• 한국식품영양과학회지
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• 제31권4호
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• pp.594-598
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• 2002

과일의 방사선 조사 여부 확인과 저장에 따른 변화를 측정하기 위해 감마선 조사로 유도된 DNA손상을 comet assay로 확인하였다. 키위, 오렌지, 배를 구입하여 0.1, 0.3, 0.5, 0.7, 1.0 kGy의 저 선량으로 조사하고 비 조사 시료와 조사 시료간의 DNA 손상정도를 tail length와 tail moment로 측정하였다. 과일 씨의 DNA를 형광 염색하여 이미지 분석기를 이용하여 comet 양상을 관찰한 결과, 모든 시료에서 비 조사 시료보다 조사시료의 tail length가 더 길었으며 조사 선량이 증가할수록 tail length가 유의적으로 길게 나타났다. Tail moment로 나타난 결과도 이와 비슷하였으나 전체적으로 tail length에 비해 그 민감도가 낮았다. 방사선 조사한 과일을 저온에서 3개월 동안 저장한 후에도 저장 전과 마찬가지로 모든 시료에서 비 조사시료보다 조사 시료의 tail length가 더 길었으며, 조사 선량이 증가할수록 tail length가 길게 나타나, 저장 후에도 comet assay를 이용해 조사 시료와 조사 시료의 방사선 조사여부를 검지할 수 있었다 따라서 본 연구결과 comet assay는 신선한 과일과 일정기간 저장한 과일의 방사선 조사여부 판별에 유용하게 사용될 수 있음을 알 수 있었다.

• 생명과학회지
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• 제21권6호
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• pp.838-844
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• 2011

본 연구는 고강도 1회성 운동 시 혈장 MDA와 SOD의 농도변화와 임파구 DNA 손상에 대한 성별의 차이를 평가하는데 목적이 있었다. 본 연구의 목적을 달성하기 위하여 남자 대학생과 여자 대학생을 대상으로 85%$VO_{2max}$ all-out 운동수행에 따른 혈장 MDA와 SOD 그리고 임파구 DNA 손상에 대한 분석을 실시하였으며, 연구 결과에 대한 결론은 다음과 같다. 85%$VO_{2max}$ all-out 운동에 따른 혈장 MDA와 SOD는 운동 종료 시 유의하게 증가하였으며, 통계적으로 유의한 차이는 나타나지 않았으나 남성이 여성에 비해 MDA는 높고 SOD는 낮은 경향을 보였다. 반면 85%$VO_{2max}$ all-out 운동에 따른 임파구 DNA 손상을 알아보기 위해 실시한 comet assay 결과 세 가지 parameter (%DNA in the tail, tail length, tail moment) 모두 운동 종료 시 유의하게 증가하였으며 남성의 %DNA in the tail과 tail length가 여성에 비해 통계적으로 유의하게 높게 나타났다. 따라서 본 연구 결과를 종합해보면 1회성 고강도 운동은 산화적 스트레스를 유발할 수 있으며 남성이 여성에 비해 산화적 손상이 더 크다고 보여진다. 그러나, DNA 손상에는 산화적 스트레스 외에도 체력, 호르몬 수치, 생활습관, 운동 강도 및 지속시간 등 여러 가지 요인들이 영향을 줄 수 있다고 보고되고 있어, 성별에 따른 DNA 손상에 대한 명확한 기전을 제시하기 위해서는 DNA 손상에 영향을 줄 수 있는 여러 요인들과의 관계를 고려한 지속적인 연구들이 필요하다고 생각된다.

• Toxicological Research
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• 제20권2호
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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.

• 한국식생활문화학회지
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• 제19권3호
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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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• 제37권9호
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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.

• 한국식생활문화학회지
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• 제19권2호
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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.

• 한국환경성돌연변이발암원학회지
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• 제21권2호
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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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• 제35권4호
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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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• 제40권8호
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• pp.708-718
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• 2007

본 연구는 비타민 E의 보충섭취가 폐경기 여성의 항산화 영양상태 및 임파구 DNA 손상을 개선할 수 있는지를 알아보고자 수행되었으며, 이중맹검법으로 실시되었고 플라시보를 위약군으로 하였다. 폐경기 여성 35명을 대상으로 6주 동안 비타민 E 보충섭취군에게는 비타민 E 캡슐($\alpha$-tocopherol 성분 400 IU/capsule)을, 위약군에게는 대두유로 만든 위약 캡슐(400 mg/capsule)을 하루 2회 섭취하도록 하였으며, 이 때 두 군의 분류는 무작위로 하였다. 항산화 영양상태를 알아보기 위해 혈장 vitamin C, $\alpha$-tocopherol, $\gamma$-tocopherol, $\alpha$-carotenoid, $\beta$-carotenoid, lycopene 농도를 측정하였고, 임파구 DNA 손상정도를 알아보기 위해서는 Comet assay를 이용하여 tail length, %DNA in tail, tail moment를 측정하였다. 비타민 E 보충섭취군에서는 혈장 vitamin C(p<0.05)와 $\alpha$-tocopherol(p<0.000)농도가 유의적으로 증가하였고, $\gamma$-tocopherol(p<0.000) 농도와 tail length(p<0.05)는 유의적으로 감소하였다. 반면, 위약군에서는 혈장 vitamin C(p<0.05)의 농도만이 유의적으로 증가하였다. 결론적으로, 본 연구는 비타민 E의 보충섭취가 혈장 항산화 상태 및 DNA 손상에 대해 부분적인 개선 효과가 있음을 보여주었다.

• 한국식품과학회지
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• 제31권4호
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• pp.906-911
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• 1999

저선량으로 조사된 곡류의 방사선 조사 여부를 판별 하는데 DNA 'Comet Assay'의 활용이 가능한지를 검토하였다. 참깨, 들깨, 밀, 보리 및 쌀을 0, 0.1, 0.3, 0.5, 0.7, 1.0 kGy로 조사하여 시료로 사용하였다. 시료에 PBS 용액을 가하여 세포 현탁액을 조제하고 agarose gel과 혼합하여 microscope slide 상에서 lysis한 후 전기영동하여 염색한 다음 DNA 절편의 이동에 의해 형성된 ‘comet’의 tail length를 측정하여 분석하였다. 0.3 kGy 이상으로 조사한 모든 시료에서 tail length가 비조사한 시료와 차이를 나타내어 조사 여부를 확실히 판별할 수 있었다. 또한 비조사된 시료에서는 tail이 없거나 아주 짧은 tail을 가진 세포들이 관찰되었으며, 조사된 시료에서는 조사선량의 증가에 따라 DNA의 손상정도가 증가함으로서 tail length가 길어지는 것이 관찰되었다. 특히 참깨, 들깨 및 밀의 경우는 0.1 kGy의 낮은 조사선량에서도 현미경을 통하여 육안으로 비조사한 시료와의 구별이 가능하였다. 따라서 DNA 'Comet Assay'는 간단하고 신속하며 비교적 저렴하게 실시할 수 있는 곡류의 방사선 조사 여부를 판별하는 데 활용할 수 있음을 확인하였다.