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Development, characterization and use of anti-orphan cytochrome P450 antibodies
Hrdinová, Johana ; Hodek, Petr (advisor) ; Koblas, Tomáš (referee)
The cytochromes P450 (P450s) are important enzymes involved in metabolic pathways, which use exogenous and endogenous substances as their substrate for various enzymatic reactions. These enzymes can also use precarcinogens as their substrate and activate them into carcinogens, which leads to a cancer development. If the P450s are induced, the cancer risk increases. Some chemopreventive compounds may induce the P450s and thus be harmful to the human body. Therefore it is necessary to pay enough attention to a study of the mechanism of action of P450s and the influence of the chemopreventive compounds on the activity of cytochromes P450. mRNA expression of most of the P450s isoforms is detected in a number of healthy (nontransformed) tissues, viz. liver, brain, heart, colon, kidney or placenta. Nevertheless there are a few P450s isoforms which mRNAs are expressed at relatively low levels in the nontransformed tissues, whereas the expression in the transformed tissues is significantly higher. One of these P450s is CYP2W1, which can be used as a prognostic marker for colorectal cancer - therefore it is useful to be able to detect a presence of this enzyme in various tissues. A detection of P450s can be accomplished by using a method Western blot. In this method, the immunodetection is achieved by using...
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Study of modified amino acid incorporation into cytochrome b5
Koberová, Monika ; Hodek, Petr (advisor) ; Pavlásková, Kateřina (referee)
A cytochrome b5 (cyt b5) can influence cytochrome P450 (CYP)-dependent reactions. In consequence of these reactions cytochrome b5 can participate in substance activation (for example drugs, carcinogens) or it can influence proportions of formed metabolites. A mechanism of cyt b5 action has not been fully explained yet. Elucidation of protein-protein interactions in monooxygenase system could explain of the mechanism of cyt b5 action. To study these interactions by using cross-linking techniques is necessary to prepare photolabile cyt b5, which after photoactivation generated higly reactive intermediates which can create a complex with nearby components of the monoogynesase system. This thesis describes how was developed the method for the production of recombinant cyt b5 with modified amino acids. Cyt b5 was expressed in a bacterial strain E. coli BL-21 (DE3) Gold. Before the expression induction, cells were transformed into the limiting medium (DMEM-LM) which did not contain L-leucine and L-methionine. The limiting medium was supplemented by deuterated amino acid d3-methyl-L-methionine and D,L-Leucine. Expressed cyt b5 was isolated and incorporation of d3-methyl-L-methionene has been verified by mass spectrometry. Cyt b5 was obtained mainly as the apoprotein (apo-cyt b5). That is why in this...
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Use of oxidation processes (AOP) for removal of micropolutants
Stříteský, Luboš ; Pešoutová,, Radka (referee) ; Hlavínek, Petr (advisor)
This thesis deals with advanced oxidation processes (AOPs) and it’s use for removal of micropollutants from wastewater. The first chapter explains the need AOPs, water quality, pollution and substances that are present in the water. Further, the first chapter outlines approach of the current legislation to micropollutants. The second chapter explains the theory and principle of operation of AOPs. This chapter is divided into two sections. The first section describes AOPs, which were tested at selected WWTP. In the second section, there are described some other AOPs. The third chapter is a literature retrieval of AOPs dealing with the removal of micropollutants. This chapter is focused on the removal of hormones by AOPs using ozone-based AOPs. The fourth chapter describes the actual testing of selected AOPs. The chapter describes selected WWTP, pilot-scale AOP unit and test results. In the last chapter there is designed and described full-scale AOP tertiary unit for removing of micropollutants. The last chapter also contains economic analysis of the proposed tertiary unit.
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Selection of suitable bioindicators for the determination of xenobiotics residuals
Musil, Jan ; Vondráčková, Ilona (referee) ; Vávrová, Milada (advisor)
Bio-indicators are organisms, which we can use to evaluate contamination of terestrial and aquatic ecosystems by xenobiotics. This thesis deals above all with bio-indicators of vegetal and animal origin, which are capable to identify and quantify persistent organic pollutants and its residual contamination. This xenobiotics comes from human activities, most by consumer and automotive industry, inconsiderate agriculture. Thanks to its unpremeditated application and waste disposal, which are contaminated with them, ecosystems are infested and persistent organic pollutant‘s residuals are persisting for decades. By the help of bio-indicators we can find the source of contamination and size up its seriousness, we can observe its development and biological effect on organisms, which live in contaminated areas. In this thesis attention is given especially to polychlorinated bifenyls (PCB), polyaromatic hydrocarbons (PAH), organochlorinated pesticides (OPC). polybrominated difenylethers (PBDE) and PCCDs/Fs, its influence on the health and its distribution in our surroundings, especially in waste dumps and forgotten pollutants sources.
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EVALUATION OF EFFECTIVITY OF PHYTOREMEDIATION METHODS
Soudek, Petr ; Petrová, Šárka ; Podlipná, Radka ; Fialová, Zuzana ; Malá, J. ; Benešová, Dagmar ; Kočí, V. ; Kafka, Z. ; Vaněk, Tomáš
Phytoremediation methods are very dynamically booming segment of remediation technologies in present time. These methods used plants and microorganisms for extraction, and in case of organics for degradation, of xenobiotics. Ability of plants involved in these detoxification reactions is depend not only on ability to uptake of xenobiotics and on enzymatic aparatus of tested plant species, but also on toxicity of accumulated contaminant and on toxicity of potential byproducts.
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Rostliny a mikroorganismy zajišťující cyklus PCB a chlorbenzoátů v životním prostředí
Macková, Martina ; Vrchotová, Blanka ; Frančová, Kateřina ; Rezek, Jan ; Beranová, K. ; Najmanová, Jitka ; Lovecká, P. ; Tříska, J. ; Vrchotová, N. ; Uhlík, Ondřej ; Sylvestre, M. ; Demnerová, K. ; Macek, Tomáš
Bacterial metabolism of plant PCB degradation products and degradation of chlorobenzoic acids by plant cells were studied.
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Interakce xenobiotik s MDR pumpami u S. cerevisiae
Hendrych, T. ; Zlámalová, Marta ; Gášková, Dana ; Pichová, Alena ; Sigler, Karel
The interactions of a number of xenobiotics with two main S. cerevisiae MDR pumps, Pdr5p and Snq2p were studied. The results showed that some compounds do not interact with MDR pumps whereas others affect one or another of the pumps, causing a loss of their ability to transport an indicator substrate out of the cells. This apparent inactivation of the pumps can be due to competition for transport with the indicator substrate or to actual (noncompetitive) inhibition of the pumps
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Následky interakcí mezi rostlinami a mikroby ovlivňujících osud xenobiotik v životním prostředí
Macková, Martina ; Vrchotová, N. ; Beranová, K. ; Najmanová, J. ; Lovecká, P. ; Kochánková, L. ; Sylvestre, M. ; Demnerová, Kateřina ; Macek, Tomáš
The effects and consequences of plant-microbe metabolic interactions influencing fate of xenobiotics in the environment.are discussed, illustrated by our results on PCB degradation by microflora in plant rhizosphere.
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