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Optimization of Experimental Settings for the Assessment of Reactive Oxygen Species Production by Human Blood
Joint Authors
Freitas, Marisa
Soares, Tânia
Rodrigues, Daniela
Sarraguça, Mafalda
Rocha, Sílvia
Lima, José L. F. C.
Ribeiro, Daniela
Fernandes, Eduarda
Source
Oxidative Medicine and Cellular Longevity
Issue
Vol. 2019, Issue 2019 (31 Dec. 2019), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2019-01-10
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
The purpose of an experimental design is to improve the productivity of experimentation.
It is an efficient procedure for planning experiments, so the data obtained can be analyzed to yield a valid and objective conclusion.
This approach has been used as an important tool in the optimization of different analytical approaches.
A D-optimal experimental design was used here, for the first time, to optimize the experimental conditions for the detection of reactive oxygen species (ROS) produced by human blood from healthy donors, a biological matrix that better resembles the physiologic environment, following stimulation by a potent inflammatory mediator, phorbol-12-myristate-13-acetate (PMA).
For that purpose, different fluorescent probes were used, as 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), 2-[6-(4′-amino)-phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (APF), and 10-acetyl-3,7-dihydroxyphenoxazine (amplex red).
The variables tested were the human blood dilution, and the fluorescent probe and PMA concentrations.
The experiments were evaluated using the Response Surface Methodology and the method was validated using specific compounds.
This model allowed the search for optimal conditions for a set of responses simultaneously, enabling, from a small number of experiments, the evaluation of the interaction between the variables under study.
Moreover, a cellular model was implemented and optimized to detect the production of ROS using a yet nonexplored matrix, which is human blood.
American Psychological Association (APA)
Soares, Tânia& Rodrigues, Daniela& Sarraguça, Mafalda& Rocha, Sílvia& Lima, José L. F. C.& Ribeiro, Daniela…[et al.]. 2019. Optimization of Experimental Settings for the Assessment of Reactive Oxygen Species Production by Human Blood. Oxidative Medicine and Cellular Longevity،Vol. 2019, no. 2019, pp.1-11.
https://search.emarefa.net/detail/BIM-1205038
Modern Language Association (MLA)
Soares, Tânia…[et al.]. Optimization of Experimental Settings for the Assessment of Reactive Oxygen Species Production by Human Blood. Oxidative Medicine and Cellular Longevity No. 2019 (2019), pp.1-11.
https://search.emarefa.net/detail/BIM-1205038
American Medical Association (AMA)
Soares, Tânia& Rodrigues, Daniela& Sarraguça, Mafalda& Rocha, Sílvia& Lima, José L. F. C.& Ribeiro, Daniela…[et al.]. Optimization of Experimental Settings for the Assessment of Reactive Oxygen Species Production by Human Blood. Oxidative Medicine and Cellular Longevity. 2019. Vol. 2019, no. 2019, pp.1-11.
https://search.emarefa.net/detail/BIM-1205038
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1205038