• Title/Summary/Keyword: DNA damages

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Molecular Coning of cDNA for Garlic Mosaic Virus Genome (마늘 모자이크 바이러스 게놈에 대한 cDNA의 클로닝)

  • 최연희
    • Journal of Plant Biology
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    • v.35 no.3
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    • pp.253-257
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    • 1992
  • Potyvirus group is the largest group among plant virus groups and damages severely plant hosts upon infectiQn. In order to investigate the mechanism by which potyviruses induce disease in plants, a cDNA clone 29-6 which is cOIlsidered to be a cDNA clone for garlic mosaic virus (GMV) was isolated. It did not hybridize to garlic latent virus genome, which is one of two major garlic viruses. Northern blot analysis shows that the genome size of garlic mosaic virus was about 9 kb. Clone 29-6 strongly hybridizes to poly(A) RNA isolated from garlic leaves, suggesting that GMV RNA is polyadenylated as other potyviruses. Nucleotide sequence analysis of cDNA clones overlapping with clone 29-6 showed that garlic plants are infected with various strains of garlic mosaic virus which are closely related to each other. other.

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XPS STUDY ON THE EFFECT OF LOW-ENERGY ELECTRON IRRADIATION ON DNA DAMAGE BY Fe3+ ION

  • Noh, Hyung-Ah;Park, Yeun-Soo;Cho, Hyuck
    • Journal of Radiation Protection and Research
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    • v.40 no.2
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    • pp.87-91
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    • 2015
  • We have employed X-ray photoelectron spectroscopy (XPS) technique to examine the combined effects of low-energy electron (LEE) irradiation and $Fe^{3+}$ ion on DNA damage. pBR322 plasmid DNA extracted from E. coli ER2420 was used for preparing DNA-$Fe^{3+}$ sample. The C1s XPS spectra were scanned for LEE-irradiated and LEE-unirradiated samples and then curve-fitted. For the samples with LEE irradiation only or with Fe ion only, no significant changes from pure DNA samples were observed - a single effect of either $Fe^{3+}$ ion or LEE irradiation did not cause a significant damage. However, when these two components were combined, the DNA damage was increased quite significantly, compared to the sum of DNA damages caused by $Fe^{3+}$ ion and by LEE irradiation independently. This observation is consistent with our previous results [Radiat. Res. 177, 775 (2012)] which was done using gel-electrophoresis technique. Partial interpretation of the observed spectrum peaks was also attempted.

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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    • v.53 no.4
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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.

Effect of Vitamin C and GSH on the Hg Induced ROS (비타민 C와 글루타치온이 수은유도 ROS 생성에 미치는 영향)

  • Kwon, Kyoung-Jin;Sheen, Yhun-Yhong
    • Environmental Analysis Health and Toxicology
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    • v.23 no.1
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    • pp.33-39
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    • 2008
  • The genotoxicity of mercury compounds have been investigated with a variety of genetic endpoints in prokaryotic and eukaryotic cells. The mercury ions are positively charged and easily form complexes with DNA by binding with negatively charged centers to cause mutagenesis. Further, the mercury ions can react with sulfhydryl (-SH) groups of proteins associated with DNA replication and alter genetic information. Another mechanism by which mercury damages DNA molecule is via its probable involvement of reactive oxygen species (ROS) and induces DNA strand breaks. In order to investigate whether the ROS production was induced by mercury, we performed ROS assay. As the result, the ROS production was significantly increased when it grows dose-dependently and time-dependently. We compared mercury alone-treated group and mercury co-treated with Vitamin C or glutathione group. As the result, the ROS production induced by mercury was decreased by Vitamin C and glutathione. Co-treated with Vitamin C and glutathione group was the most effective to lowering ROS production induced by mercury.

Baicalein and Baicalin from the Radix of Scutellaria baicalensis Georgi Inhibits Oxidative DNA Damage and Apoptosis via its Antioxidant Activity

  • Garcia, Nellie Ann S.;Jeong, Hyung-Jin
    • Korean Journal of Plant Resources
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    • v.22 no.6
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    • pp.489-497
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    • 2009
  • In this study, we evaluated and compared the protective effects of two major constituents, baicalein and baicalin, against oxidative DNA and cell damages caused by hydroxyl radical. Antioxidant properties were evaluated using DPPH and hydroxyl radicals scavenging assays and $Fe^{2+}$ chelating assay. ${\varphi}X$ 174 RFI plasmid DNA and intracellular DNA migration assay were used to evaluate the protective effect against oxidative DNA damage. Also, MTT and lipid peroxidation assays were used to evaluate their protective effects against oxidative cell damage. Both baicalein and baicalin prevented intracellular DNA and cells from oxidative damage caused by hydroxyl radical via antioxidant activities. Baicalein demonstrated a stronger antioxidant activity in scavenging DPPH radicals and chelating $Fe^{2+}$ while baicalin scavenged hydroxyl radicals more efficiently. The differences in the level of baicalein and baicalin pose a different pathological pathway for each. The antioxidant activity of baicalin was due to its ability to scavenge hydroxyl radical whilst baicalein was a stronger $Fe^{2+}$ chelator. Further investigation to compare the molecular mechanisms of antitumor activities of baicalein and baicalin is vital to anticancer research.

Formation of DNA-Protein Crosslink at Oxidized Abasic Site Mediated by Human DNA Polymerase Iota and Mitochondrial DNA Polymerase Gamma

  • Son, Mi-Young;Jun, Hyun-Ik;Goo, Sun-Young;Sung, Jung-Suk
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.1-8
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    • 2009
  • Human genomic DNA is continuously attacked by oxygen radicals originated from cellular metabolic processes and numerous environmental carcinogens. 2-deoxyribonolactone (dL) is a major type of oxidized abasic (AP) lesion implicated in DNA strand scission, mutagenesis, and formation of covalent DNA-protein crosslink (DPC) with DNA polymerase (Pol) ${\beta}$. We show here that human DNA polymerase (Pol)${\iota}$ and mitochondrial $Pol{\gamma}$ give rise to stable DNA-protein crosslink (DPC) formation that is specifically mediated by dL lesion. $Pol{\gamma}$ mediates DPC formation at the incised dL residue by its 5'-deoxyribose-5-phosphate (dRP) lyase activity, while $Pol{\gamma}$ cross links with dL thorough its intrinsic dRP lyase and AP lyase activities. Reactivity in forming dL-mediated DPC was significantly higher with $Pol{\gamma}$ than with $Pol{\iota}$. DPC formation by $Pol{\gamma}$, however, can be reduced by an accessory factor of $Pol{\gamma}$ holoenzyme that may attenuate deleterious effects of crosslink adducts on mitochondrial DNA. Comparative kinetic analysis of DPC formation showed that the rate of DPC formation with either $Pol{\iota}$ or $Pol{\gamma}$ was lower than that with $Pol{\beta}$. These results revealed that the activity of catalytic lyase in DNA polymerases determine the efficiency of DPC formation with dL damages. Irreversible crosslink formation of such DNA polymerases by dL lesions may result in a prolonged strand scission and a suicide of DNA repair proteins, both of which could pose a threat to the genetic and structural integrity of DNA.

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Protective Effect of Electrolyzed Reduced Water on the Paraquat-induced Oxidative Damage of Human Lymphocyte DNA (Paraquat에 의한 사람 임파구 DNA 손상에 대한 환원전리수의 보호효과)

  • Park, Eun-Ju;Ryoo, Kun-Kul;Lee, Yoon-Bae;Lee, Jong-Kwon;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.155-160
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    • 2005
  • Electrolyzed reduced water (ERW), showing extremely negative oxidation-reduction potential, was used to investigate the effects of paraquat-induced damages on DNA from human lymphocyte. The effect of ERW on paraquat-induced oxidative DNA damage in human lymphocytes was evaluated by Comet assay (single-cell gel electrophoresis) quantified as percentage fluorescence in tail. Comet assay has been used widely to assess the level of the DNA damage in individual cells. Lymphocytes were oxidatively challenged with various concentrations of paraquat for 30 min at $37^{\circ}C$, and were then treated with electrolyzed reduced water for 30 min. The oxidative DNA damage by paraquat, as indicated by the fluorescent tail in DNA, increased in a dose-dependent manner. However, oxidative damage of the DNA was almost completely prevented upon treatment with electrolyzed reduced water.

Evaluation of the Genetic Toxicity of Synthetic Chemicals (XIII) - Single Cell Gel Electrophoresis of Benzoyl Chloride, 2-Propyn-1-ol, and 2-Phenoxyethanol in Chinese Hamster lung Fibroblast -

  • Ryu, Jae-Chun;Kim, Youn-Jung
    • Environmental Mutagens and Carcinogens
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    • v.24 no.2
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    • pp.79-84
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    • 2004
  • Three synthetic chemicals, benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol were selected for genotoxicity testing, based on production quantity and available genotoxic data. In our previous report, benzoyl chloride induced chromosomal aberrations in Chinese hamster lung (CHL) fibroblast in vitro with and without metabolic activation, while 2-propyn-l-ol and 2-phenoxy ethanol induced only with metabolic activation. To compare the genotoxicity of chromosome aberration assay, the single cell gel electrophoresis (comet) assay subjected using CHL cells. As a result, statistically significant differences of tail moment values of benzoyl chloride, 2-propyn-1-ol, and 2-phenoxy ethanol were observed compared with control values on almost all concentrations with S9 or without S9 metabolic activation system. This results suggest that genotoxic results of the comet assay and the chromosome aberration assay show correlationship of genotoxicity in the CHL fibroblast. In summary, the positive result of chromosome aberration of benzoyl chloride, 2-propyn-l-ol, and 2-phenoxy ethanol was also induced DNA damages in comet assay with same cell line. Consequently, comet assay will be useful and more accurate tool to detect and to confirm the genotoxicity especially DNA damages in CHL fibroblast.

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Oxidative DNA Damages and Its Repair Enzyme Acitivities in the SD rats by Repeated Inhalation Exposure to the Hexavalent Chromium

  • Seung-Hee Maeng;Il-Je Yu;Hyun-Young Kim;Cheol-Hong Lim;Jong-Yoon Lee;Kwon-seob Lee;Yong-Mook Lee;Ho-Keun Chung;Hai-Won Chung
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2001.05a
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    • pp.151-151
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    • 2001
  • According to the epidemiological studies in chromium workers, hexavalent chromium is associated with the risk of lung cancer. Reactive oxygen species produced by hexavalent chromium exposure may play an important role in the carcinogenesis process. We measured one kind of oxidative DNA damages, 8-hydroxydeoxyguanosine (8-OHdG) and its repair activities as an indicators of cellular oxidative stress in rat lungs by the repeated inhalation exposure to sodium chromate.(omitted)

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Neuroprotective effects of modified Bo-Yang-Hwan-Oh-Tang in N2a neuroblastoma cells (가감보양환오탕(加減補陽還五湯)의 N2a 뇌신경세포에 대한 보호효과)

  • Lim, Kyu;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.4
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    • pp.77-84
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    • 2006
  • Objectives : To evaluate the neuroprotective effects of modified Bo-Yang-Hwan-O-Tang (BHT), we investigated the neuronal death protection effects to oxidative damages in N2a neuroblastoma cells. Methods : To study the cytotoxic effect of BHT on N2a neuronal cells, the cell viability was determined by MTT assay. To investigate the neuronal death protection of BHT, N2a neuronal cells were induced oxidative damages by H2O2, and assayed the cell viability and DNA fragmentation. We also investigated DPPH free radical scavenging effects of BHT by tube test. Results : In MTT assay, $500{\mu}g/ml$ of BHT was not showed cytotoxic effect on N2a neuronal cells. BHT protected N2a neuronal cells from H2O2-induced cell death in a dose-dependent manner. BHT also protected N2a neuronal cells from H2O2-induced DNA fragmentation. BHT scavenged DPPH free radicals in a dose-dependent manner. Conclusion : These data suggest that BHT may have strong anti-oxidant effects through the free radical scavenging and neuroprotective effects in neuronal cells.

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