• 한국식품과학회지
• /
• 제36권6호
• /
• pp.851-856
• /
• 2004

통고추의 열발광 특성(TL)분석에서 2.5 kGy 이상의 조사 시료에서 분리된 미네랄 시료는 $150^{\circ}C$ 부근에서 TL glow curve ($TL_1$)를 나타내었고, 조사선량이 증가할수록 peak intensity가 증가하였다. 재조사(1 kGy) 방법에 의한 TL ratio($TL_1/TL_2$)는 TL 분석의 신뢰도를 높여주었다. DNA comet assay에서 비조사 시료(씨)는 전형적인 intact cell을 나타내었으나 감마선 조사 시료에서는 long tail을 가진 comet을 나타내면서 선량 의존적으로 tail length가 증가되었다. 미생물학적 방법으로써 DEFT/APC값 측정에서는 조사선량에 따라 분말고추와 통고추에서 비교적 높은 상관을 보였다. 이상의 결과에서 DNA comet assay와 DEFT/APC 방법은 방사선 조사 건고추의 screening 방법으로서, TL은 확인방법으로서 적용가능성이 확인되었다.

• 한국식품과학회지
• /
• 제34권1호
• /
• pp.18-23
• /
• 2002

방사선 조사된 대두의 검지방법을 연구하기 위하여 국산 및 중국산 대두에 대하여 $0.5{\sim}4.0\;kGy$의 감마선을 조사하고 열발광(TL), 전자스핀공명(ESR) 및 DNA comet 특성을 비교 검토하였다. TL 측정에서 비조사구는 $280^{\circ}C$ 부근에서 매우 낮은 glow curve를 나타내었고, 감마선 조사구는 $200^{\circ}C$ 부근에서 조사선량에 의존적인 glow curve를 나타내었다. 재조사(re-irradiation) 방법에 의한 TL ratio$(TL_1/TL_2)$의 비교는 TL 측정 결과의 정확도를 높여 주었다. 원산지별 TL 특성 비교에서 국산을 중국산에 비해 높은 TL intensity를 나타내었다(p<0.01). 대두 껍질을 사용한 ESR 측정에서는 조사시료에서 cellulose radical 유래의 특이한 signal(g = 2.02374, 1.98715)을 보여주었고 중국산 시료의 ESR signal이 더 높게 나타났다. 비조사 대두의 DNA comet은 tail이 없거나 아주 짧은 tail을 가진 전형적인 intact cell을 나타내었다. 그러나 0.5 kGy 이상의 감마선 조사 시료에서는 조사선량에 의존적으로 comet의 tail length 증가와 더불어 comet의 크기 및 농도의 변화가 일어났다. 이상의 결과에서 TL, ESR 및 DNA comet의 분석은 대두의 방사선 조사 여부 확인을 가능하게 하였으며, 원산지별 검지특성의 차이는 거의 없었다.

• 한국환경과학회지
• /
• 제14권11호
• /
• pp.1071-1079
• /
• 2005

Using single cell gel electrophoresis, DNA single strand breaks were determined in various marine organisms. DNA damage on fish blood cells was detected to know whether there was a difference between Incheon, Pohang, Masan, and Tongyeong as a control site. Tongyeong showed the lowest DNA damage among the study areas. Mussels, transplanted to Masan Bay for one month, revealed high DNA damage at sites with high economical activity. In two weeks exposure of polychaete to Incheon sediments, higher DNA damage was detected in the sediment adjacent to Incheon harbor than open sea. These results suggested that the marine organism from the polluted area revealed a relatively high DNA damage. In addition, these areas might be contaminated with genotoxic compounds and comet assay was useful as a bioassay to detect DNA damage in marine organisms.

• Journal of Nutrition and Health
• /
• 제37권6호
• /
• pp.440-447
• /
• 2004

In this study the in vitro protective effects of several antioxidant vitamins (vitamin C, $\alpha$-tocopherol, $\beta$-carotene), fruits and vegetables (strawberry, tangerine, orange and 100％ orange juice, carrot juice), on the levels of isolated human lymphocyte DNA damage was measured using Comet assay. Comet assay has been used widely to assess the level of the DNA damage in the individual cells. Lymphocytes were pre-treated for 30 minutes with antioxidant vitamins (10, 50, 100, 500 $\mu$M) or fruits$.$vegetables (10, 100, 500, 1000 $\mu$g/ml), an4 then oxidatively challenged with 100 $\mu$M $H_2O$$_2$ for 5 min at 4$^{\circ}C$. The protective effect of antioxidant vitamins against DNA damage at a concentration of 50 $\mu$M were 50％ in vitamin C, 32％ in $\alpha$-tocopherol, whereas, fJ-carotene showed a 55％ protection at a dose as low as 10 $\mu$M. The inhibitory effects of DNA damage by strawberry, tangerine, orange, orange juices, carrot juices were 50 - 60％ with wide ranges of doses. The results of the present study indicate that most the antioxidant vitamins and fruits$.$vegetables juices produced a significant reduction in oxidative DNA damage.

• 한국양식학회지
• /
• 제16권3호
• /
• pp.196-201
• /
• 2003

Sessile organisms such as the oyster Crassostrea gigas have been given much attention as a potential biomonitoring indicator to assess the impact of toxicants on aquatic organism. In this study, we exposed cells isolated from gill of oyster (Crassostrea gigas) to hydrogen peroxide in vitro. In addition oysters were in vivo exposed to pyrene and benzo(a)pyrene at various concentrations for 2 weeks. Comet assay was used to detect DNA single strand breaks and to investigate the application of this technique as a tool for aquatic biomonitoring. Hydrogen peroxide increased DNA single strand break with increasing concentration after 30 minutes exposure in vitro. Pyrene and benzo(a)pyrene caused DNA damage only at very high concentration (100 $\mu\textrm{g}$/L or 1000 $\mu\textrm{g}$/L) at two week exposure in vivo. DNA damage was relatively higher at hemocyte than at gill. It suggested that metabolized PAHs are transferred to hemolymph from digestive gland which have a relatively high enzyme activity, and attacked the DNA of hemocyte, while gill accumulated PAHs without degrading them to their metabolites due to low enzyme activity at gill. Both in vitro and in vivo exposure experiments showed that the comet assay is an effective tool on screening whether the organism are exposed to genotoxic contaminants.

• Preventive Nutrition and Food Science
• /
• 제7권3호
• /
• pp.323-326
• /
• 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.

• Toxicological Research
• /
• 제22권4호
• /
• pp.423-429
• /
• 2006

Recent reports on the photocarcinogenicity and photogerotoxicity of many compounds led to an increasing awareness for the need of a standard approach to test for photogenotoxicity. The comet assay has been recently validated as a sensitive and specific test system for the quantification of DNA damage. Thus, the objectives of this study are to investigate the utility of photo-comet assay for detecting photo-mutagens, and to evaluate its ability to predict rodent photo-carcinogenicity. Photo-comet assays were performed using L5178Y $Tk^{+/-}$ mouse lymphoma cells on five test substances (8-methoxypsoralen, chlorpromazine, lomefloxacin, anthracene and retinoic acid) that demonstrated positive results in photocarcinogenicity tests. For the best discrimination between the test substance-mediated DNA damage and the undesirable DNA damage caused by direct UV absorption, a UV dose-response of the cells in the absence of the test substances was firstly fnalized. Out of 5 test substances, positive comet results were obtained for chlorpromazine, lomefloxacin, anthracene and retinoic acid while 8-methoxypsoralen found negative. An investigation into the predictive value of this photo-comet assay for determining the photocarcinogenicity showed that photo-comet assay has relatively high sensitivity. Therefore, the photo-comet assay with mammalian cells seems to be a good and sensitive predictor of the photocarcinogenic potential of new substances.

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

• 한국식품영양과학회지
• /
• 제30권6호
• /
• pp.1076-1081
• /
• 2001

현재 중국으로부터 수입량이 증가하고 있는 땅콩을 대상으로 국산과 중국산 시료의 TL, ESR, DNA comet(single cell gel electrophoresis) 분석을 실시하여 원산지별 특성을 비교하였다. Density separation 방법으로 추출한 미네랄의 TL측정 결과, 감마선 조사되지 않은 시료는 25$0^{\circ}C$ 부근에서 intensity가 낮은 glow curve를 나타내었고, 조사 시료는 18$0^{\circ}C$ 부근에서 아주 강한 intensity의 glow curve를 보여주었다. 첫 번째 측정된 glow curve(TL$_1$)의 normalization을 위하여 재조사 방법에 의해 TL$_2$를 측정하여 TL ratio(TL$_1$/TL$_2$)를 비교해 본 결과, 비조사 시료는 0.05 이하, 조사 시료는 0.2이상으로 방사선 조사 여부의 판별이 가능하였다. 땅콩껍질을 사용한 ESR 측정에서는 조사 유래의 특이적인 signal이 나타나지 않아 적용 가능성이 낮았다. DNA comet assay 결과, 비조사 시료는 tail이 없거나 아주 짧은 전형적인 intact cell을 나타낸 반면, 조사 시료는 long tail을 가진 comet을 나타내면서 선량 의존적으로 (r=0.761/Korean, r=0.768/Chinese) tail length가 증가하여 조사 여부의 확인이 가능하였다. 모든 실험에서 원산지별 차이는 크지 않았다. 이상의 결과로 볼 때 땅콩의 방사선 조사 여부 확인에는 TL 분석 및 DNA comet assay가 적용 가능하였다.

• Ocean and Polar Research
• /
• 제33권2호
• /
• pp.127-133
• /
• 2011

The comet assay, also called single-cell electrophoresis (SCGE) assay, is a potential sensitive monitoring tool for DNA damage in cells. The primary objective of this study was to use comet assay to ascertain if the blood cells of flounder (Pleuronichthys olivaceus) and muscle cells of clam (Saxidomus purpurata) are suitable for genotoxicity screening. This was achieved by initially exposing blood and muscle cells under in vitro conditions to the reference genotoxin hydrogen peroxide ($H_2O_2$); strong correlation between $H_2O_2$ concentration and comet values were found. Subsequently, the identification of DNA damage in isolated cells from flounder and clam was performed under in vivo exposure to benzo(a)pyrene (BaP) and tributyltin (TBT). Flounder and clam were exposed to different concentrations (1, 10, 50, 100 ${\mu}g/L$) of BaP or TBT for 4 days. Regardless of treated chemicals, blood cells of flounder were more prone to DNA breakage compared to muscle cells of clam. In conclusion, in vivo genotoxicity of BaP and TBT can be biomonitored using the comet assay. This study suggests that flounder and clam do show potential as mediums for monitoring genotoxic damage by comet assay.