|
|
Resistance mechanisms in therapy of acute myeloid leukemia
Suchá, Simona ; Čečková, Martina (advisor) ; Žák, Pavel (referee) ; Matoušková, Petra (referee)
IN ENGLISH LANGUAGE Candidate: Mgr. Simona Suchá Supervisor: Assoc. Prof. PharmDr. Martina Čečková, PhD. Title of the doctoral thesis: Resistance mechanisms in therapy of acute myeloid leukemia Acute myeloid leukemia (AML) is a hematologic cancer known for its extensive heterogeneity, poor treatment outcomes and high relapse rate. Therapy outcome is often compromised by highly resistant leukemic clones present at diagnosis, which evade chemotherapy and continue to spread the disease. Identification of their cellular features is, therefore, a key in successful targeting and eliminating of these resistant leukemic cells. AML cells can, however, develop drug resistance even overtime due to prolonged drug exposure. Extremely high adaptability of leukemic cells enables them to survive whenever therapeutic stress stimuli occur. Uncovering molecular mechanisms that cells utilize to activate their survival mode is crucial in selection of treatment leading to maximal efficacy. Based on these grounds, two main aims of this thesis were set. First, to determine clinical relevance of ABC efflux transporters in AML and to evaluate the effect of targeted agents on chemotherapy. The focus was put on agents belonging to either FLT3 inhibitors (midostaurin) or CDK4/6 inhibitors (abemaciclib, palbociclib,...
|
|
|
The effect of the tyrosine kinase inhibitor tirabrutinib on anthracycline resistance in a tumor cell line.
Lebeková, Nikola ; Wsól, Vladimír (advisor) ; Matoušková, Petra (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Nikola Lebeková Supervisor: prof. Ing. Vladimír Wsól, Ph.D. Title of diploma thesis: The effect of tyrosine kinase inhibitor tirabrutinib on anthracycline resistance in a cancer cell line Malignancies represent a broad spectrum of heterogeneous diseases caused by genetic and epigenetic alterations that negatively affect various human tissues, contributing to the development of malignancy. Chemotherapy is considered one of the main pillars in the treatment of malignant diseases. Anthracycline chemotherapeutics are widely used drugs in these indications. However, adverse effects resulting from their mechanism of action are a major obstacle. The role in the development of cardiotoxicity, as one of the adverse effects of therapy, is played by carbonyl reducing enzymes that produce toxic reduced metabolites by conferring metabolic reduction of anthracyclines. At the same time, these metabolites show significantly lower cytotoxic activity, thus contributing to the formation and further development of resistance, another severe obstacle in clinical practice. This thesis aimed to investigate the inhibitory effect of a specific tyrosine kinase inhibitor tirabrutinib at the cellular level. The...
|
|
| |
|
|
A role of microRNAs in the regulation of hepatic drug-metabolizing enzymes
Sedlmajer, Štěpán ; Smutný, Tomáš (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Štěpán Sedlmajer Supervisor: PharmDr. Tomáš Smutný, Ph.D. Title of diploma thesis: A role of microRNAs in the regulation of hepatic drug-metabolizing enzymes The liver is the main biotransformation organ in the body. Cytochromes P450 and conjugation enzymes are essential enzymes in the metabolism of drugs and other xenobiotics. Recently, there is growing evidence of post-transcriptional regulation of these enzymes through microRNA molecules. MicroRNAs are epigenetic regulators of expression, which usually bind to fully or partially complementary sequences within the 3'UTR (3' untranslated) regions of mRNA molecules. MicroRNAs have a role in interindividual variability of expression and activity of metabolic enzymes. Understanding their role and mechanisms of action may contribute to the development of new drugs and tailored pharmacotherapy. The work summarizes current selected achievements in the research on regulation of mainly hepatic cytochromes P450 and conjugation enzymes through microRNAs. The impact of RNA editing, single nucleotide polymorphisms and pseudogenes is also described.
|
|
|
The effect of zanubrutinib on tumor cell resistance to daunorubicin due to carbonyl reducing enzymes.
Havlíčková, Lucie ; Wsól, Vladimír (advisor) ; Matoušková, Petra (referee)
5 Abstract Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lucie Havlíčková Supervisor: prof. Ing. Vladimír Wsól, Ph.D. Title of diploma thesis: The effect of zanubrutinib on tumor cell resistance to daunorubicin due to carbonyl reducing enzymes Anthracycline antibiotics (ANT) are considered the first-line medication for oncologic diseases, including acute myeloid leukemia. Daunorubicin (DAUN) with a carbonyl group in position 13 is often used in chemotherapeutic treatments. Nicotinamidadenindinucleotid-phosphate dependent carbonyl reducing enzymes (CRE) create its hydroxyl metabolite daunorubicinol (DAUN-ol), which demonstrates higher toxicity on myocardial tissue and lower cytotoxicity on cancer cells, which acquire tolerance towards its effects. Overexpression of enzymes catalyzing the conversion of effective antineoplastic DAUN causes the gradual development of resistance to administered ANT. For this reason, the new pharmacotherapeutic options for preventing this process are being searched for to maintain the drug in the target cell as long as possible. One option is to use inhibitors, which lower enzymatic activity CRE and prevent unfavorable DAUN metabolism on cardiotoxic and ineffective DAUN-ol. This diploma thesis aimed to study the...
|
|
|
Antiproliferative and anthelmintic effects of fern extracts
Turoňová, Petra ; Matoušková, Petra (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Bichemical Sciences Candidate: Petra Turoňová Supervisor: doc. Ing. Petra Matoušková, PhD. Title of diploma thesis: Antiproliferative and anthelmintic effects of fern extracts Ferns belong to spore vascular plants. In this study, an antiproliferative and anthelmintic effects of selected fern extracts were investigated. The antiproliferative effect of the extracts was tested on the SW480 cell line. This cell line was originally obtained from primary adenocarcinoma of the colon. The anthelmintic effect was tested on L3 larvae Haemonchus contortus, the most common internal parasite of sheep and goats. Firstly, to determine the antiproliferative effect of the extracts, an initial screening was performed. Cytotoxicity was evaluated by measuring absorbance on a Tecan spectrophotometer. The four fern species that showed most promising antiproliferative activity were tested at three time intervals and four concetrations. H. contortus larvae L3 were exposed to fern extracts for seven days and the percentage of developed L4 larvae was detected by microscope. Athyrium distentifolium, Dryopteris aemula, Davallia canariensis and Polystichum aeculatum showed the highest antiproliferative and anthelmintic activity. In addition, Dryopteris...
|
|
|
Effect of vitamin D on ABCB1 and CYP3A4 expression in human precision-cut intestinal slices
Mazurová, Tereza ; Vokřál, Ivan (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Tereza Mazurová Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: Effect of vitamin D on the expression of ABCB1 and CYP3A4 in human intestinal slices When drugs are administered orally, their absorption is significantly affected by the intestinal barrier. This barrier expresses a variety of efflux and uptake transporters, as well as first and second-phase biotransformation enzymes. The most important efflux transporter in the intestinal barrier is P-glycoprotein, which has a broad substrate specificity. Among the first-phase biotransformation enzymes, cytochrome P450 3A4 is the most important. Their function is to protect the human body from the toxic effects of xenobiotics, including drugs. Many clinically important drugs act as substrates, inhibitors or inducers in relation to these proteins, which may result in an increased risk of drug interactions. A plethora of dietary supplements or medicines containing vitamin D can be found in the pharmacy. It is used for the proper development of bones and teeth, for the proper function of the immune system or for the treatment of osteomalacia, rickets or osteoporosis. Limited information is available on its effects on P-gp and...
|
|
|
The optimization of fenbendazole extraction from environmental samples
Brodská, Denisa ; Matoušková, Petra (advisor) ; Raisová Stuchlíková, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Denisa Brodská Supervisor: doc. Ing. Petra Matoušková, Ph.D. Consultant: Mgr. Martina Navrátilová Title of bachelor thesis: The optimization of fenbendazole extraction from environmental samples Anthelmintics are drugs used for the treatment and prophylaxis of infectious diseases caused by parasitic worms. In the Czech Republic, they are mainly used in veterinary medicine. During the biotransformation of the active substance, metabolites are formed, which are excreted together with the parent substance in faeces and urine. Fenbendazole and other similar substances can be absorbed into the soil in this way and subsequently act on non-target organisms in the environment. Fenbendazole is widely used in livestock farming, and since animal excrements are often further used for fertilization, the risk of the content of medicinal substances should be strictly monitored. For this purpose, it was necessary to develop and optimize the extraction method and also to carry out the corresponding validation of the analytical method, which will subsequently be applicable to real samples in the environment. The assumption of absorption of fenbendazole and its metabolic products, fenbendazole sulfoxide and...
|
|
|
Implementation of the diagnosis of gastrointestinal nematodes using RT-PCR.
Bezdíčková, Kristýna ; Matoušková, Petra (advisor) ; Nguyen, Thuy Linh (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Kristýna Bezdíčková Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Implementation of the diagnosis of gastrointestinal nematodes using RT-PCR Gastrointestinal nematodes can cause various diseases in farm animals. Individuals are often infected with several types of parasites at once. Since the symptoms tend to be similar, the disease is called parasitic gastroenteritis. Not only do nematodes threaten the animal health, but they can also have a major impact on the economy of the entire farm. Diagnosis is not always easy. In addition to morphological and coprological methods, biochemical or molecular analysis can be used, which tends to be more sensitive and accurate. We tried to diagnose gastrointestinal nematodes using the molecular RT-PCR method. In the thesis we focused on 6 species of nematodes (Haemonchus contortus, Teladorsagia circumcinta, Trichostrongylus spp., Nematodirus battus, Chabertia ovina and Ashworthius sidemi). The material used was feaces samples from sheep from which we isolated gDNA. The control material was gDNA isolated from H. contortus eggs and gDNA obtained from whole parasite bodies. RT-PCR was tested using the fluorescent dye SYBR Green I and...
|
|
|
New fungal fucosidases and their synthetic potential
Glozlová, Michaela ; Beránek, Martin (advisor) ; Matoušková, Petra (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Glozlová Supervisor: prof. PharmDr. Martin Beránek, Ph.D. Consultants: doc. RNDr. Pavla Bojarová, Ph.D., Mgr. Pavlína Nekvasilová Title of diploma thesis: New fungal fucosidases and their synthetic potential The literary research focuses on α-L-fucosidases, their occurrence, functions, and use in the synthesis of fucosylated compounds, which were also a part of the research, particularly their presence in the human body and involvement in pathological conditions. Furthermore, it explores the oligosaccharides found in human milk, their impact on infant health, and their synthesis, which can be facilitated through the use of α-L-fucosidases. The experimental section involves the analysis of a library of filamentous fungi with the aim of identifying a representative strain with α-L-fucosidase activity towards pNP-α-L-fucoside. Recombinant α-L-fucosidases were selected based on searches in genetic databases and prepared. The feasibility of their production in Pichia pastoris and Escherichia coli was tested, and the produced enzymes were purified by ion exchange chromatography, characterized and their regioselectivity and synthetic potential for the synthesis of fucosylated...
|
guest ::
