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Transport and biotransformation of anthelmintics in susceptible and resistant strains of Barber's pole worm (Haemonchus contortus)
Prchal, Lukáš ; Szotáková, Barbora (advisor) ; Lamka, Jiří (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lukáš Prchal Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Transport and biotransformation of anthelmintics in susceptible and resistant strains of Barber's pole worm (Haemonchus Contortus) Haemonchosis is a worldwide problem, which endanger breeding of small ruminants. This disease is caused by nematode Barber's pole worm (Haemonchus contortus), which parasites inside stomachs of host animals, causing anaemia and in many cases death. Pharmacotherapy is the most used way to prevent and/or threat animals. The use of benzimidazole anthemintics is very common, but it's failing more and more often because of resistance of parasites to pharmacotherapy. The most common reason of resistance is mutation in the gene for β-tubuline, which is target for benzimidazoles. This research focuses at the other possible mechanism of resistance, which is resistance based on enzymes and metabolism. One of the aims of this thesis was to compare the metabolism of the anthelmintic drug flubendazole in three different strains of H. contortus: the ISE strain (susceptible to common anthelmintics), the BR strain (resistant to benzimidazole anthelmintics) and the WR strain (resistant to all...
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Effect of Synthetic Magnolol Derivatives on Activity of Nuclear Receptors PPARγ and RXRα
Dymáková, Andrea ; Szotáková, Barbora (advisor) ; Trejtnar, František (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences University of Vienna Faculty of Life Sciences Department of Pharmacognosy Candidate: Andrea Dymáková Supervisors: Mag. Simone Latkolik, doc. Ing. Barbora Szotáková, Ph. D. Title of diploma thesis: Effect of Synthetic Magnolol Derivatives on Activity of Nuclear Receptors PPARγ and RXRα The nuclear receptors, peroxisome proliferator-activated receptor γ (PPARγ) and its heterodimerization partner retinoid X receptor α (RXRα) are drug targets in the treatment of diseases like the metabolic syndrome and diabetes mellitus type 2. The effort has been made to develop new agonists for PPARγ to obtain ligands with more favourable properties than currently used drugs (Berger et al. 2002, Berger et al. 2005). Magnolol was previously described as a dual agonist of PPARγ and RXRα (Fakhrudin et al. 2010, Zhang et al. 2011). Based on the bi- aryl structure of magnolol, the effort has been made to design and synthesize linked magnolol dimers. The aim of this thesis was to investigate the agonistic potential of these compounds with respect of the nuclear receptors PPARγ and RXRα in comparison to magnolol. We evaluated the ligand binding properties of the compounds and their functionality as PPARγ agonists in vitro...
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Influence of albendazole on cell proliferation in vitro
Knoppová, Kateřina ; Szotáková, Barbora (advisor) ; Jirkovská, Anna (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Kateřina Knoppová Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Supervisor specialist: RNDr. Věra Králová, Ph.D. Title of diploma thesis: Influence of albendazole on cell proliferation in vitro Albendazole (ABZ) is a benzimidazole anthelmintic widely used for the treatment of parasitic diseases in man and animals. The mechanism of action is based on inhibition of microtubule polymerization by binding of ABZ to the β-subunit of tubulin in parasites. As a result, it subsequently leads to the disruption of vital functions of the parasite and to its death. After elucidation of this mechanism, ABZ was tested as a potential anticancer agent. It has been shown that it is able to inhibit proliferation of some cancer cell lines. In this study, we examined the effect of ABZ on cell proliferation in vitro. For the purpose of our experiment we used cell lines derived from colorectal carcinoma - SW 480 and SW 620 together with a cell line NCM 460 isolated from normal colonic mucosa. The cells were treated with ABZ in the concentration range of 0.01 - 10 µM for 24, 48 and 72 hours. Cell viability was determined using WST-1 assay and Neutral Red Uptake Test (NRU). ABZ inhibited cell proliferation...
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Effect of helminthosis on host's biotransformation enzymes
Bártíková, Hana ; Szotáková, Barbora (advisor) ; Lamka, Jiří (referee)
(EN) Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Hana Bártíková Consultant: Doc. Ing. Barbora Szotáková, Ph.D. Title: The effect of parasitosis on the host's biotransformation enzymes Parasitic diseases cause significant economic losses in livestock, domestic and wild animals. World-wide spread species Haemonchus contortus parasiting in gastrointestinal tract of ruminants exhibits relatively high degree of pathogenity. To treat haemonchosis the benzimidazole anthelmintics are used. Altered activity of the enzymes participating in biotransformation of these drugs can affect the treatment. The induction of host's enzymes can contribute to the resistance of the worms to the available anthelmintics. This project studies influence of haemonchosis to the activity of sheep's biotransformation enzymes. Oxidative, conjugation and reductive enzymes have been studied both in healthy and sick sheep. Cytosolic and microsomal fractions of liver and small intestine have been used for determination of enzymatic activities. In the most of monitored oxidative enzymes the inhibition of activity has been observed. On the contrary, parasitosis has caused enzymatic induction of conjugation enzymes. The results in group of reductive enzymes don't offer...
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Effect of Synthetic Magnolol Derivatives on Activity of Nuclear Receptors PPARγ and RXRα
Dymáková, Andrea ; Szotáková, Barbora (advisor) ; Trejtnar, František (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences University of Vienna Faculty of Life Sciences Department of Pharmacognosy Candidate: Andrea Dymáková Supervisors: Mag. Simone Latkolik, doc. Ing. Barbora Szotáková, Ph. D. Title of diploma thesis: Effect of Synthetic Magnolol Derivatives on Activity of Nuclear Receptors PPARγ and RXRα The nuclear receptors, peroxisome proliferator-activated receptor γ (PPARγ) and its heterodimerization partner retinoid X receptor α (RXRα) are drug targets in the treatment of diseases like the metabolic syndrome and diabetes mellitus type 2. The effort has been made to develop new agonists for PPARγ to obtain ligands with more favourable properties than currently used drugs (Berger et al. 2002, Berger et al. 2005). Magnolol was previously described as a dual agonist of PPARγ and RXRα (Fakhrudin et al. 2010, Zhang et al. 2011). Based on the bi- aryl structure of magnolol, the effort has been made to design and synthesize linked magnolol dimers. The aim of this thesis was to investigate the agonistic potential of these compounds with respect of the nuclear receptors PPARγ and RXRα in comparison to magnolol. We evaluated the ligand binding properties of the compounds and their functionality as PPARγ agonists in vitro...
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Comparison of anthelmintics metabolism and activities of biotransformation enzymes in susceptible and resistant strains of Haemonchus contortus
Lasotová, Tamara ; Szotáková, Barbora (advisor) ; Vokřál, Ivan (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Tamara Lasotová Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Comparison of anthelmintics metabolism and activities of biotransformation enzymes in susceptible and resistant strains of Haemonchus contortus Haemonchus contortus is a predominant gastrointestinal parasite of sheep and goat. This blood-sucking parasite cause severe anaemia with mortalities, particularly in young animals. The greatest problem in pharmacotherapy of haemonchosis is a resistance to anthelmintics, which is worldwide. Biotransformation enzymes have the ability to inactivate anthelmintics via biotransformation and it contributes to development of helminths resistance. The aim of this project was to compare anthelmintics metabolism and activities of biotransformation enzymes in susceptible and resistant strains of H. contortus. For a testing three strains of H. contortus were chosen: anthelmintics-susceptible strain - ISE strain, benzimidazole-resistant strain - IRE strain, anthelmintics-resistant strain - WR strain. The results showed that the activities of reductases of carbonyl group of the model substrates are higher in resistant strains than in susceptible strain, on the contrary to...
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Effect of storage of biological material on the activity of biotransformation enzymes
Jelínková, Jana ; Szotáková, Barbora (advisor) ; Kvasničková, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Jana Jelínková Consultant: Doc. Ing. Barbora Szotáková, Ph.D. Title: Effect of storage of biological material on activity of biotransformation enzymes Effect of storage of liver and subcellullar fractions on the activities of biotransformation enzymes was studied. Activities of oxidation enzymes - cytochromes P450 (CYP1A, 2B, 2C, 3A) and flavin-containg monooxygenases (FMO), carbonyl-reducing enzymes (AKR1A1, AKR1C, 11β-HSD,CBR) and conjugation enzymes (UGT, GST) were observed. Microsomal and cytosolic subcellular fractions prepared from differently stored liver of sheep (Ovis aries) were used. (A) from fresh tissue frozen in the liquid nitrogen. (B) from liver that was stored at -80C, the temperature gradually increased to 0C and then gradually decreased back to -80C. The half of prepared microsomal and cytosolic subcellular fractions from liver stored in liquid nitrogen (A) was processed by the same storage procedure as liver (B). The second half was stored at -80C. Activities of tested biotransformation enzymes were determined toward their relatively specific substrates. The subcellular fractions were also incubated with xenobiotics (oracin, flubendazole and albendazole) and the...
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Effect of inositol hexaphosphate on adhesion and migration of cell lines SW480 and SW620
Peterková, Alena ; Szotáková, Barbora (advisor) ; Skálová, Lenka (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Alena Ježková Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Supervisor specialist: RNDr. Veronika Hanušová, Ph.D. Title of diploma thesis: Effect of inositol hexaphosphate on adhesion and migration of cell lines SW480 and SW620 Inositol hexaphosphate (IP6) is molecule, which is usually present in nature, especially in grain and legumes. It has been described several processes, which are influenced in human body by IP6, such as decrease of platelet aggregation, promoting effect on insulin secretion, hypocholesterolemic, as well as anticancer effect. Aim of this work was to study adhesion and migration properties, which are the most important processes in metastasis of cancer cells. For experiments, colorectal adenocarcinoma cell line SW480 and metastatic lymph node cancer cell line SW620, both isolated from identical patient, have been selected. The ability of migration in both cell lines was tested. The influence of IP6 in different concentrations and several time intervals on mRNA (RT-PCR) and protein level (western blot, immunofluorescence) expressions of adhesion and invasion molecules has been compared as well. Considerable ability of migration was found in metastatic cell line...
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