• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.4
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• pp.594-598
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• 2002

The changes in DNA damage were investigated during storage after irradiation. Kiwi, orange and pear were irradiated at 0.1, 0.3, 0.5, 0.7 and 1.0 kGy and stored for 3 months at 4$^{\circ}C$. The comet assay was applied to the sample seeds alt the beginning of irradiation and at the end of storage. Seeds were isolated and crushed, and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were electrophoresed for 2 min and then stained. DNA fragmentation in seeds caused by irradiation was quantified as tail length and tail moment (tail length $\times$ % DNA in tail) by comet image analyzing system. Immediately after irradiation, the differences in tail length between unirradiated and irradiated fruit seeds were significant (p<0.05) in kiwi, orange and pear seeds. With in-creasing the irradiation doses, statistically significant longer extension of the DNA from the nucleus toward anode was observed. The results represented as tail moment showed similar tendency to those of tail length, but tile latter parameter was more sensitive than the former. Similarly even 3 months after irradiation, all the irradiated fruit seeds significantly showed longer tail length than the unirradiated controls. These results indicate that the comet assay could be one of the simple methods of detecting irradiated fruit seeds. Moreover, the method could detect DNA damage even after 3 months after irradiation.

• Journal of Life Science
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• v.21 no.6
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• pp.838-844
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• 2011

The purpose of this study was to investigate gender-specific changes of plasma MDA, SOD, and lymphocyte DNA damage during high intensity exercise. In this study, 17 healthy male and 18 healthy female college students ran on a treadmill at 85%$VO_{2max}$ until the point of all-out. Blood-collecting was carried out five times (Rest, Ex-Exha, R0.5h, R4h and R24h), and with the collected blood, plasma malondialdehyde (MDA), superoxide dismutase (SOD), and lymphocyte DNA damage were analyzed. Plasma MDA and SOD concentration increased significantly at the Ex-Exha (p<0.05), and there were no significant differences in gender. For the degree of lymphocyte DNA damage, all %DNA in the tail, tail length and tail moment increased significantly at the Ex-Exha (p<0.05), and %DNA in the tail and tail length were significantly higher in the male group than in the female group (p<0.05). These results suggest that acute high intensity exercise not only causes oxidative stress but also brings about lymphocyte DNA damage. In addition, it was found that males showed higher DNA damage than females in terms of oxidative stress subject to high intensity exercise. Nevertheless, further subsequent studies are required in order to better understand the mechanism behind DNA damage varying with gender, in a way that takes into consideration physical fitness, hormonal level, exercise intensity and duration - additional factors which might affect DNA damage.

• Toxicological Research
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• v.20 no.2
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• pp.89-100
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• 2004

The comet assay (also called the single-cell gel electrophoresis assay) has been widely used for detecting DNA damage and repair in individual cells. Since the conventional methods of evaluating comet assay data using frequency statistics are unsatisfactory we developed a new quantitative measure of DNA damage/repair that is based on all information residing in the dose/time-response curves of a comet experiment. Blood samples were taken from 25 breast cancer patients before undergoing radiotherapy. The comet assay was performed under alkaline conditions using isolated lymphocytes. Tail DNA, tail length, tail moment and tail inertia of the comet were measured for each patient at four doses of $\gamma$-rays (0, 2, 4 and 8 Gy) and at four time points after irradiation (0, 10, 20 and 30 min) using 100 cells each. The resulting three-dimensional dose-time response surface was modeled by multiple regression, and the second derivative, termed 2D, on dose and time was determined. A software module was programmed in SAS/AF to compute 2D values. We applied the new method successfully to data obtained from cancer patients to be assessed for their radiation sensitivity. We computed the 2D values for the four damage measures, i.e., tail moment, tail length, tail DNA and tail inertia, and examined the pairwise correlation coefficients of 2D both on the log scale and the unlogged scale. 2D values based on tail moment and tail DNA showed a high correlation and, therefore, these two damage measures can be used interchangeably as far as DNA repair is concerned. 2D values based on tail inertia have a correlation profile different from the other 2D values which may reflect different facets of DNA damage/repair. Using the dose-time response surface, other statistical models, e.g., the proportional hazards model, become applicable for data analysis. The 2D approach can be applied to all DNA repair measures, Le., tail moment, tail length, tail DNA and tail inertia, and appears to be superior to conventional evaluation methods as it integrates all data of the dose/time-response curves of a comet assay.

• Journal of the Korean Society of Food Culture
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• v.19 no.3
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• pp.251-258
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• 2004

The changes in DNA damage were investigated during storage after irradiation. Potato, garlic were irradiated at 0.05, 0.07, 0.1 and 0.15 kGy and stored for 3 months. Ginger was irradiated at 0.01, 0.02, 0.03, 0.04 and 0.05 kGy and stored for 1 month. The comet assay was applied to the sample immediately after irradiation and at the end of storage. Samples were isolated, grounded and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were electrophoresed for 1 min. and then stained. DNA fragmentation in seeds caused by irradiation was quantified as tail length and tail moment (tail length ${\times}%$ DNA in tail) by comet image analyzing system. Right after irradiation, the differences in tail length between unirradiated and irradiated samples were significant(p<0.05) in potato, garlic and ginger. With increasing the irradiation doses, statistically significant longer extension of the DNA from the nucleus toward anode was observed. The results represented as tail moment showed similar tendency to those of tail length. Similarly in the stored samples, even 1 or 3 months after irradiation, all the irradiated samples significantly showed longer tail length than the unirradiated controls. These results indicate that the comet assay could be one of the simple methods of detecting irradiated samples. Moreover, the method could detect DNA damage even after 1 or 3 months after irradiation.

• Journal of Nutrition and Health
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• v.37 no.9
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• pp.794-800
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• 2004

Deer Antler has been known for its traditional oriental medicinal properties and has been widely used to promote growth, boost immune function, treat blood loss and chronic joint pain. Recent study showed imported (New Zealand) Deer Antler was beneficial in reducing the side effects of cancer treatments. However, there was no intervention study conducted on the effect of Korean Deer Antler on reducing the oxidative stress to patients with diabetes. One of the sensitive ways to measure endogenous oxidative stress is by measuring cellular DNA damage using single cell gel electrophoresis (COMET assay). This study was conducted to investigate the possible beneficial effect of commercial Deer Antler drink (provided by Chung-yang Deer Farm) on lymphocyte DNA damage and blood glucose of diabetic patients. Ten patients (4 men, 6 women) participated in the study and consumed 2 pouches of Deer Antler drink every day for 20 days. Blood was collected on the morning before and after the intervention for lymphocyte isolation and blood glucose analysis. Both systolic and diastolic blood pressure showed a tendency to decrease but did not reach statistical significance after the trial. Blood glucose level was not affected by the supplementation. After the intervention, over 50% reduction were noted in the cellular DNA damage, expressed as tail length (TL) and tail moment (TM: tail length ${\times}$ percent tail DNA) . Although we did not obtain beneficial effect on lowering blood glucose levels in the patients, this results suggest that Deer Antler may initially act in protecting endogenous DNA damage in short-term experiment.

• Journal of the Korean Society of Food Culture
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• v.19 no.2
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• pp.129-138
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• 2004

The changes in DNA damage were investigated during storage after irradiation. Beef, pork and chicken were irradiated at 1.0, 3.0 and 5.0 kGy and stored for 6 months at $-20^{\circ}C$. The comet assay was applied to the sample muscles at the beginning of irradiation and at the end of storage. Muscles were isolated, sliced, and the suspended cells were embedded in an agarose layer. After lysis of the cells, they were electrophoresed for 2 min. and then stained. DNA fragmentation in tissues caused by irradiation was quantified as tail length and tail moment (tail length ${\times}$ % DNA in tail) by comet image analyzing system. Right after irradiation, the differences in tail length between unirradiated and irradiated muscles were significant(p<0.05) in beef, pork and chicken. With increasing the increasing doses, statistically significant longer extension of the DNA from the nucleus toward anode was observed. Similarly even 6 months after irradiation, all the irradiated muscles significantly showed longer tail length than the unirradiated controls. The results represented as tail moment showed similar tendency to those of tail length, but the latter parameter was more sensitive than the former. These results indicate that the comet assay could be one of the simple methods of detecting irradiated muscles. Moreover, this method suggest that using comet assay, we were able to detect DNA damage differences even after 6 months after irradiation.

• Environmental Mutagens and Carcinogens
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• v.21 no.2
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• pp.128-134
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• 2001

Single cell gel electrophoresis (SCGE) assay, also called comet assay, is a rapid and sensitive method to detect DNA damage in single cell level. To evaluate the DNA damage of lymphocytes of pesticides sprayers, SCGE assay was carried out for 50 pesticides sprayer and 58 control subjects. They were interviewed with structured questionnaire to get the information about the kinds and amount of pesticide. Insecticides and fungicides were predominant among pesticides. Major components of pesticides were organophosphorus, organosulfate, cartap, carbamates, and triazole. Sprayed pesticides were classified into two groups. Group I included organophosphorus, organoarsenic, organotin, tetrazine, triazole and gramoxone, which were known to cause DNA damages. Group II pesticide were carbamates, surfactants, organosulfates, etc., which were not found as DNA damaging agents in scientific documents. Olive tail moments of 100 lymphocytes were measured by KOMET 3.1 program for each person. The means of tail moments were compared between farmers exposed to pesticides and control subjects. Farmers showed higher tail moments than control subjects (2.07$\pm$1.40 vs 1.53$\pm$0.77, p<0.05). The means of tail moments also were compared among group I sprayers (n=36), group II sprayers (n=24) and, control subject, and the means or tail moments were 3.4s$\pm$3.2o, 2.66$\pm$2.20 and 1.53$\pm$0.77 respectively. The difference between means of group I sprayers and controls was statistically significant (p<0.05). In conclusion, this study showed higher DNA damage in farmers exposed to pesticides than control subjects, and comet assay could be useful as a biological monitoring method of genotoxic pesticides for farmers.

• Journal of Preventive Medicine and Public Health
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• v.35 no.4
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• pp.275-281
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• 2002

Objectives : To evaluate the DNA damage by hair dyeing in human lymphocytes Methods : Comet assays were carried out to evaluate the DNA damage in lymphocytes by hair dyeing. Twenty subjects were selected from women volunteers whose age ranged from 55 to 67 year old. All subjects had no smoking history. Blood samples were collected before and 6 hours after hair dyeing. DNA damage was evaluated by means of the tail moments, which were quantified by a KOMET 4.0 image analysis system. Results : The tail moments before hair dyeing showed no significant differences among subjects except for the high frequency group. The mean values of the tail moments in subjects with low and high frequencies of hair dyeing were 1.39 and 1.77, respectively (p<0.05). The tail moments after hair dyeing increased significantly, The mean values of tail moments in subjects before and after hair dyeing were 1.45 and 1.79, respectively (p<0.01). However, the difference levels of DNA damage in lymphocytes before and after hair dyeing were found to be slightly lower in both the dietary supplement taking group and high frequency group. Conclusions : The high frequency group appears to have a higher level of DNA damage than the low frequency group before hair dyeing. DNA damage in lymphocytes was found to be significantly higher in the volunteers after hair dyeing. In this study, the related factors such as high frequency and taking dietary supplements appeard to reduce DNA damage in lymphocytes after hair dyeing.

• Journal of Nutrition and Health
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• v.40 no.8
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• pp.708-718
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• 2007

The purpose of this project was to evaluate whether vitamin E supplementation could improve the antioxidant status and lymphocyte DNA damage in Korean postmenopausal women. This was double blinded, placebo-controlled trial. Thirty-five subject were randomized to receive either placebo 400 mg/capsule or natural $\alpha$-tocopherol 400 IU/capsule, 2 times a day for 6 weeks. We measured plasma vitamin C, $\alpha$-tocopherol, $\gamma$-tocopherol, $\alpha$-carotenoid, $\beta$-carotenoid, lycopene concentration and tail length, %DNA in tail, tail moment in lymphocyte DNA damage index. Vitamin E supplementation group had significantly increased plasma vitamin C(p<0.05), $\alpha$-tocopherol(p<0.000), whereas $\gamma$-tocopherol(p<0.000) and tail length(p<0.05) were significantly decreased. However, placebo supplementation group also had significantly increased plasma vitamin C(p<0.05). In conclusion, our study shows that vitamin E supplementation to Korean postmenopausal women may partially improve antioxidant status and lymphocyte DNA damage.

• Korean Journal of Food Science and Technology
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• v.31 no.4
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• pp.906-911
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• 1999

This study was to determine whether DNA 'Comet Assay' can be applied to the detection of grains irradiated with low doses of Co-60 gamma radiation. Sesame, perilla, wheat, barley and rice were exposed to different doses of 0.1, 0.3, 0.5, 0.7 and 1.0 kGy. The cells isolated from the samples were embedded in a agarose gel on a microscope slide, lysed in lysis solution, and subjected to electrophoresis. DNA and its fragments migrated in the gel produced the characteristic pattern of DNA comet, of which the tail length was measured in a microscope. All the samples irradiated at 0.3 kGy and higher were applicable to detect post-irradiation by the tail length of their comets. Irradiated samples showed comets with long tails and their tail length increased with the dose, while unirradiated samples showed no or very short tails. Especially, sesame, perilla and wheat irradiated at 0.1 kGy could be distinguished from unirradiated samples by visual inspection of the slide in a microscope. Thus, DNA 'Comet Assay' might be applied to the detection of irradiated grains as a simple, inexpensive and rapid screening test.