• 제목/요약/키워드: tail DNA

검색결과 239건 처리시간 0.022초

인체 산화적 DNA손상에 대한 Human Biomonitoring도구로서 Alkaline Comet Assay의 활용 가능성 연구 (Application of the Alkaline Comet Assay for Detecting Oxidative DNA Damage in Human Biomonitoring)

  • 박은주;강명희
    • Journal of Nutrition and Health
    • /
    • 제35권2호
    • /
    • pp.213-222
    • /
    • 2002
  • The alkaline comet assay has been used with increasing popularity to investigate the level of DNA damage in biomonitoring studies within the last decade in Western countries. The purpose of this study was to evaluate the usefulness of the alkaline comet assay as a biomarker of oxidative DNA damage for monitoring in the Korean population, and also to evaluate the effect of nutritional status and lifestyle factors on H2O2 induced oxidative DNA damage measured by the alkaline comet assay in human lymphocytes. The study population consisted of 61 healthy Korean male volunteers, aged 20-28. Epidemiological background data including dietary habits, smoking habits and anthropometrical measurements were collected through personal interviews. After blood collection, the comet assay in peripheral lymphocytes and plasma lipids analysis was carried out and the results analyzed. Tail moment (TM) and tail length (TL) of the comet assay were use\ulcorner to measure DNA damage in the lymphocytes of the subjects. Statistically significant (p < 0.05) positive correlations were observed between DNA damage (TM or TL) and smoking habits expressed as cigarettes smoked per day and pack years (r = 0.311 and 0.382 for TM, r = 0.294 and 0.350 for TL, respectively). There were also significant positive correlations between DNA damage parameter and waist-hip ratio. Higher plasma triglyceride levels were associated with increased damage to DNA. There were no correlations between the consumption frequencies of vegetables and DNA damage to the subjects. However, consumption frequencies of fruit and fruit juice intake were inversely associated with the TM and TL. The results indicate that die comet assay is a simple, rapid and sensitive method for detecting lymphocyte DNA damage induced by cigarette smoking. Consumption of fruit or fruit juices could potentiall modify the damaged DNA in the human peripheral lymphocytes of young Korean men.

일부 한국 성인 남성 흡연자들의 림프구 DNA 손상의 증가 (Increased DNA Damage of Lymphocytes in Korean Male Smokers)

  • 이주현;오은하;이준영;설동근;김주자;이은일
    • Journal of Preventive Medicine and Public Health
    • /
    • 제40권1호
    • /
    • pp.16-22
    • /
    • 2007
  • Objective : The purpose of this study was to evaluate the levels of DNA damage in human lymphocytes caused by smoking and other lifestyle factors. Methods : The study population consisted of 173 normal healthy male adults from 21 to 59 years old. The demographic and lifestyle variables were obtained from administered questionnaires. The level of lymphocytic DNA damage in the peripheral blood was evaluated by the Comet assay. Statistical analyses were done by general linear model analysis and Dunnett's multiple comparison. Results : The difference in DNA damage between smokers and non-smokers was statistically significant. The means for the Tail%DNA were found to be 10.48 in the current smokers and 9.60 in the non-smokers (p < 0.05). The tail moment means were 1.58 and 1.45 (p < 0.05) for the current smokers and non-smokers, respectively. The number of cigarettes smoked per day did not result in a significant difference in the level of DNA damage among the smokers. Other lifestyle factors such as age, and drinking and exercise habits were not related to DNA damage. Conclusions : The DNA damage in the lymphocytes of smokers was found to be significantly higher than that for non-smokers. However, the number of cigarettes smoked per day was not related to DNA damage. Further study is needed to evaluate the relationship between the amount of smoking and level of damage to DNA. In addition, the status of DNA repair activities should be assessed.

Genotoxicity of Taxol and 10-Deacetyl Baccatin III Using Single Cell Gel Electrophoresis (Comet Assay) in Chinese Hamster Lung Fibroblast

  • Kim, Hyun-Joo;Kim, Kyung-Ran;Youn, Ji-Youn;Kim, Min-Hee;Ryu, Jae-Chun
    • 한국환경독성학회:학술대회논문집
    • /
    • 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
    • /
    • pp.61-61
    • /
    • 1996
  • Taxol is used as cancer therapeutic agent. It has been known as weak posotive of chromosome aberration assay in vitro in our previous results (Ryu et al., 1996) and potent clastogens in the mouse bone marrow micronucleus (Tinwell and Ashby, 1994). We performed microgel electrophoresis to determine the effect of taxol and it's precursor 10-deacetyl baccatin III(DAB) on DNA. Microgel electrophoresis is useful, rapid, simple, visual, and sensitive technique for measuring DNA breakage and repair mechanisms in mammalian 근ells. The range of concentration used for taxol were 854, 427, 213.5, 106.8, 53.4 Ug/ml, for DAB 910 ,455, 227.5 U9/ml, Cell viability always exceed 85%. We analyzed the results by using the special software of image analyzer for this comet assay (Komet 3.0). By using this image analyzer software , we can get the result as the tail moment ((mean of tail length - mean of head lengh) x tail%DNA/100). A slight increase in DNA migration was observed for taxol at the concentration of 854 Ug/m4 in the absence of S9 mixture. No increased DNA migration was observed after treatment with DAB.

  • PDF

Depletion of Cytoplasmic Tail of UL18 Enhances and Stabilizes the Surface Expression of UL18

  • Kim, Jung-Sik;Kim, Bon-Gi;Yoon, Il-Hee;Kim, Sang-Joon;Park, Chung-Gyu
    • IMMUNE NETWORK
    • /
    • 제8권4호
    • /
    • pp.130-136
    • /
    • 2008
  • Background: Human cytomegalovirus UL18, a MHC class I homologue, has been considered a natural killer (NK) cell decoy. It ligates LIR-1/ILT2 (CD85j), an NK inhibitory receptor, to prevent lysis of infected target cells. However, precise role of UL18 to NK cell cytotoxicity is yet elusive. Difficulty in clarifying the function of UL18 lies in complication in detecting UL18 mainly due to low level expression of UL18 on the surface and gradual loss of its expression. Methods: To overcome this hurdle, cDNA of cytoplasmic tail-less UL18 was constructed and expressed in swine endothelial cell (SEC). The expression level and its stability in the cell surface were monitored with FACS analysis. Results: Surface expression of UL18 is up-regulated by removing cytoplasmic tail portion from UL18F (a full sequence of UL18). SECs transfected with a cDNA of UL18CY (a cytoplasmic tail-less UL18) stably expressed UL18 molecule on the surface without gradual loss of its expression during 6 week continuous cultures. In the NK cytotoxicity assay, UL18 functions either inhibiting or activating NK cell cytotoxicity according to the source of NK cells. We found that there is individual susceptibility in determining whether the engagement of NK cell and UL18 results in overall inhibiting or activating NK cell cytotoxicity. Conclusion: In this study, we found that cytoplasmic tail is closely related to the regulatory function for controlling surface expression of UL18. Furthermore, by constructing stable cell line in which UL18 expression is up-regulated and stable, we provided a useful tool to clarify exact functions of UL18 on various immune cells having ILT2 receptor.

Comet Assay를 이용한 Flavonoids와 항산화 비타민의 인체임파구 세포 DNA 손상 보호 효과 (Protective Effect of Flavonoids on Lymphocyte DNA Damage Using Comet Assay)

  • 박유경;전은재;강명희
    • Journal of Nutrition and Health
    • /
    • 제36권2호
    • /
    • pp.125-132
    • /
    • 2003
  • The present study was attempted to investigate and compare the antioxidant potency of several well-know flavonoids, antioxidant vitamin and commercially available popular beverages. The antioxidant potency was assessed by the effect on reducing oxidative DNA damage of human lymphocytes. Cellular oxidative DNA damage was measured by SCGE (single-cell gel electrophoresis), also known as comet assay. Lymphocytes were pre-treated for 30 minutes with wide ranges of doses of apigenin, kaempferol, luteolin, myricetin, rutin, quercetin, $\alpha$-tocopherol (10,25,50,100,200,500,1000 $\mu$M) ,green tea extract or grape juice (10,50,100,250,500,1000 $\mu$g/mL) followed by a $H_2O$$_2$(100 $\mu$M) treatment for 5 min as an oxidative stimulus. The physiological function of each antioxidant substance on oxidative DNA damage was analyzed as tail moment (tail length $\times$ percentage migrated DNA in tail) and expressed as relative DNA damage score after adjusting by the level of control treatment. Cells treated with $H_2O$$_2$alone (positive control) had an extensive DNA damage compared with cells treated with phosphate buffered saline (PBS, negative control) or pre-treated with all the tested samples. Of all the six flavonoids, quercetin was the most potent antioxidant showing the lowest $ED_{50}$/ of 8.5 $\mu$g/mL (concentration to produce 50% protection of relative DNA damage). The antoxidant potency of individual flavonoids were ranked as follows in a decreasing order; luteolin (18.4 $\mu$g/mL), myricetin (19.0 $\mu$g/mL) , rutin (22.2 $\mu$g/mL) , apigenin (24,3 $\mu$g/mL) , kaempferol (25.5 $\mu$g/mL). The protective effect of $\alpha$-tocopherol was substantially lower (highest $ED_{50}$value of 55.0 $\mu$g/mL) than all the other flavonoids, while the protective effect was highest in green tea and grape juice with low ED5O value of 7.6 and 5.3, respectively. These results suggest that flavonoids, especially quercetin, and natural compounds from food product, green tea and grape juice, produced powerful anti-oxidative activities, even stronger than $\alpha$-tocopherol. Taken together, supplementation of antioxidants to lymphocytes followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.

환경재해 방지를 위한 생물정보로서의 사람 림프구 DNA 손상에 대한 방사선과 살충제의 상승작용 (Synergistic Interaction of Radiation with Pesticide on DNA Damage in Human Lymphocytes as Biological Information for Prevention of Environmental Disaster)

  • 김진규
    • 환경생물
    • /
    • 제19권1호
    • /
    • pp.19-24
    • /
    • 2001
  • 병해충을 막기 위해 농업용 살충제가 광범위하게 사용되고 있다. 농약사용에 따른 생물학적 위해가 우려되며 농약이 또 다른 환경유해요인과 인체에 상승적으로 작용할 경우 농업재해로 이어질 가능성이 있다. 다양한 인자에 의한 DNA손상을 감지하는데 유용한 단세포 겔 전기영동법을 이용하여 살충제와 방사선에 의한 사람 림프구 DNA손상을 평가하였다. 각기 다른 농도로 살충제를 10분간 전처리한 림프구와 정상 림프구에 0-2.0 Gy의 방사선으로 조사한 다음 DNA 손상도를 평가하였다. DNA가닥 절단에 대한 표식인 tail moment의 증가는 감마선에 대해서 뚜렷한 선량-반응 관계를 나타내었다. 단세포 겔 전기영동법을 통한 평가결과 권장 사용농도 이상의 살충제는 림프구에 대한 유전독성을 나타내었을 뿐 아니라 방사선과 함께 상승작용을 일으켜 림프구 DNA손상을 더욱 증가시키는 것이 확인되었다. 이와 같은 결과는 환경재해의 방재 및 예방 조치에 필요한 생물정보를 사전에 제공하는 의미를 가진다.

  • PDF

Interspecific hybridization in seahorses: artificially produced hybrid offspring of Hippocampus kuda and Hippocampus reidi

  • Han, Sang-Yun;Rho, Sum;Noh, Gyeong Eon;Kim, Jin-Koo
    • Fisheries and Aquatic Sciences
    • /
    • 제21권5호
    • /
    • pp.11.1-11.8
    • /
    • 2018
  • Interspecific hybridization experiments were conducted between the common seahorse Hippocampus kuda (male) and the slender seahorse H. reidi (female) during artificial rearing to develop a new aquarium fish with unique polyandrous mating. Molecular analysis via mitochondrial DNA (mtDNA) cytochrome b and nuclear DNA (ncDNA) ribosomal protein S7 gene supported the hybridization between the two species, and the hybrid also showed morphological characteristics of both species. Juveniles of H. kuda have dense melanophores on the whole body or only on the trunk and tail, whereas juveniles of H. reidi have thin melanophores on the whole body or present in stripes only along their prominent trunk and tail rings. However, all the hybrid juveniles had dense melanophores only on the tail, with the striped trunk rings, thus showing an intermediate pattern, and these patterns were limited to the fairly early stage of development (1-10 days old). In contrast, the two eye spines in the hybrid were apparent after 9 days old, which were not inherited from H. kuda (one eye spine), but from H. reidi (two eye spines). According to LOESS (local regression) analysis, the growth rate increased between 20 and 25 days, and the hybrids grew faster than H. kuda when they entered the explosive second phase of growth between 25 and 45 days for all the seahorses. This study highlights the hybridization between H. kuda and H. reidi may contribute to the improved taxonomic information of young seahorses.

Cloning and Nucleotide Sequence of a cDNA Encoding the Rat Triosephosphate Isomerase

  • Lee, Kyunglim;Ryu, Jiwon
    • Archives of Pharmacal Research
    • /
    • 제19권6호
    • /
    • pp.497-501
    • /
    • 1996
  • A gene coding for triosephosphate isomerase (TPI) from a rat skeletal muscle cDNA library was cloned and its nucleotide sequence was determined. The 1, 348-bp cDNA clone contains 24 bp $5^I$ noncoding region, the entire 750 bp coding region corresponding to a protein of 249 amino acids, $547bp 3^I$ noncoding region and part of a poly(A) tail. It also contains a polyadenylation signal, AATAAA, starting from 17 bp upstream of the poly(A) tail. The calculated molecular weight of rat TPI is 27.8 kDa and the net charge is +4. The deduced amino acid sequence from rat TPI CDNA sequence has 93% and 94% homology with that of mouse and human clones, respectively. The amino acids at the residue of Asn12, Lys14, His96, Glu 166, His96, His101, Ala177, Tyr165, Glu13O, Tyr2O9, and Ser212 in catalytic site are completely identical, confirming that the functional residues in TPI proteins are highly conserved throughout evolution. The most profound characteristic of rat TPI enzyme, compared with other TPIs, is that there are five cysteine substitutions at the residue of 21, 27, 159, 195 and 204. A Glu123 instead of Gly was found in rabbit, rhesus, mouse and human sequences. Through the method of RT-PCR, the mRNA transcription level of TPI gene was found to be different among various tissues and was highest in muscle.

  • PDF

열발광, 전자스핀공명 및 DNA Comet 분석에 의한 대두의 방사선 조사 여부 검지 특성 (Detection Characteristics of TL, ESR and DNA Comet for Irradiated Soybeans)

  • 이은영;정재영;노정은;조덕조;권중호
    • 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.

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

  • 이은영;정재영;조덕조;권중호
    • 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.

  • PDF