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Isolation and characterization of microsomal fraction of fungus Pleurotus ostreatus and its role in the degradation of 17α-ethinylestradiol
Valášková, Petra ; Černá, Věra (advisor) ; Hodek, Petr (referee)
A synthetic hormone 17α-ethinylestradiol (EE2) which is a component of hormonal contraception pills has been identified as a main component of the endocrine-disrupting compounds (EDc). EDc are substances that mimic natural hormones in their action. Recently their amount especially in the groundwater and the surface water has been increased, which results in a negative impact on the hormonal system especially of aquatic organisms. Since it is not easy to replace these substances from the environment by conventional techniques other possibilities of their biodegradation are examined. White rot fungi, which are able to degrade lignin in nature, have promising biodegradation abilities towards many pollutants. These fungi contain a wide range of non-specific extracellular and intracellular enzymes that play an important role in the degradation. This bachelor thesis was targeted on the study of a white rot fungus, Pleurotus ostreatus, and especially on the degradation potential of its intracellular enzymes in the biodegradation of EE2. Initially, the ability of fungi Pleurotus ostreatus to degrade EE2 in vivo was tested. During the 48 hour incubation there was replaced 95,5 % of EE2. However, the role of cytochromes P450 (CYPs) in a metabolism of EE2 was not confirmed in this experiment by reason that an...
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Antioxidant and antiinflammatory effects of bilirubin
Valášková, Petra
For a long time, bilirubin (BR) has been considered a waste molecule with potential toxic effects especially on the central nervous system. Later, it was found that BR exhibited cytoprotective effects and mildly elevated BR levels showed antioxidant, anti-inflammatory and immunomodulatory properties, however, exact mechanisms of the anti-inflammatory actions of BR have not been fully understood yet. The main aim of this study was to assess the protective effects of BR using experimental in vivo and in vitro models in relation to inflammation and oxidative stress. Partial goal was to establish validated analytical method for determination of BR and lumirubin. Gunn and heterozygous rats were treated with lipopolysaccharide (LPS, 6 mg/kg, IP) or vehicle (saline). After 12 hours, blood and organs were collected for analyses of inflammatory and hepatic injury markers. Primary rat hepatocytes were treated with BR and TNF-α, HepG2 and SH-SY5Y cell lines were treated with BR and chenodeoxycholic acid. LPS-treated Gunn rats had a significantly decreased inflammatory response and hepatic injury compared to LPS- treated normobilirubinemic controls. We found different profile of leukocytes subsets and decreased systemic mRNA expressions and concentrations of IL-6, TNF-α, IL-1β and IL-10 in Gunn rats. Hepatic mRNA...
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Antioxidant and antiinflammatory effects of bilirubin
Valášková, Petra
For a long time, bilirubin (BR) has been considered a waste molecule with potential toxic effects especially on the central nervous system. Later, it was found that BR exhibited cytoprotective effects and mildly elevated BR levels showed antioxidant, anti-inflammatory and immunomodulatory properties, however, exact mechanisms of the anti-inflammatory actions of BR have not been fully understood yet. The main aim of this study was to assess the protective effects of BR using experimental in vivo and in vitro models in relation to inflammation and oxidative stress. Partial goal was to establish validated analytical method for determination of BR and lumirubin. Gunn and heterozygous rats were treated with lipopolysaccharide (LPS, 6 mg/kg, IP) or vehicle (saline). After 12 hours, blood and organs were collected for analyses of inflammatory and hepatic injury markers. Primary rat hepatocytes were treated with BR and TNF-α, HepG2 and SH-SY5Y cell lines were treated with BR and chenodeoxycholic acid. LPS-treated Gunn rats had a significantly decreased inflammatory response and hepatic injury compared to LPS- treated normobilirubinemic controls. We found different profile of leukocytes subsets and decreased systemic mRNA expressions and concentrations of IL-6, TNF-α, IL-1β and IL-10 in Gunn rats. Hepatic mRNA...
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Antioxidant and antiinflammatory effects of bilirubin.
Valášková, Petra ; Muchová, Lucie (advisor) ; Martínková, Markéta (referee) ; Dvořák, Karel (referee)
For a long time, bilirubin (BR) has been considered a waste molecule with potential toxic effects especially on the central nervous system. Later, it was found that BR exhibited cytoprotective effects and mildly elevated BR levels showed antioxidant, anti-inflammatory and immunomodulatory properties, however, exact mechanisms of the anti-inflammatory actions of BR have not been fully understood yet. The main aim of this study was to assess the protective effects of BR using experimental in vivo and in vitro models in relation to inflammation and oxidative stress. Partial goal was to establish validated analytical method for determination of BR and lumirubin. Gunn and heterozygous rats were treated with lipopolysaccharide (LPS, 6 mg/kg, IP) or vehicle (saline). After 12 hours, blood and organs were collected for analyses of inflammatory and hepatic injury markers. Primary rat hepatocytes were treated with BR and TNF-α, HepG2 and SH-SY5Y cell lines were treated with BR and chenodeoxycholic acid. LPS-treated Gunn rats had a significantly decreased inflammatory response and hepatic injury compared to LPS- treated normobilirubinemic controls. We found different profile of leukocytes subsets and decreased systemic mRNA expressions and concentrations of IL-6, TNF-α, IL-1β and IL-10 in Gunn rats. Hepatic mRNA...
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Study of the metabolism of 17α-ethinylestradiol by cytochromes P450
Valášková, Petra ; Černá, Věra (advisor) ; Levová, Kateřina (referee)
A synthetic estrogen 17α-ethinylestradiol (EE2) is the main active component of the hormonal contraceptive pills. The rise of consumption of hormonal contraceptives has increased the risk of the back negative effects of EE2 to aquatic organisms. EE2 belongs to the endocrine disruptive compounds known for mimicking natural hormones. A more detailed examination of the transformation of this compound in vivo and in vitro can contribute to a better understanding of its negative effects. This master thesis is therefore devoted to the study of the metabolism of EE2 in two selected model organisms. The ligninolytic fungus Pleurotus ostreatus is the type of fungi with promising biodegradation ability to a lot of pollutants. These properties have led to numerous studies of the degradation potential of P. ostreatus towards EE2, with the possibility of removing this compound from the environment. EE2 has been degraded by the fungus P. ostreatus in vivo resulting in one hydroxylated metabolite, which estrogenic activity is in need for further study. In vitro studies were carried out with a microsomal fraction isolated from the mycelium of this fungus. The conversion of EE2 in vitro via CYPs dependent on NADPH has not been demonstrated, however using KHP as a cofactor, there was one metabolite of EE2 found,...
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Isolation and characterization of microsomal fraction of fungus Pleurotus ostreatus and its role in the degradation of 17α-ethinylestradiol
Valášková, Petra ; Černá, Věra (advisor) ; Hodek, Petr (referee)
A synthetic hormone 17α-ethinylestradiol (EE2) which is a component of hormonal contraception pills has been identified as a main component of the endocrine-disrupting compounds (EDc). EDc are substances that mimic natural hormones in their action. Recently their amount especially in the groundwater and the surface water has been increased, which results in a negative impact on the hormonal system especially of aquatic organisms. Since it is not easy to replace these substances from the environment by conventional techniques other possibilities of their biodegradation are examined. White rot fungi, which are able to degrade lignin in nature, have promising biodegradation abilities towards many pollutants. These fungi contain a wide range of non-specific extracellular and intracellular enzymes that play an important role in the degradation. This bachelor thesis was targeted on the study of a white rot fungus, Pleurotus ostreatus, and especially on the degradation potential of its intracellular enzymes in the biodegradation of EE2. Initially, the ability of fungi Pleurotus ostreatus to degrade EE2 in vivo was tested. During the 48 hour incubation there was replaced 95,5 % of EE2. However, the role of cytochromes P450 (CYPs) in a metabolism of EE2 was not confirmed in this experiment by reason that an...
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preparation and evaluation of economic efficiency of selected investment project
Valášková, Petra ; Hrdý, Milan (advisor) ; Pražák, Jiří (referee)
The main goal of this study is to show a preparation of an investment project aimed on heating in a concrete company and to evaluate an economic effectiveness of the individual possible variants of the project. The project is unique due to particular financing from European union's grants and due to a goal of the company to meet new emission norm. The study is divided to three parts. In the first part, a theoretical one, is described a preparation of the project and an administration process of providing the grand from European Union funds. The second chapter deals with identification of the concrete investment project and the conditions of its evaluation. Moreover, important information about investor and circumstances of the investment project realization are discussed in this chapter. Finally, the process of the concrete grant approval is analyzed. The third part evaluates economic effectiveness according to cost measures and to net present value of the individual project variants. Because of a different lifespan of project variants the cost annual equivalent and the annuity equivalent has to be used. The results of cost measures and net present value are different. This pointes to the difference between cost only view and complex cash flow view. The final overall evaluation takes into consideration the ecologic part of the project. The final resolution has to make a compromise between various measurements. The study evaluates a real project. Therefore, the data are also based on reality.
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Analysis of the Automotive industry in ČR
Valášková, Petra ; Poborský, František (advisor) ; Marek, Petr (referee)
This thesis deals with the strategic analysis. It has two parts. The first theoretical part describes the purpose, functions and strategic analysis stages. The author consider strategic analysis as an important tool with many uses. In the second part is done an automotive Industry analysis. The analysis provides an objective assessment of its current situation and determines the probable development of the industry. At first is made a description of the status and the history of the industry. Once defined the key factors. Effect of factors is analyzed in the attractiveness of the industry. The work also studies the relationship between the automotive industry in the Czech Republic and abroad. The automotive industry in the ČR is evaluated as highly developed and strongly connected with global development in the industry. The key factors are: the size of the sector, its growth potential, stage in the life cycle, structure, changeable power, cost of capital, stability of demand, technological level, the intensity of competition and legislation. The future of the automotive industry is due to the interdependence of world economies considered as poorly predictable. Great importance is attributed to energy efficiency.
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