• Title/Summary/Keyword: human lymphocyte DNA

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Effects of Reactive Oxygen Species on DNA Stability in Humnn Spermatozoa

  • Kang, Hee-Gyoo;Kim, Tai-Jeon;Bae, Hyung-Joon;Moon, Hi-Joo;Kim, Myo-Kyung;Kim, Dong-Hoon;Sungwon-Han;Lee, Ho-Joon;Yang, Hye-Young
    • Biomedical Science Letters
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    • v.7 no.4
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    • pp.181-190
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    • 2001
  • This study was designed to investigate the effects of reactive oxygen species (ROS) on DNA stability in human spermatozoa. To verify human spermatozoa were incubated with xanthine-xanthine oxidase (X 100$\mu$M-XO 50 mlU ~ 400 mIU), $H_2O_2$ (125 $\mu$M ~ 1 mM), sodium nitroprusside (SNP 0.1 $\mu$M ~ 100 $\mu$M) or lymphocyte. Otherwise, spermatozoa were incubated under low $O_2$ (5%) condition. Damage of sperm DNA was analyzed by single cell electrophoresis (Comet assay) and flow cytometry after acridine orange staining. In the presence of ROS, there was increase in DNA damage. The rate of DNA single strand breakage (9.0$\pm$1.0% ~ 46.0$\pm$4.6%) and DNA fragmentation (7.51$\pm$1.0% ~ 29.5$\pm$4.6%) were similar regardless of the kinds of ROS and exposure time. DNA damage in the lower $O_2$ condition (5%) was lower than ambient $O_2$ condition (20%). Taken together, it suggested that sperm DNA might be damaged by ROS. In the presence of ROS, increase in DNA damage and chromatin instability was obvious in spite of short exposure. Although present study reconfirmed that sperm incubation in the low concentration of ROS have the benefit m the induction of capacitation and Ah, the increase in DNA damage by ROS and possible genetic problem should be considered before the human trials.

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Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation (3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.24 no.1
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    • pp.45-53
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    • 1999
  • Fluorescence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by radiation. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to apply FISH method for high dose biological dosimetry, chromosomal abberations by radiation at doses of 1, 3, 5, and 7Gy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. The frequencies of stable translocation per cell equivalent were 0.04, 0.33, 1.22, 2.62, and 5.58 for the lymphocyte exposed to 0, 1, 3, 5, and 7Gy, respectively, and those of dicentric were 0.00, 0.06, 0.52, 1.19 and 2.44, respectively. Significantly more translocation of t(Ab), a translocated chromosome with a piece of painted acentric matrial 'b' attached to unpainted piece containing centromere 'A', than reciprocal chromosome t(Ba) was observed. The frequencies of all type of chromosome rearrangements increased with dose. From above result, FISH seemed to be useful for radiation biodosimetry by which the frequencies of various types of stable aberrations in human lymphocyte can be observed more easily than by conventional method and so will improve our ability to perform meaningful biodosimetry.

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Analysis of gamma-ray-induced DNA damage in human, mouse and rat peripheral blood lymphocytes using single-cell gel electrophoresis (단세포 전기영동법을 이용한 인체, 마우스 및 랫드 림프구의 방사선에 의해 유발된 DNA 손상 측정)

  • Oh, Heon;Jung, Uhee;Park, Hae-Ran;Kim, Sung-Ho;Jo, Sung-Kee
    • Korean Journal of Veterinary Research
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    • v.44 no.1
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    • pp.41-47
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    • 2004
  • The alkaline single-cell gel electrophoresis (SCGE) assay, called the comet assay, has been applied to detect DNA damage induced by a number of chemicals and biological factors in vivo and in vitro. The DNA damage was analysed by tail moment (TM) and tail length (TL), which were markers of DNA strand breaks in SCGE. Human, mouse and rat peripheral blood lymphocytes (PBLs) were irradiated with different doses of $^{60}Co$ ${\gamma}$-rays, e.g. 1, 2, 4, and 8 Gy at a dose rate of 1 Gy/min. A dose-dependent increase in TM (p<0.01) and TL (p<0.01) was obtained at all the radiation doses (1-8 Gy) in human, mouse and rat PBLs. Mouse PBLs were more sensitive than human PBLs which were in turn more sensitive than rat PBLs when the treated dosages were 1 and 2 Gy. However, human PBLs were more sensitive than mouse PBLs which were in turn more sensitive than rat PBLs when the irradiation dosages were 4 and 8 Gy. Data from all three species could be fitted to a linear-quadratic model. These results indicated that there may be inherent differences in the radio-sensitivity among PBLs of mammalian species.

The Effect of Ginseng Intake on the Frequency of Sister Chromatid Exchanges of Human Lymphocyte of Adult Smokers (인삼의 섭취가 흡연성인의 인체임파구 SCE 빈도수에 미치는 영향)

  • 강명희
    • Journal of Nutrition and Health
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    • v.27 no.3
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    • pp.253-262
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    • 1994
  • This study was intended to investigate the anticarcinogenic effect of ginseng previously elucidated by other researches in animal studies. The sister chromatid exchange(SCE) method of human lymphocytes was used as a biomarker. Based on the literature search and the results of our laboratory, smoking was used as a parameter elevating the SCE frequency of general human population. To evaluate the smoking and ginseng effect on SCE frequency, 98 male healthy factory workers aged 23 to 58 years were divided into 4 groups : smoker with ginseng (SG), smoker control(SC), non-smoker with ginseng(NSG), and non-smoker control(NSC) groups, according to their smoking habits and ginseng intake. The mean sponteneous SCE per cell for the SG(10.8$\pm$0.3) and SC(10.4$\pm$0.3) groups were significantly higher than the NSG(9.1$\pm$0.2) and NSC(9.3$\pm$0.3) groups(p<0.05). High frequency cells (HFCs, cells with 15 SCEs) in SG and SC groups were also greater than those in NSG and NSC groups. However, the SCE levels of the SG and SC groups were not associated with the personal smoking history and the number of cigaretts smoked per day. Ginseng intake did not show any effect on the increased SCE caused by smoking. There were no correlations of the elevated SCE among smoking and ginseng types, history of ginseng intake, and consumption frequencies of ginseng intake. These results does not support the findings of other researchers that ginseng could be a protective agent to DNA damage.

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Effects of 3-Amino benzamide and Cytosine arabinoside on the Frequencies of Dicentric and Translocation in Human Lymphocyte Induced by Radiation (3-Amino benzamide 및 Cytosine arabinoside가 방사선 조사된 림프구의 이동원 염색체 및 상호전좌 유발빈도에 미치는 영향)

  • 정해원;김수영;조윤희;김태환;조철구;하성환
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.205-210
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    • 2002
  • In irradiated human lymphocytes, translocation of chromosome has been more frequently observed than dicentric chromosome. Differences in the misrepair process leading to translocation and dicentric chromosomes may explain the above observations. In order to find out whether dicentric and translocation are originated from different mechanism, the frequencies of radiation induced translocation and dicentric in lymphocytes were examined following treatment of irradiated lymphocytes with two DNA repair inhibitors, 3AB for inhibition of poly(ADP-ribose) synthesis and Ara C for inhibition of DNA-polymerase $\alpha$. Ara C potentiated the frequencies of radiation induced dicentric and translocation. 3AB also potentiated the frequencies of radiation induced dicentric, but not translocation. These results suggest the potential differences in the mechanisms in the formation of translocation and dicentric chromosomes.

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Radioprotective effect of mefenamic acid against radiation-induced genotoxicity in human lymphocytes

  • Hosseinimehr, Seyed Jalal;Nobakht, Reyhaneh;Ghasemi, Arash;Pourfallah, Tayyeb Allahverdi
    • Radiation Oncology Journal
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    • v.33 no.3
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    • pp.256-260
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    • 2015
  • Purpose: Mefenamic acid (MEF) as a non-steroidal anti-inflammatory drug is used as a medication for relieving of pain and inflammation. Radiation-induced inflammation process is involved in DNA damage and cell death. In this study, the radioprotective effect of MEF was investigated against genotoxicity induced by ionizing radiation in human blood lymphocytes. Materials and Methods: Peripheral blood samples were collected from human volunteers and incubated with MEF at different concentrations (5, 10, 50, or $100{\mu}M$) for two hours. The whole blood was exposed to ionizing radiation at a dose 1.5 Gy. Lymphocytes were cultured with mitogenic stimulation to determine the micronuclei in cytokinesis blocked binucleated lymphocyte. Results: A significant decreasing in the frequency of micronuclei was observed in human lymphocytes irradiated with MEF as compared to irradiated lymphocytes without MEF. The maximum decreasing in frequency of micronuclei was observed at $100{\mu}M$ of MEF (38% decrease), providing maximal protection against ionizing radiation. Conclusion: The radioprotective effect of MEF is probably related to anti-inflammatory property of MEF on human lymphocytes.

Evaluation of DNA Double Strand Breaks in Human and Mouse Lymphocyte Following ${\gamma}-Irradiation$ (${\gamma}-Ray$ 조사에 따른 사람의 정상임파구와 마우스 정상임파구의 DNA Double Strand Break 발생율에 대한 비교분석)

  • Kim Tae Hwan;Kim Sung Ho;Chung In Yong;Cho Chul Koo;Ko Kyung Hwan;Yoo Seong Yul
    • Radiation Oncology Journal
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    • v.11 no.2
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    • pp.219-225
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    • 1993
  • The evaluation of radiation-induced DNA double strand breaks (DSB) was made following irradiation of human lymphocytes, murine lymphocytes and EL-4 leukemia cells over a wide dose range of $^{60}Co\;{gamma}-rays.$ In lipopolysaccharide (LPS) or phytohemagglutinin (PHA)-stimulated murine lymphocytes, the slopes of the stand scission factor (SSF) revealed that lymphocytes with LPS increased DNA DSB formation by a factor of 1.432 (p<0.005). Furthermore, strand break production was relatively inefficient in the T lymphocytes compared to the B lymuhocytes. And EL-4 leukemia cells were found to form significantly more DNA DSB to a greater extent than normal lymphocytes (p<0.005). The in vitro studies of the intrinsic radiosensitivity between human lymphocytes and murine lymphocytes showed similar phasic kinetics. However, murine lymphocytes were lower in DNA DSB formation and higher in the relative radiation dose of 10 percent DNA strand breaks at 3.5 hours following ${gamma}-irradiation$ than human lymphocytes. Though it is difficult to interpret these results, these differences may be result from environmental and genetic factors. From our data, if complementary explanations for this difference will be proposed, the differences in the dose-effect relationship for the induction of DSB between humans and mice must be related to interspecies variations in the physiological condition of the peripheral blood in vitro and not to differences in the intrinsic radiation sensitivity of the lymphocytes. These results can be estimated on the basis of dose-effect correlation enabling the interpretation of clinical response and the radiobiological parameters of cytometrical assessment.

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Antibiotic Spectrum and Mechanism of Centipedin (Centipede Scolopendra subspinipes multilans L. KOCH로부터 정제된 항균 물질 Centipedin의 항균 Spectrum 및 작용 Mechanism 연구)

  • Kim, Ki-Tae;Hong, Sa-Weon;Won, Ho-Shik;Kim, Hyo-Joon;Park, Kyung-Bae;Cho, Key-Seung
    • Korean Journal of Microbiology
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    • v.34 no.1_2
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    • pp.31-36
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    • 1998
  • A broad spectrum of antibiotic action was studied with Centipedin purified from centipede Scolopendra subspinipes multilans L. Koch aganist gram-positive, gram-negative bacteria and fungi. The minimal inhibitory concentrations(MICs) were determined in liquid medium. The significant antibiotic activity was obtained aganist gram-negative Klebsiella pneumoniae ATCC 8308 responsible for causing infection at lung and intestine. The MIC value against Klebsiella pneumoniae ATCC 8308 was $2{\mu}g/ml$, and this Centipedin was active against Proteus vulgaris NRRL B-123. In addition, it has been shown that Centipedin blocks procaryotic RNA transcription and a little of DNA replication system in vitro. Centipedin did not exhibit any significant cytotoxicity against animal cells such as human blood leukemia (HL-60) and mouse B lymphocyte myeloma cell.

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Exploiting cDNA Microarray-Based Approach Combined with RT-PCR Analysis to Monitor the Radiation Effect: Antioxidant Gene Response of ex vivo Irradiated Human Peripheral Blood Lymphocyte

  • Sung, Myung-Hui;Jun, Hyun-Jung;Hwang, Seung-Yong;Hwang, Jae-Hoon;Park, Jong-Hoon;Han, Mi-Young;Lee, U-Youn;Park, Eun-Mi;Park, Young-Mee
    • Environmental Mutagens and Carcinogens
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    • v.22 no.3
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    • pp.142-148
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    • 2002
  • Although ionizing radiation (IR) has been used to treat the various human cancers, IR is cytotoxic not only to cancer cells but to the adjacent normal tissue. Since normal tissue complications are the limiting factor of cancer radiotherapy, one of the major concerns of IR therapy is to maximize the cancer cell killing and to minimize the toxic side effects on the adjacent normal tissue. As an attempt to develop a method to monitor the degree of radiation exposure to normal tissues during radiotherapy, we investigated the transcriptional responses of human peripheral blood lymphocytes (PBL) following IR using cDNA microarray chip containing 1,221 (1.2 K) known genes. Since conventional radiotherapy is delivered at about 24 h intervals at 180 to 300 cGy/day, we analyzed the transcriptional responses ex-vivo irradiated human PBL at 200 cGy for 24 h-period. We observed and report on 1) a group of genes transiently induced early after IR at 2 h, 2) of genes induced after IR at 6 h, 3) of genes induced after IR at 24 h and on 4) a group of genes whose expression patters were not changed after IR. Since Biological consequences of IR involve generation of various reactive oxygen species (ROS) and thus oxidative stress induced by the ROS is known to damage normal tissues during radiotherapy, we further tested the temporal expression profiles of genes involved in ROS modulation by RT-PCR. Specific changes of 6 antioxidant genes were identified in irradiated PBL among 9 genes tested. Our results suggest the potential of monitoring post-radiotherapy changes in temporal expression profiles of a specific set of genes as a measure of radiation effects on normal tissues. This type of approach should yield more useful information when validated in in vivo irradiated PBL from the cancer patients.

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The Effect of Carrot Juice Supplementation on Plasma Antioxidant Status and Lymphocyte DNA Damage in Korean Male Smokers

  • Lee, Hye-Jin;Jeon, Eun-Jae;Kim, Jung-Shin;Park, Yoo-Kyoung;Kang, Myung-Hee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.10b
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    • pp.176-176
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    • 2003
  • Cigarette smoke is a complex mixture of thousands of compounds, many of which are known or suspected human carcinogens. A preventive effect of the consumption of vegetables or fruits on various types of cancer has been confirmed from numerous studies.(omitted)

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