• Title/Summary/Keyword: DNA protection

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Protection of ROS-induced cytotoxicity and DNA damage by the extract of Alpinia of ficinarum (양강추출물의 활성산소종 유도 세포독성과 DNA 손상에 대한 방어효과)

  • 이승철;신경승;허문영
    • Journal of Food Hygiene and Safety
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    • v.17 no.2
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    • pp.106-116
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    • 2002
  • The 70% ethanol extract of Alpinia officinarum and its major flavonoid, galangin showed strong antioxidative effect on the lipid peroxidation of ethyl linolate with Fenton's reagent and free radical scavenging effect to DPPH radical generation. However, they did not reveal any pro-oxidant effect on bleomycin-Fe(III) dependent DNA degradation. They also showed the protective effect against $H_2O$$_2$, KO$_2$ or UV-induced cytotoxicity in mammalian cells. They also showed the suppressive effect of DNA damage induced by $H_2O$$_2$ or KO$_2$ with dose-dependent manner in single cell gel electrophoresis(SCGE) assay. On the other hand, they have an anticlastogenic effect against adriamycin-induced micronucleated reticulocyte in peripheral blood of mice. These results suggest that the mechanism of inhibition by 70% ethanol extract of Alpinia officinarum and galangin against reactive oxygen species (ROS)-induced genotoxicity or cytotoxicity is due, at least partly, to their antioxidative and free radical scavenging properties without pro-oxidant effect. All these results indicate that 70% ethanol extract of Alpinia officinarum and galangin may be useful for protection against ROS-induced cytotoxicity and DNA damage.

Protection of Peroxynitrite-Induced DNA Damage by Dietary Antioxidants

  • Moon Hye-Kyung;Yang Eun-Sun;Park Jeen-Woo
    • Archives of Pharmacal Research
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    • v.29 no.3
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    • pp.213-217
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    • 2006
  • The present study was undertaken to test the hypothesis that dietary antioxidants protect DNA damage induced by peroxynitrite, a potent physiological inorganic toxin. The present study showed that dietary antioxidants such as (-)-epigallocatechin gallate, quercerin, rutin, resveratrol, and ursolic acid inhibit single strand breaks in supercoiled plasmid DNA induced by 3-morpholinosydnomine N-ethylcarbamide (SIN-1), a generator of peroxynitrite through the reaction between nitric oxide and superoxide anion. The formation of 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in calf thymus DNA by SIN-1 was also inhibited by dietary antioxidants. When U937 cells were incubated with 1 mM SIN-1 bolus, a significant increase of 8-OH-dG level was observed. However, oxidative DNA damage was significantly lower in the cells pre-treated with dietary antioxidants when cells were exposed to SIN-1.

Evaluation of protective effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes in the single cell gel electrophoresis assay (단세포 겔 전기영동법을 이용한 사람 림프구 DNA 손상에 대한 복숭아씨 추출물의 방사선 방어효과 평가)

  • Kim, Jin-Kyu;Park, Tae-Won;Lee, Chang-Joo;Chai, Young-Gyu
    • Journal of Radiation Protection and Research
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    • v.24 no.2
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    • pp.93-99
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    • 1999
  • The alkaline single-cell gel electrophoresis (SCGE) assay, called the comet assay, has been applied to the detection of DNA damage from a number of chemical and biological factors in vivo and in vitro. The comet assay is a novel method to assess DNA single-strand breaks, alkali-labile sites in individual cells. The effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes was evaluated by the SCGE assay. The lymphocytes, with or without pretreatment of the extracts, were exposed to 0, 0.1, 0.3, 0.5, 1.0 and 2.0 Gy of $^{60}Co$ gamma ray. Significantly increased tail moment, which was a marker of DNA strand breaks in the comet assay, showed an excellent dose-response relationship. The treatment of the peach kernel extracts reduced the DNA damage by 30 % in irradiated groups as compared to that in non-treated control groups. The result indicates that the extracts shows radioprotective effect on lymphocyte DNA when assessed by the comet assay.

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Tomato Yellow Leaf Curl China Virus Impairs Photosynthesis in the Infected Nicotiana benthamiana with βC1 as an Aggravating Factor

  • Farooq, Tahir;Liu, Dandan;Zhou, Xueping;Yang, Qiuying
    • The Plant Pathology Journal
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    • v.35 no.5
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    • pp.521-529
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    • 2019
  • Tomato yellow leaf curl China virus is a species of the widespread geminiviruses. The infection of Nicotiana benthamiana by Tomato yellow leaf curl China virus (TYLCCNV) causes a reduction in photosynthetic activity, which is part of the viral symptoms. ${\beta}C1$ is a viral factor encoded by the betasatellite DNA ($DNA{\beta}$) accompanying TYLCCNV. It is a major viral pathogenicity factor of TYLCCNV. To elucidate the effect of ${\beta}C1$ on plants' photosynthesis, we measured the relative chlorophyll (Chl) content and Chl fluorescence in TY-LCCNV-infected and ${\beta}C1$ transgenic N. benthamiana plants. The results showed that Chl content is reduced in TYLCCNV A-infected, TYLCCNV A plus $DNA{\beta}$ (TYLCCNV A + ${\beta}$)-infected and ${\beta}C1$ transgenic plants. Further, changes in Chl fluorescence parameters, such as electron transport rate, $F_v/F_m$, NPQ, and qP, revealed that photosynthetic efficiency is compromised in the aforementioned N. benthamiana plants. The presense of ${\beta}C1$ aggravated the decrease of Chl content and photosynthetic efficiency during viral infection. Additionally, the real-time quantitative PCR analysis of oxygen evolving complex genes in photosystem II, such as PsbO, PsbP, PsbQ, and PsbR, showed a significant reduction of the relative expression of these genes at the late stage of TYLCCNV A + ${\beta}$ infection and at the vegetative stage of ${\beta}C1$ transgenic N. benthamiana plants. In summary, this study revealed the pathogenicity of TYLCCNV in photosynthesis and disclosed the effect of ${\beta}C1$ in exacerbating the damage in photosynthesis efficiency by TYLCCNV infection.

Lysinabacillus fusiformis and Paenibacillus alvei Obtained from the Internal of NasutitermesTermites Revealed Their Ability as Antagonist of Plant Pathogenic Fungi

  • Fitriana, Yuyun;Tampubolon, Desi Apriani Teresa;Suharjo, Radix;Lestari, Puji;Swibawa, I Gede
    • The Plant Pathology Journal
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    • v.38 no.5
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    • pp.449-460
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    • 2022
  • This study was performed to reveal phenotypic characters and identity of symbiont bacteria of Nasutitermes as well as investigate their potential as antagonist of plant pathogenic fungi. Isolation of the symbiont bacteria was carried out from inside the heads and the bodies of soldier and worker termite which were collected from 3 locations of nests. Identification was performed using phenotypic test and sequence of 16S ribosomal DNA (16S rDNA). Antagonistic capability was investigated in the laboratory against 3 phytopathogenic fungi i.e., Phytophthora capsici, Ganoderma boninense, and Rigidoporus microporus. Totally, 39 bacterial isolates were obtained from inside the heads and the bodies of Nasutitermes. All the isolates showed capability to inhibit growth of P. capsici, however, 34 isolates showed capability to inhibit growth of G. boninense and 32 isolates showed capability to inhibit growth of R. microporus. Two bacterial strains (IK3.1P and 1B1.2P) which showed the highest percentage of inhibition were further identified based on their sequence of 16S rDNA. The result showed that 1K3.1P strain was placed in the group of type strain and reference strains of Lysinibacillus fusiformis meanwhile 1B1.2P strain was grouped within type strain and reference strains Paenibacillus alvei. The result of this study supply valuable information on the role of symbiont bacteria of Nasutitermes, which may support the development of the control method of the three above-mentioned phytopathogenic fungi.

DNA Binding Specificity of Proteus mirabilis Transcription Regulator (Proteus mirabilis 전사 조절 단백질의 DNA 결합 특성)

  • Gang, Jong-Back
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.158-162
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    • 2011
  • Amino acid sequence alignment shows that $\underline{P}$roteus $\underline{m}$irabilis $\underline{t}$ranscription $\underline{r}$egulator (PMTR) has cystein sequence homology at metal binding domain to CueR (copper resistance) protein, which conserves two cysteins (Cys 112 and Cys 120 in PMTR). Gel shift assay revealed that PMTR protein bound to promoter region of Escherichia coli copA (copper-translocating P-type ATPase) and Proteus mirabilis atpase (putative copper-translocating P-type ATPase) genes except that of E. coli zntA (zinc-translocating P-type ATPase) gene. DNase I protection experiment indicated that PMTR protein protected the region over -35 box and close to -10 box. DNase I hypersensitive bases were shown at C and A bases of labeled template strand and at G and C bases of labeled non-template strand of DNA. These hypersensitive bases were appeared in other metalloregulatory proteins of MerR family, which suggests protein-induced DNA bending.

Mechanism of Sulfonylurea Herbicide Resistance in Broadleaf Weed, Monochoria korsakowii (광엽잡초 물옥잠의 Sulfonylurea 제초제에 대한 저항성 작용기작)

  • Park, Tae-Seon;Lhm, Yang-Bin;Kyung, Kee-Sung;Lee, Su-Heon;Park, Jae-Eup;Kim, Tae-Wan;Kim, Kil-Ung
    • The Korean Journal of Pesticide Science
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    • v.7 no.4
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    • pp.239-247
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    • 2003
  • This experiment was carried out to study the resistant mechanism of sulfonylurea(SU) herbicides to Monochoria korsakowii occurring in the rice fields of Korea. The activity of acetolactate synthase(ALS), absorption and translocation of $[^{14C}]$bensulfuron-methyl, and DNA sequence of ALS genes were studied. The apparent SU resiatance to Monochoria korsakowii was confirmed in greenhouse testes. Fresh weight accumulation$(GR_{50})$ in the resistant biotype was about 5- to 64-fold higher in the presence of six SU herbicides compared to the susceptible biotype. The ALS activity isolated from the resistant biotype to herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity$(I_{50})$ was 14- to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of $[^{14C}]$bensulfuron uptake and translocation. However, the DNA sequence from the resistant biotype differed from that of the susceptible biotype by single nucleotide substitution at three amino acid each in the middle region excluding the ends of ALS genes. We found three point mutations causing substitution of serine for threonine at amino acid 168, arginine for histidine at amino acid 189, and a aspartic acid for phenylalanine at amino acid 247, respectively, in the resistant biotype.

Molecular Characteristics of Phytophthora katsurae Using PCR-SSCP Analysis (PCR-SSCP 분석에 의한 Phytophthora katsurae의 분자생물학적 특성)

  • Lee, Sun-Keun;Jang, Ha-Na;Lee, Dong-Hyeon;Lee, Sang-Hyun;Lee, Sang-Yong;Lee, Jong-Kyu
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.169-176
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    • 2011
  • Phytophthora katsurae is the fungus responsible for chestnut ink disease. The objectives of this study were to determine if a single-strand conformation polymorphism (SSCP) analysis of rDNA-ITS region, elongation factor 1 alpha gene and ${\beta}$-tubulin gene could be used for rapid identification and genetic diversity of P. katsurae, and to assess the potential use of the SSCP technique as a diagnostic tool for P. katsurae. Each regions amplified by PCR using primers designed to overlap the genus Phytophthora were characterized for the Phytophthora species. PCR products were denatured and electrophoresed for SSCP analysis. P. katsurae isolates showed an unique pattern in SSCP analysis and were easily distinguished from other Phytophthora species used as the control. This indicates that SSCP analysis is an useful technique for distinguishing Phytophthora species from genetically close relatives, and show that the SSCP analysis of each region is an efficient detection tool for P. katsurae. But PCR-SSCP analysis of single-gene may have difficulty in distinguishing P. katsurae from other Phytophthora species. Therefore, PCR-SSCP analysis of multi-genes can be useful for rapid and effective identification of P. katsurae.

A Duplex PCR for Detection of Phytophthora katsurae Causing Chestnut Ink Disease (밤나무 잉크병균, Phytophthora katsurae의 검출을 위한 Duplex PCR)

  • Lee, Dong-Hyeon;Lee, Sun-Keun;Kim, Hye-Jeong;Lee, Sang-Hyun;Lee, Sang-Yong;Lee, Jong-Kyu
    • Research in Plant Disease
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    • v.18 no.2
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    • pp.73-79
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    • 2012
  • Phytophthora katsurae is a fungal pathogen responsible for chestnut ink disease. We designed two duplex primer sets (SOPC 1F/1R+KatI 3F/5R, SOPC 1-1F/1-1R+KatI 3F/5R) to detect P. katsurae. SOPC 1F/1R and SOPC 1-1F/1-1R primer pairs were designed for sequence characteristic amplification regions (SCAR) marker, and KatI 3F/5R primer pair was used for P. katsurae-specific primer designed from internal transcribed spacer (ITS) region. To assess the sensitivity of duplex PCR, genomic DNA was serially diluted 10-fold to make the final concentrations from 1 mg/ml to 1 ng/ml. The sensitivity for two primer sets were 1 ${\mu}g/ml$ and 100 ng/ml, respectively. To find detection limits for zoospores of P. katsurae, each zoospore suspension was serially diluted 10-fold to make the final concentrations from $1{\times}10^6$ to $1{\times}10^2$ cells/ml, and then DNA was extracted. The limits of detection for all of two primer sets were $1{\times}10^5$ cells/ml. All of two primer sets were specific to P. katsurae in PCR detection and did not produce any P. katsurae-specific PCR amplicons from other 16 Phytophthora species used as the control. This study shows that duplex PCR using two primer sets might be a useful tool for rapid and efficient detection of P. katsurae.