In order to assess the redox status of the treated cells and possible formation of reactive metabolites, determination of reduced glutathione (GSH) and oxidized glutathione
(GSSG) was performed as follows.
We report a rapid and fully automated HPLC method for determining total (tGSH), reduced (GSH), and oxidized glutathione
(GSSG) in whole blood.
Caption: Figure 2: The relationships between individual plasmatic AROS production rate ([micro]mol x [min.sup.-1]) and A of (a) TAC (mM), (b) TBARS ([micro]M), (c) PC (nmol x [mg.sup.-1] protein), (d) 8-iso-PGF2[alpha] (ng x [mg.sup.-1] creatinine), (e) 8-OH-dG (ng x [mg.sup.-1] creatinine), (f) oxidized glutathione
([micro]molx [L.sup.-1]), and total (g) Cys ([micro]mol x [L.sup.-1]), (h) CysGly ([micro]mol x [L.sup.-1]), and (i) Hcy ([micro] mol x [L.sup.-1]) recorded in 50 km (full symbols) and in 100 km (empty symbols) race are shown.
Abbreviations: HPE, Hypericum perforatum extract; GSH, GSSG, reduced, oxidized glutathione
; CE, capillary electrophoresis; ROS, reactive oxygen species
The reduced glutathione/ oxidized glutathione
ratio (GSH/GSSG) is used to evaluate oxidative stress status in biological systems, and alterations of this ratio have been demonstrated in aging, cancer, HIV replication, and cardiovascular diseases (15).
Tertbutyl hydroperoxide (tBHP), 2,4-dinitrophenylhydrazine (DNPH), urea, oxidized glutathione
(GSSG), nicotinamide adenine di-nucleotide phosphate (NADPH), 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB), 2-vinyl pyridine, glutathione reductase, ethyl acetate, Bradford reagent, mercury orange, and 2,7-dichlorofluorescein diacetate (DCF-DA) were obtained from Sigma-Aldrich (St.
Enzymatic method for quantitative determination of nanogram amounts of total oxidized glutathione
: applications to mammalian blood and other tissues.
In the presence of GR and NADPH, the oxidized glutathione
(GSSG) is immediately converted to the reduced form (GSH) with concomitant oxidation of NADPH to [NADP.sup.+].
Abbreviations tGSH: Total glutathione GSSG: Oxidized glutathione
MDA: Malondialdehyde GSH-Px: Glutathione peroxidase HCV: Hepatitis C virus HIV-1: Human immunodeficiency virus-1 FBDs: Former blood donors DCs: Dendritic cells NRTIs: Nucleoside reverse transcriptase inhibitors NNRTIs: Nonnucleoside reverse transcriptase inhibitors ALT: Alanine aminotransferase AST: Aspartate aminotransferase [gamma]-GT: [gamma]-Glutamyl transpeptidase APRI: Aspartate aminotransferase to platelet ratio index FIB-4: Fibrosis index based on four factors ESLD: End-stage liver disease HAART: Highly active antiretroviral therapy.
Benzo(a)pyrene [B(a)P], 7,12-dimethylbenzanthracene (DMBA), 1-Chloro-2,4-dinitrobenzene (CDNB), 5,5'-dithiobis-2-nitrobenzoic acid (DTNB), reduced glutathione (GSH), oxidized glutathione
(GSSG), pyrogallol, 2,6-dichlorophenolindophenol (DCPIP), potassium ferricyanide, triton X-100, ethylenediamine tetracetic acid (EDTA), bovine serum albumin (BSA), sodium pyruvate, thiobarbituric acid (TBA), reduced nicotinamide adenine dinucleotide (NADH) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) were obtained from Sigma Chemical Co.
In another low glutathione pool (initial CK activity, 171 U/L), supplementation with reduced glutathione (final concentration, 3.0 [micro]mol/L) and oxidized glutathione
(Sigma Chemicals; final concentrations, 1.5 and 3.0 [micro]mol/L) was examined.
Other antioxidant proteins include enzymes that mediate biosynthesis of glutathione and proteins that recycle oxidized glutathione
, peroxiredoxins, and glutathione peroxidases (glutaredoxins and thioredoxins) [20, 29].