Three biomarkers of oxidative stress were measured from first morning urine samples22 (link). All analyses were conducted using commercial ELISA kits in accordance with manufacturer procedure in Yale Reproductive Ecology Lab by the same researcher (8-OHdG, Enzo Life Sciences, catalogue ADI-EKS-350; Cu-Zn SOD, Enzo Life Sciences, catalog ALX-850-033; TBARS, Cayman Chemical Company, catalogue 10009055).
8-OHdG is a modified nucleotide base and a by-product of repaired DNA damage that is excreted in urine. Levels of 8-OHdG depict the amount of oxidative damage accumulated in cellular DNA through repaired lesions on the guanine base pair caused by ROS. Cu–Zn SOD catalyzes the dismutation of superoxide anion radicals to free oxygen and hydrogen peroxide. Levels of Cu–Zn SOD represent a primary line of enzymatic defense produced in the cytoplasm to neutralize ROS. Levels of TBARS reflect peroxidation of cellular lipids resulting from oxidative stress.
The average inter-assay variability for 8-OHdG, Cu–Zn SOD, and TBARS analysis was 14%, and intra-assay variability was 5%. All results were corrected for urine concentration based on samples’ specific gravity measured with refractometer (model PAL-10S ATAGO, U.S.A., Inc).
8-OHdG is a modified nucleotide base and a by-product of repaired DNA damage that is excreted in urine. Levels of 8-OHdG depict the amount of oxidative damage accumulated in cellular DNA through repaired lesions on the guanine base pair caused by ROS. Cu–Zn SOD catalyzes the dismutation of superoxide anion radicals to free oxygen and hydrogen peroxide. Levels of Cu–Zn SOD represent a primary line of enzymatic defense produced in the cytoplasm to neutralize ROS. Levels of TBARS reflect peroxidation of cellular lipids resulting from oxidative stress.
The average inter-assay variability for 8-OHdG, Cu–Zn SOD, and TBARS analysis was 14%, and intra-assay variability was 5%. All results were corrected for urine concentration based on samples’ specific gravity measured with refractometer (model PAL-10S ATAGO, U.S.A., Inc).
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