantibodies, which can lead to DNA damage. The aim of this study was to investigate, through comet assay, whether the level of DNA damage in SLE patients is different from that of healthy subjects. Twenty-five adult SLE patients with SLEDAI up to ten, and 25 healthy subjects were paired according to age, gender and Body Mass Index (BMI). Other anthropometric variables were also assessed. comet assay was assessed as the marker of oxidative stress described as DNA Damage (DD) percentage. Waist Circumference (WC), Hip Circumference (HC) and BMI were also performed. Exclusion criteria for patients and controls comprised smoking and other chronic disorders. Level of damage index was remarkably higher in SLE patients than in controls, and no significant differences between the groups were found for age, BMI, WC and HC. No stratification concerning gender was performed, since there were just two males per group. No correlation was observed between BMI and DD (%). DD increased in SLE, which reflects the oxidant/antioxidant imbalance in these patients. These findings support an association between oxidative stress and SLE. This stronger correlation observed in patients with low disease activity may be useful in elucidating the mechanisms of disease pathogenesis.

关键词： Systemic Lupus Erythematosus DNA Damage comet assay Oxidative Stress