• Title/Summary/Keyword: DNA-level

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Changes in Polyamine Level and Chloroplast DNA Methylation in Chlamydomonas reinhardtii (Chlamydomonas의 Polyamine 함량변화와 엽록체 DNA Methylation)

  • 이순희
    • Journal of Plant Biology
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    • v.37 no.1
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    • pp.101-109
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    • 1994
  • Relationship between polyamine level and DNA methylation in the absence or presence of MGBG(l mM), which is an enzyme-activated reversible inhibitor of SAMDC, has been investigated during gametogenesis of Chlamydomonas. In the absence of MGBG, polyamine levels decreased in Chlamydomonas 137C(+) and 137C(-) during gametogenesis. And polyamine level of 137C(+) was 2-5 times as much as that of 137C(-) and showed a significant decrease unlike that of 137C(-). In vitro, MGBG inhibited ctDNA methylation of 137C(+) by 20-30% but did not inhibited that of 137C(-). Also, MGBG inhibited DNA methylase by 60% in vitro. The results obtained in the present work suggest the possibility that the changes of polyamine level may be associated with ctDNA methylation during gametogenesis of Chlamydomonas.omonas.

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Non-invasive Biological Monitoring of DNA Adducts Formed at Workers Handling 3,3-Dichlorobenzidine(DCB) by Using GC/MS

  • Lee, Jin-Heon
    • Journal of Environmental Health Sciences
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    • v.29 no.4
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    • pp.21-26
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    • 2003
  • We examine the metabolites(DCB and acetyl DCB) extracted from exfoliated urothelial cells of 33 workers who employed DCB-handling industries. The characteristics of workers submitted urine, whose age, working years and smoking persons were 41.9$\pm$11.1, 8.7$\pm$5.5 and 25(32.0%), respectively. DNA adduct was isolated from the exfoliated urothelial cells by applying $^{32}$ p-postlabeling procedure. Metabolites(DCB and acetyl DCB) were extracted from DNA adducts by hydrolyzing and N-glycosylase. Concentrations of DCB and acetyl DCB were 28.6$\pm$5.25 ng/g DNA, and 17.0$\pm$3.73 ng/g DNA, respectively. The regression between DCB level and exposure years of workers is y = 1.668 + 2.588x(p = 0.005, $r^2$= 0.394). The regression between acetyl DCB level and exposure years of workers is y = 8.071 + 1.325x(p = 0.076, $r^2$= 0.222). Smoking workers are significantly higher than non-smoking workers on DCB and acetyl DCB level(p = 0.065 and 0.021, respectively). DCB level was 33.9$\pm$7.14 ng/g DNA on smokers, and 23.1$\pm$9.97 ng/g DNA on non-smokers. Acetyl DCB was 25.1$\pm$5.27 ng/g DNA on smokers, and 8.92$\pm$7.22 ng/g DNA on non-smokers.

A Visualization Tool for Computational Analysis of DNA Methylation Level Using Bisulfite Sequencing Data

  • Tae, Hong-Seok
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.136-137
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    • 2011
  • Methylation of cytosine is a post-synthesis modification that does not affect the primary DNA sequence but greatly influences gene expression level and phenotypes of an organism. As high-throughput sequencing of bisulfite-treated DNA is the most efficient method to identify methylated sites, several tools to map sequencing reads on a reference are available. But tools to visualize and to interpret the methylation level of methylation sites are currently insufficient. Herein, we present a novel tool to visualize the methylation level of CpG sites.

INDUCTION OF MITOCHONDRIAL DNA DELETION BY IONIZING RADIATION IN HUMAN LUNG FIBROBLAST IMR-90 CELLS

  • Eom, Hyeon-Soo;Jung, U-Hee;Park, Hae-Ran;Jo, Sung-Kee
    • Journal of Radiation Protection and Research
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    • v.34 no.2
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    • pp.49-54
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    • 2009
  • Mitochondrial DNA (mtDNA) deletion is a well-known marker for oxidative stress and aging and also contributes to their unfavorable effects in cultured cells and animal tissues. This study was conducted to investigate the effect of ionizing radiation (IR) on mtDNA deletion and the involvement of reactive oxygen species (ROS) in this process in human lung fibroblast (IMR-90) cells. Young IMR-90 cells at population doubling (PD) 39 were irradiated with $^{137}Cs$ $\gamma$-rays and the intracellular ROS level was determined by 2',7'-dichlorofluorescein diacetate (DCFH-DA) and mtDNA common deletion (4977bp) was detected by nested PCR. Old cells at PD 55 and $H_2O_2$-treated young cells were compared as the positive control. IR increased the intracellular ROS level and mtDNA 4977 bp deletion in IMR-90 cells dose-dependently. The increases of ROS level and mtDNA deletion were also observed in old cells and $H_2O_2$-treated young cells. To confirm the increased ROS level is essential for mtDNA deletion in irradiated cells, the effects of N-acetylcysteine (NAC) on IRinduced ROS and mtDNA deletion were examined. 5 mM NAC significantly attenuated the IR-induced ROS increase and mtDNA deletion. These results suggest that IR induces the mtDNA deletion and this process is mediated by ROS in IMR-90 cells.

The outcome of perinatal prophylaxis for HBeAg positive mothers according to the maternal HBV-DNA levels at the delivery time (HBeAg 양성 산모의 분만 직후 HBV-DNA 수치에 따른 주산기 예방조치의 결과)

  • Jeong, On;Kim, Jong-Hyun
    • Clinical and Experimental Pediatrics
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    • v.50 no.4
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    • pp.348-354
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    • 2007
  • Purpose : Perinatal hepatitis B viral infection is decreasing; however, 10% of babies to HBeAg positive mothers still become chronic carriers despite perinatal prophylaxis. Although, the cause of prophylaxis failure is still unclear, an importance of maternal HBV-DNA level at the delivery time has been suggested. This study was established to certify if it would be a useful predictable factor for the outcome of perinatal prophylaxis. Methods : Twenty-nine HBeAg positive mothers whose babies had known outcomes of prophylaxis were selected. To determine the amount of maternal HBV-DNA, a quantitative PCR was performed with the WHO International Standard for HBV DNA NAT assays. Results : The mean logarithm HBV-DNA level of mothers with failed outcomes was significantly higher than that of mothers with succeessful outcomes (7.99 vs. 6.72, P=0.015). The predictable maternal HBV-DNA cut-off level to prophylaxis outcome was $2.83{\times}10^7copies/mL$ (100 pg/mL). None out of the case 16 (0%) who had below this level, and 5 out of 13 (38.5%) who had above this level of maternal HBV-DNA failed in perinatal prophylaxis. Conclusion : Mothers with higher levels of HBV-DNA at delivery time would be prone to a worse outcome of prophylaxis using the conventional approach. Perinatal prophylaxis failure rate can be reduced, if we try to introduce more potent prophylactic treatment into the cases with this risk factor.

Complementary DNA Cloning and Nucleotide Sequence Analysis of Coat Protein Gene from TMV Tomato Strain (토마토에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석)

  • 이청호;이영기;강신웅;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.18 no.2
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    • pp.101-106
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    • 1996
  • Tobacco mosaic virus (TMV) tomato strain was isolated from tomato "Seo-Kwang" in Korea. The virion was purified by density gradient centrifugation, and total viral RNA was isolated from the purified particles. Coat protein (CP) cDNA of the virus was synthesized by RT-PCR, and the purified cDNA fragment was subcloned to pBluescript II SK-. The analysis of nucleotide sequence showed that this cDNA was 693 nucleotides long from the insert of clone p1571 and p1572 which contain complete codons of the viral coat protein gene (474 nucleotides) and 3' untranslated region. The nucleotides of coat protein encoding cDNA of the strain were 6 nucleotides less than that of TMV common strain isolated from tobacco plant in Korea. The CP gene showed 70% maximum homology with that of the common strain in the nucleotide level and 86% maximum homology in amino acid level.cid level.

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Identification of Radiation-Sensitive Gene in U937 Cell by using cDNA-Chip Composed of Human Cancer Related Gene (U937 세포에서 발암관련 유전자들로 구성된 DNA chip을 이용한 방사선 감수성 유전자들의 선발)

  • 김종수;김인규;강경선;윤병수
    • Environmental Mutagens and Carcinogens
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    • v.22 no.1
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    • pp.54-59
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    • 2002
  • We have used cDNA microarray hybridization to identify gene regulated in response to gamma-irradiation in U-937 cell. The cDNA-chip was composed entirely of 1,000 human cancer related gene including apoptosis and angiogenesis etc. In gamma-irradiated U-937 cell, highly charged protein, ribosomal protein L32, four and a half LIM domains 3, lipocalin 2 (oncogene 24p3) and interleukin 15, ataxia telangiectasia mutated (includes complementation groups A, C and D) genes showed increased level of its transcription, and cell division cycle 25A, dihydrofolate reductase, topoisomerase (DNA) II beta(180kD), kinase suppressor of ras and strarigin genes showed reduced level of its transcription compared to untreated U-937 cell. The significant change of level of transcription was not found in well-known ionizing radiation(IR)-responsive gene, such as transcription factor TP53 and p53 related gene, except ataxia telangiectasia mutated gene.

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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.

A Case Report of the Use of Traditional Korean Medicine to Treat a Patient with Chronic Hepatitis B Whose HBV-DNA Level Failed to Drop Below Undetectable Levels Following Long-term Treatment with Entecavir (Entecavir와 한의약 치료를 병행한 후 HBV-DNA가 음전된 만성 B형 간염 환자 증례 보고)

  • Han, Chang-woo
    • The Journal of Internal Korean Medicine
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    • v.38 no.2
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    • pp.284-288
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    • 2017
  • The patient presented to the clinic with the complaint of severe fatigue. The patient had been diagnosed with chronic hepatitis B a number of years earlier. Although he had used entecavir, an antiviral agent, for over two years, his HBV-DNA level had not dropped below undetectable levels. The fatigue seemed to be associated with chronic hepatitis B. Traditional Korean medicine (TKM) therapy for chronic hepatitis B was administered in conjunction with entecavir and at the same dose. The excessive fatigue gradually decreased following the treatment. On the 28th day, laboratory tests revealed that the patient's bilirubin level was slightly lower and that his HBV-DNA level had dropped below undetectable levels. The addition of TKM therapy may have contributed to the HBV-DNA clearance. No similar cases have been reported in Korea. Herein, we summarize the patient's progress.

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

  • Lee, Joo-Hyun;Oh, Eun-Ha;Lee, June-Young;Sul, Dong-Geun;Kim, Joo-Ja;Lee, Eun-Il
    • Journal of Preventive Medicine and Public Health
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    • v.40 no.1
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    • pp.16-22
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    • 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.