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The changes in expression of carbonyl reductases during life cycle of nematode
Ňorková, Kateřina ; Skálová, Lenka (advisor) ; Szotáková, Barbora (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Kateřina Ňorková Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The changes in expression of carbonyl reductases during life cycle of nematode Barber 's pole worm (Haemonchus contortus, from the family Trichostrongylidae) is a nematode attacking the mucosa of the gastrointestinal tract of small ruminants. Infections caused by this nematode are manifested by anemia and edema. Anthelmintics, drugs against parasitic worms, are used for prophylaxis and treatment of these infections. However, a huge problem is the worldwide growing resistance to all available anthelmintics. Therefore, it is very important to study the possible mechanisms of resistance and factors that contribute to the development of resistance. In recent years, one of the mechanisms of resistance to anthelmintics has been found to be increased expression and activity of some enzymes of metabolizing drugs. This work dealt with the monitoring of changes in the expression of selected genes from the detrinth of Dehydrogenas/Reductas with a short chain (SDR). Expression of 20 SDR genes was determined using the quantitative PCR method in all developmental nematode stages (eggs, larvaes, adults) of...
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Analysis of MHC-II bound peptides of dendritic cells
Pavlosková, Jana ; Szotáková, Barbora (advisor) ; Balonová, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Jana Pavlosková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Consultant: Ing. Marek Link, Ph.D. Title of diploma thesis: Analysis of MHC-II bound peptides of dendritic cells Antigen presentation is an important process for the immune system, which is mediated by antigen presenting cells. These cells can capture and process antigen material and present antigenic peptides to T lymphocytes. The immunopeptidome represents a collection of peptides displayed by MHC molecules on the surface of antigen presenting cells. Analysis of immunopeptidome plays a crucial role in identifying peptides to which the organism responds. This knowledge is important for developing new vaccines, medication, and disease diagnostics. This thesis focuses on the analysis of MHC-II bound peptides of dendritic cells, with the aim of verifying and demonstrating the suitability of methodological procedures for immunopeptidome analysis. The study was conducted on the mouse dendritic cells. Additionally, the cells were pulsed with the protein antigen ovalbumin to model foreign antigen binding and presentation. An immunoaffinity resin protein G agarose with immobilized antibody targeting I-Ab was successfully created for...
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Expression and activities of antioxidant enzymes in the in vivo models of non-alcoholic fatty liver disease
Sochová, Gabriela ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Gabriela Sochová Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Advisor: Mgr. Gabriela Svobodová Title of diploma thesis: Expression and activities of antioxidant enzymes in the in vivo models of non-alcoholic fatty liver disease Non-alcoholic fatty liver disease is one of the most common liver diseases. This disease is caused by an imbalance between fat intake and fat degradation. Lipid accumulation can be caused by several factors (genetic predisposition, diet, metabolic diseases, etc.). If these factors act for a long enough period, the disease can lead to liver fibrosis or even cirrhosis. The aim of this diploma thesis was to compare the expression and activity of the antioxidant enzymes glutathione-S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD), and catalase in two models of NAFLD (dietary model (K) - high fat, fructose, and cholesterol diet (FFC); chemical model - induced by monosodium glutamate (MSG)). Furthermore, the effect of LiPR31 and liraglutide on the expression and activity of antioxidant enzymes were investigated in these models. Expression and activity were determined in 20,000×g supernatant of mouse liver. RT-qPCR was used...
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Determination of ganciclovir in lung transplant recipients
Hrach, Filip ; Szotáková, Barbora (advisor) ; Kotaška, Karel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Filip Hrach Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Consultant: doc. Ing. Eva Klapková, Ph.D. Title of diploma thesis: Determination of ganciclovir in lung transplant recipients Ganciclovir is a nucleoside analogue that is widely used for the treatment of infections caused by herpes viruses, including cytomegalovirus. Patients after lung transplantation are always given valganciclovir (ganciclovir prodrug) prophylactically to prevent cytomegalovirus infection. The aim of this thesis was to establish a reliable, sensitive and rapid analytical method for the determination of ganciclovir and to validate this method. Furthermore, to determine ganciclovir levels in patients after lung transplantation. The final objective was to evaluate the importance of therapeutic monitoring of ganciclovir. A liquid chromatography-tandem mass spectrometry method was established for the determination of ganciclovir. Validation parameters necessary for bioanalytical methods have been established.All validation parameters met the conditions for successful validation.Valganciclovir is used for oral administration of antiviral drugs and ganciclovir for intravenous administration. Patients after lung...
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Study of the effect of albendazole in the environment on the antioxidant system of red clover (Trifolium pratense)
Navrátilová, Gabriela ; Szotáková, Barbora (advisor) ; Langhansová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Gabriela Mikešková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Study of the effect of albendazole in the environment on the antioxidant system of red clover (Trifolium pratense) Today, anthelmintics are among the essential drugs for the therapy and prophylaxis of infectious diseases caused by parasitic worms. However, these substances may be further released into the environment by the excrements of treated animals and affect non-target organisms, such as plants, which have the ability to absorb and accumulate various substances. In this study, we focused on the effect of the often used anthelmintic albendazole on the antioxidant system of red clover (Trifolium pratense), which is commonly found in meadows and pastures. The effect of low concentrations of albendazole available in the environment on the antioxidant enzymes catalase, superoxide dismutase, ascorbate peroxidase, peroxidase, glutathione peroxidase, glutathione reductase and glutathione S-transferase was studied. In addition to antioxidant enzymes, we also monitored the concentration of plant photosynthetic pigments chlorophyll a, b and carotenoids. The results were evaluated in control and...
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The use of new ex vivo tests to find ferns with anthelmintic effect
Bednářová, Klára ; Skálová, Lenka (advisor) ; Szotáková, Barbora (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Klára Bednářová Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The use of new ex vivo tests to find ferns with anthelmintic effect Haemonchus contortus is one of the worldwide spread nematode parasitizing in the gastrointestinal tract of small ruminants. The nematodes cause a disease called haemonchosis. A typical symptom of the disease is anemia resulting from the sucking of blood by nematodes. The disease also reduces productivity and may result in the death of the host. Only a few drugs are used for the treatment of haemonchosis, the number of newly introduced drugs is low. Nowadays, the emerging resistance to anthelmintics is a growing problem due to the high H. contortus adaptivity as well as the incorrect use of synthetic drugs. In addition, the synthetic drugs pollute the environment, which is why there is an effort to find herbal products with an anthelmintic effect. Anthelmintic activity has been demonstrated in some Asian and African ferns, so we decided to test the European fern species and evaluate their anthelmintic activity. Anthelmintic activity against nematodes, specifically H. contortus, was tested using egg hatch test and ATP bioluminescence assay in...
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The effect of selected herbicides on UGTs expression and activity of antioxidant enzymes in Arabidopsis thaliana
Rysová, Samanta ; Matoušková, Petra (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Samanta Rysová Supervisor: doc. Ing. Petra Matoušková, Ph.D. Consultant: RNDr. Radka Podlipná, Ph.D. Title of diploma thesis: The effect of selected herbicides on UGTs expression and activity of antioxidant enzymes in Arabidopsis thaliana Herbicides are widely used to control weeds in agriculture. Although their use is essential to ensure crop production, their effect occurs to be reduced due to the development of herbicide resistence. Therefore it is important to clarify the mechanism of herbicide resistence, so that herbicides can be used effectively and new substances can be developed if necessary. At the same time, herbicides acts as an abiotic stress factor on the plant, thus affecting the antioxidant apparatus of plant cells. The aim of this work was to investigate the effect of selected herbicides - tribenuron-methyl (TBM), pinoxaden (PNX) and sulcotrione (SLC), on the expression of selected UGTs that could play a role in herbicide resistence. Secondary, we want to find out the effect of TBM on the activity of selected antioxidant enzymes - ascorbate peroxidase (APX), catalase (CAT) and superooxide dismutase (SOD). Arabidopsis thaliana was used as a model plant and the experiment was...
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Single nucleotide polymorphisms in the pathogenesis of mature B-cell neoplasms
Wagnerová, Andrea ; Szotáková, Barbora (advisor) ; Beránek, Martin (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Andrea Wagnerová Supervisor: prof. Ing. Barbora Szotáková, Ph.D., Mgr. Kateřina Černá Pilátová, Ph.D. Title of diploma thesis: Single nucleotide polymorphism in the patogenesis of mature B-cell neoplasms Chronic lymphocytic leukemia (CLL) is a disease characterized by clonal proliferation of B-lymphocytes. CLL is a lymphoproliferative disorder with a number of defined risk factors, but its etiopathogenesis remains unresolved to some degree. Over the past 10-15 years, there have been several studies that primarily suggest a genetic predisposition to the disease. The influence of external factors on the etiology of CLL has not been demonstrated. There is also a defined asymptomatic condition associated with CLL called monoclonal B-lymphocytosis, which is often referred to as a precursor of CLL. Immunophenotyping of lymphocytes by flow cytometry was performed in subjects of the control group (100 subjects) and the CLL/MBL group (patients diagnosed with CLL or MBL, 50 subjects). All subjects in the control and CLL/MBL groups were screened for selected genetic polymorphisms (rs17483466, rs735665, rs7176508, rs872071, rs13397985) that have been associated with the occurrence of CLL and MBL in other...
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Optimization of liver spheroids preparation
Vojtěchová, Tereza ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Tereza Vojtěchová Supervisor: assoc. prof. PharmDr. Iva Boušová, Ph.D. Advisor: PharmDr. Martin Ambrož, Ph.D. Title of diploma thesis: Optimization of liver spheroids preparation Various in vitro liver models play an irreplaceable role in preclinical drug evaluation. One of them is multicellular hepatic spheroids. These are 3D cell aggregates that are formed by the process of cellular aggregation. They are used in drug development and represent a promising tool for identifying new targets and drug screening for various liver diseases. The aim of this thesis was to introduce and optimize the preparation of liver spheroids by the "forced-floating" method. The liver of male and female Wistar rats was used for hepatocyte isolation and spheroid preparation. Due to the low viability of isolated hepatocytes, we tried to optimize the isolation procedure. Nevertheless, we managed to obtain only a small number of hepatocytes with sufficient viability for subsequent formation of spheroids. The spheroids were incubated with various lipogenic reagents for 7 and 14 days and their viability was monitored by measuring adenosine triphosphate (ATP) production. Incubation of spheroids with lipogenic substrates...
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The use of new ex vivo tests to find synthetic compounds with anthelmintic effect
Mičundová, Magdalena ; Skálová, Lenka (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Magdalena Mičundová Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The use of new ex vivo tests to find synthetic compounds with anthelmintic effect Haemonchus contortus is a parasitic nematode causing disease called haemonchosis in sheep and goats. This worldwide spread parasite feeds on the host's bloodstream, causing anemia and edema that can lead to death of the animal. Anthelmintic drugs are used for haemonchosis treatment, but recently their efficacy has decreased due to development of drug resistance in nematodes. Therefore, it is necessary to look for new drugs that will be effective in resistant nematode strains. In this work the anthelmintic effect of 13 new benzyloxyamide derivates (BLK) was studied in H. contortus eggs and adults. An egg hatch test, in which BLK8 was the most effective compound, was used first. A new method of bioluminiscence determination of adenosine triphosphate (ATP) was used in adults of H. contortus to monitor the effect of BLK analogues on viability of adult nematodes. The efficacy of BLK analogues was compared with the effect of classical anthelmintic levamisole. It is obvious from the results that several BLK analogues (especially...
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