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Study of expression of biotransformation enzymes and transport proteins in kidneys
Felešová, Martina ; Smutná, Lucie (advisor) ; Kuchařová, Monika (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Martina Felešová Supervisor: PharmDr. Lucie Smutná, PhD. Title of diploma thesis: Study of expression of biotransformation enzymes and transport proteins in kidneys Xenobiotics are a part of the everyday life of living organisms. Their source is, among other things, diet, environment, administered medicines, food colorings and others. The living organisms have developed complex metabolic processes that eliminate these substances from the body in order to protect themselves against their negative influence. Within this work, we specifically focused on the effect of the murine Car agonist - TCPOBOP on influencing the expression of biotransformation and transport genes. The essence was the isolation of mRNA from the murine kidneys using the classic phenol-chloroform method, followed by the transcription of mRNA into cDNA. In the last step, the RT-PCR reaction was performed. The results are graphs showing how the administration of TCPOBOP affected the expression of the monitored genes. It is clear from these graphs that TCPOBOP has some effect on the expression of target genes. In the case of Cyp3a11, Cyp2b10, Slc22a2 and Ugt3a2, there was a several-fold increase in expression after...
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In vitro cytotoxicity study of selected antimycobacterial substances on human hepatocytes
Klimová, Miroslava ; Kuchařová, Monika (advisor) ; Smutná, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Student: Miroslava Klimová Supervisor: Mgr. Monika Kuchařová, Ph.D. Consultant: Mgr. Pavel Bárta, Ph.D. Title of diploma thesis: In vitro cytotoxicity study of selected antimycobacterial substances on human hepatocytes Clinical research is constantly progressing, new, missing drugs are being searched for, more effective substances are being synthesized, adverse effects are being minimized. The aim of this thesis was to determine the effect of ten substances (JZ195, JZ186, MH17, JZ178, MH9, MH18, JZ177, JZ170, JZ172, JZ174) in the HepG2 cell line and evaluate their cytotoxic potential. The tested substances were derived from the structure of pyrazine-2- carboxamide and synthesized within the Department of Organic and Bioorganic Chemistry at the Faculty of Pharmacy in Hradec Králové, Charles University and developed for the purpose of having promising effect on mycobacterial strains. Cytotoxicity was evaluated in vitro by CellTiter96® AQueous One Solution Cell Proliferation Assay, commercially available colorimetric method based on reduction of the tetrazolium salt to a violet-coloured formazan which directly reflects the number of viable cells. We assessed the hepatotoxic effect using the IC50...
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Miniaturized and fast method for solubility and level of supersaturation determinations of drug nanocrystals
Paľová, Romana ; Nováková, Veronika (advisor) ; Kuchařová, Monika (referee)
Title of thesis: Miniaturized and fast method for solubility and level of supersaturation determinations of drug nanocrystals Author: Romana Paľová Department: Biophysics and Physical Chemistry Supervisor: Doc. Veronika Nováková, Ph.D. Specialized supervisor: Assoc. Prof. Leena Peltonen, Ph.D. As a formulation strategy for improving the bioavailability of poorly water-soluble drugs, drug nanocrystals can be employed. Several studies have shown that drug nanocrystals exhibit higher dissolution rate compared to conventional drug formulations. However, when orally administered, various conditions in gastrointestinal tract, such as changes in pH may result in problematic absorption. In this work, the preparation of drug nanosuspension of itraconazole (ITZ), as model poorly water-soluble compound, was carried out. The nanosuspension was prepared by using wet milling method of preparation, with suitable stabilizers added (Poloxamer 407 and Hydroxypropyl methylcellulose). Hydrochloric acid buffer pH 1.2 was chosen to create an environment similar to gastric fluids. In order to investigate changes in solubility of ITZ nanosuspension, UV spectrophotometer was used. Measurement was performed at the wavelength 551 nm, where dissolved ITZ does not absorb light and no change in absorbance occurred....
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Determination of fatty acids in human tissues
Turňová, Ivana ; Kuchařová, Monika (advisor) ; Nováková, Veronika (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biophysics and Physical Chemistry Candidate: Ivana Turňová Supervisor of Diploma Thesis: Mgr. Monika Kuchařová, Ph.D. Consultant: prof. MUDr. Zdeněk Zadák, CSc. Title of Diploma Thesis: Determination of fatty acids in human tissues The fatty acids are non-negligible component of lipids as one of the basic nutrients. This thesis in its theoretical part presents above all the group of polyunsaturated fatty acids (PUFA), which are important structural units of the cell membranes, they are also the precursors to several significant biologically active substances. In the human organism PUFA participate in many physiological and pathological processes this way, where they cause the large spectrum of actions. Onwards the thesis describes the gas chromatography method that is used in chemical analysis of lipids for the fatty acid determination as the gold standard. Experimentally, the fatty acid representation was determined in the blood, muscle, heart, liver and kidney among 26 cadavers divided into two groups according to the presence/absence of inflammatory process at the time of death. The data obtained were statistically analysed. The results were descriptively evaluated and there were indicated possible explanations of differences...
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