• Title/Summary/Keyword: human lymphocyte DNA

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Effects of Smoking and Age on SCE Frequency Reflecting DNA Damage of Human Lymphocytes in Elderly Koreans (노인의 흡연상태와 나이가 SCE 빈도수로 본 임파구 DNA 손상에 미치는 영향)

  • 이정희;강명희
    • Journal of Nutrition and Health
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    • v.36 no.8
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    • pp.851-858
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    • 2003
  • Sister chromatid exchange (SCE) has recently become a common cytogenic assay system for detecting exposure to chemical mutagens and carcinogens. One application of SCE is the monitoring of populations believed to have been exposed to such agents. A cross-sectional study of SCE frequency in peripheral blood lymphocytes from 45 Koreans aged 61 to 84 years was conducted. The effect of cigarette smoking and age on SCE was assessed by different degrees of smoking status such as smokers (n = 14), ex-smokers (n = 16) and non-smokers (n = 15). Mean spontaneous SCE per cell for the smokers (11.5 $\pm$ 1.1) was significantly higher (p < 0.05) than that for the non-smokers (8.8 $\pm$ 0.3). However, mean SCE frequencies per cell for the ex-smokers (10.3 $\pm$ 0.6) were not significantly different from those of the smokers or the non-smokers. The smokers showed an increased number of high SCE frequency cells (HFCs) when compared to the ex-smokers and non-smokers (p < 0.05). The mean SCE frequencies of the non-smokers showed a statistically significant increase (p < 0.05) with the subject's age. These results show that age and smoking habits contribute a great deal in setting a higher degree of basal DNA damage in elderly Koreans, and smoking appeared to be a more significant damaging factor than age.

Genotoxic and Anti-Genotoxic Effects of Vanillic Acid Against Mitomycin C-Induced Genomic Damage in Human Lymphocytes In Vitro

  • Erdem, Merve Guler;Cinkilic, Nilufer;Vatan, Ozgur;Yilmaz, Dilek;Bagdas, Deniz;Bilaloglu, Rahmi
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.4993-4998
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    • 2012
  • Vanillic acid, a vegetable phenolic compound, is a strong antioxidant. The aim of the present study was to determine its effects on mitomycin C-induced DNA damage in human blood lymphocyte cultures in vitro, both alone and in combination with mitomycin C (MMC). The cytokinesis block micronucleus test and alkaline comet assay were used to determine genotoxic damage and anti-genotoxic effects of vanillic acid at the DNA and chromosome levels. MMC induced genotoxicity at a dose of $0.25{\mu}g/ml$. Vanillic acid ($1{\mu}g/ml$) significantly reduced both the rates of DNA damaged cells and the frequency of micronucleated cells. A high dose of vanillic acid ($2{\mu}g/ml$) itself had genotoxic effects on DNA. In addition, both test systems showed similar results when tested with the negative control, consisting of dimethyl sulfoxide (DMSO) in combination with vanillic acid ($1{\mu}g/ml$)+MMC. In conclusion, vanillic acid could prevent oxidative damage to DNA and chromosomes when used at an appropriately low dose.

Evidence for VH Gene Replacement in Human Fetal B Cells

  • Lee, Jisoo;Cho, Young Joo;Lipsky, Peter E.
    • IMMUNE NETWORK
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    • v.2 no.2
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    • pp.79-85
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    • 2002
  • Background: In contrast to evidences of Ig H chain receptor editing in transformed cell lines and transgenic mouse models, there has been no direct evidence that this phenomenon occurs in human developing B cells. Methods: $V_HDJ_H$ rearrangements were obtained from genomic DNA of individual $IgM^-$ B cells from liver and $IgM^+B$ cells from bone marrow of 18 wk of gestation human fetus by PCR amplification and direct sequencing. Results: We found three examples of H chain receptor editing from $IgM^+$ and $IgM^-human$ fetal B cells. Two types of $V_H$ replacements were identified. The first involved $V_H$ hybrid formation, in which part of a $V_H$ gene from the initial VDJ rearrangement is replaced by part of an upstream $V_H$ gene at the site of cryptic RSS. The second involved a gene conversion like replacement of CDR2, in which another $V_H$ gene donated a portion of its CDR2 sequence to the initial VDJ rearrangement. Conclusion: These data provide evidence of receptor editing at the H chain loci in developing human B cells, and also the first evidence of a gene conversion event in human Ig genes.

Radiation induced Chromosome aberration in human Iymphocyte detected by Fluorescence in sifu hybridization (FISH(Fluorescence in situ hybridization)를 이용하여 분석한 방사선에 의해 유발된 림프구의 염색체 이상)

  • 정해원;손은희;기혜성;하성환
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.88-96
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    • 1996
  • Fluorescence in situ hybridization (FISH) with the DNA probe for human chromosome 4 was used to analyse in vitro radiation induced chromosome rearrangement in peripheral lymphocyte. Translocations, dicentrics, acentrics and color junctions involving the painted chromosome were scored according to the Protocol for Aberration Identification and Nomenclature Terminology (PAINT) system. The frequency of chromosome rearrangements including reciprocal translocation, dicentric, acentric fragment and color junction increased with radiation dose. The frequency of dicentric chromosome reduced by the fixation time following irradiation, whereas that of translocation was relatively persistent. The applicability of FISH for scoring stable translocation for biological dosimetry was demonstrated.

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Sister Chromatid Exchanges in Lymphocytes on Normal Human Blood Culture with Mercury chloride or Methylmercury Chloride (Mercury chloride 및 Methylmercury chloride가 정상인(正常人)의 혈액배양(血液培養)에서 임파구(淋巴球)의 자매염색분체교환(姉妹染色分體交換)에 미치는 영향(影響))

  • Koh, Dai-Ha
    • Journal of Preventive Medicine and Public Health
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    • v.17 no.1
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    • pp.245-250
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    • 1984
  • Reciprocal exchanges of DNA in sister chromatids (SCEs) are induced by various carcinogens and mutagens, although the quantitative relationship between the number of mutations and SCEs induced varies among chemicals. Nevertheless, the analysis of SCEs production by various agents often proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity. Mercury, even if which has no evidences for mutagenicity and carcinogenicity, is reported to exert some cytotoxic effects, such as chromosomal aberrations or bad influences to ovulation and reproduction in experimental animals, etc.. In this study, tests for sister chromatid exchanges have been carried out on normal human lymphocytes in whole blood culture to add mercury chloride ($HgCl_2$) or methylmercury chloride ($CH_3\;HgCl$) for 72 hr. The results indicate the dose-dependent relationship between the frequencies of SCEs and the concentrations of $HgCl_2,\;CH_{3}HgCl$ and 5-bromo-2'-deoxyuridine (BrdU). Lymphocyte proliferation has depressed in the higher concentration of mercury.

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Human HS1BP3 induces cell apoptosis and activates AP-1

  • Shi, Taiping;Xie, Jieshi;Xiong, Ying;Deng, Weiwei;Guo, Jinhai;Wang, Feng;Ma, Dalong
    • BMB Reports
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    • v.44 no.6
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    • pp.381-386
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    • 2011
  • In the present study, we characterized the function of HS1-binding protein 3 (HS1BP3), which is mutated in essential tremor and may be involved in lymphocyte activation. We found that HS1BP3 localized to the mitochondria and endoplasmic reticulum partially. Overexpression of HS1BP3 induced apoptosis in HEK293T and HeLa cell lines. When these cell lines were transfected with HS1BP3, they exhibited nuclear DNA condensation, externalization of phosphatidylserine (PS), and cleavage of poly ADP ribose polymerase (PARP). Furthermore, suppression of HS1BP3 or HS1 expression attenuates HS1BP3 induced apoptosis. In addition, HS1BP3 enhanced activator protein 1 (AP-1)-mediated transcription in a dose-dependent manner. Therefore, we conclude that HS1BP3 regulates apoptosis via HS1 and stimulates AP-1-mediated transcription.

Evaluation of protective effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes in the single cell gel electrophoresis assay (단세포 겔 전기영동법을 이용한 사람 림프구 DNA 손상에 대한 복숭아씨 추출물의 방사선 방어효과 평가)

  • Kim, Jin-Kyu;Park, Tae-Won;Lee, Chang-Joo;Chai, Young-Gyu
    • Journal of Radiation Protection and Research
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    • v.24 no.2
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    • pp.93-99
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    • 1999
  • The alkaline single-cell gel electrophoresis (SCGE) assay, called the comet assay, has been applied to the detection of DNA damage from a number of chemical and biological factors in vivo and in vitro. The comet assay is a novel method to assess DNA single-strand breaks, alkali-labile sites in individual cells. The effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes was evaluated by the SCGE assay. The lymphocytes, with or without pretreatment of the extracts, were exposed to 0, 0.1, 0.3, 0.5, 1.0 and 2.0 Gy of $^{60}Co$ gamma ray. Significantly increased tail moment, which was a marker of DNA strand breaks in the comet assay, showed an excellent dose-response relationship. The treatment of the peach kernel extracts reduced the DNA damage by 30 % in irradiated groups as compared to that in non-treated control groups. The result indicates that the extracts shows radioprotective effect on lymphocyte DNA when assessed by the comet assay.

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Transactivation potential of the C-terminus of human ALG-2 (Human ALG-2 C-말단의 전사활성화 능력 분석)

  • Kim, Keun-Soo;Kim, Eun-Hee
    • The Journal of Natural Sciences
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    • v.11 no.1
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    • pp.89-94
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    • 1999
  • ALG-2 (apoptosis linked gene-2) is a 22 kDa calcium-binding protein necessary for apoptosis induced by various stimuli in lymphocyte. The transactivation of human ALG-2 was assessed in yeast as a fusion protein with the DNA binding domains (DBDs) of LexA. The C-terminal of hALG-2 (93-191 amino acid) exhibited transacitivation of the reporter gene, LacZ, whereas the full-length hALG-2 (1-91 amino acid) and its N-terminal (1-98 amino acid) did not. The transactivation of LacZ reporter was driven more strongly (more than 2.7-fold increase) by the C-terminus of hALG-2 than by the B42, as a positive control for transactivation. Hence, our data suggested a possible regulatory role of the N-termini of hALG-2 upon transactivation.

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Genotoxicity and Identification of Differentially Expressed Genes of Formaldehyde in human Jurkat Cells

  • Kim, Youn-Jung;Kim, Mi-Soon;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.1 no.4
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    • pp.230-236
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    • 2005
  • Formaldehyde is a common environmental contaminant found in tobacco smoke, paint, garments, diesel and exhaust, and medical and industrial products. Formaldehyde has been considered to be potentially carcinogenic, making it a subject of major environmental concern. However, only a little information on the mechanism of immunological sensitization and asthma by this compound has been known. So, we performed with Jurkat cell line, a human T lymphocyte, to assess the induction of DNA damage and to identify the DEGs related to immune response or toxicity by formaldehyde. In this study, we investigated the induction of DNA single strand breaks by formaldehyde using single cell gel electrophoresis assay (comet assay). And we compared gene expression between control and formaldehyde treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity ($IC_{30}$) of formaldehyde was determined above the 0.65 mM in Jurkat cell in 48 h treatment. Based on the $IC_{30}$ value from cytotoxicity test, we performed the comet assay in this concentration. From these results, 0.65 mM of formaldehyde was not revealed significant DNA damages in the absence of S-9 metabolic activation system. And the one differentially expressed gene (DEG) of formaldehyde was identified to zinc finger protein 292 using $GeneFishing^{TM}$ method. Through further investigation, we will identify more meaningful and useful DEGs on formaldehyde, and then can get the information on the associated mechanism and pathway with immune response or other toxicity by formaldehyde exposure.

Study of Signaling Pathway on Apoptotic Cell Death Induced by Extract of Ailanthus altissima in Human Jurkat Lymphocytes (저근백피(樗根白皮) 추출물에 의한 급성 림프성 백혈병 Jurkat Lymphocytes의 세포고사 유도 및 신호기전 연구)

  • Lee, Ki Ouk;Kim, Ae Wha;Lim, Kyu Sang;Yun, Young Gab
    • Herbal Formula Science
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    • v.25 no.3
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    • pp.349-362
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    • 2017
  • Objectives : We investigated whether the components of Ailanthus altissima induced apoptotic cell death in Jurkat acute lymphoblastic leukemia (ALL) cells. Methods : Regulation of cell proliferation is a complex process involving the regulated expression and/or modification of discrete gene products, which control transition between different stages of the cell cycle. Results : Upon treatments with Ailanthus altissima, the concentration-dependent inhibitions of cell viability were observed as compared to untreated control group. The capability of Ailanthus altissima to induce apoptosis was associated with proteolytic cleavage of specific target proteins such as poly(ADP-ribose)polymerase (PARP) and beta-catenin proteins suggesting the possible involvement of caspases. Ailanthus altissima also caused apoptosis as measured by cell morphology and DNA fragmentation. Conclusions : These results indicate that the increase of apoptotic cell death by Ailanthus altissima may be due to the inhibition of cell cycle in human Jurkat lymphocytes. Conclusively, these current and further findings will provide novel approaches to understanding and treating major diseases.