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

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.

Correlation between human health and reactive oxygen species produced in blood: a long-term chemiluminescence and fluorescence analysis

“…PMA was used for stimulation in this study. The stimulant induced CL and dFL/dt were shown to be proportional to the instantaneous generation of O 2 −· and OCl − , respectively 9 . Therefore, for simplicity, we denote these parameters as CL-O 2 −· and FL-OCl − hereafter.…”

“…The band-rejection filters in the set are optimized for efficiently blocking excitation light and transmitting MCLA-derived CL (Em-Max 465 nm) and APF-derived FL (Em-Max 515 nm). Separation of CL and FL submitted to a single PMT was attained by repeating on/off of the excitation lights at a high-enough speed 8 , 9 , 14 .…”

“…* The total O 2 −· produced after stimulation is proportional to CL AUC 9 , 14 , and, since FL is the cumulative amount of OCl − , the total OCl − produced after stimulation is equal to FL MAX − FL BASE POINT (‘a−b’ in Fig. S3 ).…”

“…Previously we developed a simultaneous chemiluminescence (CL) and fluorescence (FL) monitoring system CFL-C2000 8 . By improving the system, we have recently developed CFL-P2200 9 and demonstrated that it can semi-automatically detect ROS and hROS, i.e., O 2 −· and OCl − elicited in a small amount of blood samples by using CL and FL detecting reagents. This newer system was recently applied to studying the correlative relationships between O 2 −· /OCl − and diseases, such as Alzheimer's disease 10 , arteriosclerosis 11 , and hypertension 12 with the rodent models.…”

Changes in Whole-Blood microRNA Profiles during the Onset and Treatment Process of Cerebral Infarction: A Human Study

“…However, it remains uncertain whether the five miRNAs are the cause or the result of cerebral infarction. Our future task is to measure inflammation markers and reactive oxygen species [28] in the blood of patients, which may provide some information regarding the relationship between these five miRNAs and the onset or treatment process of cerebral infarction. Table 5.…”