• 제목/요약/키워드: SCGE

검색결과 48건 처리시간 0.028초

DNA damage와 Apoptosis를 정량화하는 단세포전기영동법 (Single Cell Gel Electrophoresis (comet assay) to Detect DNA Damage and Apoptosis in Cell Level)

  • 류재천;김현주;서영록;김경란
    • 한국환경성돌연변이발암원학회지
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    • 제17권2호
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    • pp.71-77
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    • 1997
  • The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

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Application of the SCGE Assay for Detecting Induced DNA Damage in Plant Leaves

  • Kim, Jin Kyu;Song, Hi Sup;Kim, Do Young;Gichner, Tomas
    • 한국환경생물학회:학술대회논문집
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    • 한국환경생물학회 2003년도 학술대회
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    • pp.68-73
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    • 2003
  • The possibility of using the alkaline protocol of the single cell gel electrophoresis (SCGE) assay as a method for detecting induced DNA damage has been studied for six major plants. The EMS was applied as a model genotoxic agent on young excised leaves of the tested crops for 18 h at 26$^{\circ}C$ in the dark. With increasing concentrations of 0 to 10 mM EMS, the DNA damage, expressed by the averaged median tail moment values, significantly increased in nuclei of all plants studied. As the results, no correlation between the diameter of nuclei and sensitivity to EMS treatment was observed. The data obtained demonstrate the feasibility of using the SCGE assay for detecting induced DNA damage in plants.

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감마선 조사가 커피박 추출물의 색도 및 생리활성에 미치는 영향 (Effect of gamma irradiation on the color values and physiological properties of spent coffee ground extraction)

  • 송하연;김혜민;김우식;양미소;변의홍;장범수;최대성;변의백
    • 한국식품과학회지
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    • 제49권5호
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    • pp.544-549
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    • 2017
  • 본 연구는 폐자원인 커피박의 산업적 이용도를 증진시키기 위하여 감마선 조사를 이용하여 커피박 추출물의 색도 개선에 관하여 평가하기 위해 수행되었다. 커피박 추출물로부터 유용성 성분을 얻기 위하여 70% 에탄올로 추출한 후 감마선 조사를 한 결과 30, 50 kGy의 조사선량에서 커피박 추출물의 색도가 개선된 것을 확인하였다. 30, 50 kGy로 감마선 조사된 커피박 추출물에서 성분변화가 일어났지만, 항산화능력과 미백활성능에는 0 kGy 커피박 추출물 처리군과 비교했을 때 유의적인 변화가 없는 것을 확인하였다. 이는 다른 성분들이 감마선 조사에 의해 감소하였지만, 퀸산이 상당량 증가함으로써 성분변화로 인한 커피박 추출물의 생리활성능 감소를 보완한 것으로 보이며, 이상의 결과로 감마선 조사가 커피박 추출물의 진한 갈색도를 개선함으로써 커피박 추출물이 화장품 소재로 활융될 수 있는 부가가치를 부여 할 수 있으며, 이러한 폐기물의 재활용을 통해 커피박 폐기로 인한 환경문제 및 경제적 문제점을 개선하는데 기여할 수 있을 것으로 판단된다.

Single-cell Gel Electrophoresis Assay에 의한 대합에서의 In Vivo 유전독성 평가 (Use of the In Vivo Single-cell Gel Electrophoresis Assay for Evaluating Genotoxicity in Clam)

  • 김일용;현창기
    • Toxicological Research
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    • 제20권3호
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    • pp.225-232
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    • 2004
  • The suitability of the single cell gel electrophoresis (SCGE) assay as a test for the monitoring of genotoxicity of aquatic environment was evaluated. The SCGE assay was employed to detect DNA damage induced in clam (Spisula sachalinensis) exposed to a direct mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) or an indirect mutagen, benzo[a]pyrene (B[a]P). The cells of gill and digestive glands were isolated from clam by homogenization, which was the optimized cell dissociation method, and the level of DNA damage was assessed and expressed as mean tail length. In the gill cells, significant dose- and time-dependent increase was observed in the mean tail length at the concentration from 0.01 to 0.5 ppm MNNG for 96 h. The linear correlation between relative dam-age index (RDI) values was suggested to provide criteria of genotoxicity monitoring for direct acting mutagen. The dose- and time-dependent responses of the digestive glands cells were less sensitive than those of the gill cells. In contrast, the genotoxic response resulting from the exposure of 0.01~1.0 ppm B[a]P to clam revealed a higher sensitivity in the digestive glands cells than the gill cells. The comparison between the time profiles of genotoxic responses in clam and carp, the latter had been obtained in our previous study, indicated that the metabolism of genotoxic compounds in the two aquatic organisms were quite different each other. We conclude that the SCGE assay has the potential as a screening test for routine genotoxicity monitoring of aquatic organisms because of its higher sensitivity and simplicity.

Evaluation of Protective Effects of Houttuynia cordata on H2O2-Induced Oxidative DNA Damage Using an Alkaline Comet Assay in Human HepG2 Cells

  • Hah, Dae-Sik;Kim, Chung-Hui;Ryu, Jae-Doo;Kim, Eui-Kyung;Kim, Jong-Shu
    • Toxicological Research
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    • 제23권1호
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    • pp.25-31
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    • 2007
  • To evaluate the protective effect of Houttuynia cordata on hydrogen peroxide-induced oxidative DNA damage in HepG2 cell line, we used an alkaline single-cell gel electrophoresis (SCGE; comet assay). The DNA damage was analyzed by tail moment (TM) and tail length (TL), which used markers of DNA strand breaks in SCGE. The $100{\mu}g/ml$ of methanolic extract of Houttuynia cordata root showed significant protective effects (p < 0.01) against hydrogen peroxide-induced DNA damage in HepG2 cells and increased cell viability against hydrogen peroxide. The results of this study indicate that Houttuynia cordata root methanol extract acts as a potential antioxidant, and exhibits potential anticancer properties, which may provide a clue to find applications in new pharmaceuticals for oxidative stability.

The Regional Economic Impacts of Taiwan High Speed Rail

  • Huang, Hank C.C.;Hsu, Tao Hsin;Lin, Cynthia M.T.
    • International Journal of Railway
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    • 제1권1호
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    • pp.12-19
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    • 2008
  • Starting her business operation on January 5 2007, Taiwan High Speed Rail(THSR) shapes a new time-space frame for Taiwan western corridor, where more than 90% of national population lives around and more than 95% gross domestic product created from. Comparing with the four-hour traveling time by highway before 2007, THSR reduces the time required to one and half hours from Taipei to Kaohsiung. It will not only benefit the communication along the island from north to south, but also change the location advantages/disadvantages for all cities in these regions. Therefore, this paper establishes a spatial computable general equilibrium model(SCGE Model) to simulate the economic effect of High Speed Rail(HSR). This SCGE model divides Taiwan economy into fifteen geographic regions and thirteen industries. Each region has three sectors: household sector, transportation sector, and industries sector. Following the behavior function of economic theories, the general equilibrium can be achieved simultaneously. Thus, gross regional product (GRP), capital formation, employment income and welfare/utility level can be all observed by calculating the different economic result between cases with-/ without-HSR. Besides, this model presents the social welfare benefit from HSR operation, the polarization phenomenon among regions and within certain region, unbalance distribution of welfare along the HSR line, and industries development divergence among regions etc. These major findings should be useful for regional development policy making.

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The Regional Economic Impacts of Taiwan High Speed Rail

  • Huang, Hank C.C.;Hsu, Tao Hsin;Lin, Cynthia M.T.
    • 한국철도학회:학술대회논문집
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    • 한국철도학회 2007년도 추계학술대회 논문집
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    • pp.1896-1912
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    • 2007
  • Starting her business operation on January 5 2007, Taiwan High Speed Rail (THSR) shapes a new time-space frame for Taiwan western corridor, where more than 90% of national population lives around and more than 95% gross domestic product created from. Comparing with the four-hour traveling time by highway before 2007, THSR reduces the time required to one and half hours from Taipei to Kaohsiung. It will not only benefit the communication along the island from north to south, but also change the location advantages/disadvantages for all cities in these regions. Therefore, this paper establishes a spatial computable general equilibrium model (SCGE Model) to simulate the economic effect of High Speed Rail (HSR). This SCGE model divides Taiwan economy into fifteen geographic regions and thirteen industries. Each region has three sectors: household sector, transportation sector, and industries sector. Following the behavior function of economic theories, the general equilibrium can be achieved simultaneously. Thus, gross regional product (GRP), capital formation, employment income and welfare/utility level can be all observed by calculating the different economic result between cases with-/ without-HSR. Besides, this model presents the social welfare benefit from HSR operation, the polarization phenomenon among regions and within certain region, unbalance distribution of welfare along the HSR line, and industries development divergence among regions etc. These major findings should be useful for regional development policy making.

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환경재해에 관한 생물정보로서의 이온화 방사선과 살충제의 인체 위해성 비교 연구 (Comparative Study on Human Risk by Ionizing Radiation and Pesticide as Biological Information about Environmental Disaster)

  • 김진규;현성희
    • Journal of Radiation Protection and Research
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    • 제26권4호
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    • pp.385-392
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    • 2001
  • 환경독성물질이나 생물 위해 요소의 환경내 준위가 일정 수준 이상일 경우 환경재해가 유발될 수 있다. 이온화 방사선의 산업적 의료적 이용이 점차 증가하고 있으며 병해충을 막기 위한 살충제 사용의 점진적 증가로 인해 이들에 의한 재해 가능성을 배제할 수 없다. 이들 재해 요인들에 의한 인체 위해도를 비교하기 위하여 단세포 겔 전기영동법 (SCGE)을 이용하여 사람 림프구 DNA 손상에 미치는 방사선과 살충제의 영향을 각각 평가하였다. 각기 다른 농도로 살충제를 10분간 처리한 림프구에 대한 SCGE 분석을 실시하였고 또한 $0{\sim}2.0Gy$의 방사선을 조사한 림프구에 대한 SCGE 분석을 실시하여 DNA 손상도를 평가하였다. DNA 손상도는 감마선에 대해서 뚜렷한 선량-반응 관계를 나타내었을 뿐 아니라 살충제에 대해서도 명확한 농도-반응 관계를 나타내었다. 파라치온은 농업권장 사용농도인 $1mg{\ell}^{-1}$에서도 림프구에 대해 강한 유전독성을 나타내는데 이러한 유전독성은 0.1 Gy의 감마선에 의해 유발되는 DNA 손상에 상응하며 $2mg{\ell}^{-1}$의 파라치온은 임상적 증상을 야기할 가능성이 있는 전신 외부피폭 방사선량인 0.25 Gy에 상응하는 세포손상을 유발하였다. 이와 같은 연구를 통해 방사선과 살충제의 인체 위해도를 비교할 수 있는 실험적 자료와 환경재해 예방에 필요한 생물정보를 제공할 수 있다.

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인삼지용성성분의 지질과산화 및 산화적 DNA손상에 대한 억제효과 (Protective effect of Ginseng Petroleum Ether Extract Against Lipid Peroxidation and Oxidative DNA Damage)

  • 허문영
    • 한국식품위생안전성학회지
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    • 제12권4호
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    • pp.315-320
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    • 1997
  • Panax ginseng C.A. Meyer has been extensively used in the traditional oriental medicine as a restorative, tonic and prophylatic agent. This study was devised to develop a chemopreventive agent from panax ginseng to be able to suppress the genotoxicity and oxidative damage by ractive oxygen species, which are involved with cancer or aging. Ginseng petroleum ether extract (GPE) and one of its fraction, P2, showed an antioxidative effect on the lipid peroxidiphenyl-2-picryl hydrazil (DppH) radical generation. They also showed the suppressive effect of H2O2 or KO2 induced DNA damage by single cell gel electrophoresis (SCGE). Results from our study indicate that GPE and P2 are capable of protecting lipid peroxidation, and oxidative DNA damage. Therefore, GPE and P2 may be useful chempreventive agents which are involved with cancer and aging.

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Free Radical Involvement in the DNA Damaging Activity of Fumonisin Bl

  • Lee, Wan-Hee;Lee, Kil-Soo
    • Toxicological Research
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    • 제17권4호
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    • pp.249-253
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    • 2001
  • Fumonisin B1, a mycotoxin, is thought to induce esophageal cancer in humans and apoptosis in animal cells by inhibiting ceramide synthase. Dumonisin Bl may also generate reactive oxygen species directly or indirectly, leading to DNA damage and lipid peroxidation. In this study, a DNA fragmentation assay, dichlorofluorescein (DCF) analysis, and single cell gel electrophoresis (SCGE) were used to investigate the involvement of cellular free radicals, specifically hydrogen peroxide, in the DNA damaging activity of fumonisin B1. From an in vitro DNA fragmentation assay, E. coli DNA, damage by fumonisin Bl was increased by the addition of superxide dismutase (SOD) and decreased by catalase. SCGE and DCF analysis in vivo showed that the nuclear DNA damage and intracellular free radicals in cultured rat hepatocytes treated with fumonisin B1 were increased with the concentration of fumonisin Bl . DNA damage and free radical generation were inhibited by the addition of catalase. Fumonisin Bl , in the presence of SOD, produces hydrogen peroxide causing oxidative DNA damage and protein malfunction, leading to genotoxicity and cytotoxicity of the toxin.

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