Abstract
Oxidative stress has been implicated in various disorders and controlling it would be important for healthy life. We have developed a new optical system for easily and accurately measuring oxidative stress in whole blood. It is optimized for simultaneously detecting reactive oxygen species (ROS) and highly reactive ROS (hROS), elicited mostly by white blood cells in a few microliters of blood. Results obtained by using this system show at least four important findings. 1) chemiluminescence of MCLA was confirmed to be attributable to O2-•. 2) PMA-stimulated cells released O2-• longer and more slowly than fMLP-stimulated ones. 3) fluorescence produced by APF oxidation was confirmed to be attributable to hROS, mostly OCl-, produced by myeloperoxidase. 4) the generation of OCl- was found to be a slower process than the O2-• generation. We also conducted pilot studies of oxidative stress in healthy volunteers.
Highlights
Oxidative stress is the term describing an imbalance between oxidant production, potentially leading to oxidative injury, and antioxidant capacity of the cells, bodily fluids and tissues [1,2]
The previous system (CFL-C2000) [10,11] used a cuvette (10 mm square) as the sample container (Panel a in S1 Fig). Such cuvettes loaded with colored samples containing light absorbing substances such as blood are susceptible to strong light absorptions
The highest peak area of the CLMCLA signals was obtained with 750-fold dilution of whole blood in cuvettes and the corresponding data of the samples on a glass slide was obtained with 500-fold dilution
Summary
Oxidative stress is the term describing an imbalance between oxidant production, potentially leading to oxidative injury, and antioxidant capacity of the cells, bodily fluids and tissues [1,2]. Afterwards, the definition of oxidative stress has been re-defined as “an imbalance between oxidants and antioxidants in favor of the oxidants, leading to a disruption of the redox signaling and control, and/or molecular damage” [3]. It is important to monitor produced oxidants, collectively called reactive oxygen species (ROS), at an early stage for preventing such diseases. All phagocytic cells, including white blood cells (WBC) have a well-characterized superoxide radical (O2-) generating plasma membrane oxidase capable of producing the large amounts of ROS required for its function in host defense [6].
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