• 제목/요약/키워드: tail DNA

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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
    • 생태와환경
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    • 제41권4호
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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
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 1997년도 제20회 화학물질의 환경독성과 건강영향
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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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Thermoluminescence, DNA Comet 및 DEFT/APC 분석에 의한 방사선처리 건고추의 저장 중 검지 특성 (Identification Characteristics of Irradiated Dried Red Pepper during Storage by Analysis of Thermoluminescence, DNA Comet, and DEFT/APC)

  • 김병근;권중호
    • 한국식품과학회지
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    • 제36권6호
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    • pp.851-856
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    • 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.

First Record of Acrobeloides nanus (Cephalobidae: Rhabditida: Nematoda) from Korea

  • Kim, Taeho;Kim, Jiyeon;Bae, Yeon Jae;Park, Joong-Ki
    • Animal Systematics, Evolution and Diversity
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    • 제32권4호
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    • pp.258-265
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    • 2016
  • Acrobeloides nanus (de Man, 1880) Anderson, 1968 belonging to the family Cephalobidae Filpijev, 1934 (Cephalobomorpha) is newly reported from South Korea. This species is distinguished from other Acrobeloides species by its low and blunt labial probolae, five lateral incisures with middle incisure extending to the tail tip, and bluntly rounded tail. In this study, details of morphological characters of A. nanus is described and illustrated based on optical and scanning electron microscopy. In addition, molecular sequence data of the D2-D3 region of 28S rDNA, 18S rDNA and mitochondria DNA cox1 region from this species are provided as DNA barcode sequences.

Red beet(Beta vulgaris L.) leaf supplementation improves antioxidant status in C57BL/6J mice fed high fat high cholesterol diet

  • Lee, Jeung-Hee;Son, Chan-Wook;Kim, Mi-Yeon;Kim, Min-Hee;Kim, Hye-Ran;Kwak, Eun-Shil;Kim, Se-Na;Kim, Mee-Ree
    • Nutrition Research and Practice
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    • 제3권2호
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    • pp.114-121
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    • 2009
  • The effect of diet supplemented with red beet (Beta vulgaris L.) leaf on antioxidant status of plasma and tissue was investigated in C57BL/6J mice. The mice were randomly divided into two groups after one-week acclimation, and fed a high fat (20%) and high cholesterol (1%) diet without (control group) or with 8% freeze-dried red beet leaf (RBL group) for 4 weeks. In RBL mice, lipid peroxidation determined as 2-thiobarbituric acid-reactive substances (TBARS value) was significantly reduced in the plasma and selected organs (liver, heart, and kidney). Levels of antioxidants (glutathione and $\beta$-carotene) and the activities of antioxidant enzyme (glutathione peroxidase) in plasma and liver were considerably increased, suggesting that antioxidant defenses were improved by RBL diet. Comet parameters such as tail DNA (%), tail extent moment, olive tail moment and tail length were significantly reduced by 25.1%, 49.4%, 35.4%, and 23.7%, respectively, in plasma lymphocyte DNA of RBL mice compared with control mice, and indicated the increased resistance of lymphocyte DNA to oxidative damage. In addition, the RBL diet controlled body weight together with a significant reduction of fat pad (retroperitoneal, epididymal, inguinal fat, and total fat). Therefore, the present study suggested that the supplementation of 8% red beet leaf in high fat high cholesterol diet could prevent lipid peroxidation and improve antioxidant defense system in the plasma and tissue of C57BL/6J mice.

Paraquat에 의한 사람 임파구 DNA 손상에 대한 환원전리수의 보호효과 (Protective Effect of Electrolyzed Reduced Water on the Paraquat-induced Oxidative Damage of Human Lymphocyte DNA)

  • 박은주;류근걸;이윤배;이종권;이미영
    • Applied Biological Chemistry
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    • 제48권2호
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    • pp.155-160
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    • 2005
  • 본 연구에서는 매우 낮은 음의 산화환원전위를 나타내는 환원전리수가 paraquat에 의한 사람 임파구 DNA의 손상에 미치는 영향을 Comet assay를 사용하여 조사하였다. 환원전리수 처리에 의해서 paraquat에 의해 산화적으로 손상된 DNA가 회복되는 정도를 손상된 DNA로 인한 꼬리부분의 형광광도의 %비율로 나타내었다. Comet assay는 개별 세포의 DNA의 산화적 손상을 측정하는데 널리 사용되고 있다. 사람 임파구에 다양한 농도의 paraquat을 $37^{\circ}C$에서 30분간 처리한 후, 환원전리수를 첨가하여 30분간 반응시켰다. 그 결과 paraquat에 의한 임파구 DNA의 손상은 paraquart 농도증가에 의존적으로 증가하였다. 그러나 환원전리수를 처리한 결과 DNA의 산화적 손상이 paraquat 미처리 대조군 수준으로 거의 다 복구되었다.

Evaluation of Protective Effects of Houttuynia cordata on H2O2-Induced Oxidative DNA Damage Using an Alkaline Comet Assay in Human HepG2 Cells

  • Hah, Dae-Sik;Kim, Chung-Hui;Ryu, Jae-Doo;Kim, Eui-Kyung;Kim, Jong-Shu
    • Toxicological Research
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    • 제23권1호
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    • pp.25-31
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    • 2007
  • To evaluate the protective effect of Houttuynia cordata on hydrogen peroxide-induced oxidative DNA damage in HepG2 cell line, we used an alkaline single-cell gel electrophoresis (SCGE; comet assay). The DNA damage was analyzed by tail moment (TM) and tail length (TL), which used markers of DNA strand breaks in SCGE. The $100{\mu}g/ml$ of methanolic extract of Houttuynia cordata root showed significant protective effects (p < 0.01) against hydrogen peroxide-induced DNA damage in HepG2 cells and increased cell viability against hydrogen peroxide. The results of this study indicate that Houttuynia cordata root methanol extract acts as a potential antioxidant, and exhibits potential anticancer properties, which may provide a clue to find applications in new pharmaceuticals for oxidative stability.

염모제 사용에 의한 인체림프구의 DNA 손상 변화 (Effects of Hair Dyeing Application on the DNA Damage in Human Lymphocytes)

  • 김영철;심미자;권정숙
    • Environmental Analysis Health and Toxicology
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    • 제19권1호
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    • pp.101-107
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    • 2004
  • To ascertain the effects of hair dyeing application on the DNA damage in human lymphocytes, a mixture of permanent black colored hair dye with the same amount of oxidant containing 6% hydrogen peroxide was used. A hair dyeing with contacting the scalp (conventional dyeing) and a hair dyeing with 3 to 4mm away from the scalp (alternative dyeing) were applied to each If young healthy women. Blood was taken from the brachial vein at two sampling times, just before and 6 hours after the hair dyeing, and tail extent moment(TEM) and tail length (TL) were measured by using a comet assay. After dyeing, TL was significantly increased in both conventional dyeing group and alternative dyeing group compared with before dyeing as an average of 47% and 28%, respectively, and TL for conventional dyeing group was higher than alternative dyeing group as an average of 1.2 fold. After dyeing, TEM was significantly increased in both conventional dyeing group and alternative dyeing group compared with before dyeing as an average of 192% and 76%, respectively, and TEM for conventional dyeing group was significantly higher than alternative dyeing group as an average of 1.7 fold. Therefore, alternative dyeing application was induced to lower lymphocyte DNA damage than conventional dyeing application, and TEM was appeared to be a more sensitive tool for the measurement of lymphocyte DNA damage than TL in this study.

식용 버섯의 조리방법에 따른 Jurkat 세포주 DNA 손상 보호 효과 (Protective Effect of Edible Mushrooms (Pleurotus ostreatus, Flammulina velutipes, Lentinula edodes) according to Different Cooking Methods on DNA Damage of Jurkat Cell Line)

  • 조윤정;김경희;육홍선
    • 한국식품영양학회지
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    • 제28권1호
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    • pp.34-39
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    • 2015
  • 본 연구는 식용 버섯의 조리방법에 따른 항산화 생리활성의 평가를 위해 수행되었으며, 산화적 스트레스에 의한 DNA 손상 감소 효과를 통해 조리방법을 달리한 버섯 추출물의 유전독성학적 방호효과를 살펴보았다. Human lymphocyte에 조리방법을 달리한 3가지 버섯(느타리, 팽이, 표고)의 추출물을 처리하고, hydrogen peroxide($H_2O_2$)로 산화적 손상을 준 후, DNA 감소 효과를 Comet assay로 평가한 결과, 모든 시료군에서 산화적 손상에 의한 DNA 손상 감소 효과를 나타냈다. 3가지 버섯 모두 비조리군이 조리군보다 높은 효과를 나타냈는데, 이는 조리과정에 의한 페놀성 화합물의 감소로 인한 것으로 보이며, 조리군 중에서 볶기와 전이 비교적 낮은 DNA 손상 감소 효과를 나타낸 것은 조리 시 첨가되었던 대두유의 가열 산화에 의한 것으로 사료된다. 결론적으로, 조리된 버섯은 생버섯에 비해 산화적 스트레스에 의한 DNA 손상 감소효과가 낮으나, 양성 대조군과 비교하였을 때 손상을 유의적으로 감소시킨 것으로 나타났다. 또한, 본 연구에서 사용한 네 가지 조리법(굽기, 데치기, 볶기, 전) 중 DNA 손상 감소에 효과적인 조리법은 대두유를 사용하지 않은 굽기와 데치기인 것으로 판단된다.

방사선 조사된 고불포화지방산 식이를 섭취한 Mouse의 산화적 스트레스 (Oxidative Stress of Mouse Fed Irradiated Diet Containing High Unsaturated Fatty Acid)

  • 서대영;박선영;강명희;서광선;이선영
    • Journal of Nutrition and Health
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    • 제39권7호
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    • pp.599-609
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    • 2006
  • In this study, we investigated the in vivo effect by intake of the irradiated foods such as mackerel and sesame seed which are high in unsaturated fatty acid through TBARS (thiobarbituric acid reactive subtance) and the tissue pathological and genotoxicological test. Thirty two ICR mice are divided into four groups, one non-irradiated (control) group and three irradiated (5, 10, 20 kGy, respectively) groups. Sesame seed and pulverized mackerel in modified AIN93M diet were mixed together then divided into four identical parts. Three parts of them were irradiated by doses of 5, 10, and 20 kGy. These experimental diet were fed to each group for 4, 8 and] 6 weeks. The results of the study were as follows: No significant differences in weight gain were found in each group. Peroxide value of the irradiated diet was higher than that of the non-radiated one and also increased according to the storage period. TBA values in plasma, liver, kidney and Peyer's patch were not significantly different among 4 groups. DNA% in tail, tail length (TL) and tail moment (TM) values of the blood lymphocyte in 4, 8 and 16 week groups and the liver in 16 week group were much measured over the control DNA % in tail of kidney of 8 week group was significantly than the control and TL and TM of 8 week and TM of 16 week groups showed a tendency of higher values. By Peyer's DNA % in tail of 8 week group, DNA % in tail and TM of 16 week groups increased significantly over the control, Ultrastructural examination shows myeline figures and swollen mitochondria in parietal cells and intestinal epithelial cells of 8 and 16 weeks groups. After this study, we need further investigations on the safety of highly consumed foods which contain high contents of unsaturated fatty acid, largely imported and which are possible to be irradiated.