• Title/Summary/Keyword: DNA oxidation

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Inhibitory Effect of Extract from Acanthocoris sordidus on Oxidative Damage (꽈리허리노린재(Acanthocoris sordidus) 추출물이 산화적 손상에 미치는 억제 효과)

  • Park, Young Mi;Lim, Jae Hwan;Lee, Jong Eun;Seo, Eul Won
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1078-1084
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    • 2014
  • Here, we showed that Acanthocoris sordidus extract inhibited both cell and DNA damage caused by oxidative stress. In a radical scavenging assay, the scavenging activity of the A. sordidus extract against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radicals was 48.9% and 37.8%, respectively, that of ascorbic acid, which was used as a positive control. The ferrous iron chelating activity of the A. sordidus extract was 80.0% compared to that when ethylenediaminetetraacetic acid (EDTA) was used a control. To verify the inhibitory effect of the extract on oxidative cell damage induced by reactive oxygen species (ROS), a lipid peroxidation assay was performed. The results showed that peroxidation was completely inhibited in an extract-treated group compared to a radical-treated group. The level of p21 protein expression was 68.1% that of a control sample. The DNA cleavage-inhibiting property of the A. sordidus extract-treated group was 53.3% that of a control group. Moreover, the phosphorylation of the H2AX protein was reduced to 39.0% of that treated with radical agents, indicating that the extract might inhibit the DNA damage that causes radical oxidation. Taken together, our findings suggest that the A. sordidus extract is effective not only in repressing oxidation by free oxygen radicals and hydroxyl radicals but also in decreasing cell and DNA damage caused by oxidative stress.

Cloning of the Large Subunit of Replication Protein A (RPA) from Yeast Saccharomyces cerevisiae and Its DNA Binding Activity through Redox Potential

  • Jeong, Haeng-Soon;Jeong, In-Chel;Kim, Andre;Kang, Shin-Won;Kang, Ho-Sung;Kim, Yung-Jin;Lee, Suk-Hee;Park, Jang-Su
    • BMB Reports
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    • v.35 no.2
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    • pp.194-198
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    • 2002
  • Eukaryotic replication protein A (RPA) is a single-stranded(ss) DNA binding protein with multiple functions in DNA replication, repair, and genetic recombination. The 70-kDa subunit of eukaryotic RPA contains a conserved four cysteine-type zinc-finger motif that has been implicated in the regulation of DNA replication and repair. Recently, we described a novel function for the zinc-finger motif in the regulation of human RPA's ssDNA binding activity through reduction-oxidation (redox). Here, we show that yeast RPA's ssDNA binding activity is regulated by redox potential through its RPA32 and/or RPA14 subunits. Yeast RPA requires a reducing agent, such as dithiothreitol (DTT), for its ssDNA binding activity. Also, under non-reducing conditions, its DNA binding activity decreases 20 fold. In contrast, the RPA 70 subunit does not require DTT for its DNA binding activity and is not affected by the redox condition. These results suggest that all three subunits are required for the regulation of RPA's DNA binding activity through redox potential.

Effects of Diet with Added Butterbur (Petasites japonicus Maxim) on the Plasma Lipid Profiles and Antioxidant Index of Mice (머위(Petasites japonicus Maxim) 첨가 식이가 마우스 혈장 지질 수준 및 항산화 지표에 미치는 영향)

  • Oh Sang-Hee;Yang Yun-Hyung;Kwon Oh-Yoon;Kim Mee-Ree
    • Journal of the East Asian Society of Dietary Life
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    • v.16 no.4
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    • pp.399-407
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    • 2006
  • We evaluated the effects of butterbur (Petasites japonicus Maxim) addition to the diet on lipid profiles and antioxidant biomarkers such as total glutathionine, TBARS value, carbonyl value, GPx, GR, SOD and paraoxonase activity in the plasma or liver of mice. The distribution of body fat deposition, total cholesterol (TC) contents, and atherogenic index in the plasma were significantly decreased in the butterbur group. The levels of GSH and the activity of GR and SOD were significantly higher in the liver of the butterbur group than in that of the control group. Lipid oxidation of the liver and kidney and protein oxidation of the liver and heart were decreased in the butterbur group. Additionally, the DNA damage, as determined using the comet assay (single cell gel assay) with alkaline electrophoresis and as quantified by measuring the tail length (TL), was decreased in the butterbur group. The results of the present study showed that a diet with added butterbur exerts degenerative disease-protective effects on oxidative DNA damage and lipid peroxidation.

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Antimicrobial and Antioxidative Effects of Roselle (Hibiscus sabdariffa L.) Flower Extract and Its Fractions on Skin Microorganisms and Oxidation

  • Kang, Pil-Sung;Seok, Jae-Hwan;Kim, Yo-Han;Eun, Jae-Soon;Oh, Suk-Heung
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.409-414
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    • 2007
  • Antimicrobial and antioxidative effects of 14 different herbal flower extracts on skin microorganisms and oxidation were tested in this research. Herbal flower extracts were prepared with 70% ethanol. Among the herbal flower extracts, roselle (Hibiscus sabdariffa L.) flower extract showed the highest antimicrobial activity against Staphylococcus epidermidis as determined by a paper disc method. The seventy % ethanol extract of roselle flower was fractionated by sequential hexane, chloroform, ethyl acetate, n-butanol, and water fractionation. The growth of S. epidermidis, Streptomyces collinus, Streptomyces coeruleoprunus, Salmonella enteritidis, Vibrio parahaemolyticus, and Malassezia pachydermatis was most efficiently inhibited by ethyl acetate fraction of roselle flower extract as determined by a paper disc method and growth inhibition curves. In addition, the ethyl acetate fraction, water fraction and butanol fraction showed free radical scavenging and DNA cleavage inhibition activities. These results demonstrate that roselle flowers hold antimicrobial and antioxidative activities against skin microorganisms and oxidants.

Molecular Cloning and M13 Subcloning of Genes Encoding Catechol Dioxygenases

  • Kim, Young-Soo;Choi, Bong-Soo;Min, Kyung-Rak
    • Archives of Pharmacal Research
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    • v.15 no.1
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    • pp.48-51
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    • 1992
  • Achromobacter xylosoxidans KF701 and Pseudomonas putida (NAH7) were significantly different in degradative capability of aromatic compounds including benzoates, biphenyls, and naphthalene. However, both of the bacterial strains can grown on catechol as the sole carbon and energy source. Catechol 2, 3-dioxygenase gene for naphthalene oxidation or biphenyl oxidation was cloned into Escherichia coli HB 701. A E. coli HB 101 clone containing catechol 2, 3-dioxygenase gene from P. putida (NAH7) contains a recombinant plasmid with 3.60kb pBR322 and 6-kb insert DNA. Another E. coli HB101 clone containing catechol 2, 3-dioxygenase gene from A. xylosoxidans KF 701 has a recombinant plasmid with 4.4kb pBR322 and 10-kb insert DNA. Physical maps of the recombinant plasmids were constructed, and catechol 2, 3-dioxygenase gene in the recombinant plasmide was further localized and subcloned int M13. The cloned-catechol 2, 3-dioxygenase game products were identified as yellow bands on nondenaturaing polyacrylamide gel after electrophoresis followed by activity staining with catechol solution.

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Antioxidant Activities of Garlic (Allum sativum L.) with Growing Districts

  • Lee, Eun-Jung;Kim, Kwan-Soo;Jung, Hye-Young;Kim, Deuk-Ha;Jang, Hae-Dong
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.123-130
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    • 2005
  • Hydrogen-donating capacity, scavenging activity of reactive oxygen including superoxide anion radical and hydrogen peroxide, metal-chelating activity, and reducing power of garlic extracts were investigated. All tested garlic extracts exhibited in vitro antioxidant activities, with Uiseong extract showing highest hydrogen-donating and hydrogen peroxide-scavenging activities, and reducing power, followed by Seosan and Samchek extracts, in proportion to total thiosulfinate contents. Higher scavenging activity of superoxide anion radical was observed in Uiseong than Seosan and Samchek extracts. Metal-chelating activity increased in order of Uiseong < Seosan < Samchek, showing inverse relations to total thiosulfinate content. Garlic extracts of Uiseong and Seosan showed weak prooxidant activities and that of Samchek showed strong antioxidant activity against $Cu^{+2}$-induced human LDL oxidation. Protective effects on peroxyl and hydroxyl radical-induced DNA strand damages were observed in all tested garlic cloves. These results indicate growing conditions of garlic cloves affect total thiosulfinate content and antioxidant activities.

LITHOAUTOTROPHIC NITROGEN REMOVAL WITH ANAEROBIC GRANULAR SLUDGE AS SEED BIOMASS AND ITS MICROBIAL COMMUNITY

  • Ahn, Young-Ho;Lee, Jin-Woo;Kim, Hee-Chul;Kwon, Soo-Youl
    • Environmental Engineering Research
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    • v.11 no.4
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    • pp.173-180
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    • 2006
  • Autotrophic nitrogen removal and its microbial community from a laboratory scale upflow anaerobic sludge bed reactor were characterized with dynamic behavior of nitrogen removal and sequencing result of molecular technique (DNA extraction, PCR and amplification of 16S rDNA), respectively. In the experiment treating inorganic wastewater, the anaerobic granular sludge from a full-scale UASB reactor treating industrial wastewater was inoculated as seed biomass. The operating results revealed that an addition of hydroxylamine would result in lithoautotrophic ammonium oxidation to nitrite/nitrate, and also hydrazine would play an important role for the success of sustainable nitrogen removal process. Total N and ammonium removal of 48% and 92% was observed, corresponding to nitrogen conversion of 0.023 g N/L-d. The reddish brown-colored granular sludge with a diameter of $1{\sim}2\;mm$ was observed at the lower part of sludge bed. The microbial characterization suggests that an anoxic ammonium oxidizer and an anoxic denitrifying autotrophic nitrifier contribute mainly to the nitrogen removal in the reactor. The results revealed the feasibility on development of high performance lithoautotrophic nitrogen removal process with its microbial granulation.

Antioxidative Properties of Sachil-Tang Extract

  • Yi, Hyo-Seung;Moon, Jin-Young
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.4
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    • pp.872-882
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    • 2009
  • Sachil-Tang (SCT) has been traditionally used as a prescription of spasm of the esophagus by stress, pectoralgia and oppressive feeling of the chest in Oriental medicine. This study was carried out to investigate the antioxidant activities of the ethanol extract of SCT and its inhibitory effect on intracellular oxidation and vascular cell adhesion molecule-1 expression in human umbilical vein endothelial cells (HUVECs) using various methods. The SCT extract showed a strong inhibitory effect on free radical generating model systems, including DPPH radical, superoxide anions, hydroxyl radical, peroxynirite and nitric oxide. Besides, the SCT extract exhibited a strong inhibitory effect on lipid peroxidation in rat liver homogenate induced by $FeCl_2$-ascorbic acid, and protected plasmid DNA against the strand breakage in a Fenton's reaction system. The SCT extract also inhibited copper-mediated oxidation of human low-density lipoprotein (LDL), and repressed relative electrophoretic mobility of LDL. Furthermore, the SCT extract protected intracellular oxidation induced by various free radical generators and inhibited expression of vascular cell adhesion molecule-1 (VCAM-1) in HUVECs. These results suggest that SCT can be an effective natural antioxidant and a possible medicine of atherosclerosis.

Antioxidant Effects of 2,3,6-tribromo-4,5-dihydroxybenzyl Methyl Ether (TDB) from the Red Alga, Symphyocladia latiuscula

  • Park, Hye-Jin;Kim, Hyeung-Rak;Choi, Jae-Sue
    • Fisheries and Aquatic Sciences
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    • v.12 no.2
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    • pp.86-89
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    • 2009
  • 2,3,6-Tribromo-4,5-dihydroxybenzyl methyl ether (TDB) from the methanolic extract of the red alga Symphyocladia latiuscula exhibits major antioxidant activity. In this study, the activity of TDB against oxidative damage in deoxyribose and DNA was investigated in vitro for potential applications in preventing mutagenesis caused by DNA damage. TDB inhibited the oxidation of deoxyribose at concentrations of up to $1{\mu}g$/mL in the presence of $Fe^{+3}$-EDTA/$H_2O_2$. Furthermore, TDB showed no pro oxidant activity as determined by absence of the reduction of bleomycin-$Fe^{+3}$ to bleomycin-$Fe^{+2}$, which leads to DNA damage. Based on these results, TDB demonstrated considerable antioxidant activity without prooxidant properties.

The Molecular Mechanism of Safrole-induced DNA Adducts and its Role to Oral Carcinogenesis

  • Liu, Tsung-Yun
    • Environmental Mutagens and Carcinogens
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    • v.23 no.3
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    • pp.99-102
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    • 2003
  • IARC classified areca quid as a human carcinogen. Areca quid chewed in Taiwan includes Piper betle inflorescence, which contains high concentrations of safrole (15 mg/fresh weight). Safrole is a documented rodent hepatocarcinogen, and chewing areca quid may contribute to human exposure (420 $\mu$m in saliva). The carcinogenicity of safrole is mediated through 1'-hydroxysafrole formation, followed by sulfonation to an unstable sulfate that reacts to form DNA adducts. Using human liver microsomes and Escherichia coli membranes expressing bicistronic human P450s, CYP2E1 and CYP2C9 were identified as the main P450s involved in the activation of safrole. We have demonstrated the presence of stable safrole-dGMP adducts in human oral tissues following areca quid chewing using $^{32}$ P-postlabeling and HPLC mass spectrometry methods. By studying 88 subjects with a known AQ chewing history and 161 matched controls, we have demonstrated that the presence of safrole-DNA adducts in peripheral blood cells was correlated to AQ chewing, and CYP2E1 seemed to play an important role in the modulation of safrole-DNA adduct formation. We have also shown that safrole can form stable safrole-DNA adducts as well as oxidative damages in rodent liver. However, the stable safrole-DNA adducts may represent a more significant initial lesion as compared to the rapidly repaired safrole-induced 8-hydroxy-2'-deoxyguanosine. This oxidative DNA damage is mediated through the formation of hydoryxchavicol, the major safrole metabolite in human urine. Hydroxychavicol may have gone through two-electron oxidation to the o-quinone; then via one-electron reduction to semiquinone radicals to generate oxidative DNA damage. However, these reactive metabolites can be efficiently conjugated by GSH. These data suggest that safrole may contribute to the initiation of oral carcinogenesis through safrole-DNA adduct and not oxidative DNA damage. In addition, CYP2E1 may modulate this adduct formation.

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