|
|
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...
|
|
|
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...
|
|
|
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...
|
|
|
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...
|
|
|
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...
|
|
|
Effect of prenylated flavonoids on biotransformation enzymes in vitro
Pataráková, Paula ; Szotáková, Barbora (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Paula Pataráková Supervisor: Prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Effect of prenylated flavonoids on biotransformation enzymes in vitro Prenylated flavonoids xanthohumol (XH), isoxanthohumol (IXH), 6-prenylnaringenin (6PN) and 8-prenylnaringenin (8PN) are flavanones and chalcones occurring in hops and their characteristic feature is prenyl chain on A ring. As these substances, present in beer and dietary supplements, are foreign compounds for organism, they can affect drug- metabolizing enzymes, including carbonyl reducing enzymes. The aim of our study was to find out, whether XH, IXH, 6PN, and 8PN have any impact on cytosolic carbonyl reductase 1 (CBR1) and aldo-keto reductase 1C subfamily (AKR1C). Viability test revealed that prenylated flavonoids in lower concentrations do not affect or even increase the viability of primary rat hepatocytes, but higher concentrations are toxic. AKR1C activity was increased in 8PN treated hepatocytes, the activity of CBR1 was significantly increased in IXH and 8PN treated samples. An increase in expression of AKR1C3 in IXH and 6PN treated samples, and decrease in XH and 8PN samples was observed. Expression of CBR1 was at the detection...
|
|
|
The activity of biotransformation enzymes in mouflons with dicrocoeliosis and in healthy mouflons
Urbánková, Marie ; Szotáková, Barbora (advisor) ; Gavelová, Martina (referee)
Charles University Faculty of Pharmacy Department of Biochemical Sciences Hradec Králové, Czech Republic Field: Patobiochemistry and xenobiochemistry Candidate: Mgr. Marie Urbánková Consultant: Ing. Barbora Szotáková, Ph.D. Title: The activity of biotransformation enzymes in mouflons with dicrocoeliosis and in healthy mouflons Abstract In vitro activities of cytochromes P450 (7-alkyl/aryloxyresorufin dealkyl(aryl)ases, testosterone hydroxylase/oxidase, 7-methoxy-4-trifluoromethyl-coumarin demethylase), flavine monooxygenases (toward thiobenzamide), reductases of carbonyl group (toward oracin, 4-pyridine-carboxaldehyde, 1-acenaphthenol, DL-glyceraldehyde, ketoprofene, naloxone, daunorubicin, metyrapone) and cojnugation enzymes (p-nitrophenol-UDP- glukuronosyl transferase, UGT, 1-chloro-2,4-dinitrobenzene glutathione-S-transferase, GST) in old, female mouflons (Ovis musimon, Bovidae), both in healthy ones and those suffered from dicrocoeliosis were studied and compared. This parasitary disease is caused by Dicrocoelium dendriticum. Various methods were used, especially spectrophotometry, spectrofluorimetry, and method based on incubation of enzymes with relative specific substrate followed with HPLC analysis of biotransformation products. The amount of proteins was determined using methods with reduction of...
|
|
|
Effect of Nanoparticles on Plant Metabolism
Cyrusová, Tereza ; Szotáková, Barbora (advisor) ; Hnilička, František (referee) ; Smrček, Stanislav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Tereza CYRUSOVÁ Supervisor: prof. Ing. Barbora SZOTÁKOVÁ, Ph.D. Consultant: RNDr. Radka PODLIPNÁ, Ph.D. Title of Doctoral Thesis: Effect of Nanoparticles on Plant Metabolism Along with the expansion of nanotechnologies, which are used in many disciplines, the field of nanotoxicology has recently been developed. It deals with the negative effects of engineered nanoparticles on the environment and biota. Considerable attention has already been paid to the toxicity of nanoparticles to plants, however, the mechanisms responsible for their effects are not fully understood yet. At the same time, the possible positive effect of nanoparticles on plants has been investigated in recent years. The aim of our work was to study and evaluate the effect of nanoparticles on the growth of selected plant species and to evaluate their phytotoxicity. We focused on the study of nanoparticles in the form of metal oxides. The content of accumulated metals in germinating plants of mustard (Sinapis alba L.), in in vitro cultures of true-fox sedge (Carex vulpina L.) and the concentration of metal ions released into the culture media were measured by atomic absorption spectrometry (Cu, Fe, Mn, Zn). and inductively...
|
|
|
The use of xCELLigence in the study of irinotecan effects on the colorectal carcinoma cells in vitro
Vatahová, Lenka ; Szotáková, Barbora (advisor) ; Hašková, Pavlína (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Vatahová Supervisor: RNDr. Věra Králová, Ph.D. Title of diploma thesis: The use of xCELLigence in the study of effects of irinotecan in colorectal cancer cells Irinotecan is one of the cytostatic drugs, administered either alone or in combination with 5- fluorouracil (5-FU). Its mechanism of action lies in blocking topoisomerase I resulting in a single-threaded irreparable disruption, which is lethal for the cell. In our study, we examined the use of xCELLigence in the study of effects of irinotecan in colorectal cancer cells. xCELigence, which allows tracking of cells in real time was compared with 2 other methods for determination cell viability, specifically with the WST-1 assay and the Brilliant Blue (BB) assay. Experiments were performed on three cell lines derived from colorectal carcinoma- SW 620, SW 480, and HT 29. The cells were treated with irinotecan in concentration ranges from 5 to 100 µg/ml for 24, 48 and 72 hours. We compared the sensitivity of individual methods and found, that the values differed according to the cell lines. In all cells we calculated the IC 50 value at the time of 72 hours of treatment with irinotecan. And while we have shown that the...
|
|
|
Modulation of biotransformation and antioxidant enzymes by selected natural compounds
Lněničková, Kateřina ; Szotáková, Barbora (advisor) ; Anzenbacherová, Eva (referee) ; Trejtnar, František (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Kateřina LNĚNIČKOVÁ Supervisor: prof. Ing. Barbora SZOTÁKOVÁ, Ph.D. Title of Doctoral Thesis: MODULATION OF BIOTRANSFORMATION AND ANTIOXIDANT ENZYMES BY SELECTED NATURAL COMPOUNDS Public interest in various dietary supplements containing herbs, herbal extracts or isolated active compounds has increased significantly over past decades. Consumption of these supplements increases worldwide and they are often consumed in unreasonably high doses, as they are generally considered as safe. Upon the intake to organism, these compounds are, as other xenobiotics, modified mostly by xenobiotic-metabolizing enzymes and they could influence these enzymes at the same time. Potential modulation of xenobiotic-metabolizing enzymes' activity (induction or inhibition) can seriously affect pharmacokinetics of concomitantly administered drugs. Knowledge of the possible impact of natural compounds on the xenobiotic-metabolizing enzymes is essential for their safe use. The aim of this doctoral thesis was to study the effects of selected herbal extracts and their active chemical constituents on the activity and expression of xenobiotic-metabolizing and antioxidant enzymes. We have focused on the study of effects of...
|
guest ::
