Alkaline comet assay

Lebailly P, Vigreux C, Lechevrel C, et al. 1998. DNA damage in mononuclear leukocytes of farmers measured using the alkaline comet assay Modifications of DNA damage levels after a one-day field spraying period with selected pesticides. Cancer Epidemiol Biomarkers Prev 7 929-940. 

M. Venturi, R. J. Hambly, B. Glinghammar, J. J. Rafter and I. R. Rowland, Genotoxic activity in human faecal water and the role of bile acids a study using the alkaline comet assay. Carcinogenesis, 1997,18(12), 2353. 

Utilising the alkaline comet assay Jolly et al showed that exposing the nontumourigenic squamous oesophageal cell line, Het-IA to deoxycholate at... 

Sardas S, Aygun N, Gamli M, et al Use of alkaline comet assay (single cell gel electrophoresis technique) to detect DNA damage in lymphocytes of operating room personnel occupationally exposed to anaesthetic gases. Mutat Res 418(2-3) 93-100, 1998... 

Ondeger 0, Zorlu AF, Basaran N (1999) Use of the alkaline comet assay to monitor DNA damage in technicians exposed to low-dose radiation. J Occup Environ Med 41 693-698 Valenzuela MT, Nunez Ml, Guerrero M, Villalobos M, de Almodovar JMR (2000) Capillary electrophoresis of DNA damage after irradiation apoptosis and necrosis. J Chromatogr A, 871 321 -330 van Eldik R, Meyerstein D (2000) High-pressure radiolysis as a tool in the study of transition metal reaction mechanisms. Acc Chem Res 33 207-214... 

Studies using the alkaline comet assay have not found potassium iodide to produce DNA damage. 

Pfuhler, S. and Wolf, H. W. (1996) Detection of DNA crosslinking with the alkaline comet assay. Environ. Mol. Mutagen. 27,196-201. 

Moretti M, Villarini M, SimonucciS, et al. Effects of co-exposure to extremely low frequency (ELF) magnetic fields and benzene of benzene metabolites determined in vitro by the alkaline comet assay. Toxicol Lett 2005 157(2) 119 28. 

Hartmann, A., Kiskinis, E., Fjallman, A., and Suter, W. (2001). Influence of cytotoxicity and compound precipitation on test results in the alkaline comet assay. Mutat Res 497, 199-212. 

A., Speit, G., Thybaud, V., and Tice, R. R. (2003). Recommendations for conducting the in vivo alkaline Comet assay. 4th International Comet Assay Workshop. Mutagenesis 18, 45-51. 

More recently, Poul et al (2004) tested iodate alongside bromate in two different in vitro tests - the alkaline Comet assay and the cytokinesis-block micronucleus assay. Iodate concentrations of up to lOmM failed to induce primary DNA damage as determined in the Comet assay, while bromate significantly increased damage at concentrations of 2.5 mM and above. The results of the Comet assay could be confirmed by those obtained in the second test system, which detects not only clastogenic events, but also... 

Ansari, K.M., A. Dhawan, S.K. Khanna, and M. Das. 2005. In vivo DNA damaging potential of sanguinarine alkaloid, isolated from argemone oil, using alkaline Comet assay in mice. Food Chem. Toxicol. 43(1) 147-153. 

Combining the data on cell survival and amount of DNA damage obtained by the alkaline comet assay, one can conclude that mES cells are indeed more sensitive than differentiated or somatic cells, as fewer lesions lead to similar cell toxicity. In agreement with this, upon UV-C treatment, in mES cells only half of photoproducts (cyclobutane pyrimidine dimers and (6-4) photoproducts) are induced (Van Sloun 1999) and at the same time a higher level of cell death is observed compared to somatic cells (de Waard et al. 2008). Therefore the same amount of DNA lesions, induces a higher level of cell death in mES cells compared to somatic cells. Moreover, by using cell survival to compare the sensitivity of cell types, the sensitivity is underestimated. However, to enable a sound comparison of the sensitivity of mES cells and somatic cells, one should perform the appropriate genotoxicity assays at the appropriate dose (Boxl). 

Box 1 Commonly-used genotoxicity assays Alkaline comet assay... 

It has been shown that mES have a high level of spontaneously induced DNA strand breaks as detected by alkaline comet assay. However, global chromatin decondensation seems involved rather than high levels of DNA strand break formation (Banath et al. 2009). 

Pyrimethamine (FYR) Oral treatment of pregnant mice with 50mg PYR/kg E13, alkaline comet assay 6h after treatment E13, alkaline comet assay 16h after treatment - 18 and 28 times higjher DNA migrationi in head and body, respectively -3 and 19 times higher DNA migration in head and body, respectively Tsuda et al 1996... 

Moller, P. (2006). The alkaline comet assay towards validation in biomonitoring of DNA damaging exposures. Basic Clin Pharmacol Toxicol. Vol. 98 No. 4 pp. 336-345 ISSN 1742-7835... 

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