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Preparation and characterization of synthetic mRNA coding for pancreatic transcription factors
Loukotová, Šárka ; Hodek, Petr (advisor) ; Jirák, Daniel (referee)
Diabetes mellitus type I is severe autoimmune disease which is caused by destruction of insulin-producing β-cells in pancreas. Diabetic patients are dependent on external usage of insulin during their whole life. Nowadays the only treatment of diabetes type I is transplantation of entire pancreas or isolated Langerhans islets. Due to the fact that this kind of treatment is very demanding and limited availability of suitable donors, the researchers are intensively working on development of new alternative ways how to produce the insulin-producing cells. One of the possible approaches on producing insulin-positive cells is transdifferentiation of pancreatic exocrine cells via transcription factors. In this diploma thesis, the transdifferentiation of exocrine cells AR42J was carried out with in vitro synthesized mRNA encoding transcription factors Pdx1, Ngn3 and MafA. The primary mRNA structure was optimized in order to prepare highly stable mRNA which is correctly translated into the protein. The main stabilizing elements in mRNA structure include 3' and 5' untranslated region derived from highly stable β-globin mRNA. In order to verify the function of synthetic mRNA the immunofluorescence staining of transcription factors has been investigated. Synthetic mRNAs encoding transcription factors Pdx1,...
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Unjustified Enrichment in Business Law
Loukotová, Šárka ; Horáček, Vít (advisor) ; Elek, Štefan (referee)
Unjustified Enrichment in Business Law Abstract The aim of my thesis is to analyse the concept of unjustified enrichment in connection with the commercial law. In fact this topic is usually described in the civil law studies and at the same time the unjustified enrichment is minor topic compared to the obligations arising from the contracts. This is the reason why I have chosen this issue describe and focus on the business aspects. The thesis is composed of seven chapters. Chapter one as an introduction defines basic legislation acts, which I am dealing with throughout the thesis and roughly describes there are changes in connection with the extensive recodification of the Czech private law. Chapter two is subdivided into two parts. Part one focuses on the origin of the unjustified enrichment in Ancient Rome as the base for the civil law. Part two provides an outline of past legislation within the territory of the Czech Republic until its establishment. Chapter three concentrates on the legislation effective until 31 December 2013. It illustrates the approach to decision-making by the Supreme Court including the decision-making in connection with the limitation of rights according to Commercial Code. Chapter four analyses the changes after New Civil Code came into force in relation to commercial law. Here...
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Volatilization of inorganic chlorides by haloperoxidases of soil microorganisms and plants
Loukotová, Šárka ; Stiborová, Marie (advisor) ; Moserová, Michaela (referee)
Biogeochemical cycle of chlorine, particularly formation of organically bound chlorine (Clorg), is still known only in outlines. In continental ecosystems chloride acts as stress factor, and also as source of chlorine; it originates from precipitation, which comes from marine cloud masses. Clorg are formed naturally by biotical and abiotical way. The biological factors are microorganisms, soil enzymes, plants and even animals. Halogenation processes undergo in marine environment as well, the main difference is the presence of bromine besides chlorine, therefore the reaction and formation of brominated compounds. One group of Clorg represents volatile chlorinated hydrocarbons (VOCl). Several VOCl can react with atmospheric ozone, consequently causing its depletion. The best known sources of VOCl are soil, the microorganisms living in it, and their exoenzymes released into it. The heme peroxidases in the group EC 1.11.1.X can perform halogenation of organic compounds. Hydrogen peroxide or organic peroxides are the substrates of this reaction in which hypochlorous acid is generated. The halogen in reactive form then carries chlorine onto the organic compound. To study enzymatic mediation of chlorination processes, we chose commercially available enzymes (chloroperoxidase from Caldariomyces fumago and...
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