EPR-Spin Trapping and Flow Cytometric Studies of Free Radicals Generated Using Cold Atmospheric Argon Plasma and X-Ray Irradiation in Aqueous Solutions and Intracellular Milieu

PloS One
Hidefumi UchiyamaTakashi Kondo

Abstract

Electron paramagnetic resonance (EPR)-spin trapping and flow cytometry were used to identify free radicals generated using argon-cold atmospheric plasma (Ar-CAP) in aqueous solutions and intracellularly in comparison with those generated by X-irradiation. Ar-CAP was generated using a high-voltage power supply unit with low-frequency excitation. The characteristics of Ar-CAP were estimated by vacuum UV absorption and emission spectra measurements. Hydroxyl (·OH) radicals and hydrogen (H) atoms in aqueous solutions were identified with the spin traps 5,5-dimethyl-1-pyrroline N-oxide (DMPO), 3,3,5,5-tetramethyl-1-pyrroline-N-oxide (M4PO), and phenyl N-t-butylnitrone (PBN). The occurrence of Ar-CAP-induced pyrolysis was evaluated using the spin trap 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS) in aqueous solutions of DNA constituents, sodium acetate, and L-alanine. Human lymphoma U937 cells were used to study intracellular oxidative stress using five fluorescent probes with different affinities to a number of reactive species. The analysis and quantification of EPR spectra revealed the formation of enormous amounts of ·OH radicals using Ar-CAP compared with that by X-irradiation. Very small amounts of H atoms were detected wherea...Continue Reading

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Citations

Apr 18, 2016·Archives of Biochemistry and Biophysics·Mati Ur RehmanTakashi Kondo
Dec 12, 2018·The Journal of Small Animal Practice·M C NolffA Meyer-Lindenberg
Jan 25, 2019·Critical Reviews in Food Science and Nutrition·Kexin ZhangBrijesh K Tiwari
Apr 6, 2019·Environmental Science and Pollution Research International·Aleksandra Djukić-VukovićLjiljana Mojović
Feb 12, 2020·Physical Chemistry Chemical Physics : PCCP·Arpad Mihai RostasStefan Weber
Jul 28, 2020·Cancers·Gabriella Pasqual-MeloSander Bekeschus
Jan 14, 2020·Redox Biology·Sander BekeschusRajesh Kumar Gandhirajan
Jan 23, 2021·Archives of Biochemistry and Biophysics·Yasumasa OkazakiShinya Toyokuni
Apr 4, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Helena MotalnBoris Rogelj
May 9, 2021·Archives of Biochemistry and Biophysics·Yasumasa OkazakiShinya Toyokuni
Oct 6, 2018·Analytical Chemistry·Yury GorbanevAnnemie Bogaerts

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Methods Mentioned

BETA
electron paramagnetic resonance
flow cytometry
X-ray
deamination
Fluorescence

Software Mentioned

GraphPad Prism

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