• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.206-212
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• 1995

A strain BL-29, which produces a extracellular lytic enzyme on E. coli was isolated from the soil. The strain was identified as belonging to the genus Bacillus sp. The lytic enzyme was purified to homogeneity by ion exchange chromatography and gel filtration. Specific activity of the purified enzyme was 28, 850 U/mg protein and yield of the enzyme was 5$%$. The purified enzyme showed a single band on SDS-PAGE and its molecular weight was estimated to be 31, 000 by SDS-polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum temperature and pH were $55^{\circ}C$ and pH 10.0, respectively. The enzyme was stable at $45^{\circ}C$ but enzyme activity was reduced by up to 50$%$ when the temperature was raised to $55^{\circ}C$ for 15 min. Stable range of pH was from 5.0 to 11.0. but Enzyme activity was inhibited by lead-acetate, mercuric chloride, ethylene glycol-bis-[$\beta$-aminoethyl ether]-N, N, $N^1, $N^1$-tetraacetic acid (EGTA), and ethylenediamine tetraacetic acid (EDTA), but not affected considerably by treatment with other chemical reagents.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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• pp.182-182
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• 2001

Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develope as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay and single cell gel electrophoresis (Comet) assay.(omitted)

• 한국환경과학회지
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• 제14권11호
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• pp.1071-1079
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• 2005

Using single cell gel electrophoresis, DNA single strand breaks were determined in various marine organisms. DNA damage on fish blood cells was detected to know whether there was a difference between Incheon, Pohang, Masan, and Tongyeong as a control site. Tongyeong showed the lowest DNA damage among the study areas. Mussels, transplanted to Masan Bay for one month, revealed high DNA damage at sites with high economical activity. In two weeks exposure of polychaete to Incheon sediments, higher DNA damage was detected in the sediment adjacent to Incheon harbor than open sea. These results suggested that the marine organism from the polluted area revealed a relatively high DNA damage. In addition, these areas might be contaminated with genotoxic compounds and comet assay was useful as a bioassay to detect DNA damage in marine organisms.

• 약학회지
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• 제51권4호
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• pp.239-245
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• 2007

In order to evaluate the genotoxicity of airborne particulate matter extracted with dichloromethane (APE), the rat microsome mediated (S-9) or DNA repair enzyme treated Comet assays were performed using the single cell gel electrophoresis in A549 human lung carcinoma cells. It was found that the cells interacting with APE showed more DNA single-strand breaks relative to untreated cells. The genotoxicity of APE was increased with the treatment of S-9 mixture. Microsome mediated DNA damage was inhibited by CYP1Al inhibitor, quercetin. The APE also showed oxidative DNA damage evaluated by endonuclease III treatment. Oxidative DNA damage of APE was inhibited by antioxidants such as vita- min C and Trolox. We also found that the vegetables or fruits extract may reduce APE-induced genotoxicity by their anti- oxidant activity and CYP1A1 inhibition.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2001년도 International Symposium on Dietary and Medicinal Antimutgens and Anticarcinogens
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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)

• 한국식품영양과학회지
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• 제29권5호
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• pp.843-848
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• 2000

콩류의 방사선 조사 여부를 DNA comet assay로 확인하였다. 대두, 강남콩, 그리고 팥을 각각 0.1, 0.3, 0.5, 0.7, 그리고 1.0 kGy의 저선량으로 조사하고 비조사 시료와 조사시료간의 DNA 손상정도를 현미경으로 육안 검사 및 comet의 tail 길이를 측정하여 상호 비교하였다. 모든 시료에서 비조사 시료보다 조사시료의 tail 길이가 더 길었으며 조사 선량이 증가할수록 tail 길이가 길게 나타났다. 비조사 시료에서도 손상된 세포의 comet 모양의 핵이 관찰되기는 하였으나 대부분 비손상된 세포의 원형모양의 핵이 관찰되었으며 조사된 시료에서는 comet 모양의 핵이 주로 관찰되는 경향이어서 비조사 시료와 조사 시료간에 comet 양상을 비교할 수 있었다. 한편 100개의 세포를 무작위로 선택하여 comet tail 길이를 측정하고 통계분한 결과, 비조사 시료와 조사시료간의 유의적인 차이가 있었고 조사선량이 증가할수록 tail 길이가 유의적으로 증가함을 확인하였다. DNA comet assay로 간단하고 비교적 저렴한 비용을 짧은 시간 내에 콩류의 방사선 조사 여부를 확인할 수 있었다.

• 동아시아식생활학회지
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• 제4권3호
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• pp.59-65
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• 1994

이 실험에서는 소의 유즙에 있는 성장인자를 세포배양을 통해 확인하고 크로마토그래피를 사용하여 분리 정제하였다. 5%의 우유를 포함하는 배지를 FBS(fetal bovine serum)를 포함한 배지와 비교했을때 Africal green monkey kidney cell의 성장 촉진 효과가 비슷하게 나타났으며, 우유의 성장인자를 분리 정제한 결과 FBS배지에서보다 2,000배 이상의 성장 촉진 효과가 나타났다. 이 성장인자는 hydrophobic column(phenyl-sepharose)과 gel-filtration column(sephadex G-100)을 사용하여 분리했으며 그 결과 milk-deriven growth factor는 phenyl-sepharose에서 50% ethylene glycol step에서 용출되므로 hydrophobic protein임이 증명되었고 size exclusion column chromatography로 부터 분자량이 100,000-15,0000 범위의 고분자 물질임이 밝혀졌다. 또한 우유의 성장인자는 매우 다양하며 그중 본 실험으로부터 정제된 물질이 주요 성장인자로 밝혀졌다.

• Toxicological Research
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• 제29권1호
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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.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
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• pp.61-61
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• 1996

Taxol is used as cancer therapeutic agent. It has been known as weak posotive of chromosome aberration assay in vitro in our previous results (Ryu et al., 1996) and potent clastogens in the mouse bone marrow micronucleus (Tinwell and Ashby, 1994). We performed microgel electrophoresis to determine the effect of taxol and it's precursor 10-deacetyl baccatin III(DAB) on DNA. Microgel electrophoresis is useful, rapid, simple, visual, and sensitive technique for measuring DNA breakage and repair mechanisms in mammalian 근ells. The range of concentration used for taxol were 854, 427, 213.5, 106.8, 53.4 Ug/ml, for DAB 910 ,455, 227.5 U9/ml, Cell viability always exceed 85%. We analyzed the results by using the special software of image analyzer for this comet assay (Komet 3.0). By using this image analyzer software , we can get the result as the tail moment ((mean of tail length - mean of head lengh) x tail%DNA/100). A slight increase in DNA migration was observed for taxol at the concentration of 854 Ug/m4 in the absence of S9 mixture. No increased DNA migration was observed after treatment with DAB.

• 약학회지
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• 제54권6호
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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.