• Title/Summary/Keyword: SCGE

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

  • 류재천;김현주;서영록;김경란
    • Environmental Mutagens and Carcinogens
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    • v.17 no.2
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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
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2003.11a
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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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Effect of gamma irradiation on the color values and physiological properties of spent coffee ground extraction (감마선 조사가 커피박 추출물의 색도 및 생리활성에 미치는 영향)

  • Song, Ha-Yeon;Kim, Hye-Min;Kim, Woo Sik;Yang, Mi-So;Byun, Eui-Hong;Jang, Beom-Su;Choi, Dae Seong;Byun, Eui-Baek
    • Korean Journal of Food Science and Technology
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    • v.49 no.5
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    • pp.544-549
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    • 2017
  • The spent coffee grounds (SCG) are considered valuable by-products because they contain various bioactive compounds. The SCG extraction (SCGE) was irradiated at doses ranging between 30 and 50 kGy. The deep dark-brown color of SCGE was changed to a bright yellow color by gamma irradiation. The content of the bioactive compounds of gamma-irradiated SCGE was analyzed by high-performance liquid chromatography. Interestingly, the content of quinic acid was increased by gamma irradiation, whereas other compounds were decreased. Although the contents of bioactive compounds were changed by gamma irradiation, the biological activities (radical scavenging activity and whitening effects) of SCGE were unaffected. Our findings suggest that gamma irradiation can effectively improve the color values of SCGE without the loss of biological activities. Consequently, gamma irradiation can be a useful tool for improving the utilization of SCGE in the cosmetic industry.

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

  • Kim Il-Yang;Hyun Chang-Kee
    • Toxicological Research
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    • v.20 no.3
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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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    • v.23 no.1
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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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    • v.1 no.1
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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.
    • Proceedings of the KSR Conference
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    • 2007.11a
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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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Comparative Study on Human Risk by Ionizing Radiation and Pesticide as Biological Information about Environmental Disaster (환경재해에 관한 생물정보로서의 이온화 방사선과 살충제의 인체 위해성 비교 연구)

  • Kim, Jin-Kyu;Hyun, Soung-Hee
    • Journal of Radiation Protection and Research
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    • v.26 no.4
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    • pp.385-392
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    • 2001
  • Environmental risk factors such as ionizing radiations, heavy metals, and pesticides can cause environmental disasters when they exist in excess. The increases in use of ionizing radiation and agricultural pesticide are somewhat related to the possibility of the disaster. The risk of radiation and pesticide was evaluated by means of the single cell gel electrophoresis (SCGE) assay on the human blood lymphocytes. The lymphocytes were irradiated with $0{\sim}2.0Gy$ of $^{60}Co$ gamma ray. Another groups of lymphocytes were exposed to various concentrations of parathion. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed a clear dose- or concentration-response relationship. Parathion of a recommended concentration for agricultural use ($1mg {\ell}^{-1}$ ) has a strong cytotoxic effect on lymphocytes, which is equivalent to damage induced by 0.1 Gy of ${\gamma}$-ray. Furthermore, $2mg{\ell}^{-1}$ of parathion can give rise to DNA damage equivalent to that induced by 0.25 Gy at which the radiation-induced damage can start to develop into clinical symptoms. The comparative results of this study can provide an experimental basis and biological information for the prevention of environmental disaster.

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

  • 허문영
    • Journal of Food Hygiene and Safety
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    • v.12 no.4
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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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    • v.17 no.4
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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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