• Title/Summary/Keyword: lymphocyte DNA

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The Influence of Smoking and Low Dose Radiation Exposure to the Damage of the Lymphocyte DNA (흡연과 낮은 방사선 피폭량이 Lymphocyte DNA 손상에 미치는 영향)

  • Shin Heuyn-Kil;Kim Yun-Joo;Kwon Eun-Hye;Yook Jin-Young;Choi Soo-Yong
    • Environmental Analysis Health and Toxicology
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    • v.18 no.4
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    • pp.237-242
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    • 2003
  • Single cell gel electrophoresis (SCGE) was used to the experiment with the variation on the amount of smoking and low dose radiation exposure to find how much the Lymphocyte DNA was damaged, and especially for whom smoke a lot(about 20 or more than 20 cigarettes a day) it was found to be highly damaged. While, the damage of 'not more than 20 cigarettes a day' was found to be not so much significant as like for whom smoke about or more than 20 cigarettes a day And, according to the different amount of the radiation exposure, the Lymphocyte DNA was found to be considerably damaged for 0-13m Sv (P<0.01), it was not able to prove the relationship between the DNA damage and the radiation exposure.

Lymphocyte DNA Damage and Anti-Oxidative Parameters are Affected by the Glutathione S-Transferase (GST) M1 and T1 Polymorphism and Smoking Status in Korean Young Adults (흡연 여부에 따른 Glutathione S-transferase (GST) M1 및 T1 유전자 다형성이 우리나라 젊은 성인의 임파구 DNA 손상과 항산화 영양상태 지표들 간의 관련성에 미치는 영향)

  • Han, Jeong-Hwa;Lee, Hye-Jin;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.44 no.5
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    • pp.366-377
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    • 2011
  • Glutathione S-transferase (GST) is a multigene family of phase II detoxifying enzymes that metabolize a wide range of exogenous and endogenous electrophilic compounds. GSTM1 and GSTT1 gene polymorphisms may account for inter-individual variability in coping with oxidative stress. We investigated the relationships between the level of lymphocyte DNA and antioxidative parameters and the effect on GST genotypes. GSTM1 and GSTT1 were characterized in 301 young healthy Korean adults and compared with oxidative stress parameters such as the level of lymphocyte DNA, plasma antioxidant vitamins, and erythrocyte antioxidant enzymes in smokers and non smokers. GST genotype, degree of DNA damage in lymphocytes, erythrocyte activities of superoxide dismutase, catalase, and glutathione peroxidase (GSH-Px), and plasma concentrations of total radical-trapping antioxidant potential (TRAP), vitamin C, ${\alpha}$- and ${\gamma}$-tocopherol, ${\alpha}$- and ${\beta}$-carotene, and cryptoxanthin were analyzed. Lymphocyte DNA damage assessed by the comet assay was higher in smokers than that in non-smokers, but the levels of plasma vitamin C, ${\beta}$-carotene, TRAP, erythrocyte catalase, and GSH-Px were lower than those of non-smokers (p < 0.05). Lymphocyte DNA damage was higher in subjects with the GSTM1- or GSTT1-present genotype than those with the GSTM1-present or GSTT1- genotype. No difference in erythrocyte antioxidant enzyme activities, plasma TRAP, or vitamin levels was observed in subjects with the GSTM1 or GSTT1 genotypes, except ${\beta}$-carotene. Significant negative correlations were observed between lymphocyte DNA damage and plasma levels of TRAP and erythrocyte activities of catalase and GSH-Px after adjusting for smoking pack-years. Negative correlations were observed between plasma vitamin C and lymphocyte DNA damage only in individuals with the GSTM1-present or GSTT1- genotype. The interesting finding was the significant positive correlations between lymphocyte DNA damage and plasma levels of ${\alpha}$-carotene, ${\beta}$-carotene, and cryptoxanthin. In conclusion, the GSTM1- and GSTT1-present genotypes as well as smoking aggravated antioxidant status through lymphocyte DNA damage. This finding confirms that GST polymorphisms could be important determinants of antioxidant status in young smoking and non-smoking adults. Consequently, the protective effect of supplemental antioxidants on DNA damage in individuals carrying the GSTM1- or GSTT1-present genotypes might show significantly higher values than expected.

The effect of carrot juice, ${\beta}$-carotene supplementation on lymphocyte DNA damage, erythrocyte antioxidant enzymes and plasma lipid profiles in Korean smoker

  • Lee, Hye-Jin;Park, Yoo-Kyoung;Kang, Myung-Hee
    • Nutrition Research and Practice
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    • v.5 no.6
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    • pp.540-547
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    • 2011
  • High consumption of fruits and vegetables has been suggested to provide some protection to smokers who are exposed to an increased risk of numerous cancers and other degenerative diseases. Carrot is the most important source of dietary ${\beta}$-carotene. Therefore, the objective of this study was to investigate whether carrot juice supplementation to smokers can protect against lymphocyte DNA damage and to compare the effect of supplementationof capsules containing purified ${\beta}$-carotene or a placebo (simple lactose). The study was conducted in a randomized and placebo-controlled design. After a depletion period of 14 days, 48 smokers were supplemented with either carrot juice (n = 18), purified ${\beta}$-carotene (n = 16) or placebo (n = 14). Each group was supplemented for 8 weeks with approximately 20.49 mg of ${\beta}$-carotene/day and 1.2 mg of vitamin C/day, as carrot juice (300 ml/day) or purified ${\beta}$-carotene (20.49 mg of ${\beta}$-carotene, 1 capsule/day). Lymphocyte DNA damage was determined using the COMET assay under alkaline conditions and damage was quantified by measuring tail moment (TM), tail length (TL), and% DNA in the tail. Lymphocyte DNA damage was significantly decreased in the carrot juice group in all three measurements. The group that received purified ${\beta}$-carotene also showed a significant decrease in lymphocyte DNA damage in all three measurements. However, no significant changes in DNA damage was observed for the placebo group except TM (P = 0.016). Erythrocyte antioxidant enzyme was not significantly changed after supplementation. Similarly plasma lipid profiles were not different after carrot juice, ${\beta}$-carotene and placebo supplementation. These results suggest that while the placebo group failed to show any protective effect, carrot juice containing beta-carotene or purified ${\beta}$-carotene itself had great antioxidative potential in preventing damage to lymphocyte DNA in smokers.

Effects of the NADPH Oxidase p22phox C242T Polymorphism on Endurance Exercise Performance and Oxidative DNA Damage in Response to Aerobic Exercise Training

  • Paik, Il-Young;Jin, Chan-Ho;Jin, Hwa-Eun;Kim, Young-Il;Cho, Su-Youn;Roh, Hee-Tae;Suh, Ah-Ram;Suh, Sang-Hoon
    • Molecules and Cells
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    • v.27 no.5
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    • pp.557-562
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    • 2009
  • We examined the effects of the NADPH oxidase p22phox C242T polymorphism on endurance exercise performance and oxidative DNA damage in response to acute and chronic exercises. One hundred three subjects were recruited, among which 26 healthy subjects (CC: 12, TC: 12, and TT: 2) were studied during rest, exercise at 85% $VO_2max$, and recovery before and after 8 weeks of treadmill running. Lymphocyte DNA damage increased significantly in response to exercise (p < 0.05). There were no significant differences in plasma MDA, SOD concentrations and lymphocyte DNA damage between CC genotype and T allele group, but significant endurance training differences were observed. Endurance training increased exercise time to exhaustion in both the CC genotype and T allele groups (p < 0.05) but no significant difference was found between groups. The results of the current study with young, healthy, Korean men are interpreted to mean that 1) the majority had the CC genotype of the NADPH oxidase p22phox C242T polymorphism (82.5%: CC, 15.5%: TC, 1.9%: TT), 2) acute exercise increased lymphocyte DNA damage, 3) endurance training significantly increased exercise time to exhaustion, and alleviated lymphocyte DNA damage, and 4) The NADPH oxidase p22phox C242T polymorphism, however, did not alter lymphocyte DNA damage or exercise performance at rest, immediately after exercise, or during recovery.

Gender-Specific Changes of Plasma MDA, SOD, and Lymphocyte DNA Damage during High Intensity Exercise (고강도 운동 시 성별에 따른 혈장 MDA, SOD 및 임파구 DNA 손상 변화)

  • Cho, Su-Youn;Chung, Young-Soo;Kwak, Yi-Sub;Roh, Hee-Tae
    • Journal of Life Science
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    • v.21 no.6
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    • pp.838-844
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    • 2011
  • The purpose of this study was to investigate gender-specific changes of plasma MDA, SOD, and lymphocyte DNA damage during high intensity exercise. In this study, 17 healthy male and 18 healthy female college students ran on a treadmill at 85%$VO_{2max}$ until the point of all-out. Blood-collecting was carried out five times (Rest, Ex-Exha, R0.5h, R4h and R24h), and with the collected blood, plasma malondialdehyde (MDA), superoxide dismutase (SOD), and lymphocyte DNA damage were analyzed. Plasma MDA and SOD concentration increased significantly at the Ex-Exha (p<0.05), and there were no significant differences in gender. For the degree of lymphocyte DNA damage, all %DNA in the tail, tail length and tail moment increased significantly at the Ex-Exha (p<0.05), and %DNA in the tail and tail length were significantly higher in the male group than in the female group (p<0.05). These results suggest that acute high intensity exercise not only causes oxidative stress but also brings about lymphocyte DNA damage. In addition, it was found that males showed higher DNA damage than females in terms of oxidative stress subject to high intensity exercise. Nevertheless, further subsequent studies are required in order to better understand the mechanism behind DNA damage varying with gender, in a way that takes into consideration physical fitness, hormonal level, exercise intensity and duration - additional factors which might affect DNA damage.

Efficacy of a DNA Vaccine Carrying Eimeria maxima Gam56 Antigen Gene against Coccidiosis in Chickens

  • Xu, Jinjun;Zhang, Yan;Tao, Jianping
    • Parasites, Hosts and Diseases
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    • v.51 no.2
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    • pp.147-154
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    • 2013
  • To control coccidiosis without using prophylactic medications, a DNA vaccine targeting the gametophyte antigen Gam56 from Eimeria maxima in chickens was constructed, and the immunogenicity and protective effects were evaluated. The ORF of Gam56 gene was cloned into an eukaryotic expression vector pcDNA3.1(zeo)+. Expression of Gam56 protein in COS-7 cells transfected with recombinant plasmid pcDNA-Gam56 was confirmed by indirect immunofluorescence assay. The DNA vaccine was injected intramuscularly to yellow feathered broilers of 1-week old at 3 dosages (25, 50, and $100{\mu}g/chick$). Injection was repeated once 1 week later. One week after the second injection, birds were challenged orally with $5{\times}10^4$ sporulated oocysts of E. maxima, then weighed and killed at day 8 post challenge. Blood samples were collected and examined for specific peripheral blood lymphocyte proliferation activity and serum antibody levels. Compared with control groups, the administration of pcDNA-Gam56 vaccine markedly increased the lymphocyte proliferation activity (P<0.05) at day 7 and 14 after the first immunization. The level of lymphocyte proliferation started to decrease on day 21 after the first immunization. A similar trend was seen in specific antibody levels. Among the 3 pcDNA-Gam56 immunized groups, the median dosage group displayed the highest lymphocyte proliferation and antibody levels (P<0.05). The median dosage group had the greatest relative body weight gain (89.7%), and the greatest oocyst shedding reduction (53.7%). These results indicate that median dosage of DNA vaccine had good immunogenicity and immune protection effects, and may be used in field applications for coccidiosis control.

PCR에 의한 HIV의 진단

  • Kang, Chun
    • The Microorganisms and Industry
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    • v.18 no.2
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    • pp.26-30
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    • 1992
  • HIV는 ELISA나 WB에 의해 항체가 검출되기 수개월 혹은 수 년전에도 proviral DNA 상태로 감염된 세포의 chromosome내에 존재하는 것이 주지의 사실이다. 그동안 Southern blot, in situ hybridization등에 의해 이 proviral DNA를 검출하려는 연구가 진행되어 왔으나 lymphocyte $10^{4}$-$10^{6}$개 중 1개가 감염되어 있으며 lymphocyte chromosomal DNA에 비해 viral DNA의 양이 미량이므로 검출하기에는 민감도가 낮은 문제점이 있다. 본 고에서는 근래 개발되어 널리 사용되고 있는 polymerase chain reaction(PCR)을 이용한 HIV의 진단에 관해 살펴보고자 한다.

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Comparison of the Protective Effect of Antioxidant Vitamins and Fruits or Vegetable Juices on DNA Damage in Human Lymphocyte Cells Using the Comet Assay (Comet Assay를 이용한 항산화 비타민과 과일.야채즙의 인체 임파구 세포 DNA 손상 감소 효과 비교)

  • 전은재;박유경;김정신;강명희
    • Journal of Nutrition and Health
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    • v.37 no.6
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    • pp.440-447
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    • 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.

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

  • Park, Eun-Ju;Ryoo, Kun-Kul;Lee, Yoon-Bae;Lee, Jong-Kwon;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.155-160
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    • 2005
  • Electrolyzed reduced water (ERW), showing extremely negative oxidation-reduction potential, was used to investigate the effects of paraquat-induced damages on DNA from human lymphocyte. The effect of ERW on paraquat-induced oxidative DNA damage in human lymphocytes was evaluated by Comet assay (single-cell gel electrophoresis) quantified as percentage fluorescence in tail. Comet assay has been used widely to assess the level of the DNA damage in individual cells. Lymphocytes were oxidatively challenged with various concentrations of paraquat for 30 min at $37^{\circ}C$, and were then treated with electrolyzed reduced water for 30 min. The oxidative DNA damage by paraquat, as indicated by the fluorescent tail in DNA, increased in a dose-dependent manner. However, oxidative damage of the DNA was almost completely prevented upon treatment with electrolyzed reduced water.

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

  • Kim Young-Chul;Sim Mi-Ja;Kwon Chong-Suk
    • Environmental Analysis Health and Toxicology
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    • v.19 no.1
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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.