• Title/Summary/Keyword: damage of DNA

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단세포전기영동법(single Cell Gel Electrophoresis Assay)을 이용한 농약 살포자의 DNA손상 평가 (Evaluation of DNA damage in Pesticide Sprayers using Single Cell Gel Electrophoresis)

  • 이연경;이도영;이은일;이동배;류재천;김해준;설동근
    • 한국환경성돌연변이발암원학회지
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    • 제21권2호
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    • pp.128-134
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    • 2001
  • Single cell gel electrophoresis (SCGE) assay, also called comet assay, is a rapid and sensitive method to detect DNA damage in single cell level. To evaluate the DNA damage of lymphocytes of pesticides sprayers, SCGE assay was carried out for 50 pesticides sprayer and 58 control subjects. They were interviewed with structured questionnaire to get the information about the kinds and amount of pesticide. Insecticides and fungicides were predominant among pesticides. Major components of pesticides were organophosphorus, organosulfate, cartap, carbamates, and triazole. Sprayed pesticides were classified into two groups. Group I included organophosphorus, organoarsenic, organotin, tetrazine, triazole and gramoxone, which were known to cause DNA damages. Group II pesticide were carbamates, surfactants, organosulfates, etc., which were not found as DNA damaging agents in scientific documents. Olive tail moments of 100 lymphocytes were measured by KOMET 3.1 program for each person. The means of tail moments were compared between farmers exposed to pesticides and control subjects. Farmers showed higher tail moments than control subjects (2.07$\pm$1.40 vs 1.53$\pm$0.77, p<0.05). The means of tail moments also were compared among group I sprayers (n=36), group II sprayers (n=24) and, control subject, and the means or tail moments were 3.4s$\pm$3.2o, 2.66$\pm$2.20 and 1.53$\pm$0.77 respectively. The difference between means of group I sprayers and controls was statistically significant (p<0.05). In conclusion, this study showed higher DNA damage in farmers exposed to pesticides than control subjects, and comet assay could be useful as a biological monitoring method of genotoxic pesticides for farmers.

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Superoxide dismutase의 활성차이에 따른 식물세포의 paraquat에 대한 반응과 핵 DNA 손상 검정 (Nucleus-DNA Damage and Different Response of Plant Cells to Paraquat in Relation to Enzyme Activity of Superoxide Dismutase.)

  • 권순태;이명현;오세명;정도철;김길웅
    • 생명과학회지
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    • 제14권4호
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    • pp.614-619
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    • 2004
  • This study was undertaken to investigate the different responses of cultured plant cells to paraquat treatment and nucleus-DNA damage in relation to enzyme activity of superoxide dismutase (SOD). Furthermore, this study was also carried out to understand the antioxidative mechanism of plant cells to environmental stress. We selected two different species of plant cultured cells, Ipomoea batatas as high-SOD species and Lonicera japonica as low-SOD species. The total activity and specific activity of SOD in a chlorophyllous cell of I. batatas were 3,736 unit/gㆍfresh weight and 547 unit/mgㆍprotein, respectively, and those in L. japonica were 23 unit/gㆍfresh weight and 13 unit/mgㆍprotein, respectively SOD activity in chlorophyllous I. batatas cells reached its maximum level at 10 to 15 days after subculture, whereas that in L. japonica remained at a very low SOD level during the whole period of subculture. In comparison to L. japonica, I. batatas, a high-SOD species, showed high tolerance to paraquat 10 and 50 mg/l treatment in terms of cell viability and electrolyte leakage. Based on the result of comet assay, the nucleus-DNA damage of two species by paraquat 50 mg/l treatment was not significantly different. However, I. batatas cells repaired their damaged DNA more effectively than the cells of the low-SOD species, L. japonica.

Free Radical Scavenging Activity and Protective Ability of Methanolic Extract from Duchesnea indica Against Protein Oxidation and DNA Damage

  • Hu, Weicheng;Shen, Wei;Wang, Myeong-Hyeon
    • Preventive Nutrition and Food Science
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    • 제14권4호
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    • pp.277-282
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    • 2009
  • The antioxidant potency of methanolic extract of Duchesnea indica (MDI; Indian strawberry) was investigated by employing various established in vitro systems, such as total phenolic content, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, reducing power assay, metal chelating assay, superoxide radical scavenging activity and protective ability of DNA damage and protein oxidation. MDI inhibited metal chelating by 75.57% at 2 mg/mL, scavenged 50% DPPH free radical at 29.13 ${\mu}$g/mL, and eliminated approximately 46.21% superoxide radical at the concentration of 1 mg/mL. In addition, MDI showed strong ability on reducing power, DNA damage protection and protein oxidation protection. Overall, results suggested that MDI might be beneficial as a potent antioxidant and effectively employed as an ingredient in food applications.

Suppressive Effect of Aqueous Extract of Red-Ginseng on the Herbicide-induced DNA Damage and Hemolysis

  • Seo, Yoo-Na;Lee, Mi-Young
    • Journal of Applied Biological Chemistry
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    • 제53권4호
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    • pp.202-206
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    • 2010
  • The effects of aqueous extracts of red ginseng on the damage of DNA and erythrocyte by herbicides were evaluated using comet assay and hemolysis assay. Notably, the oxidative DNA damage and erytbrocyte hemolysis by 2,4-D (2,4-dichlorophenoxyacetic acid) and 2,4,5-T (2,4,5-trichlorophenoxyacetic acid) were significantly suppressed by red ginseng treatment. Moreover, red ginseng could suppress significantly paraquat-induced oxidative DNA damage and hemolysis. These suppressive effects of red ginseng on the herbicide-induced damages might be due to the antioxidant components.

Characterization of gltA::luxCDABE Fusion in Escherichia coli as a Toxicity Biosensor

  • Ahn, Joo-Myung;Kim, Byoung-Chan;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제11권6호
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    • pp.516-521
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    • 2006
  • The use of gltA gene, as a new biomarker for environmental stress biomonitoring, was investigated because of its key position as the first enzyme of the tricarboxylic acid (TCA) cycle. A recombinant bioluminescent Escherichia coli strain, EBJM2, was constructed using a plasmid carrying the citrate synthase (gltA) promoter transcribing the Photorhabdus luminescens IuxCDABE genes (gltA::luxCDABE). The responses from this strain were studied with five different classes of toxicants: DNA damage chemicals, phenolics, oxidative-stress chemicals, PAHs, and organic solvents. EBJM2 responded strongly to DNA damage chemicals, such as mitomycin C (MMC) and methyl-nitro-nitrosoguanidine (MNNG) and nalidixic acid with the strongest responses. In contrast, tests with several compounds from the other four classes of toxicants gave no significant response. Therefore, EBJM2 was found to be sensitive to DNA damage chemicals.

Exposure to Triclosan Induces Mortality through Oxidative Stress and DNA Damage in the Java Medaka Oryzias javanicus

  • Seong Duk Do;Jae-Sung Rhee
    • 한국해양생명과학회지
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    • 제9권1호
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    • pp.33-40
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    • 2024
  • To understand the detrimental effects of triclosan on Java medaka (Oryzias javanicus) embryos, fertilized embryos were exposed to different concentrations (1, 10, 50, 100, 200, 400, 600, 800, and 1,000 ㎍ l-1) of triclosan until hatching. Then, we examined the survival rate and developmental parameters as well as alterations in antioxidant constituents and DNA damage markers. The results showed dose-dependent mortality, hatching delays, and developmental abnormalities in the embryos. Additionally, there were significant increases in oxidative stress parameters and antioxidant responses, along with elevated DNA damage. These findings suggest that sublethal concentrations of triclosan induce toxic effects through oxidative stress on Java medaka embryos, as evidenced by changes in in vivo parameters and biochemical constituents.

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

  • 이길수;공석경;최수영
    • Biomolecules & Therapeutics
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    • 제11권2호
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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.

Comet assay를 이용한 갯지렁이 (Perinereis aibuhitensis)의 혈구세포에 대한 유전독성 평가 (Evaluation of Genotoxicity in Blood Cells of a Polychaetous Worm (Perinereis aibuhitensis), Using Comet Assay)

  • 서진영;성찬경;최진우;이창훈;류태권;한기명;김기범
    • Environmental Analysis Health and Toxicology
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    • 제20권4호통권51호
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    • pp.333-341
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    • 2005
  • In order to know whether polychaetes could be used as an appropriate organism for the detection of genotoxicity, DNA strand breaks were evaluated in blood cells of a nereidae worm (Perinereis aibuhitensis) exposed to various aquatic chemical pollutants (e.g. Cd, Pb, Pyrene, Benaor[a]pyrene). Hydrogen peroxide increased DNA strand breaks up to the highest concentration (10 $\mu$M). Higher concentration than 0.1 $\mu$M showed a significantly more DNA damage than control. Cadmium and lead also showed higher DNA damage than control, over 1.0 and 1 $\mu$g/L, respectively. In case of pyrene, DNA damage was detected even at 0.001 $\mu$g/L. However, DNA damage decreased due to apoptosis at the highest concentration of pyrene and Pb. This study suggested that the polythaetous blood cells could be used effectively for screening genotoxic contaminants in the environment.

UV에 의해 손상된 DNA 회복에 미치는 cobaltous chloride의 효과 (Effect of Cobaltous Chloride on the Repair of UV-induced DNA Damage)

  • 김국찬;김영진;이강석
    • Journal of Radiation Protection and Research
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    • 제20권2호
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    • pp.71-78
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    • 1995
  • 본 연구에서는 유전자 손상회복에 관여하는 단백질을 이용하여 돌연변이 생성을 억제시키는 물질로서 알려진 cobaltous chloride가 유전자 손상회복에 미치는 영향을 연구하므로서 방사선으로 인한 손상방지 및 방사선 방어효과에 대한 적용가능성을 평가하였다. Cobaltous chloride가 RecA 단백질의 기능에 미치는 영향을 조사한 결과 RecA 단백질에 의한 DNA strand exchange 반웅에 있어 cobaltous chloride 처리로 RecA 단백질이 $_{ss}DNA$로 부터 SSB 단백질과 더 효과적으로 경쟁함으로써 안정된 $RecA-_{ss}DNA$ complex의 형성을 유도하고, 증가된 ATPase활성에 의한 ATP 가수분해로 손상된 DNA의 회복이 촉진될 수 있다는 사실을 입증 해주고 있다. 또한 RecA단백질은 UV에 의해 손상된 supercoiled DNA에 더 효과적으로 결합됨이 관찰되었으며 UV 선량과도 상관관계가 있음을 확인하였다. 따라서 이와 같은 연구결과들은 방사선으로 인한 유전적인 손상방지 및 방사선 방어효과에 관한 연구에 적용될 수 있을 것으로 기대된다.

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Molecular Cloning and Characterization of a recA-like Gene Induced by DNA Damage from a Fluorescent Pseudomonas sp.

  • Ok Bong Kim;Na Young Kim;Jae Hoon Jeong;Si Wouk Kim;Hye Gwang Jeong;Seong Myeong Yoon;Jong Kun Park;Jung Sup Lee
    • Animal cells and systems
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    • 제3권2호
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    • pp.229-236
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    • 1999
  • The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

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