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Modulation of expression and activity of selected plant detoxifying enzymes by anthelmintics
Graňáková, Patrícia ; Szotáková, Barbora (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Patrícia Graňáková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Consultant: RNDr. Radka Podlipná, Ph.D. Title of diploma thesis: Modulation of expression and activity of selected plant detoxifying enzymes by anthelmintic Anthelmintics represent risk to environment as they may impact non-target organisms including plants, which come into contact with these pharmaceuticals in fields by fertilization with dung from treated animals or in pastures by excrements of treated animals. After uptake, these substances can increase the production of reactive oxygen species in plants, with the risk of oxidative stress and plant damage, and also affect the antioxidant enzymes. The aim of this work was to investigate the effect of two widely used anthelmintics ivermectin and fenbendazole on the activity and expression of selected antioxidant enzymes in soybean (Glycine max). Soybean was cultivated in a greenhouse and watered with a 10 µM solution of the selected drug. The changes of activity and gene expression of antioxidant enzymes were measured in root, leaf, pod and seed samples. Results showed that both anthelmintics caused significant decrease of superoxide dismutase, ascorbate peroxidase, glutathione...
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Environmental risks of anthelmintics
Dojčanská, Nikoleta ; Szotáková, Barbora (advisor) ; Karlíčková, Jana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Nikoleta Dojčanská Supervisor: prof. Ing. Barbora Szotáková, Ph.D Title of diploma thesis: Environmental risks of anthelmintics Anthelmintics are drugs used in human and veterinary medicine for the treatment and prophylaxis of infectious diseases caused by parasites- helminths. They are admittedly beneficial to the treated organisms. However, their overall effect on the ecosystem has not yet been sufficiently explored. Anthelmintics, together with their metabolites, enter the environment in multiple ways. One possibility is directly through urine and excrements of treated livestock or indirectly by leakage of contaminated manure into surface and groundwater. Such pollution has an effect on various animal organisms and plants that have the ability to uptake xenobiotics and remove them through metabolic pathways. The aim of this work was to investigate the effect of antihelmintics ivermectin and fenbendazole on soybean plants (Glycine max). We investigated the concentration of total phenolic compounds spectrophotometrically using the Folin-Ciocalteu method. Using the colorimetric method of aluminum chloride, we found the concentration of flavonoids in soybean plants. The concentration of isoflavons-...
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Toxicity of pulegone and menthofuran and their effect on miRNA expression in human liver slices
Lancošová, Bibiána ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bibiána Lancošová Supervisor: doc. PharmDr. Boušová Iva, Ph.D Title of diploma thesis: Toxicity of pulegone and menthofuran and their effect on miRNA expression in human liver slices Monoterpenes are secondary plant metabolites, characterised by their odour, which have a positive impact on human body and are used not only in cosmetic and food industry, but also in medication. R-pulegone (PUL) and its metabolite R-menthofuran (MF) are the most represented in plants of genus Mentha (Mint), with higher doses of these substances exerting certain risk of hepatotoxicity. The aim of this thesis was to assess the concentration, at which the viability of liver cells decreases and also to observe the changes in individual microRNA expression as a result of influence of PUL, MF and paracetamol, as the reference compound. Five samples of human liver obtained from patients of both genders and at age of 57 to 73 years represented biological material for hepatotoxicity testing. The samples were stored in Euro-Collins solution and subsequently cut into precision-cut liver slices with thickness of 150- 170 µm, placed to 12-well plate and put into incubation medium with the tested substances for 24 hours. The...
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Effect of albendazol on the expression of selected enzymes in Haemonchus contortus
Božíková, Martina ; Matoušková, Petra (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Martina Božíková Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of albendazol on the expression of selected enzymes in Haemonchus contortus Haemonchus contortus is the dominant gastrointestinal pathogen of small ruminants that can cause weakening or even death of the host organism. Losses are mainly recorded in young animals, which subsequently reflects on the farm economy itself. Therefore, prevention and therapy still play the most important role. On the other hand, the increased ability of worms to survive this treatment poses a serious problem. One of the possibilities that might be causing this problem is the increased activity of enzymes responsible for the metabolism of xenobiotics. The aim of this master's thesis was to investigate the effect of albendazole (ABZ) on the gene expression of selected biotransformation enzymes; UDP-glycosyltransferases (UGTs). For this purpose, two strains of H. contortus - anthelmintics-susceptible ISE (Inbred Susceptible Edinburgh) and anthelmintics-resistant WR (White River) were selected. One group of the parasites was cultivated for 12 hours in a media containing albendazole, the other group, which served as control, was...
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The effect of natural compounds on transport by OATP drug transporters
Zemčíková, Lucie ; Trejtnar, František (advisor) ; Mičuda, Stanislav (referee) ; Szotáková, Barbora (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: MSc. Lucie Zemčíková Supervisor: Assoc. Prof. PharmDr. František Trejtnar, CSc. Title of doctoral thesis: The effect of natural compounds on transport by OATP drug transporters OATP membrane transporters belong to carrier proteins responsible for transporting certain drugs (e.g. hypolipidemics from the group of statins) and other xenobiotics across the biological membranes and tissue barriers within the body. These transport proteins play an important role in pharmacokinetic processes such as absorption, distribution and elimination. The potential modulation of their transport function by natural compounds commonly present in plant food or food supplements may result in the changes of the concentration of their substrate (drug) in the cells and body fluids, that may affect the effect and toxicity of these drugs. The aim of this study was to obtain data on the interactions of selected natural compounds with human transporters OATP2B1 and OATP1A2 and their ability to affect drug transport mediated by these transporters. These two OATP transporters are involved in the drug uptake especially in organ barriers important for pharmacokinetics. For the study natural compounds from the group of...
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Effect of albendazole on the activity of selected enzymes in tapeworm Hymenolepis diminuta
Krejzová, Andrea ; Vokřál, Ivan (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Andrea Krejzová Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: Effect of albendazole on the activity of selected enzymes in tapeworm Hymenolepis diminuta The efficacy of anthelmintics used to treat diseases caused by helminths is not always sufficient, and in some cases, we are directly facing resistance to these drugs. Helminths, including tapeworms, are able to defend against the toxic effect of anthelmintics using several mechanisms. Xenobiotic metabolizing enzymes and transport proteins belong to these mechanisms. When xenobiotic metabolizing enzymes are induced, the efficacy of therapy may be significantly reduced. The effect of xenobiotic metabolizing enzymes on the drug resistance development has been already described in number of helminths. In tapeworms this information is still missing. Main aim of this study was to determine effect of drug albendazole on the activity of selected xenobiotic metabolizing enzymes in rat tapeworm (Hymenolepis diminuta). Tapeworms were incubated with albendazole (1 μM and 10 μM) for 24 hours. Then activities of selected enzymes in cytosol-like, microsome-like and mitochondria-like fractions were determined. This study is focused on...
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Detection of epithelial-mesenchymal transition markers (EMT) in cells in vitro
Špaček, Petr ; Szotáková, Barbora (advisor) ; Trejtnar, František (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Petr Špaček Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Consultant: RNDr. Věra Králová, Ph.D. Title of diploma thesis: Detection of epithelial-mesenchymal transition (EMT) markers in cells in vitro Epithelial-mesenchymal transition (EMT) is a process during which motile mesenchymal-like cells develop from non-motile parent epithelial cells. Physiologically, EMT plays important roles during embryonic development and wound healing. Loss of control over this mechanism can lead to fibrosis and cancer progression. Motile mesenchymal-like cells can pass through the basal lamina, get into the blood vessels and spread to distant tissues. Transition is regulated by EMT biomarkers. The biomarkers comprise wide spectrum of proteins, including cell surface proteins (E-cadherin, N-cadherin), cytoskeletal proteins (vimentin), microRNA (miR 200) and transcription factors (Snail, Twist). In this study, expression of EMT biomarkers was evaluated using RT-PCR and Western blotting. The ability to migrate was assessed using real-time analysis with the x-CELLigence system. Two known triggers of EMT, the StemXVivo™ EMT Inducing Media Supplement (IS) and TGF-β, were compared in human oral cancer cell lines DOK and...
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Effect of parasite on activity of biotransformation and antioxidant enzymes in host intestine
Mládková, Zuzana ; Szotáková, Barbora (advisor) ; Skálová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Zuzana Mládková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Effect of parasite on activity of biotransformation and antioxidant enzymes in host intestine Hymenolepis diminuta, known as a rat tapeworm, is commonly used in science as a model of Cestoda for studying physiology, biochemistry and drug metabolism in tapeworms. Recently, Hymenolepis diminuta has been studied for helminth-based therapy for a potential treatment of inflammatory bowel disease. The aim of this study was to determine how H. diminuta influences the activity of detoxification enzymes of the host intestine. At first 6 male rats (Wistar breed) were infected by cysticercoids of Hymenolepis diminuta previously isolated from the beetles Tenebrio molitor (intermediate host). At the same time the physiological saline solution was administered to the control group of 6 male rats. All rats were housed for 2 months in animal quarters with 12 h light/dark cycle. Thereafter, the tapeworms were removed from their intestines. Intestinal mucosa containing metabolic active enzymes was isolated. Subsequently, the subcelullar fractions were prepared and used for in vitro experiment. The activity of enzymes was...
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Influence of glutamate-induced obesity on detoxification enzymes contains glutathione
Duchačová, Kateřina ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Kateřina Duchačová Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of rigorous thesis: Influence of glutamate-induced obesity on detoxification enzymes contains glutathione Obesity and related comorbidities occur more frequently in the population and belong to the most studied contemporary health topics. Aging of the organism is much discussed topic as well. Various animal models are used to observe changes in the organism at the organ, tissue, cellular and molecular levels. Comparison of specific activities and protein expressions of glutathione peroxidase (GPx) and glutathione reductase (GR), and specific activity of glutathione-S-transferase (GST) in the liver of obese mice and healthy controls according to the pathological state, aging and sex was the aim of this study. Mice with monosodium glutamate (MSG)-induced obesity, osmolal controls (NaCl) and mice without treatment of both sexes and at 8 and 20 weeks of age were used. Specific enzyme activities were determined spectrophotometrically. Protein expressions were determined using polyacrylamide gel electrophoresis followed by immunoblotting. The biggest obesity-related changes in enzyme activity were observed in GST, which activity...
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Liver slices - model system for studying the effect of sesquiterpenes on detoxifying enzymes
Slavíčková, Zlata ; Szotáková, Barbora (advisor) ; Ambrož, Martin (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Zlata Slavíčková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Liver slices - model system for studying the effect of sesquiterpenes on detoxifying enzymes Sesquiterpenes are naturally occurring substances, mainly in higher plants as secondary metabolites. Together with monoterpenes, represent the main components of plant essential oils. Sesquiterpenes are biologically active compounds, among their biological activities belong anti-inflammatory, anti-carcinogenic, antioxidant, antibacterial and antimycotic effects. The aim of this diploma thesis was to find out the influence of three structurally similar acyclic sesquiterpenes cis-nerolidol, trans-nerolidol, and farnesol on the activity of selected biotransformation enzymes - aldo-keto reductase (AKR1A1, AKR1C9), carbonyl reductase 1 (CBR1), NAD(P)H-quinone oxidoreductase 1 (NQO1), cytochrome P450 (CYP1A1, CYP1A2, CYP2B, CYP3A), sulfotransferase (SULT), glutathione-S-transferase (GST), and UDP-glucuronosyltransferase (UGT) in rat liver. The influence of sesquiterpenes on selected enzymes was studied in precision-cut liver slices. The precision-cut tissue slices were obtained from rat liver (Rattus norvegicus, tribe...
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