Rapid on-site dual optical system to measure specific reactive oxygen species (O2-• and OCl-) in a tiny droplet of whole blood

PLoS One. 2018 Aug 1;13(8):e0200573. doi: 10.1371/journal.pone.0200573. eCollection 2018.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aniline Compounds / chemistry
  • Area Under Curve
  • Diet
  • Exercise
  • Fluoresceins / chemistry
  • HL-60 Cells
  • Humans
  • Hypochlorous Acid / blood*
  • Imidazoles / chemistry
  • Luminescent Measurements
  • Neutrophils / metabolism
  • Peroxidase / metabolism
  • Pyrazines / chemistry
  • ROC Curve
  • Singlet Oxygen / blood*

Substances

  • Aniline Compounds
  • Fluoresceins
  • Imidazoles
  • Pyrazines
  • aminophenyl fluorescein
  • 2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyrazin-3-one
  • Singlet Oxygen
  • Hypochlorous Acid
  • Peroxidase

Grants and funding

This work was supported by the Council for Science, Technology and Innovation (CSTI), Cross-ministerial Strategic Innovation Promotion Program (SIP), and “Technologies for creating next-generation agriculture, forestry and fisheries” (Funding agency: NARO Bio-oriented Technology Research Advancement Institution). KK, KT, AH, TM, MH, NM, and HT are employed by Hamamatsu Photonics K.K. YW is employed by Healthcare Systems Co. Ltd. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.