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Biotranformation of fenbendazole in soya (Glycine max)
Martínková, Lenka ; Raisová Stuchlíková, Lucie (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Martínková Supervisor: RNDr. Lucie Raisová Stuchlíková, Ph.D. Title of diploma thesis: Biotransformation of fenbedazole in soya (Glycine max) Veterinary drugs are used in large amounts in modern husbandry for treatment and prevention of diseases in animals. Anthelmintics administered to animals enter into environment primarily through its excretion in faeces or urine. Following excretion, drugs may persist in the environment and impact non-target organisms. Plants are able to uptake xenobiotics, including drugs, and detoxify them via biotransformation. However, only drug biotransformation into non-toxic and stable metabolites and their consequent accumulation in plants represent drug detoxification. For that reasons, knowledge of biotransformation pathways of drug in plants is very important. Soybean plants, eventually seeds, are further used in agriculture as feed for cattle and absorbed anthelmintics including their metabolites can enter the food chain. The results showed that fenbendazole entered plant and enzymatic systems of plant were able to biotransform fenbendazole via several reactions. We found differences in the metabolites between the roots, leaves, seeds and pods of soybean.
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Study of protein-protein interaction of DHRS7 enzyme by pull-down assay
Hermanová, Kateřina ; Novotná, Eva (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Kralove Department of Biochemical Sciences Candidate: Kateřina Káchová Supervisor: RNDr. Eva Novotná, Ph.D. Title of diploma thesis: Study of protein-protein interaction of DHRS7 enzyme by pull-down assay Dehydrogenase/reductase (SDR family) member 7 (DHRS7) is one of the less studied enzymes of SDR superfamily. It has been proven that this enzyme is in vitro involved in reductive metabolism of various compounds, such as steroids, retinoids and xenobiotics. Recently results pointing out to possible role of this enzyme in the pathogenesis of prostate cancer or other diseases has been published. It would be suitable to better characterize this enzyme to clarifying its patho/physiological role in the organism. Because protein-protein interactions seem to be important for the function of proteins, the aim of this study was to identify interaction partners of the DHRS7, and thus contribute to the improvement of understanding of this enzyme. For our experiments, pull-down assay, in vitro method was utilized. The first step was immobilization of DHRS7 enzyme (bait protein) to suitable carrier (His Mag Sepharose Ni particles and nonmagnetic Protino Ni-IDA particles). Subsequently, the carrier with immobilized DHRS7 was incubated with the lysate of Hep G2...
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IN VITRO assays for investigating nucleic acid delivery.
Mihaličoková, Dajana ; Jirkovská, Anna (advisor) ; Matoušková, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové, Department of Biochemical Sciences University of Vienna, Faculty center for Pharmacy, Department of Pharmaceutical Chemistry, Laboratory of MacroMolecular Cancer Therapeutics Candidate: Dajana Mihaličoková Supervisor (Charles University): PharmDr. Anna Jirkovská, Ph.D. Supervisor (University of Vienna): Univ.Prof. Dipl. Ing. Dr. Manfred Ogris Co-supervisor (University of Vienna): Dr. Haider Sami, Ph.D. Title of diploma thesis: In vitro assays for investigating nucleic acid assay Keywords: transfection, splice correction, BCA assay, polyplexes One of the most important tasks of biochemical research is to find out the right way how to cure cancer, genetic disorders and other illnesses which are still not curable. Towards this, gene therapy is emerging as a potential treatment owing to its ability to deliver genetic material inside the cell. Reporer gene based transfection process can be used to study gene expression. Transfection is mediated by vectors, either of viral or non-viral origin. Non-viral vectors offer several advantages over the viral counterparts like easier to synthesize, relatively cheap and the most important is their non-immunogenicity. Cationic polymers based on polyethylenimine form complexes with plasmid DNA reffered to as...
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Effect of doxorubicin on selected mice microRNAs
Dugasová, Lucia ; Matoušková, Petra (advisor) ; Jansová, Hana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lucia Dugasová Supervisor: doc. Ing. Petra Matošková Ph.D. Title of diploma thesis: Effect of doxorubicin on selected mice microRNAs Doxorubicin (DOX) belongs to the anthracycline antibiotics used by patients suffering from cancer. Its undesirable effect is cardiotoxicity. Prognostic of cardiotoxicity depends on its early detection, and therefore many studies examine the properties of microRNAs that could be appropriate biomarkers for heart failure in the future. MicroRNAs are involved in almost all processes in the human body, in development, differentiation, proliferation and apoptosis. They are also located in the extracellular space, they are stable and have many other positive properties. My diploma thesis is focused on monitoring changes in microRNA levels that can occur in heart damage by doxorubicin. The cardiac tissues and plasma of mice treated with doxorubicin were used as samples and compared with control samples. Samples were processed and evaluated by real-time quantitative PCR (qPCR). A statistically significant increase in expression in heart tissue occurred only in miR-34a and decrease in miR-205 level. Plasma mice samples showed a statistically significant increase in...
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Effect of imatinib on selected mice microRNAs
Heglasová, Silvia ; Matoušková, Petra (advisor) ; Skarková, Veronika (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Silvia Heglasová Supervisor: Doc. Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of imatinib on selected mice microRNAs Imatinib (IMB) belongs to low molecular weight tyrosine kinase inhibitors (TKI). One of the undesirable effects of IMB is cardiotoxicity. Early diagnosis of heart damage is important for improving human health. MicroRNAs (miRNAs) are intensively studied as potential early biomarkers of cardiac damage. They are not only intracellular but also circulating freely in the bloodstream. Plasma miRNAs are stable, resistant to repeated freezing and methods of their detection are very sensitive. In my diploma thesis I have studied changes in the expression of miRNAs in plasma in the IMB induced heart damage. I have analyzed circulating plasma miRNAs and also miRNA levels in mice hearts. Selected miRNAs were measured by quantitative real-time PCR (qPCR). Plasma Troponin T levels were also measured as classical biomarker of cardiotoxicity. Troponin T levels in the IMB-treated plasma were very variable. This fact points to different responses of individuals to IMB. Changes in expression of observed miRNA in plasma affected by IMB were not significant. Statistically...
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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 impact of selected prenylflavonoids on the effect of anticancer therapy in cancer cell lines
Pešková, Klára ; Matoušková, Petra (advisor) ; Skálová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Klára Pešková Supervisor: Ing. Petra Matoušková, Ph.D. Title of diploma thesis: The impact of selected prenylflavonoids on the effect of anticancer therapy in cancer cell lines Prenylflavonoids 6-prenylnaringenin and 8-prenylnaringenin and flavonoid naringenin are substances with anti-proliferative properties. Prenylflavonoids are present mostly in hop and in beer. Main source of naringenin are primarily citrus fruits. In this diploma thesis the effect of compounds was tested by neutral red uptake test in cell lines SW480 and SW620. These substances were tested individually and then in combination with cytostatic agent oxaliplatin. Oxaliplatin is used in treatment of cancer disease. 6-prenylnaringenin was the most effective compound in both cell lines. Statistical evaluation of the effect was carried out by Graphpad using the method One-way ANOVA in comparison with control. Combination index was determined by software program Compusyn using the method by Chou-Talalay.
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Synthesis of epitestosterone and 4-hydroxyestrone glucuronides using recombinant human UDP-glucuronosyltransferases.
Kašparová, Michaela ; Skálová, Lenka (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Michaela Kašparová Supervisors: Prof. RNDr. Lenka Skálová, Ph.D., Erkka Järvinen, M.Sc. (Pharm.) Title of diploma thesis: Synthesis of epitestosterone and 4-hydroxyestrone glucuronides using recombinant human UDP-glucuronosyltransferases Steroid hormones constitute an important part of the human endocrine system and are involved in a variety of physiological and pathological processes. Phase I and phase II biotransformation reactions convert these lipophilic, biologically active compounds to inactive, water-soluble metabolites that are readily excretable into bile or urine. One of the most common phase II biotransformation reactions is conjugation with glucuronic acid that is enabled by the catalytic activity of UDP-glucuronosyltransferases (UGTs). This study is focused on development and optimization of an enzymatic method for producing conjugates of glucuronic acid and two naturally occurring steroid hormones, epitestosterone and 4-hydroxyestrone. Recombinant human UGT2B7 was employed as a catalyst for the production of epitestosterone 17-glucuronide and 4-hydroxyestrone 4- glucuronide while synthesis of 4-hydroxyestrone 3-glucuronide was accomplished by UGT1A10 isoform. The synthesis reactions...
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