Search

Your search keyword '"Dušinská M"' showing total 13 results
Start Over Author "Dušinská M" Journal toxicology letters
13 results on '"Dušinská M"'

1. P22-23: Expediting the uptake of Next-Generation Risk Assessment (NGRA) into EU chemicals legislation under PARC

Author

Herzler, M., Apruzzese, I., Bandow, N., Busquet, F., Cavoski, A., Colbourne, J., Di Consiglio, E., Dusinska, M., Flingelli, G., Holden, L., Hornek-Gausterer, R., Kaiser, A., Lee, R., Vicente, J. Lobo, L⊘fstedt, M., Longhin, E., Mouaziz, H., Namorado, S., von der Ohe, P., Rolfheim-Bye, S., Rousselle, C., Tcheremenskaia, O., Tsitsimpikou, C., Uhl, M., and Viegas, S.

Published
2023
Full Text
View/download PDF

13. DNA alkylation lesions and their repair in human cells: modification of the comet assay with 3-methyladenine DNA glycosylase (AlkD).

Author

Hašplová K, Hudecová A, Magdolénová Z, Bjøras M, Gálová E, Miadoková E, and Dušinská M

Subjects
Alkylation, Cell Line, DNA chemistry, DNA Adducts chemistry, DNA Adducts drug effects, Humans, In Vitro Techniques, Lymphocytes enzymology, Time Factors, Comet Assay methods, DNA drug effects, DNA Damage, DNA Glycosylases pharmacology, DNA Repair, Methyl Methanesulfonate pharmacology, Mutagens pharmacology
Abstract

3-methyladenine DNA glycosylase (AlkD) belongs to a new family of DNA glycosylases; it initiates repair of cytotoxic and promutagenic alkylated bases (its main substrates being 3-methyladenine and 7-methylguanine). The modification of the comet assay (single cell gel electrophoresis) using AlkD enzyme thus allows assessment of specific DNA alkylation lesions. The resulting baseless sugars are alkali-labile, and under the conditions of the alkaline comet assay they appear as DNA strand breaks. The alkylating agent methyl methanesulfonate (MMS) was used to induce alkylation lesions and to optimize conditions for the modified comet assay method with AlkD on human lymphoblastoid (TK6) cells. We also studied cellular and in vitro DNA repair of alkylated bases in DNA in TK6 cells after treatment with MMS. Results from cellular repair indicate that 50% of DNA alkylation is repaired in the first 60 min. The in vitro repair assay shows that while AlkD recognises most alkylation lesions after 60 min, a cell extract from TK6 cells recognises most of the MMS-induced DNA adducts already in the first 15 min of incubation, with maximum detection of lesions after 60 min' incubation. Additionally, we tested the in vitro repair capacity of human lymphocyte extracts from 5 individuals and found them to be able to incise DNA alkylations in the same range as AlkD. The modification of the comet assay with AlkD can be useful for in vitro and in vivo genotoxicity studies to detect alkylation damage and repair and also for human biomonitoring and molecular epidemiology studies., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Published
2012
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results