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피부에 폭로된 폐가솔린엔진오일의 표적장기 DNA adducts 형성과 케로신의 세척효과에 관한 연구
이진헌,그린탈라스카 한국산업위생학회 1998 한국산업보건학회지 Vol.8 No.2
Used gasoline engine oils(UGEO) are carcinogenic in long term studies and capable of increasing the number of carcinogen-DNA adducts in short term studies when dermally applied to mice. The carcinogenic risk of UGEO has been attributed to the concentration of polycyclic aromatic hydrocarbons(PAH) which accumulate in the lubricating system during the combustion of gasoline. When dermally exposed to UGEO, the use of hand cleanser was commonly recommended for removing it. But generally workers who dermally exposed oils, use kerosene as cleaner which make skin trouble. During this study, female mice aged 4-6 weeks were utilized to evaluate the efficiency of kerosene, as solvent-based cleanser, following dermal exposure to UGEO. DNA adduct were detected at skin and lung tissues by using the ^(32)P-postlabeling method. Washing with cleansers were done at two different interval times following dermal application of UGEO. The total DNA adducts in skin and lung tissues were statistically significantly increased in positive control groups, and of which the total adduct level in skin tissues was statistically significant higher than those in lung tissues(p=0.005). When washing kerosene, the DNA adduct level in skin tissues was statistically significantly decreased(p=0.0001). But DNA adducts in lung tissue was statistically increased(p=0.0039), and that washed at 8hr post exposure was more severly increase(p$lt;0.05). The slope of regression between DNA adducts of lung between skin tissues was 1.0802. In conclusion, skin cleaning with kerosene facilitates passage of carcinogens to the lungs of animals dermally treated with used gasoline engine oils(UGEO).
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근로자의 뇨중 상피세포에서 32P - postlabeling 에 의한 발암물질의 DNA adducts 측정방법에 대한 연구
이진헌,노재훈,그린 탈라스카 한국산업위생학회 2000 한국산업보건학회지 Vol.10 No.1
Carcinogen-DNA adduct analysis has potential for biomonitoring the earliest effects of exposure to many chemical carcinogens. They are the covalent reaction products of electrophiles and nucleophilic sites on DNA and the initial damage to DNA induced by many carcinogens. So many researchers begin to use them as biomarker for monitoring the earliest exposure of carcinogens and develop the effective analytical techniques about them. Randerath, Gupta and coworkers(1981, 1982) has also developed a ^(32)P-postlabelling method as one among them A major project for biomonitoring workers with carcinogen-DNA adducts is to develop noninvasive samples instead of tissues of target organs such as baldder and lung. This study use the exfoliated urothelial cells in urine for examine benzidine-DNA. adducts. The content of exfoliated urothelial cells is not enough to significantly measure DNA content with spectrophotometer, and require the another way. So firstly washing the collected cells with PBS and 70% ethanol arid centrifuge them for removing the crystals in urine, which block the isolation of DNA adducts. And then, measure the total nucleotide after ^(32)P-postlabelling for calculating RAL. [γ-^(32)P]ATP using for ^(32)P-postlabelling, can synthesize with [^(32)P]H₃PO₄, and reagent and enzyme mixture (RM, EM), which is very economic in case of requiring a lot of them. Chromatography was composed of two steps. First step was to separate adduct ones from unadducted nucleotide, and secondary step was separate each adduct, which were performed with 4 kinds of solvents and different directions on TLC. With this procedure, we measure the DNA adducts in exfoliated urothelial cells of workers who were employed in benzidine and benzidine-dye company. RAL of adducts were 89.0 × 10^7 and 57.0 × 10^7 in them. In conclusion, we can significantly measure the DNA adduct in exfoliated urothelial cells by using the above ^(32)P-postlabelling procedures, and use them to be biomonitoring workers who exposed carcinogens.
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근로자의 뇨중 상피세포에서 <sup>32</sup>P-postlabeling에 의한 발암물질의 DNA adducts측정방법에 대한 연구
이진헌,노재훈,그린 탈라스카,Lee, Jin Heon,Roh, Jaehoon,Talaska, Glenn 한국산업보건학회 2000 한국산업보건학회지 Vol.10 No.1
Carcinogen-DNA adduct analysis has potential for biomonitoring the earliest effects of exposure to many chemical carcinogens. They are the covalent reaction products of electrophiles and nucleophilic sites on DNA and the initial damage to DNA induced by many carcinogens. So many researchers begin to use them as biomarker for monitoring the earliest exposure of carcinogens and develop the effective analytical techniques about them. Randerath, Gupta and coworkers(1981, 1982) has also developed a $^{32}P$-postlabelling method as one among them. A major project for biomonitoring workers with carcinogen-DNA adducts is to develop non-invasive samples instead of tissues of target organs such as baldder and lung. This study use the exfoliated urothelial cells in urine for examine benzidine-DNA adducts. The content of exfoliated urothelial cells is not enough to significantly measure DNA content with spectrophotometer, and require the another way. So firstly washing the collected cells with PBS and 70% ethanol and centrifuge them for removing the crystals in urine, which block the isolation of DNA adducts. And then, measure the total nucleotide after $^{32}P$-postlabelling for calculating RAL. $[{\gamma}-^{32}P]ATP$ using for $^{32}P$-postlabelling, can synthesize with $[^{32}P]H_3PO_4$, and reagent and enzyme mixture (RM, EM), which is very economic in case of requiring a lot of them. Chromatography was composed of two steps. First step was to separate adduct ones from unadducted nucleotide, and secondary step was separate each adduct, which were performed with 4 kinds of solvents and different directions on TLC. With this procedure, we measure the DNA adducts in exfoliated urothelial cells of workers who were employed in benzidine and benzidine-dye company. RAL of adducts were $89.0{\times}10^7$ and $57.0{\times}10^7$ in them. In conclusion, we can significantly measure the DNA adduct in exfoliated urothelial cells by using the above $^{32}P$-postlabelling procedures, and use them to be biomonitoring workers who exposed carcinogens.
ScienceON
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