• Title/Summary/Keyword: Comet

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DNA Strand Breaks in Mitotic Germ Cells of Caenorhabditis elegans Evaluated by Comet Assay

  • Park, Sojin;Choi, Seoyun;Ahn, Byungchan
    • Molecules and Cells
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    • v.39 no.3
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    • pp.204-210
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    • 2016
  • DNA damage responses are important for the maintenance of genome stability and the survival of organisms. Such responses are activated in the presence of DNA damage and lead to cell cycle arrest, apoptosis, and DNA repair. In Caenorhabditis elegans, double-strand breaks induced by DNA damaging agents have been detected indirectly by antibodies against DSB recognizing proteins. In this study we used a comet assay to detect DNA strand breaks and to measure the elimination of DNA strand breaks in mitotic germline nuclei of C. elegans. We found that C. elegans brc-1 mutants were more sensitive to ionizing radiation and camptothecin than the N2 wild-type strain and repaired DNA strand breaks less efficiently than N2. This study is the first demonstration of direct measurement of DNA strand breaks in mitotic germline nuclei of C. elegans. This newly developed assay can be applied to detect DNA strand breaks in different C. elegans mutants that are sensitive to DNA damaging agents.

Genetic Toxicity Test of 1,2-Dibromoethane by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Kim, Ki-Y.;Kim, Ji-H.;Kwon, Kyoung-J.;Go, Seo-Y.;Min, Kyung-N.;Lee, Woo-S.;Park, Sue-N.;Shee, Yhun-Y.
    • Biomolecules & Therapeutics
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    • v.14 no.4
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    • pp.246-252
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    • 2006
  • 1,2-Dibromoethane(DBE) has been widely used as a soil fumigant, an additive to leaded gasoline and an industrial solvent. In this study, we have carried out in vitro genetic toxicity test of 1,2-dibromoethane and microarray analysis of differentially expressed genes in response to 1,2-dibromoethane. 1,2-Dibromoethane showed mutations in base substitution strain TA1535 both with and without exogenous metabolic activation. 1,2-Dibromoethane showed mutations in frame shift TA98 both with and without exogenous metabolic activation. 1,2-Dibromoethane showed DNA damage based on single cell gel/comet assay in L5178Y cells both with and without exogenous metabolic activation. 1,2-Dibromoethane increased micronuclei in CRO cells both with and without exogenous metabolic activation. Microarray analysis of gene expression profiles in L5178Y cells in response to 1,2-dibromoethane selected differentially expressed 241 genes that would be candidate biomarkers of genetic toxic action of 1,2-dibromoethane.

Assessment of DNA Damage using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and Toxic Effects in Chickens by T-2 Toxin Treatment (T-2 toxin을 투여한 닭에서 Comet assay 방법을 이용한 DNA 손상 평가와 독성)

  • Hah Dae-Sik;Heo Jung-Ho;Lee Kuk-Cheon;Cho Myung-Heui;Kim Kuk-Hun;Kim Chung-Hui;Lue Jae-Du;Lee Seung-Hwan;Kim Gon-Sup;Kim Eui-Gyung;Kim Jong-Shu
    • Toxicological Research
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    • v.22 no.2
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    • pp.75-85
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    • 2006
  • This study was designed to evaluate the possible DNA damaging effects of T-2 toxin using an alkaline single cell gel electrophoresis (SCGE) comet assay and also to investigate toxic effects in chickens. A total of 20 chickens were used in these experiments. Graded concentrations of dietary T-2 toxin (0, 4, 8, and $16{\mu}g/g$ of diet) were given to groups of 5 broiler chickens. In comet assay, The DNA damage was analysed by the tail extent moment (TEM) and tail length (TL), which were used as markers of DNA strand breaks in SCGE. A significant dose-dependent increase in the extent of DNA migration as well as in the percentage of cells with tails was observed after treatment with T-2 toxin (P<0.05). Treatment with the low T-2 toxin ($4{\mu}/g$ of diet) induced a relatively low level of DNA damage in comparison with the high T-2 toxin ($16{\mu}/g$ of diet) group. The growth rate was significantly reduced by concentrations of 8, and $16{\mu}/g$ of diet (P < 0.05). The feed conversion ratio were significantly affected by any concentrations (P < 0.05). The relative weight of the spleen, and lung was decreased by the growth inhibitory concentrations. The bursa of Fabricius, thymus, and kid- ney were decreased in relative weight by concentrations of $16{\mu}/g$ of diet. The relative weight of the liver and heart were unaffected. The hemoglobin (Hb), hematocrit (HCT), and mean corpuscular hemoglobin (MCH) were decreased at concentration of $16{\mu}/g$ of diet. As compared with control chickens, there was no marked change in serum components except uric acid in T-2 treated chickens. All lymphoid tissues retained atrophic and lymphoid cell depletion throughout the three weeks trial.

Evaluation of Oxidative DNA Damage Using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay, and the Protective Effects of N-Acetylcysteine Amide on Zearalenone-induced Cytotoxicity in Chang Liver Cells

  • Kang, Changgeun;Lee, Hyungkyoung;Yoo, Yong-San;Hah, Do-Yun;Kim, Chung Hui;Kim, Euikyung;Kim, Jong Shu
    • Toxicological Research
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    • v.29 no.1
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    • pp.43-52
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    • 2013
  • Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by several species of Fusarium that are found in cereals and agricultural products. ZEN has been implicated in mycotoxicosis in farm animals and in humans. The toxic effects of ZEN are well known, but the ability of an alkaline Comet assay to assess ZEN-induced oxidative DNA damage in Chang liver cells has not been established. The first aim of this study was to evaluate the Comet assay for the determination of cytotoxicity and extent of DNA damage induced by ZEN toxin, and the second aim was to investigate the ability of N-acetylcysteine amide (NACA) to protect cells from ZEN-induced toxicity. In the Comet assay, DNA damage was assessed by quantifying the tail extent moment (TEM; arbitrary unit) and tail length (TL; arbitrary unit), which are used as indicators of DNA strand breaks in SCGE. The cytotoxic effects of ZEN in Chang liver cells were mediated by inhibition of cell proliferation and induction of oxidative DNA damage. Increasing the concentration of ZEN increased the extent of DNA damage. The extent of DNA migration, and percentage of cells with tails were significantly increased in a concentration-dependent manner following treatment with ZEN toxin (p < 0.05). Treatment with a low concentration of ZEN toxin (25 ${\mu}M$) induced a relatively low level of DNA damage, compared to treatment of cells with a high concentration of ZEN toxin (250 ${\mu}M$). Oxidative DNA damage appeared to be a key determinant of ZEN-induced toxicity in Chang liver cells. Significant reductions in cytolethality and oxidative DNA damage were observed when cells were pretreated with NACA prior to exposure to any concentration of ZEN. Our data suggest that ZEN induces DNA damage in Chang liver cells, and that the antioxidant activity of NACA may contribute to the reduction of ZEN-induced DNA damage and cytotoxicity via elimination of oxidative stress.

Inhibitory effect of Korean mistletoes on the oxidative DNA damage (한국산 겨우살이의 산화적 DNA 손상 억제작용)

  • Lee, So-Jin;Lee, Mi-Kyoung;Choi, Geun-Pyo;Kim, Na-Young;Roh, Seong-Kyu;Heo, Moon-Young;Kim, Jong-Dai;Lee, Hyeon-Yong;Lee, Jin-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.2
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    • pp.89-96
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    • 2003
  • Korean mistletoes extracts were investigated for in vitro antioxidation activity, with 1,1-diphenyl-2-picrylhydrazine(DPPH), and an inhibitory effect on oxidative DNA damage by using comet assay. The Korean mistletoes were 4 different kinds classified by their host plants (Korean Viscum sp. in Quercus acutissima Carr., Korean Viscum sp. in Castanea crenata, Korean Viscum sp. in Betula platyphylla, and Korean Viscum sp. in Salix koreensis). The samples were extracted with ethanol, and fractonationed with n-butanol, ethyl acetate, chloroform, n-hexane, and second distilled water. Among them, ethyl acetate fraction from Korean Viscum sp. in Betula platyphylla showed the strongest activities to electron donating ability on 1,1-diphenyl-2-picrylhydrazyl(DPPH) and the protective effect on oxidative DNA damage.

Production of an Interactive Media Artwork with motif of Characteristics of comets (혜성의 특성을 모티브로 한 인터랙티브 미디어 아트 제작)

  • Lee, O-Jung;Kim, Hyung-Gi
    • Journal of Digital Contents Society
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    • v.18 no.8
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    • pp.1529-1535
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    • 2017
  • Due to its unique formation and mystery phenomena, comets is a major motif that inspires various creative fields. Media art that has been developed and expanded in step with the development of modern scientific technology has expressed the unique characteristics of comets such as their structure in the art as a motif, using technologies. This study describes the process of producing "Near by", an distance-recognition interactive media art. offering the audience a chance to create a tail of a comet as they close to it. changes the color of a LED structure representing the nucleus of a comet as the audience approaches it. At the same time, a video of a comet's tail is projected. Through this interaction, the audience will renew their perception on comets and emotionally interact with the art. This work capturing not only the formative characteristics of a comet, but also its forming process as a motif. Such composite way of expressing a motif could be presented as a possibility of extending the expression of the natural phenomenon in the future media art field.

Effect of Microsurgical Varicocelectomy on Human Sperm Nucleus DNA Integrity (미세술기를 이용한 정계정맥류절제술이 정자 핵 내 DNA Integrity에 미치는 효과)

  • Kim, Gi-Young;Lee, Jae-Seok;Chi, Hee-Jun;Kim, Jong-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.3
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    • pp.245-251
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    • 2010
  • Objective: Human sperm nucleus DNA damage may negatively affect pregnancy outcome, and the spermatozoa of infertile men have more DNA damage than that of fertile men. The aim of this study was to evaluate the effect of microsurgical varicocelectomy on human sperm nucleus DNA integrity. Methods: We reviewed the medical records of 18 subfertile male patients who underwent microsurgical varicocelectomy at our hospital from April 2006 to April 2007. Varicocele was diagnosed by physical examination and Doppler ultrasound. Standard semen analysis was performed in 18 patients before and 4 months after microsurgical varicoceletcomy using a computer assisted semen analyzer. Sperm nucleus DNA integrity was assessed by a single-cell gel electrophoresis (comet assay). Results: No recurrence of varicocele was observed after 4 months later. The DNA fragmentation index improved after varicocelectomy compared with pre-operatively (19.3 versus 13.7%, respectively, p<0.05). Semen analysis parameters (total count, concentration, motile sperm, viability, strict morphology) increased after varicocelectomy, but the difference did not reach statistical significance. Conclusion: Our data suggest that microsurgical varicocelectomy can improve semen analysis parameters and human sperm nucleus DNA integrity in infertile men with varicocele.

Investigation of in vitro Antigenotoxic Effect of Korean Mistletoe(Viscum album coloratum) Using Comet Assay (Comet assay에 의한 한국산 겨우살이(Viscum album coloratum)의 항유전독성 규명)

  • Park, Jong-Heum;Ji, Seung-Taek;Hyun, Chang-Kee;Chin, Koo-Bok;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.32 no.2
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    • pp.461-468
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    • 2000
  • To investigate the antigenotoxicity of Korean mistletoe using Comet assay, the crude extract was divided into 4 fractions, i.e. fraction I (MW range over 14,000), fraction II $(8,000{\sim}14,000)$, fraction III $(3,500{\sim}8,000)$, and fraction IV (below 3,500) by molecular weight fractionation. In the non-tumoral 3T3 cells, fraction IV could reduce DNA damage induced by MNNG in a dose dependent pattern while fraction I and III which were known to contain lectins and viscotoxins, respectively, did not show the activity. By heat treatment, the antigenotoxic activity of faction IV, though was gradually diminished according to heating time, was found to be maintained significantly. From the Sephadex G-25 gel filtration chromatography, a more purified fraction responsible for the activity of faction IV was obtained from the latter part of total elute. Therefore, it was concluded that the antigenotoxic components of Korean mistletoe were water soluble substances of MW below 1,000 and there is a possibility of utilization as a material of functional foods for chemoprevention.

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