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Type of publication: Konferencijų tezės nerecenzuojamuose leidiniuose / Conference theses in non-peer-reviewed publications (T2)
Field of Science: Biologija / Biology (N010)
Author(s): Dainauskaitė, Viktorija Skaidrutė;Rodaitė-Riševičienė, Raminta;Pakhomov, Andrei G;Saulis, Gintautas
Title: Generation of hydrogen peroxide by high-voltage pulses in cell-free media
Is part of: Smart Bio: international conference, 18-20 May 2017, Kaunas : abstracts book. Kaunas : Vytautas Magnus University, 2017
Extent: p. 78-78
Date: 2017
Keywords: Reactive oxygen species;Amplex Red;Electrochemical reactions
ISBN: 9786098104424
Abstract: Electroporation offers a number of applications in biology, oncology, immunology, and biotechnology. However, when a high-voltage is applied to the electrolyte solution, besides membrane permeabilization, various electrolysis reactions occur at the electrode-solution interfaces. One of the results of these electrochemical reactions is generation of reactive oxygen species (ROS). Although ROS formation following electroporation with long (μs–ms) electric pulses or nsPEF exposure has been reported earlier, no detailed analysis of this process has been carried out yet. In this study, generation of hydrogen peroxide as a result of the exposure of a cell-free media by high-voltage pulses has been studied. Generation of hydrogen peroxide in hightly buffered solution HB1, Dulbecco's Modified Eagle's medium (DMEM), Phosphate buffered saline (PBS), and distilled water was studied. The cuvette with stainless-steel electrodes was used. Hydrogen peroxide was detected with AmplexRed® (10- Acetyl-3,7-dihydroxyphenoxazine), which has a great specificity, stability, and selectivity. AmplexRed is nonfluorescent until, in the presence of horseradish peroxidase, it reacts with hydrogen peroxide to produce highly fluorescent resorufin. 50 μM of the AmplexRed dye were added to the medium and treated with high-voltage pulses. Fluorescence intensity in a solution was measured using TECA7 GeniosPro spectrophotometer (Tecan Group, Männedorf, Switzerland). Treatment of HB1 medium by a single square-wave electric pulse with the duration of 0.5–2 ms and the amplitude of 100– 400 V (0.5-2.0 kV/cm) significantly increased dye emission. The AmplexRed fluorescence intensity was also dependent on the number of pulses, the conductivity, and composition of the medium (PBS, H2O, DMEM, HB1).[...]
Affiliation(s): Biologijos katedra
Gamtos mokslų fakultetas
Vytauto Didžiojo universitetas
Appears in Collections:Universiteto mokslo publikacijos / University Research Publications

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