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Optimization of Oxidative Stress Diagnostic Methods
Mercová, Miroslava ; Macáková, Kateřina (advisor) ; Mladěnka, Přemysl (referee)
67 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Mgr. Miroslava Mercová Consultant: Ing. Kateřina Macáková, PhD. Consultant - specialist: doc. RNDr. Miroslav Pohanka, PhD. Title of Thesis: Optimization of oxidative stress diagnostic methods Many methods have been developed to measure markers related to oxidative stress, but only little attention has been paid to their optimization for sample handling. Sample processing and storage can reduce stability of detected markers and therefore influence the results obtained. In this experiment we studied the effect of freezing - thawing cycles on the stability of some oxidative stress markers. The objective was to evaluate the process of repeated freezing as a potential risk factor that can interfere with the precision of measurements. Wistar rat tissue homogenates (brain, liver, heart, kidney, spleen) were frozen (-80 řC) and thawed at room temperature for five cycles. TBARS, FRAP, DPPH-test, GSH levels and GR activity were measured after the every freeze - thaw cycle. The measured data show changes in levels of the assessed markers. Their stability was affected in all of the tested samples, already after the first cycle. In many cases, we found strong linear dependence between...
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Optimization of Oxidative Stress Diagnostic Methods
Mercová, Miroslava ; Macáková, Kateřina (advisor) ; Mladěnka, Přemysl (referee)
67 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Mgr. Miroslava Mercová Consultant: Ing. Kateřina Macáková, PhD. Consultant - specialist: doc. RNDr. Miroslav Pohanka, PhD. Title of Thesis: Optimization of oxidative stress diagnostic methods Many methods have been developed to measure markers related to oxidative stress, but only little attention has been paid to their optimization for sample handling. Sample processing and storage can reduce stability of detected markers and therefore influence the results obtained. In this experiment we studied the effect of freezing - thawing cycles on the stability of some oxidative stress markers. The objective was to evaluate the process of repeated freezing as a potential risk factor that can interfere with the precision of measurements. Wistar rat tissue homogenates (brain, liver, heart, kidney, spleen) were frozen (-80 řC) and thawed at room temperature for five cycles. TBARS, FRAP, DPPH-test, GSH levels and GR activity were measured after the every freeze - thaw cycle. The measured data show changes in levels of the assessed markers. Their stability was affected in all of the tested samples, already after the first cycle. In many cases, we found strong linear dependence between...
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Evaluation of oxidative stress markers after intoxication with nerve agents and treatment with acetylcholinesterase reactivators
Mercová, Miroslava ; Opletal, Lubomír (advisor) ; Mladěnka, Přemysl (referee)
53 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Botany and Ecology Candidate: Miroslava Mercová Consultant: Doc. RNDr. Lubomír Opletal, CSc. Title of Thesis: Evaluation of oxidative stress markers after intoxication with nerve agents and treatment with acetylcholinesterase reactivators In this study, the influence of soman poisoning and treatment with atropine and some oximes on redox status in rat erythrocytes, liver and plasma were investigated. Male Wistar rats were divided into eight groups, each of six animals. The first group was exposed to atropine only, the second group to one LD50 of soman and atropine. The groups 3-7 were treated with soman, atropine and one of named oximes (HI-6, trimedoxime, K203) or with their combinations (HI-6 + trimedoxime, HI-6 + K203). The control (eighth) group was treated with saline solution only. To study the markers of oxidative stress the TBARS method (thiobarbituric acid reactive substances), glutathion reductase activity and the FRAP (ferric reducing antioxidant power) method were chosen. There was plasma protein content and acetylcholinesterase activity assessed, too. The experimental data showed significant alterations in followed parameters. The presence of oxidative stress is offset by enhanced...
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