• Title/Summary/Keyword: 환경 DNA

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Genotoxicity (DNA damage) on Blood Cells of Parrot Fish (Oplegnathus fasciatus) Exposed to Acidified Seawater Making of CO2 (이산화탄소로 산성화된 해수에 노출된 돌돔(Oplegnathus fasciatus) 혈구세포에 대한 유전독성(DNA 손상))

  • Choi, Tae Seob;Lee, Ji-Hye;Sung, Chan-Gyoung;Lee, Jung-Suk;Park, Young-Gyu;Kang, Seong-Gil
    • Journal of Environmental Science International
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    • v.23 no.3
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    • pp.483-492
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    • 2014
  • DNA damage such as genotoxicity was identified with comet assay, which blood cell of a marine parrot fish (Oplegnathus fasciatus) was exposed to an acidified seawater, lowered pH gradient making of $CO_2$ gas. The gradient of pH were 8.22, 8.03, 7.81, 7.55 with control as HBSS solution with pH 7.4. DNA tail moment of fish blood cell was $0.548{\pm}0.071$ exposed seawater of pH 8.22 condition, on the other hand, DNA tail moment $1.601{\pm}0.197$ exposed acidified seawater of pH 7.55 lowest condition. The approximate difference with level of DNA damage was 2.9 times between highest and lowest of pH. DNA damage with decreasing pH was significantly increased with DNA tail moment on blood cell of marine fish (ANOVA, p < 0.001). Ocean acidification, especially inducing the leakage of sequestered $CO_2$ in geological structure is a consequence from the burning of fossil fuels, and long term effects on marine habitats and organisms are not fully investigated. The physiological effects on adult fish species are even less known. This result shown that the potential of dissolved $CO_2$ in seawater was revealed to induce the toxic effect on genotoxicity such as DNA breakage.

The Molecular Mechanism of Safrole-induced DNA Adducts and its Role to Oral Carcinogenesis

  • Liu, Tsung-Yun
    • Environmental Mutagens and Carcinogens
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    • v.23 no.3
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    • pp.99-102
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    • 2003
  • IARC classified areca quid as a human carcinogen. Areca quid chewed in Taiwan includes Piper betle inflorescence, which contains high concentrations of safrole (15 mg/fresh weight). Safrole is a documented rodent hepatocarcinogen, and chewing areca quid may contribute to human exposure (420 $\mu$m in saliva). The carcinogenicity of safrole is mediated through 1'-hydroxysafrole formation, followed by sulfonation to an unstable sulfate that reacts to form DNA adducts. Using human liver microsomes and Escherichia coli membranes expressing bicistronic human P450s, CYP2E1 and CYP2C9 were identified as the main P450s involved in the activation of safrole. We have demonstrated the presence of stable safrole-dGMP adducts in human oral tissues following areca quid chewing using $^{32}$ P-postlabeling and HPLC mass spectrometry methods. By studying 88 subjects with a known AQ chewing history and 161 matched controls, we have demonstrated that the presence of safrole-DNA adducts in peripheral blood cells was correlated to AQ chewing, and CYP2E1 seemed to play an important role in the modulation of safrole-DNA adduct formation. We have also shown that safrole can form stable safrole-DNA adducts as well as oxidative damages in rodent liver. However, the stable safrole-DNA adducts may represent a more significant initial lesion as compared to the rapidly repaired safrole-induced 8-hydroxy-2'-deoxyguanosine. This oxidative DNA damage is mediated through the formation of hydoryxchavicol, the major safrole metabolite in human urine. Hydroxychavicol may have gone through two-electron oxidation to the o-quinone; then via one-electron reduction to semiquinone radicals to generate oxidative DNA damage. However, these reactive metabolites can be efficiently conjugated by GSH. These data suggest that safrole may contribute to the initiation of oral carcinogenesis through safrole-DNA adduct and not oxidative DNA damage. In addition, CYP2E1 may modulate this adduct formation.

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Protective Effects of Ginkgo Biloba Leaf Extract(GBE) against 1,2,4-benzenetriol Induced Toxicity in Vitro (Ginkgo biloga 잎 추출물의 1,2,4-benzenetriol에 대한 항산화 효과에 대한 연구)

  • 이영준;김태연;정해원
    • Journal of Environmental Health Sciences
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    • v.27 no.1
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    • pp.124-130
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    • 2001
  • Ginkgo biliba has been used for bronchitis and asthma in oriental countries and its leaf extract(GBE) contains 24% ginkgoflavone glycoside and 6% terpenoid. Flavonoids and terpenoids are known to have various antioxidant effects such as scavenging of free radicals and chelation of transtional metals. Antioxidant effect of GBE against 1,2,4-benzenetriol(BT), one of toxic metabolites of benzene, was demonstrated throughbsister chromatid exchange(SCE) analysis, single cell gel electrophoresis(SCGE) analysis, DNA cleavage assay and lipid peroxidation production analysis. The means of SCE frequencies at 10, 25 and 50$\mu$M concentration of BT were 7.72, 8.02, 9.22 respectively. In addition of GBE with concentration of 50, 200 and 500$\mu\textrm{g}$/$m\ell$, SCE frequencies were decreased significantly.(p<0.05) According to SCGE analysis, BT induced DNA damage in a dose-dependent manner at concentration of 10 and 50 $\mu$m and the DNA damage induced by BT was significantly protected by GBE(p<0.001). No genotoxicity was observed by GBE treatment alone on DNA cleavage. The effect of BT on lipid peroxidation product, Malondiadehyde(MDA), was increased with concentration of BT(10 and 50 $\mu$M) and reduction in MDA was noted when GBE was added. From above results it is suggested that GBE could protect the cell and DNA from pro-oxidant effect by reactive oxigen species induced by BT.

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DNA Damage and Micronuclei Induced by Di (2-ethylhexyl) phthalate in Human Breast Carcinoma MCF-7 cells (Di(2-ethylhexyl) phthalate에 의해 유도된 DNA손상과 소핵 형성)

  • 김종원;한의식;박미선;엄미옥;김인숙;전혜승;정해관;심웅섭;오혜영
    • Environmental Mutagens and Carcinogens
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    • v.21 no.1
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    • pp.34-43
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    • 2001
  • Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

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Seasonal Change of RNA/DNA Ratio and Blood Characteristics of Black Sea Bream Acanthopagrus schlegeli Habituated in Geojae Costal Area, Kyungnam Province, Korea (경남 거제해역에 서식하는 감성돔(Acanthopagrus schlegeli)의 연간 RNA/DNA 및 혈액학적 특성 변화)

  • Kim, Su-Kyoung;Shim, Na-Young;Lee, Do-Hyun;Kim, Dae Hyun;Yoon, Seong-Jong
    • Journal of Environmental Science International
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    • v.22 no.1
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    • pp.37-45
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    • 2013
  • The monthly variations of blood characteristics and RNA/DNA of black sea bream, Acanthopagrus schlegeli, habituated in Geojae costal area were analysed to determine health condition of natural stocks in terms of gonad maturation and spawning season from March 2010 to February 2011. Spawning season determinated by gonadosomatic index is from June to August. RNA/DNA ratio of black sea bream muscle was strongly correlated with spawning season. During the gonad maturation RNA/DNA ratio in dorsal muscle tissue was decreased contrast to rapid increase during spawning season. Blood composition factors increased in terms of gonad maturation are aspartate aminotransferase, cholesterol, triglyceride, total protein, glucose, globulin, alkaline phosphatase and inorganic phosphate. Other blood factors increased during spawning season are alanine aminotransferase, blood urea nitrogen, uric acid and lactate dehydrogenase.

Development of Exposure Biomarkers for Endocrine Disrupting Chemicals Using DNA Microarray (DNA 마이크로어레이를 이용한 내분비장애물질 노출지표 개발)

  • Yang, Mi-Hi
    • Environmental Analysis Health and Toxicology
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    • v.20 no.4 s.51
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    • pp.327-332
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    • 2005
  • 장기간 노출 시 발암 등 인체 유해성을 갖는 환경유래 내분비장애물질(endocrine disrupting chemicals, EDCs)에 대한 선택적이고 민감한 노출지표를 개발하기 위하여 본 연구에서는 DNA microarray를 이용하였다. 피험자는 아직 특별한 질환을 갖지 않는 18세 이상 연령, 성을 맞춘 EDCs고농도 노출군(N = 16)과 저농도군(N = 16)으로 구성되었다. 노출정도 구분은 10년 이상 거주지가 K산업폐기물 소각장과 2.5 km 반경 내, 외 인지에 따라 고노출군,저노출군으로 구분하였다. 피험자의 말초혈에서 total RNA를 분리, 각 군당 B인씩 pool로 cDNA를 합성하여 oligonucleotide DNA 칩에 적용하였다. 유전자발현의 차이를 GenePixPro 4.0 software를 이용하여 분석하였다. 총 3장의 칩을 이용하여 공통적으로 저노출군보다 고노출군에서 2배 이상 발현의 증가를 보인 유전자는 plasminogen activator(PLAT)등 12종이 관찰되었고, l/2이하로 발현의 감소를 보인 유전자는 kallikrein 3 (KLK3)등 29종이었다. 이 들 유전자는 PLAT등 면역계 반응에 관여하는 유전자 및 apoptosis, transport, G protein, chromatin, 암화, 발생 (development), 대사 등에 관여하는 유전자들이었다. 그러므로 KLK3등 본 연구에서 발굴한 유전자는 향후 확대된 인구에서 본 연구 결과의 확인을 통하여 EDCs특이적 노출지표로써, 나아가 암 등 EDCs관련 질병의 기전 및 병인학을 구명하는데 이용가치가 높다고 사료된다.

DNA Analysis of mtDNA COI Gene in the Sharp-toothed Eel (Muraenesox cinereus Forskal) from Yeosu, Jinhae, Jeju, Goseoung, Jangheung and Haenam Populations in Korea Using PCR-aided RFLP

  • Oh, Taeg-Yun;Jeong, Sun-Beom;Cho, Eun-Seob
    • Journal of Environmental Science International
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    • v.20 no.4
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    • pp.551-554
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    • 2011
  • The production of the sharp-toothed eel by commercial catch off waters of Korea is annually declined after 1978. This study was carried out to obtain the stock management of the sharp-toothed eel using the PCR-aided RFLP method. The mtDNA COI gene was amplified using species-specific primers and PCR product was observed to 700 bp. Amplified DNA fragments were treated with six kinds of restriction enzymes (BaeHI, EcoRI, PstI, Ksp22, HinfI and HaeIII). The treatment of HaeIII showed a distinct PCR product between Yeosu/Jinhae/Jeju/Goseoung and Jangheung/Haenam populations that were observed from 300 to 400 bp in reference to 100 bp molecular marker. However, DNA fragment within populations had an identical pattern. The phylogenetic homology is 82% between two populations inferred from RFLP PCR product pattern using NTsysPC ver. 2.1. The use of HaeIII plays an important role in discriminating populations. It is thought that adults after over-wintering in the southern part of Jeju migrate to the Yeosu, Jinhae and Goseoung regions to spawn instead of to southwestern waters. Individuals within populations showed a relatively active genetic mixing and migration regardless of geography. However, the genetic ancestor of Jangheung and Haenam populations is appeared to be more adjacent to China or Japan than Jeju.

Effect of the Internal Shape of eDNA Filter Case made by the PBF method Metal 3D Printer on Water Sampling Performance (PBF 방식 금속 3D프린터로 제작된 환경DNA 필터 케이스의 내부 형상이 포집 성능에 미치는 영향)

  • Lee, Seung-Min;Park, Se-Hyun;Kwak, Ihn-Sil;Kim, Hyoung-Ho;Kwak, Tae-Soo
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.20 no.8
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    • pp.74-79
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    • 2021
  • This study focuses on designing a filter case using a water pump for application in eDNA filtering systems. Filter cases, channel type and net type were designed based on the flow field and made using a 3D printer for metal. Flow analysis was conducted for each filter case, and the results were consistent with the pressure experiment results. Furthermore, the water sampling performance test showed that the channel-type filter case exhibited a high flow rate and low pressure through the filter. The eDNA extraction experiment showed that the channel type exhibited improved capture ability compared to the net type.

Isolation of HRD3 gene, a homologous RAD3 gene from fission yeast Schizosaccharomyces pombe

  • Choi, In-Soon;Jin, Yong-Hwan;Park, Sang-Dai
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.77-82
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    • 1996
  • The RAD3 gene of Saccharomyces cerevisiae is required for excision repair and is essential for cell viability. RAD3 encoded protein possesses a single stranded DNA-dependent ATPase and DNA-RNA helicase activies. To examine the extent of conservation of structure and function of RAD3 during eukaryotic evolution, we have cloned the RAD3 homolog, HRD3, from the distantly related yeast Schizosaccharomyces pombe. Here, we report the partial cloning and characterization of HRD3 gene (Homologous of RAD3 gene) which was isolated by PCR amplification using conserved domain of Saccharomyces cerevisiae RAD3 gene. Chromosomal DNA isolated from S. pombe had similar restriction patterns to those from S. cerevisiae, as determined by Southern blot analysis. The 2. 8 kb transcript of mRNA was identified by Northern hybridization. The level of transcript did not increase upon UV-irradiation, suggesting that the HRD3 gene in S. pombe is not UV-inducible.

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