|
|
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...
|
|
|
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...
|
|
|
Characterization of carbonyl-reducing enzymes in barber's pole worm
Kunzová, Lucie ; Szotáková, Barbora (advisor) ; Novotná, Eva (referee)
Charles University, Faculty of Pharmacy in Hradci Králové Department of Biochemical Sciences Candidate: Lucie Kunzová Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Consultant: Mgr. Nikola Rychlá Title of Thesis: Characterization of carbonyl-reducing enzymes in barber's pole worm Barber's pole worm (Haemonchus contortus), one of the most pathogenic parasites of small ruminants as sheep and goats, has developed resistance against all used anthelmintic drugs. Detoxifying enzymes, which represent the main defense system against harmful xenobiotics, can also contribute to the development of drug resistance, and they are not fully explored in this organism. This research is focused on the study of carbonyl-reducing enzymes in barber's pole worm using specific substrates and inhibitors. Subcellular fractions (cytosol and microsomes) were prepared from adult worms for following in vitro experiments. Protein concentration and reductase activity towards different substrates were determined in each fraction. Substrates were chosen based on knowledge of human biotransformation enzymes. The highest activity in cytosol has been detected for glyceraldehyde (8,22 ± 0,82 [nmol/min/mg]) and menadione (4,63 ± 0,46 [nmol/min/mg]), in microsomal fraction for menadione (4,38 ± 0,36 [nmol/min/mg]) and metyrapone (1,92...
|
|
|
Characterization of carbonyl-reducing enzymes in barber's pole worm
Kunzová, Lucie ; Szotáková, Barbora (advisor) ; Novotná, Eva (referee)
Charles University, Faculty of Pharmacy in Hradci Králové Department of Biochemical Sciences Candidate: Lucie Kunzová Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Consultant: Mgr. Nikola Rychlá Title of Thesis: Characterization of carbonyl-reducing enzymes in barber's pole worm Barber's pole worm (Haemonchus contortus), one of the most pathogenic parasites of small ruminants as sheep and goats, has developed resistance against all used anthelmintic drugs. Detoxifying enzymes, which represent the main defense system against harmful xenobiotics, can also contribute to the development of drug resistance, and they are not fully explored in this organism. This research is focused on the study of carbonyl-reducing enzymes in barber's pole worm using specific substrates and inhibitors. Subcellular fractions (cytosol and microsomes) were prepared from adult worms for following in vitro experiments. Protein concentration and reductase activity towards different substrates were determined in each fraction. Substrates were chosen based on knowledge of human biotransformation enzymes. The highest activity in cytosol has been detected for glyceraldehyde (8,22 ± 0,82 [nmol/min/mg]) and menadione (4,63 ± 0,46 [nmol/min/mg]), in microsomal fraction for menadione (4,38 ± 0,36 [nmol/min/mg]) and metyrapone (1,92...
|
|
| |
|
|
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...
|
|
|
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...
|
|
|
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...
|
|
|
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...
|
|
|
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...
|
guest ::
