International Journal of Environmental Sciences

Volume 3 Issue 1 2012- July 2012        Pages: 75- 83               <<Previous    Next>>

Determination of oxidative stress in human lymphocytes exposed to Isocyanates

Author Information:

Saxena. A
Assistant Professor, Biotechnology department, IASCA, ITM Universe, Gwalior (MP), India

ABSTRACT

In present work it was found that during environmental stress, ROS levels can be increased, resulting  in  damage  to  cellular  structures  where  Glutathione  levels  get diminished. The dye having high affinity for glutathione fluoresced Blue when bound to reduced or oxidized glutathione whereas unbound dye remained non-fluorescent. Thus, decrease in glutathione level could be detected in apoptotic cells. Similarly, measurement of 8-oxo-dG indicated oxidative damage where an 8-oxo-dG antibody bound to the 8-oxo- dG in the sample, standard/ prebound to the wells. Anti-8-oxo-dG bound to 8-oxo-dG washed away while those captured by the immobilized 8-oxo-dG were detected by a secondary antibody: HRP conjugate. The intensity of yellow color was inversely proportional to the concentration of 8-oxo-dG. Another measure of oxidative stress is CM- H2DCFDA; a cell permeant, non-fluorescent indicator, which is redox sensitive dye until removal of the acetate groups by intracellular esterases, and oxidation occurred within the cell.

Keywords: Oxidative stress, Isocyanates, Lymphocytes, Elisa

DOI: 10.6088/ijes.2012030131009

© 2012 Copyright by the authors, licensee Integrated Publishing Association.This is an open access article distributed under the Creative Commons Attribution License (3.0) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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