• Title/Summary/Keyword: single-cell gel electrophoresis (SCGE)

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Antigenotoxic Effect of Dominant Bacterial Isolates from Kimchi in vitro (김치로부터 분리된 우세 균주들의 in vitro 항유전 독성효과)

  • Choi, Jun-Won;Park, Jong-Heum;Ji, Seung-Taek;Choi, Ok-Byung;Shin, Heuyn-Kil
    • Korean Journal of Food Science and Technology
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    • v.31 no.4
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    • pp.1071-1076
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    • 1999
  • This study was carried out to isolate the colonies of dominant fermented bacteria from Kimchi (Korean native fermented foodstuffs) and investigate their inhibitory potentials on mutagenesis and carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) as direct carcinogen. For this purpose, single cell gel electrophoresis technique (SCGE assay, or comet assay) which is a sensitive and rapid technique for detecting the presence of DNA strand breaks in individual cells was used. DNA damages of Kimchi isolates were compared with that of the positive control, MNNG. Among 3 isolates from Tongbaechu Kimchi, two isolates B-1 and B-2 showed antigenotoxicities (p<0.01). All 4 isolates from Yulmu Kimchi had antigenotoxicities (p<0.05). Also, 3 of 5 isolates from Chonggak Kimchi and 2 of 9 isolates from Kaktugi were antigenotoxic (p<0.05 and p<0.01, respectively).

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Evaluation of DNA Damage and Repair Kinetics in the Earthworm (Eisenia fetida) Exposed to Radiation and Mercury (방사선과 수은에 의해 유도된 Eisenia fetida 체강세포의 DNA 손상 및 수복 평가)

  • Ryu, Tae-Ho;Nili, Mohammad;An, Kwang-Guk;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.68-73
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    • 2011
  • The single cell gel electrophoresis (SCGE) assay is a microelectrophoretic technique for assessments of DNA damage at the level of the individual eukaryotic cell. The SCGE assay, due to its simplicity, sensitivity and need of a few cells, has advantages compared to other genomic damage assays such as sister chromatid exchange, chromosomal aberration and micronucleus test. In this study, investigated were the levels of DNA damage and the repair kinetics in the coelomocytes of Eisenia fetida treated with HgCl2 and ionizing radiation by means of the SCGE assay. For detecting DNA damage and repair in coelomocytes, earthworms (E. fetida) were irradiated with six doses of ${\gamma}$-rays (0, 2.5, 5, 10, 20 and 50 Gy) and in vivo exposed to mercuric chloride at 0, 80 and 160 mg $kg^{-1}$ for 48 hours. Then the Olive tail moments were measured during 0~12 hours after irradiation and 0~72 hours after Hg treatment. The results showed that the more the oxidative stress was induced by mercury and radiation, the longer the repair time was required. Also, the results suggest that the SCGE assay may be used as an important tool for comparison of the sensitivity of different species to oxidative stresses.

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.

Protective Effects of a Herb, Artemisia capillaris, Against Radiation-induced DNA Damage (방사선 유도 DNA 손상에 대한 인진쑥의 방어효과)

  • Jo, Sung-Kee;Oh, Heon;Cheon, Eui-Hyun;Jeong, U-Hee;Cho, Nam-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.22-27
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    • 2004
  • In the present study, the protective effects of Artemisia capillaris (AC) on the DNA damage induced by $^{60}$ Co ${\gamma}$-rays were evaluated using alkaline single-cell gel electrophoresis (SCGE, comet assay) in the mouse peripheral lymphocytes and micronuclei (MN) formation test in the Chinese hamster ovary (CHO) cells. We also investigated the effect of AC on 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation in the mouse liver and thymus exposed to ${\gamma}$-ray, The tail moment and the frequency of MN, which were markers of DNA damage in the SCGE and MN formation test, were decreased in the groups treated with AC extract before exposure to 200 cGy of ${\gamma}$-ray. We also observed its activities, lowering 8-OHdG level, an index of oxidative DNA damage, in the groups treated with AC extract before whole body ${\gamma}$-irradiation (800 cGy). It is plausible that scavenging of free radicals by AC may have played an important role in providing the protection against the radiation-induced damage to the DNA. These results indicated that AC protects the DNA damage induced by ${\gamma}$-rays and might be a useful radioprotector, especially since it is a relatively nontoxic product.

MUTAGENICITY AND ANTIMUTAGENICITY OF CATECHIN AND TANNINS FROM THE BARK OF HAMAMELIS VIRGINIANA L. IN HUMAN Hep G2 CELLS

  • Sundermann, Volker-Mersch;Andreas Dauer;Evelyne Lhoste;Siegfried Knasmuller
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.10a
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    • pp.95-96
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    • 2001
  • The mutagenic and antimutagenic activities of catechin, hamamelitannin and two proanthocyanidin fractions prepared from the bark of Hamamelis virginia L. - a commonly used medicinal herb - were investigated in a human derived hepatoma (Hep G2) cell line using the single cell gel electrophoresis (SCGE, syn. Comet assay) for the detection of DNA-migration. The cells possess different phase I and phase II enzymes involved in the biotransformation of xenobiotics.(omitted)

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13-Week Oral Gavage Toxicity with Sophora Japonica Linne Seed Extract in Sd Rats

  • Lee, Jae-Hyung;Kim, Il-Yong;Hyun, Chang-Kee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.10b
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    • pp.133-133
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    • 2003
  • In recent years. attention has focused on the application of the alkaline single cell gel electrophoresis (SCGE or Comet) assay in environmental mutagenesis. To evaluate the suitability of the assay as a monitoring. technique, the DNA damages in liver cells and erythrocytes of carp (Cyprinus carpio) exposed to benzo[${\alpha}$]pyrene (B[${\alpha}$]P) were estimated comparatively with the in vivo Comet assay and the micronucleus test (MNT).(omitted)

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Protective Effect of the 70% Ethanolic Extract of Alpinia officinarum and Galangin Against $KBrO_3$-induced DNA and Chromosomal Damage in Mice (Galangin 및 양강추출물의 $KBrO_3$ 유도 DNA 및 염색체 손상에 대한 보호효과)

  • Yang, Hye-Eun;Heo, Moon-Young
    • YAKHAK HOEJI
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    • v.54 no.6
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    • pp.423-428
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    • 2010
  • The aim of this study was to evaluate the in vivo effect of galangin and the 70% ethanolic extract of Alpinia officinarum (AO) toward $KBrO_3$-induced DNA and chromosomal damage in mice. Galangin and AO inhibited the formation of 8-hydroxy-2'-deoxyguanosine (8-OH2'dG) as an indicator of DNA oxidative damage in the liver cell. Galangin and AO showed the inhibitory effect on the formation of DNA single strand break in the splenocyte by single cell gel electrophoresis (SCGE) assay and also inhibited micronucleated reticulocyte (MNRET) formation of peripheral blood in tail blood of mice. Vit-E revealed antigenotoxic effects in DNA and chromosome levels, but galangin was more potent active compound compare to vit-E under our experimental conditions. The results suggest that the extract of Alpinia officinarum containing galangin can modify the oxidative DNA and chromosomal damage and may act as chemopreventive agent against oxidative stress in vivo.

The Protective Effects of Ganoderma lucidum on the DNA Damage and Mutagenesis (DNA손상 및 돌연변이에 대한 명지버섯의 방어효능)

  • 이길수;공석경;최수영
    • Biomolecules & Therapeutics
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    • v.11 no.2
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    • pp.139-144
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    • 2003
  • Ganoderma lucidum is commonly known as medically potent mushroom, which has been widely used in China and other oriental countries for the treatment of various diseases, including cancer. In this report, we investigated the anti-oxidant and protective effect of Ganodema lucidum extract (GLE) against the DNA damage induced by free radical and U.V. In the assay of cell growth inhibition, the inhibitory cell growth rate induced by hydroxyl radical was dose-dependently decreased by GLE. This results support that GLE has a detoxifying activity against cytotoxicity of hydroxyl radical in E. coli cell. GLE also protected ColE1 plasmid DNA damage in the concentration of 200$\mu\textrm{g}$ per reaction on the DNA fragmentation assay. The nuclear tailing by hydrogen peroxide in single cell gel electrophoresis(SCGE) was decreased by GLE in the concentration of 50$\mu\textrm{g}$/ml. These data indicate that Ganoderma lucidum has an anti-oxidative activity to hydrogen peroxide. The mutation rate after irradiation of U.V. was reduced by 50$\mu\textrm{g}$/ml GLE and total number of Rif (Rifampicin) resistant mutants was decreased in a concentration dependent manner when added the GLE exogenously in a culture media. According to the results, it is likely that GLE has not only an anti-oxidative activity to hydroxyl radical but also an anti-mutagenic activity to U.V. mutagenesis.

Multiple-biometric Attributes of Biomarkers and Bioindicators for Evaluations of Aquatic Environment in an Urban Stream Ecosystem and the Multimetric Eco-Model (도심하천 생태계의 수환경 평가를 위한 생지표 바이오마커 및 바이오인디케이터 메트릭 속성 및 다변수 생태 모형)

  • Kang, Han-Il;Kang, Nami;An, Kwang-Guk
    • Journal of Environmental Impact Assessment
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    • v.22 no.6
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    • pp.591-607
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    • 2013
  • The objectives of the study were to evaluate the aquatic environment of an urban stream using various ecological parameters of biological biomarkers, physical habitat quality and chemical water quality and to develop a "Multimetric Eco-Model" ($M_m$-E Model) for the ecosystem evaluations. For the applications of the $M_m$-E model, three zones including the control zone ($C_Z$) of headwaters, transition zone ($T_Z$) of mid-stream and the impacted zone ($I_Z$) of downstream were designated and analyzed the seasonal variations of the model values. The biomarkers of DNA, based on the comet assay approach of single-cell gel electrophoresis (SCGE), were analyzed using the blood samples of Zacco platypus as a target species, and the parameters were used tail moment, tail DNA(%) and tail length (${\mu}m$) in the bioassay. The damages of DNA were evident in the impacted zone, but not in the control zone. The condition factor ($C_F$) as key indicators of the population evaluation indicator was analyzed along with the weight-length relation and individual abnormality. The four metrics of Qualitative Habitat Evaluation Index (QHEI) were added for the evaluations of physical habitat. In addition, the parameters of chemical water quality were used as eutrophic indicators of nitrogen (N) and phosphorus (P), chemical oxygen demand (COD) and conductivity. Overall, our results suggested that attributes of biomarkers and bioindicators in the impacted zone ($I_Z$) had sensitive response largely to the chemical stress (eutrophic indicators) and also partially to physical habitat quality, compared to the those in the control zone.

Comparative Study on Human Risk by Ionizing Radiation and Pesticide as Biological Information about Environmental Disaster (환경재해에 관한 생물정보로서의 이온화 방사선과 살충제의 인체 위해성 비교 연구)

  • Kim, Jin-Kyu;Hyun, Soung-Hee
    • Journal of Radiation Protection and Research
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    • v.26 no.4
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    • pp.385-392
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    • 2001
  • Environmental risk factors such as ionizing radiations, heavy metals, and pesticides can cause environmental disasters when they exist in excess. The increases in use of ionizing radiation and agricultural pesticide are somewhat related to the possibility of the disaster. The risk of radiation and pesticide was evaluated by means of the single cell gel electrophoresis (SCGE) assay on the human blood lymphocytes. The lymphocytes were irradiated with $0{\sim}2.0Gy$ of $^{60}Co$ gamma ray. Another groups of lymphocytes were exposed to various concentrations of parathion. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed a clear dose- or concentration-response relationship. Parathion of a recommended concentration for agricultural use ($1mg {\ell}^{-1}$ ) has a strong cytotoxic effect on lymphocytes, which is equivalent to damage induced by 0.1 Gy of ${\gamma}$-ray. Furthermore, $2mg{\ell}^{-1}$ of parathion can give rise to DNA damage equivalent to that induced by 0.25 Gy at which the radiation-induced damage can start to develop into clinical symptoms. The comparative results of this study can provide an experimental basis and biological information for the prevention of environmental disaster.

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