• Title, Summary, Keyword: comet assay

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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.

Use of comet assay as a bioassay in marine organisms exposed to genotoxicants (유전독성물질로 오염된 해양생물의 생물검정법으로서 comet assay 이용)

  • Kim Gi-Beum;An Joon-Gun;Kim Jae-Won
    • Journal of Environmental Science International
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    • v.14 no.11
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    • pp.1071-1079
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    • 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.

Antioxidative properties of traditional herbal medicines and the application of comet assay on antioxidative study

  • Szeto, Yim Tong;Wong, Kam Shing;Kalle, Wouter;Pak, Sok Cheon
    • CELLMED
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    • v.3 no.3
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    • pp.22.1-22.10
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    • 2013
  • Traditional Chinese medicine (TCM) in single herb or formula prescription has been used for thousands of years. Many of them possess antioxidant activity and the activity may contribute the therapeutic effect. This paper would review the relationship of traditional herbal medicine and antioxidant with particular reference to ginseng. This medicinal herb has been used worldwide with extensive tonic effect. The comet assay, a technique for DNA protecting and damaging investigation would be introduced and the application of comet assay on TCM would be discussed.

Detection of Irradiated Grains Using the DNA ‘Comet Assay’ (DNA ‘Comet Assay’를 이용한 곡류의 방사선 조사 여부 확인)

  • Kim, Choong-Ki;Yang, Jae-Seung;Lee, Hae-Jung
    • 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.

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The Effects of Antioxidant Vitamins Via Comet and Mitogenic Assay in Mice Exposed to $AFB_1$ ($AFB_1$에 노출된 마우스에서 Comet Assay와 Mitogenic Assay에 의한 항산화 비타민의 효과)

  • Park, Seon-Ja
    • Journal of Korean Biological Nursing Science
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    • v.3 no.2
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    • pp.21-33
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    • 2001
  • The objective of this study was to examine the effects of antioxidant vitamins on the cellular oxidant damage by observing the mitogenicity in the mouse spleen and the strand breaks of DNA in mouse blood induced by $AFB_2$. Intraperitoneal(i.p.) injections of vitamin C(VC) of 10 mg/kg and vitamin E(VE) of 63.8 mg/kg were repeatedly administered to male ICR mice of 6 weeks old at intervals of 4 times every 2 days. After one hour vitamin treatments, $AFB_1$ of 0.4 mg/kg was injected into the $AFB_2$ plus vitamin treated groups in the same way. On the other hands, into the $AFB_2$ only treated group, only $AFB_2$ was injected without vitamins in the same method as above. The results of the experiment are as follows ; as regard to comet assay, DNA strand breaks were clearly present and they formatted a typical comet tail in the mice blood of the $AFB_2$ only treated groups. However, comet tails apparently disappeared in $AFB_2$ plus antioxidant vitamins treated groups since oxidant damage was controlled in an almost similar level to the control group. Mitogenicity of the spleen also showed a similar tendency as before, and these differences were more remarkably observed in the reaction against Con-A, which is a T-cell mitogen. In these data, the statistical significance was p<0.01. The LDL and VLDL levels were 408.72, 504.47 mg/dl respectively in the $AFB_2$ only treated groups. Compared with the $AFB_1$ only treated groups, those of $AFB_2$ plus antioxidant vitamin treated groups decreased to 272.06(VC), 305.28 mg/dl(VE), respectively. On the other hand, HDL levels were diminished to 32.60, 29.60 mg/dl in $AFB_2$ only treated groups, compared to 42.23, 41.14 mg/dl in the $AFB_2$ plus antioxidant vitamins treated groups. But, blood glucose levels were not statistically significant.

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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.

Detection Characteristics of TL, ESR and DNA Comet for Irradiated Peanuts by Origins (TL, ESR및 DNA Comet분석에 의한 원산지별 땅콩의 방사선 조사 검지 특성)

  • 이은영;정재영;조덕조;권중호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.30 no.6
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    • pp.1076-1081
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    • 2001
  • Gamma-irradiated peanuts, Korean and Chinese origins, were investigated on detection properties by thermoluminescence (TL), electron spin resonance (ESR), and DNA comet assay (single cell gel electrophoresis). TL measurement showed that the non-irradiated sample revealed a glow curve with low intensity at about 25$0^{\circ}C$, while the irradiated samples showed higher intensity around at 18$0^{\circ}C$. TL ratio (TL$_1$/TL$_2$) of area for TL$_1$ glow curve to TL$_2$ was below 0.05 for the non-irradiated sample and 0.2 or more for the irradiated ones, thus identifying each other. ESR spectroscopy for the irradiated peanuts using outer skin showed negligible signals induced by irradiation, indicating ESR is little applicable to the detection of irradiated peanuts. In DNA comet assay, the non-sample had no or very short tails, whereas the irradiated samples revealed the cells with long tails. Significance in the increase of their lengths depending on irradiation dose (r=0.761/Korean, r=0.768/Chinese) was also found. There was no remarkable difference in detection properties by origins of samples in all determinations, It is concluded that TL analysis or DNA comet assay is suitable for detection of irradiated peanuts and a combined method is recommendable for enhancing the reliability of detection results.

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Enhanced Prediction of Potential Rodent Carcinogenicity by Utilizing Comet Assay and Apoptotic Assay in Combination

  • Lee, Michael
    • Proceedings of the Korean Society of Toxicology Conference
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    • pp.95-95
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    • 2003
  • The comet assay has been recently validated as a sensitive and specific test system for the quantification of DNA damage. with 11 substances that demonstrated positive results in at least one test among 4 standard short-term genotoxicity tests, and to evaluate its ability to predict rodent carcinogenicity.(omitted)

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Genotoxicity on Structural Derivatives of Sophoricoside, a Component of Sophora Japonica, in Bacterial and Mammalian Cells

  • Ryu, Jae-Chun;Kim, Youn-Jung;Kim, Mi-Soon;Kim, Min-Ji;Sarma, Sailendra Nath;Jung, Sang-Hun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.179-188
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    • 2005
  • To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.