• Title/Summary/Keyword: DNA in tail

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Effects of lymphocyte DNA damage levels in Korean plant food groups and Korean diet regarding to glutathione S-transferase M1 and T1 polymorphisms (건강한 성인의 glutathione S-transferase M1과 T1 유전자 다형성에 따른 한식에서의 식물성 식품군과 한식의 DNA 손상 감소 효과)

  • Kim, Hyun-A;Lee, Min-Young;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.10-24
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    • 2017
  • Purpose: GST (glutathione S-transferase) M1 and T1 gene polymorphisms are known to affect antioxidant levels. This study was carried out to evaluate genetic susceptibility by measuring the effect of DNA damage reduction in the Korean diet by vegetable food according to GST gene polymorphisms using the ex vivo method with human lymphocytes. Methods: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups. A total of 84 foods, which constituted more than 1% of the total intake in each food group, were finally designated as a vegetable food in the Korean diet. The Korean diet applied in this study is the standard one-week meals for Koreans (2,000 Kcal/day) suggested by the 2010 Dietary Reference Intakes for Koreans. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. Results: In the Korean food group, the DNA damage protective effect of GSTM1 and GSTT1 was found to be greater in mutant type and wild-type, respectively. and the DNA damage protective effect according to the combined genotype of GSTM1 and GSTT1 was different depending on the food group. On the other hand, in Korean Diet, the DNA damage protective effect appeared to be larger in GSTM1 wild-type than in mutant type and was found to not be affected by GSTT1 genotype. Conclusion: These results can be used as basic data to demonstrate the superiority of the antioxidant function of Korean dietary patterns and food groups. Furthermore, it may be a starting point to begin research on customized antioxidant nutrition according to individual genes.

Detection of Irradiated Astragalus membranaeus Bunge and Havenia duzcis Thumb Using DNA Comet Assay

  • Yi, Jin-Hee ;Song, Kyung-Bin
    • Preventive Nutrition and Food Science
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    • v.7 no.3
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    • pp.323-326
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    • 2002
  • Ionizing radiation can be used to sanitize herbs contaminated by various microorganisms. However, health concerns related to irradiation damage to complex molecules in plants necessitate that methods be developed to monitor such damage. To elucidate DNA damage of herbs caused by irradiation, the DNA comet assay was used for Astragalus membranaceus Bunge and Havenia dulcis Thumb, irradiated at 1, 5, 7, and 10 kGy. With increasing irradiation doses, the tails of comets became longer with average tail length increasing from 17 (non-irradiated) to 124 (10 kGy) $\mu$m in Astragalus membranaceus Bunge. Above 7 kGy, some of the tails were separated from the heads of comets. Distribution patterns of the tail length of In comets selected randomly in the irradiated herbs were analyzed to quantify the DNA damage. These results clearly suggest that the DNA comet assay is an effective and inexpensive tool for the detection of irradiation damage to DNA in herbs.

Effects of 60-Hz Magnetic Fields on DNA Damage Responses in HT22 Mouse Hippocampal Cell Lines

  • Mun, Gil-Im;Lee, Seungwoo;Kim, Nam;Lee, Yun-Sil
    • Journal of electromagnetic engineering and science
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    • v.15 no.3
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    • pp.123-128
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    • 2015
  • Previously, we investigated extremely low-frequency magnetic fields (ELF-MFs) on diverse DNA damage responses, such as phosphorylated H2AX (${\gamma}H2AX$), comet tail moments, and aneuploidy production in several non-tumorigenic epithelial or fibroblast cell lines. However, the effect of ELF-MF on DNA damage responses in neuronal cells may not be well evaluated. Here, we investigated the effects of ELF-MF on the DNA damage responses in HT22 non-tumorigenic mouse neuronal cells. Exposure to a 60-Hz, 2 mT ELF-MF did not produce any increased ${\gamma}H2AX$ expression, comet tail moments, or aneuploidy formation. However, 2 mT ELF-MF transiently increased the cell number. From the results, ELF-MF could affect the DNA damage responses differently, depending on the cell lines.

Protective Effect of Yellow-Green Vegetable Juices on DNA Damage in Chinese Hamster Lung Cell Using Comet Assay (Comet Assay를 이용한 케일, 명일엽, 당근, 돌미나리 녹즙의 Chinese Hamster Lung 세포 DNA 손상 보호 효과)

  • 전은재;김정신;박유경;김태석;강명희
    • Journal of Nutrition and Health
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    • v.36 no.1
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    • pp.24-31
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    • 2003
  • The present study was attempted to investigate the antioxidant capacity of popular yellow-green vegetable juices (kale, Angelica keishei, carrot, small water dropwort) and to investigate the effect of vegetable juices on protecting oxidative damage to DNA in cultured Chinese hamster lung (CHL) cells. Antioxidant capacity was analyzed by TRAP assay (Total radical-trapping antioxidant potential). Cellular DNA dmamage was measured by SCGE (single-cell gel electrophoresis, also known as comet assay. Cells incubated in medium with PBS (negative control) or with various concentration of the freeze dried green juices (25, 50, 100, 250 $\mu\textrm{g}$/$m\ell$) resuspended in PBS were treated with $H_2O_2$ (200 ${\mu}{\textrm}{m}$) as an oxidative stimulus for 5 min at 4$^{\circ}C$. The physiological function of each vegetable juice on oxidative DNA damage was analyzed and expressed as tail moment (tail length X percentage migrated DNA in tail) . Kale juice had the highest TRAP value suggesting that kale has the highest antioxidant capacity followed by Angelica keishei, small water dropwort and carrot. Cells treated with $H_2O_2$ had extensive DNA damage compared with cells treated with PBS or pre-treated with vegetable juice extracts. All green juices inhibited $H_2O_2$-induced DNA damage with kale being the most effective juice among the tested juices. These results indicate that green juice supplementation to CHL cells followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species. (Korean J Nutrition 36(1) : 24-31, 2003)

The effect of carrot juice, ${\beta}$-carotene supplementation on lymphocyte DNA damage, erythrocyte antioxidant enzymes and plasma lipid profiles in Korean smoker

  • Lee, Hye-Jin;Park, Yoo-Kyoung;Kang, Myung-Hee
    • Nutrition Research and Practice
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    • v.5 no.6
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    • pp.540-547
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    • 2011
  • High consumption of fruits and vegetables has been suggested to provide some protection to smokers who are exposed to an increased risk of numerous cancers and other degenerative diseases. Carrot is the most important source of dietary ${\beta}$-carotene. Therefore, the objective of this study was to investigate whether carrot juice supplementation to smokers can protect against lymphocyte DNA damage and to compare the effect of supplementationof capsules containing purified ${\beta}$-carotene or a placebo (simple lactose). The study was conducted in a randomized and placebo-controlled design. After a depletion period of 14 days, 48 smokers were supplemented with either carrot juice (n = 18), purified ${\beta}$-carotene (n = 16) or placebo (n = 14). Each group was supplemented for 8 weeks with approximately 20.49 mg of ${\beta}$-carotene/day and 1.2 mg of vitamin C/day, as carrot juice (300 ml/day) or purified ${\beta}$-carotene (20.49 mg of ${\beta}$-carotene, 1 capsule/day). Lymphocyte DNA damage was determined using the COMET assay under alkaline conditions and damage was quantified by measuring tail moment (TM), tail length (TL), and% DNA in the tail. Lymphocyte DNA damage was significantly decreased in the carrot juice group in all three measurements. The group that received purified ${\beta}$-carotene also showed a significant decrease in lymphocyte DNA damage in all three measurements. However, no significant changes in DNA damage was observed for the placebo group except TM (P = 0.016). Erythrocyte antioxidant enzyme was not significantly changed after supplementation. Similarly plasma lipid profiles were not different after carrot juice, ${\beta}$-carotene and placebo supplementation. These results suggest that while the placebo group failed to show any protective effect, carrot juice containing beta-carotene or purified ${\beta}$-carotene itself had great antioxidative potential in preventing damage to lymphocyte DNA in smokers.

Multiple-biometric Attributes of Biomarkers and Bioindicators for Evaluations of Aquatic Environment in an Urban Stream Ecosystem and the Multimetric Eco-Model (도심하천 생태계의 수환경 평가를 위한 생지표 바이오마커 및 바이오인디케이터 메트릭 속성 및 다변수 생태 모형)

  • Kang, Han-Il;Kang, Nami;An, Kwang-Guk
    • Journal of Environmental Impact Assessment
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    • v.22 no.6
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    • pp.591-607
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    • 2013
  • The objectives of the study were to evaluate the aquatic environment of an urban stream using various ecological parameters of biological biomarkers, physical habitat quality and chemical water quality and to develop a "Multimetric Eco-Model" ($M_m$-E Model) for the ecosystem evaluations. For the applications of the $M_m$-E model, three zones including the control zone ($C_Z$) of headwaters, transition zone ($T_Z$) of mid-stream and the impacted zone ($I_Z$) of downstream were designated and analyzed the seasonal variations of the model values. The biomarkers of DNA, based on the comet assay approach of single-cell gel electrophoresis (SCGE), were analyzed using the blood samples of Zacco platypus as a target species, and the parameters were used tail moment, tail DNA(%) and tail length (${\mu}m$) in the bioassay. The damages of DNA were evident in the impacted zone, but not in the control zone. The condition factor ($C_F$) as key indicators of the population evaluation indicator was analyzed along with the weight-length relation and individual abnormality. The four metrics of Qualitative Habitat Evaluation Index (QHEI) were added for the evaluations of physical habitat. In addition, the parameters of chemical water quality were used as eutrophic indicators of nitrogen (N) and phosphorus (P), chemical oxygen demand (COD) and conductivity. Overall, our results suggested that attributes of biomarkers and bioindicators in the impacted zone ($I_Z$) had sensitive response largely to the chemical stress (eutrophic indicators) and also partially to physical habitat quality, compared to the those in the control zone.

Genotoxicity (DNA damage) on Blood Cells of Parrot Fish (Oplegnathus fasciatus) Exposed to Acidified Seawater Making of CO2 (이산화탄소로 산성화된 해수에 노출된 돌돔(Oplegnathus fasciatus) 혈구세포에 대한 유전독성(DNA 손상))

  • Choi, Tae Seob;Lee, Ji-Hye;Sung, Chan-Gyoung;Lee, Jung-Suk;Park, Young-Gyu;Kang, Seong-Gil
    • Journal of Environmental Science International
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    • v.23 no.3
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    • pp.483-492
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    • 2014
  • DNA damage such as genotoxicity was identified with comet assay, which blood cell of a marine parrot fish (Oplegnathus fasciatus) was exposed to an acidified seawater, lowered pH gradient making of $CO_2$ gas. The gradient of pH were 8.22, 8.03, 7.81, 7.55 with control as HBSS solution with pH 7.4. DNA tail moment of fish blood cell was $0.548{\pm}0.071$ exposed seawater of pH 8.22 condition, on the other hand, DNA tail moment $1.601{\pm}0.197$ exposed acidified seawater of pH 7.55 lowest condition. The approximate difference with level of DNA damage was 2.9 times between highest and lowest of pH. DNA damage with decreasing pH was significantly increased with DNA tail moment on blood cell of marine fish (ANOVA, p < 0.001). Ocean acidification, especially inducing the leakage of sequestered $CO_2$ in geological structure is a consequence from the burning of fossil fuels, and long term effects on marine habitats and organisms are not fully investigated. The physiological effects on adult fish species are even less known. This result shown that the potential of dissolved $CO_2$ in seawater was revealed to induce the toxic effect on genotoxicity such as DNA breakage.

Detection of Irradiated Fruits Using the DNA Comet Assay (DNA Comet Assay를 이용한 과일의 방사선 조사 확인)

  • Oh, Kyong-Nam;Park, Jun-Young;Kim, Kyeung-Eun;Yang, Jae-Seung
    • Korean Journal of Food Science and Technology
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    • v.32 no.3
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    • pp.531-537
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    • 2000
  • The simple microgel electrophoresis of single cells, a 'comet assay', on fruit seeds enabled the rapid identification of irradiated fruits by comparing the intact non-irradiated cells and the damaged cells of irradiated fruits. Grapes and plums were irradiated with 0.1, 0.5, 0.7, 1.0 kGy and strawberries, peaches, apples, and nectarines were irradiated with only 1.0 kGy. Seeds were isolated, crushed, and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were subjected to microgel electrophoresis for 2 minutes, and then stained. The DNA radiation-induced fragmentation of all the fruits stretched and migrated out of the cells forming a tail toward the anode giving the appearance of a comet, while the undamaged cells appeared as intact nuclei without tails. Grape and plum seeds irradiated at 0.5 kGy and higher showed significant increases in tail length. With increasing the irradiation doses, longer extention of the DNA from the nucleus toward the anode was observed. Strawberry, peach, apple, and nectarine seeds irradiated with 1.0 kGy also showed the longer tails than non-irradiated ones. DNA comet assay as a rapid and inexpensive screening technique could be an officially validated method for the detection of irradiated fruits.

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Comet Assay as a New DNA-Level Approach for Aquatic Ecosystem Health Assessments

  • Sung, Min-Sun;Lee, Sang-Jae;Lee, Jae-Hoon;Park, Sun-Young;Ly, Sun-Yung;An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.41 no.4
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    • pp.466-471
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    • 2008
  • Little is known about DNA-level and physiological levels for health assessments of stream or river environments. Recently, comet assay, so called Single Cell Gel Electrophoresis (SCGE) is introduced for assessments of DNA damage in the medical science, food science and mammal toxicology. The comet assay is known as a biomarker which is one of the best barometers in assessing the DNA damage by oxidative stress. In this study, we conducted the comet assay using sentinel species, Zacco platypus, as one of the pre-warning alarm systems for the aquatic ecosystem health assessments and also applied it to Gap Stream as a model system. Tail extent moments in the S1 and S2 were 5.20 and 9.90 respectively and the moment was 19.89 in the S3. Statistical ANOVA in the tail moments showed a significant difference (n=75, p<0.05) between S1 and S3. Also, the proportions of DNA in the tail were 14.47, 23.64, and $30.04{\mu}m$ in the upstream (control site), midstream, downstream sites, respectively. Our results in the downstream were accord with previous studies of individual-level, population-level, and community-level in Gap Stream. Our results suggest that the comet assay may be used as an important tool for diagnosing ecological health of aquatic ecosystems in the level of DNA.

Mechanism study on DNA damage and Apoptosis induced by heak shock using Comet Assay

  • Seo, Young-Rok;Han, Sung-Sik;Kim, L. O′Neill;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 1997.12a
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    • pp.101-101
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    • 1997
  • Comet assay, single cell gel electrophoresis has been known as useful, rapid, simple, visual, and sensitive technique for measuring the DNA breakage in mammalian ce1ls. For evaluation of DNA damage using comet assay, early studies reported a change in comet length and intensity with DNA damage using simple visual technique, such as fluorescence microscopy with eyespiece. In recent, some workers are observing and analyzing nucleotide of comets using quantitative fluorescence image analysis system to estimate 'tail moment', which is defined as the product of the tail length and the fraction of total DNA in tail. Our laboratory also adopted the image analysis software for qualification. In addition, many of the practical features of comet assay render it potentially attractive as useful tool for molecular toxicology and carcinogenesis, because the system is already showing considerable promise as rapid predictor in both in vitro and in vivo experimental designs. Recently, the comet assay becomes a attractive technique to study of apoptosis, because apoptotic fragmentation of nuclear DNA into nucleosomal sizes can be evaluated by the comet assay. So, we attempted to apply the comet assay to studying the effect of various stress on the apoptosis-sensitive cell lines. Particularly, focusing on the hyperthermic apoptosis, we could find that heat shock(44˚C for 60 minutes) was sufficient to induced apoptosis in these cell lines. But using the highly sensitive comet assay, we could not detect DNA breaks immediately after heat shock.

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