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COLORIMETRIC ANALYSIS OF BIOCHEMICAL MARKERS USING PHOTOGRAPHY
Mackurová, Kateřina ; Pohanka, Miroslav (advisor) ; Vlček, Vítězslav (referee)
University of Pardubice Faculty of Chemical Technology in Pardubice Performed at: University of defence in Brno, Fakulty of Military Health Sciences in Hradec Králové, Department of Molecular Pathology and Biology Student: Kateřina Mackurová Leader of diploma thesis: pplk. prof. RNDr. Miroslav Pohanka, Ph.D., DSc. Consultant: Mgr. Šárka Štěpánková, Ph.D. Title of diploma thesis: Colorimetric Analysis Of Biochemical Markers Using Photography Keywords: colorimetry, acetylcholinesterase, tacrine, photometric method Acetylcholine, which is responsible for synaptic transmission, is nowadays considered to be the main neurotransmitter in the brain. It is degraded into choline and acetate by two enzymes - acetylcholinesterase (AChE) (EC 3.1.1.7) and butyrylcholinesterase (BChE) (EC 3.1.1.8) AChE inhibitors represent an interesting subject of current scientific research since they belong to the main treatments for Alzheimer's disease or myasthenia gravis. The aim of the thesis was to verify the photometric equipment while determining the reaction of AChE with the substrate on pH papers, to determine the kinetic parameters AChE, and to optimize the method for the determination of the takrin inhibitor. Furthermore, the goal was to determine the influence of selected matrices on AChE using the photometric...
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Following of acetylcholinesterase-pesticide interaction using biosensor technology
Drobík, Oto ; Opletalová, Veronika (advisor) ; Vlček, Vítězslav (referee)
FOLLOWING OF ACETYLCHOLINESTERASE-PESTICIDE INTERACTION USING BIOSENSOR TECHNOLOGY Oto Drobík Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Organophosphorus and carbamate pesticides are toxic substances used in agriculture as insecticides. These substances are highly toxic for humans as acute poisoning can lead to death via inhibition of acetylcholinesterase. After acetylcholinesterase inhibition, accumulation of acetylcholine follows. In compliance with the thesis aims, functional electrochemical biosensor was constructed. It was based on a standard screen printed amperometric sensor with three electrodes. Herein sensors with platinum and carbon working electrode were tested. Suitable method of electrochemical assay was chosen and some the basic conditions were optimized. Kinetic parameters of enzyme reaction were assessed, too. Various methods of acetylcholinesterase capturing onto sensor were tested and compared one to each other. Paraoxon-methyl and diisopropylfluorophosphate were chosen as the representative of pesticides. Limit of detections for the both pesticides were estimated to be 10-6 - 10-5 mol/l. The prepared biosensors were compared with the standard Ellman's method based on spectrophotometry and the...
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Criminological Aspects of Hate Crime
Vlček, Vítězslav ; Kopečný, Zdeněk (advisor) ; Galovcová, Ingrid (referee)
Criminological Aspects of Hate Crime Abstract Hate crime refers to a set of crimes committed for a prejudicial motive against a group of people for their essential personality characteristics such as nationality, religion, or sexual orientation. Topicality of the issue stems from the fact that related social phenomena such as racism, xenophobia, radicalisation, or discrimination of minorities are far from being satisfactorily dealt with even in Western society. The fundamental goal of this thesis is to provide the reader with a comprehensive criminological view of hate crimes in the territory of the Czech Republic. Main findings include the following: the concept of hate crime is not unified in theory nor legislation in terms of protected characteristics, which negatively translates into research and policy of hate crime. The quantity of registered crime is in the order of less than 200 crimes per year, its trend is downward, and its structure is predominantly non-violent; unfortunately, it is also accompanied by high latency. The Czech perpetrator is usually male and a repeat offender, in early adulthood, with a basic education and without a factual relationship to extremist groups. Victims of violent forms of hate crime are, in principle, suffering more intense harm (especially psychological) than victims...
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Following of acetylcholinesterase-pesticide interaction using biosensor technology
Drobík, Oto ; Opletalová, Veronika (advisor) ; Vlček, Vítězslav (referee)
FOLLOWING OF ACETYLCHOLINESTERASE-PESTICIDE INTERACTION USING BIOSENSOR TECHNOLOGY Oto Drobík Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Organophosphorus and carbamate pesticides are toxic substances used in agriculture as insecticides. These substances are highly toxic for humans as acute poisoning can lead to death via inhibition of acetylcholinesterase. After acetylcholinesterase inhibition, accumulation of acetylcholine follows. In compliance with the thesis aims, functional electrochemical biosensor was constructed. It was based on a standard screen printed amperometric sensor with three electrodes. Herein sensors with platinum and carbon working electrode were tested. Suitable method of electrochemical assay was chosen and some the basic conditions were optimized. Kinetic parameters of enzyme reaction were assessed, too. Various methods of acetylcholinesterase capturing onto sensor were tested and compared one to each other. Paraoxon-methyl and diisopropylfluorophosphate were chosen as the representative of pesticides. Limit of detections for the both pesticides were estimated to be 10-6 - 10-5 mol/l. The prepared biosensors were compared with the standard Ellman's method based on spectrophotometry and the...
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In vitro reactivation of cholinesterases previously inhibited by pesticides
Drtinová, Lucie ; Opletalová, Veronika (advisor) ; Vlček, Vítězslav (referee)
In vitro reactivation of choliesterases after pesticides inhibition Organophosphate pesticides are toxic substances supplied commercially as insecticides in order to protect agricultural commodities, for suppression of epidemic events etc. The use of pesticides in the Czech Republic is strongly regulated by law similarly as in the whole EU. The regulation is aimed at monitoring and regulating the use of products containing toxic pesticides. In this diploma work, I measured the efficacy of previously synthesized oxime reactivators using the standard spectrophotometric method and multi-channel spectrophotometer. After experimental testing, the ability of selected pesticides to inhibit acetylcholinesterase (AChE) was assessed. I choose tree pesticides that are capable of binding to AChE without any metabolic activation. It was paraoxon ethyl, paraoxon methyl, and DFP. Subsequently, I experimentally observed the ability of 78 selected oximes reactivators to restore AChE activity in vitro. I proved that the DFP inhibited AChE can be reactivated with high efficacy using monopyridinium reactivators but bispyridinium reactivators are also suitable. In contrast to DFP, paraoxon ethyl as well as paraoxon methyl inhibited AChE was reactivated with only bispyridinium reactivators. Number of oxime groups isn not...
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