Refinement of the Dichlorofluorescein Assay for Flow Cytometric Measurement of Reactive Oxygen Species in Irradiated and Bystander Cell Populations

2008 ◽  
Vol 169 (4) ◽  
pp. 460-468 ◽  
Author(s):  
Kurt Hafer ◽  
Keisuke S. Iwamoto ◽  
Robert H. Schiestl
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Cell Populations ◽  
Oxygen Species ◽  
Bystander Cell ◽  
Flow Cytometric ◽  
Flow Cytometric Measurement

Flow Cytometric Detection of Reactive Oxygen Species

BIO-PROTOCOL ◽  
2013 ◽  
Vol 3 (8) ◽  
Author(s):  
Hsin-Yi Chang ◽  
Hsuan-Cheng Huang ◽  
Tsui-Chin Huang ◽  
Pan-Chyr Yang ◽  
Yi-Ching Wang ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Flow Cytometric

scholarly journals Intradialytic granulocyte reactive oxygen species production: a prospective, crossover trial.

1993 ◽  
Vol 4 (2) ◽  
pp. 178-186 ◽  
Author(s):  
J Himmelfarb ◽  
K A Ault ◽  
D Holbrook ◽  
D A Leeber ◽  
R M Hakim
Keyword(s):  
Reactive Oxygen Species ◽  
Reactive Oxygen ◽  
Fold Increase ◽  
Ros Production ◽  
Oxygen Species ◽  
Susceptibility To Infection ◽  
Flow Cytometric ◽  
Ros Formation ◽  
Dialysis Membranes ◽  
Species Production

By the use of flow cytometric techniques, this prospective, randomized crossover study was designed to analyze intradialytic granulocyte reactive oxygen species (ROS) formation in whole blood with complement-activating and noncomplement-activating hollow fiber membranes. Dialysis with a complement-activating membrane resulted in a 6.5-fold increase in granulocyte hydrogen peroxide production 15 min after dialysis initiation and remained significantly elevated (P < 0.01) through the first 30 min with this membrane in comparison to both predialysis values and simultaneous values with a noncomplement-activating membrane. Further studies demonstrated that blood obtained at 15 min with a complement-activating membrane generated significantly less granulocyte ROS production in response to Staphylococcus aureus incubation than blood obtained either predialysis or at the same time in dialysis with a noncomplement-activating membrane. Both complement-activating and noncomplement-activating dialysis membranes caused slightly decreased granulocyte responsiveness to phorbol myristate acetate. It was concluded that hemodialysis with complement-activating membranes results in increased granulocyte ROS production and decreased responsiveness to S. aureus challenge during the dialysis procedure. These results document the potential role of ROS in hemodialysis-associated pathology and susceptibility to infection.

Flow-cytometric analysis of reactive oxygen species in cancer cells under treatment with brassinosteroids

Steroids ◽  
2017 ◽  
Vol 117 ◽  
pp. 11-15 ◽  
Author(s):  
Pyotr A. Kisselev ◽  
Olesya V. Panibrat ◽  
Aliaksei R. Sysa ◽  
Marina V. Anisovich ◽  
Vladimir N. Zhabinskii ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cancer Cells ◽  
Flow Cytometric Analysis ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Flow Cytometric

Effects of Resveratrol, Berberine and Their Combinations on Reactive Oxygen Species, Survival and Apoptosis in Human Squamous Carcinoma (SCC-25) Cells

2019 ◽  
Vol 19 (9) ◽  
pp. 1161-1171 ◽  
Author(s):  
Magdalena Skonieczna ◽  
Dorota Hudy ◽  
Aleksandra Poterala-Hejmo ◽  
Tomasz Hejmo ◽  
Rafal J. Buldak ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Combination Treatment ◽  
Cellular Localization ◽  
Reactive Oxygen ◽  
Squamous Carcinoma ◽  
Oxygen Species ◽  
Combined Application ◽  
Flow Cytometric ◽  
Species Survival ◽  
Cellular Reactive Oxygen Species

Background: Levels of cellular Reactive Oxygen Species (ROS) influence the oxidized/reduced states of cellular proteins, and create redox-signaling pathways that can activate transcription factors, kinases, and phosphatases. ROS levels can be increased radically by external factors, including ionizing and UV radiation or exposure to chemical compounds. These increased ROS levels can, in turn, lead to oxidative damage of DNA. Natural plant treatments against cancer can modulate these processes by inducing or decreasing ROS production. Methods: Here we report new observations that squamous carcinoma (SCC-25) cells, exposed to 24 hours of combined resveratrol and berberine treatment, contain increased ROS levels. Using flow cytometry, for drug activity characteristics, an accumulation of ROS was observed. A combination of different dyes, CellROX Green (Life Technologies) and DCFH-DA (Sigma), allowed for flow cytometric estimation of levels of cellular ROS as well as cellular localization. Results: Live staining and microscopic observations confirmed the accumulation of ROS in SCC-25 cells following a combination treatment at concentrations of 10μg/ml. Additionally, the cytotoxicity of the compounds was significantly improved after their combined application. Additive effects were observed for doses lower than the calculated IC50 of berberine [IC50=23µg/ml] and resveratrol [IC50=9µg/ml]. Viability (MTS) assays and analysis of isobolograms revealed a significant impact on cell viability upon combination treatment. Conclusion: These results suggest that administration of berberine, in the presence of resveratrol, could be decreased even to 50% (half the IC50 for berberine) for cancer treatment.

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