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Detection of Total Reactive Oxygen Species in Adherent Cells by 2',7'-Dichlorodihydrofluorescein Diacetate Staining
Published on: June 23, 2020
Oxidative stress: potential of distinct peroxide determination systems
Meinrad Lindschinger1, Karl Nadlinger, Norbert Adelwöhrer
1Institute for Nutritional and Metabolic Diseases, Schwarzl Outpatient Clinic, Lassnitzhöhe, Austria. office@lindschinger.at
Abstract:
When reactive oxygen species attack biological structures, peroxides, which are short-lived oxidative intermediates, are generated. We evaluated the potential of two different, commercially available peroxide activity assays (Pox-Act and d-ROMS) to see whether the results were associated with the clinical condition of subjects who were participating in a routine health care program. Furthermore, we determined the total antioxidant status (TAS) and the titer of autoantibodies against oxidized low-density lipoprotein (oLAb) to verify the hydroperoxide measurements. Subjects with medical conditions (hereafter referred to as patients) had significantly increased serum peroxide levels compared to healthy subjects. The d-ROMS kit indicated that 86% of subjects had an increased level of total peroxides. Although the assays had a significant correlation (p<0.001), 34% of the subjects had an increased total peroxide concentration in the Pox-Act assay that was clearly associated with clinical symptoms. Furthermore, the sensitivity of the Pox-Act assay was 35 times higher than that of the d-ROMS kit. In subjects with medical conditions, there was a trend toward a decreased TAS and a slightly increased oLAb titer in comparison to healthy subjects, but this was not statistically significant. The Pox-Act assay seems to be a valuable tool for the determination of total peroxides, while the results from the d-ROMS kit should be considered with caution.
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