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The effect of fern extracts on cell line SW480
Zacharská, Andrea ; Matoušková, Petra (advisor) ; Skálová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Student: Andrea Zacharská Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of fern extracts on the SW480 cell line Cancer is one of the leading causes of death in the world. Although we have several drugs available for the treatment of cancer, none of them are completely effective and safe. The fundamental problem is toxicity, which experts try to eliminate by looking for other treatment alternatives like plants and their secondary metabolites with antiproliferative and antioxidant effects against free radicals, that can cause cancer. Our work is focused on the antioxidant, as well as the prooxidant effect of 4 species of ferns (Athyrium filix - femina, Dryopteris dilatata, Dryopteris borreri, Blechnum spicant), whose extracts of different concentrations were studied as potentially beneficial substances against oxidative stress on the SW480 cell line. We used the fluorescent probe H2DCF-DA to detect the production of reactive oxygen species (ROS) in cells. In the initial setup the potential prooxidant effect was monitored alongside with the terc- butyl hydroperoxide solution as a positive control. The results demonstrated that these test substances did not induce the ROS...
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The effect of thujone and piperitone on the expression of biotransformation enzymes in liver slices.
Štětková, Lenka ; Skálová, Lenka (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lenka Štětková Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The effect of thujone and piperitone on the expression of biotransformation enzymes in liver slices Monoterpenes are natural substances that are widely distributed in the essential oils of many plant species. Thujone and piperitone are found in certain plant species that can be part of dietary supplements, herbal preparations or used in foods as flavourings. The aim of this diploma thesis was to find out how thujone and piperitone affect the expression of selected biotransformation enzymes from the cytochrome P450 superfamily, the sulfotransferase and the UDP-glucuronosyltransferase superfamilies. Precision-cut liver slices obtained from the livers of 3 human donors were used for these experiments. Slices were incubated with thujone or piperitone for 24 hours at 10 μM and 50 μM concentrations. After incubation, the amount of mRNA and protein was quantified. The obtained results showed considerable interindividual variability. CYP1A2 gene expression was significantly increased in one donor sample after incubation with thujone (50 μM) and piperitone (10 μM, 50 μM). Piperitone (50 μM) increased the amount of mRNA of...
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The effect of selected herbicides on UGTs expression and activity of antioxidant enzymes in Arabidopsis thaliana
Rysová, Samanta ; Matoušková, Petra (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Samanta Rysová Supervisor: doc. Ing. Petra Matoušková, Ph.D. Consultant: RNDr. Radka Podlipná, Ph.D. Title of diploma thesis: The effect of selected herbicides on UGTs expression and activity of antioxidant enzymes in Arabidopsis thaliana Herbicides are widely used to control weeds in agriculture. Although their use is essential to ensure crop production, their effect occurs to be reduced due to the development of herbicide resistence. Therefore it is important to clarify the mechanism of herbicide resistence, so that herbicides can be used effectively and new substances can be developed if necessary. At the same time, herbicides acts as an abiotic stress factor on the plant, thus affecting the antioxidant apparatus of plant cells. The aim of this work was to investigate the effect of selected herbicides - tribenuron-methyl (TBM), pinoxaden (PNX) and sulcotrione (SLC), on the expression of selected UGTs that could play a role in herbicide resistence. Secondary, we want to find out the effect of TBM on the activity of selected antioxidant enzymes - ascorbate peroxidase (APX), catalase (CAT) and superooxide dismutase (SOD). Arabidopsis thaliana was used as a model plant and the experiment was...
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Monitoring changes in microRNA expression in cervical cancer
Štveráková, Tereza ; Matoušková, Petra (advisor) ; Skarková, Veronika (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Kralove Department of Biochemical Sciences Author: Bc. Tereza Štveráková Supervisor: doc. Ing. Petra Matoušková, Ph.D. Consultant: Ing. Helena Kovaříková, Ph.D. Title of diploma thesis: Monitoring changes in microRNA expression in cervical cancer Cervical cancer is the fourth most common cancer among women globally. Prevention of cervical cancer is based on routine screening using quality diagnostic tests. This diploma thesis focused on searching for sensitive and specific diagnostic markers that would detect the early stage of carcinogenesis. Suitable candidates for a diagnostic markers are microRNAs (miRNAs), which negatively post-transcriptionally regulate gene expression. In carcinogenesis, miRNAs are deregulated and may play a significant role in cancer development. The aim of this diploma thesis was to identify significantly deregulated miRNAs that could be used as diagnostic markers of cervical cancer. In this diploma thesis, the combination of miR-423-5p and miR-25-3p was selected as the endogenous miRNA control based on the results of real-time PCR analysis and the results of the RefFinder algorithm. After that, the relative expression of selected miRNAs (miR-10b-5p, miR-145-5p a miR-182-5p) was determined by real-time PCR and...
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The effects of topoisomerase II beta on the sensitivity of the cancer cells to the antineoplastics
Jaščevská, Nikola ; Jirkovská, Anna (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Nikola Jaščevská Supervisor: PharmDr. Anna Jirkovská, Ph.D. Title of diploma thesis: The effects of topoisomerase II beta on the sensitivity of the cancer cells to the antineoplastics Topoisomerase II (TOP II) is a cellular enzyme responsible for solving topological problems of double-stranded DNA. Alpha and beta isoforms of TOP II are different gene products having similar catalytic activities. The expression of TOP IIα is cell-cycle dependent, peaking in G2/M phase, while TOP II isoform is expressed constitutively throughout the cell cycle. It is therefore present also in non-proliferating differentiated cells. Anthracycline antibiotics are an old class of anticancer drugs, belonging to TOP II poisons. Although their clinical usefulness is high, the incidence of side effects (especially myelotoxicity and cardiotoxicity) may limit the therapy. The key role of TOP II inhibition, which is present also in cardiomyocytes, has been increasingly discussed. Dexrazoxane, the only clinically used cardioprotective, leads to depletion of TOP II in cardiomyocytes, which may explain its cardioprotection. Although TOP II was previously shown to be dispensable for cellular proliferation, its possible...
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The role of UDP-glycosyltransferase in development of drug resistance in parasitic nematodes
Dimunová, Diana ; Matoušková, Petra (advisor) ; Bílková, Zuzana (referee) ; Kašný, Martin (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Diana Dimunová Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of Doctoral Thesis: The role of UDP-glycosyltransferases in drug resistance in parasitic nematodes The diseases caused by parasitic nematodes represent a serious problem, which threatens livestock's health, because pharmacotherapy is complicated by widespread anthelmintic resistance. Understanding of the mechanisms of parasite drug resistance and defense strategies is important to maintaining the effectiveness of currently used anthelmintics and developing new approaches to controlling these infections. The ability of parasites to inactivate anthelmintics through their metabolism, which is provided by biotransformation enzymes, may contribute to the development of drug resistance. The UDP-glycosyltransferases (UGT) superfamily can protect parasites from the toxic actions of anthelmintics by modifying drugs to inactive glycoside metabolites. These metabolites have been identified in benzimidazole metabolism to an increased extent in a resistant strain of H. contortus, which suggests the involvement of UGTs in anthelmintic resistance. In the genome of this parasitic nematode, 32 genes encoding UGTs divided into 15 families have...
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Plasmid constuction for expression of UDP-glucosyltransferase from Haemonchus contortus
Machalová, Barbora ; Matoušková, Petra (advisor) ; Novotná, Eva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Barbora Wengrynová Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Plasmid construction for expression of UDP-glukosyltransferase from Haemonchus contortus The barber's pole worm (Haemonchus contortus), like other organisms, uses biotransformation enzymes to metabolize exogenous and endogenous substances. Important representatives of these enzymes are also UDP-glycosyltransferases, which catalyze the formation of a covalent bond between hexose activated by uridine diphosphate and the metabolized compound, thus helping to eliminate undesirable substances. Increased attention to these enzymes is due to the involvement of UDP-glycosyltransferases in the development of resistance to anthelmintics, which are used to treat hemonchosis. The study of their properties and functions can significantly help in solving resistance issues. UGTs are divided into several families and subfamilies depending on sequence similarities. This work focuses on three representatives of UGT in the Haemonchus contortus, UGT24C1, UGT365B6 a UGT368B2. The theoretical part of the diploma thesis includes knowledge about the Haemonchus contortus, hemonchosis and its treatment, biotransformation enzymes and...
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The role of UDP-glycosyltransferase in development of drug resistance in parasitic nematodes
Dimunová, Diana ; Matoušková, Petra (advisor) ; Bílková, Zuzana (referee) ; Kašný, Martin (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Diana Dimunová Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of Doctoral Thesis: The role of UDP-glycosyltransferases in drug resistance in parasitic nematodes The diseases caused by parasitic nematodes represent a serious problem, which threatens livestock's health, because pharmacotherapy is complicated by widespread anthelmintic resistance. Understanding of the mechanisms of parasite drug resistance and defense strategies is important to maintaining the effectiveness of currently used anthelmintics and developing new approaches to controlling these infections. The ability of parasites to inactivate anthelmintics through their metabolism, which is provided by biotransformation enzymes, may contribute to the development of drug resistance. The UDP-glycosyltransferases (UGT) superfamily can protect parasites from the toxic actions of anthelmintics by modifying drugs to inactive glycoside metabolites. These metabolites have been identified in benzimidazole metabolism to an increased extent in a resistant strain of H. contortus, which suggests the involvement of UGTs in anthelmintic resistance. In the genome of this parasitic nematode, 32 genes encoding UGTs divided into 15 families have...
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Bioinformatic aspects of nuclear receptors in pharmacotherapy
Škoda, Josef ; Pávek, Petr (advisor) ; Kollár, Peter (referee) ; Matoušková, Petra (referee)
IN ENGLISH LANGUAGE Candidate: Mgr. Josef Škoda Supervisor: Prof. PharmDr. Petr Pávek, PhD. Title of the doctoral thesis: Bioinformatical aspects of nuclear receptors in pharmacotherapy The liver plays a central role in energy homeostasis via processing post-prandial excess energy into storage molecules and reusing stored energy via gluconeogenesis during the fasting period. This tight energy balance is maintained by a myriad of regulating processes. Dysfunction of metabolic control is a key event of severe diseases in today's population, starting with obesity, insulin resistance, and metabolic syndrome, proceeding to end-stage complications in type 2 diabetes, whole organ malfunctions, or even tumor diseases. Master players in metabolic regulation are nuclear receptors (NR) activated by endogenous stimuli or scavenging for nutritional or toxicological signals. NRs regulate gene transcription activation and therefore maintain liver metabolic plasticity. In this thesis, modern molecular biology approaches were used to study ligands of NRs and the effects of their treatment. After the typical activation of a NR, hundreds of genes are regulated, which is beyond the ability to study with conventional biology methods. For this purpose, omics methods are an ideal solution. They are characterized by...
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The effect of zanubrutinib on tumor cell resistance to daunorubicin due to carbonyl reducing enzymes.
Havlíčková, Lucie ; Wsól, Vladimír (advisor) ; Matoušková, Petra (referee)
5 Abstract Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lucie Havlíčková Supervisor: prof. Ing. Vladimír Wsól, Ph.D. Title of diploma thesis: The effect of zanubrutinib on tumor cell resistance to daunorubicin due to carbonyl reducing enzymes Anthracycline antibiotics (ANT) are considered the first-line medication for oncologic diseases, including acute myeloid leukemia. Daunorubicin (DAUN) with a carbonyl group in position 13 is often used in chemotherapeutic treatments. Nicotinamidadenindinucleotid-phosphate dependent carbonyl reducing enzymes (CRE) create its hydroxyl metabolite daunorubicinol (DAUN-ol), which demonstrates higher toxicity on myocardial tissue and lower cytotoxicity on cancer cells, which acquire tolerance towards its effects. Overexpression of enzymes catalyzing the conversion of effective antineoplastic DAUN causes the gradual development of resistance to administered ANT. For this reason, the new pharmacotherapeutic options for preventing this process are being searched for to maintain the drug in the target cell as long as possible. One option is to use inhibitors, which lower enzymatic activity CRE and prevent unfavorable DAUN metabolism on cardiotoxic and ineffective DAUN-ol. This diploma thesis aimed to study the...
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