DETECTION OF DNA-PSORALEN PHOTOADDUCTS in situ |
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Authors: | ZOFIA ZAREBSKA† MARIA JARZABEK-CHORZELSKA ‡ GENOWEFA RZSA WIESLAW GLISKI MARIA PAWIKA TADEUSZ CHORZELSKI STEFANIA JABLOKA |
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Institution: | Department of Dermatology. Academy of Medicine Institute of Biochemistry and Biophysics. Polish Academy of Sciences, Warsaw. Poland;?Institute of Biochemistry and Biophysics. Polish Academy of Sciences, Warsaw. Poland |
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Abstract: | Abstract— An immunological method, with the use of specific immune serum, has been developed for detection of 8-methoxypsoralen (8-MOP) photoadducts to DNA, formed in situ in cell nuclei, after combined treatment with 8MOP and UV-A irradiation (Zarçbska et al. , 1978). Lymphocytes fixed on slides or in suspension, and cryostat sections of different mammalian tissues, served as antigenic substrate, after treatment with 8-MOP and UV-A in vitro. Specific fluorescence in these substrates was detected in the nuclei after treatment with 30 ? 140 kJ/m2 UV-A in the presence of 0.1-0.3 μg/cm2 8-MOP. PHA-stimulated-lymphocytes appeared to be the most sensitive substrate. However, hairless mice treated with high doses of UV-A in vivo , 70 ? 360 kJ/m2 did not reveal a specific fluorescence of epidermal nuclei, unless a high local concentration of 8-MOP was attained. The apparent discrepancy in the level of photoadduct detection between the in vitro and in vivo treated specimens was explained by the low number of DNA-8-MOP-photoadducts formed in vivo under these experimental conditions. The relevance of these findings to the role of DNA-8-MOP-photoadducts formed during PUVA photochemotherapy is discussed. |
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