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The effect of mebendazole on proliferation, migration and adhesion of oral cancer cells in vitro
Stupková, Eva ; Skálová, Lenka (advisor) ; Boušová, Iva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Eva Stupková Supervisor: RNDr. Věra Králová, Ph.D. Title of diploma thesis: Effect of mebendazole on proliferation, migration and adhesion of oral carcinoma cells in vitro Mebendazole is one of commonly used anthelmintics in both human and veterinary medicine. The mechanism of its action is based on microtubule synthesis inhibition. This thesis is focused on mebendazole effect on cancer cells. Especially, we studied the effect of mebendazole on proliferation, migration and adhesion of squamous-cell carcinoma (SCC) cells, very aggressive and highly invasive cancerous disease. Our experiments ran on two cancerous cell lines (DOK, PE/CA-PJ15) and on non-cancerous gingival fibroblasts. We used WST-1 method for proliferation change assessment and xCELLigence RTCA DP Analyzer for measuring the effect on migration and adhesion of SCC cells. In our experiments, mebendazole caused decrease of SCC proliferation, migration and it also had effect on adhesion, in clinically achievable concentrations.
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Biological activity of potential antioxidants in L-6 rat myoblasts
Drahošová, Vlasta ; Boušová, Iva (advisor) ; Dršata, Jaroslav (referee)
Charles University in Prague University Roma Tre Faculty of Pharmacy in Hradec Králové Department of Sciences Department of Biochemical Sciences Laboratory of Physiology Candidate: Vlasta Drahošová Supervisor: Doc. PharmDr. Iva Boušová, Ph.D. Prof. Sandra Incerpi Title of diploma thesis: Biological activity of potential antioxidants in L-6 rat myoblasts This diploma thesis is based on the results of research conducted at the Department of Sciences, University of Roma Tre in Rome under the supervision of Professor Sandra Incerpi. Team of the Laboratory of physiology has been involved in the research of various natural substances with antioxidant and cytostatic properties for many years. Antioxidants are able to eliminate reactive oxygen species (ROS), which exert irreplaceable physiological effects in the body, but they are also a cause of certain diseases progression and aging due to the damage of various biomolecules. In this work, I studied the antioxidant activity of six synthetic derivatives of 4-methylcoumarin (AB-3, AB-4, AB-7, AB-8, AB-11, AB-12), which were synthesized and characterized by prof. Virinder Parmar (University of Delhi, India). Study was conducted in the L-6 rat myoblasts cell line (determination of intracellular ROS using dichlorofluorescein assay, cell proliferation assay)...
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Oxidative damage to proteins
Bártová, Jana ; Boušová, Iva (advisor) ; Zemanová, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Jana Bártová Supervisor: PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: Oxidative damage to proteins Hypochlorous acid is a strong oxidant, which is used by phagocytic cells to kill ingested bacteria. Surrounding structures including proteins may be damaged in case the content of a phagocyte is spilled to the vicinity. Catechins belong to a group of flavonoids whose antioxidant properties were demonstrated in many studies. My diploma thesis was focused on a potential effect of eight naturally occurring derivatives of catechin towards oxidative damage of human serum albumin (HSA) by hypochlorous acid. I used two spectrophotometric (determination of carbonyl groups and chloramines) and three electrophoretic methods (native PAGE, SDS-PAGE, immunoblotting). Dependence of an antioxidant effect on its concentration (0-3 mM) was observed during determination of chloramines and obtained data were used for calculation of EC50 values of individual catechins using program GraphPad Prism. The most effective compound was catechin whose EC50 was 4.8 µM. Results suggest that antioxidant ability decreases with increasing number of hydroxyl groups in the molecule of flavonoid. Carbonyl groups are a...
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Effect of prenylated flavonoids on biotransformation enzymes in vitro
Pataráková, Paula ; Szotáková, Barbora (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Paula Pataráková Supervisor: Prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Effect of prenylated flavonoids on biotransformation enzymes in vitro Prenylated flavonoids xanthohumol (XH), isoxanthohumol (IXH), 6-prenylnaringenin (6PN) and 8-prenylnaringenin (8PN) are flavanones and chalcones occurring in hops and their characteristic feature is prenyl chain on A ring. As these substances, present in beer and dietary supplements, are foreign compounds for organism, they can affect drug- metabolizing enzymes, including carbonyl reducing enzymes. The aim of our study was to find out, whether XH, IXH, 6PN, and 8PN have any impact on cytosolic carbonyl reductase 1 (CBR1) and aldo-keto reductase 1C subfamily (AKR1C). Viability test revealed that prenylated flavonoids in lower concentrations do not affect or even increase the viability of primary rat hepatocytes, but higher concentrations are toxic. AKR1C activity was increased in 8PN treated hepatocytes, the activity of CBR1 was significantly increased in IXH and 8PN treated samples. An increase in expression of AKR1C3 in IXH and 6PN treated samples, and decrease in XH and 8PN samples was observed. Expression of CBR1 was at the detection...
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Modification of selected biotransformation enzymes during ageing (in rat)
Vyskočilová, Erika ; Boušová, Iva (advisor) ; Svobodová, Hana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Erika Vyskočilová Supervisor: PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: MODIFICATION OF SELECTED BIOTRANSFORMATION ENZYMES DURING AGEING (IN RAT) Ageing is an inevitable biological process characterized by progressive functional decline of all organ systems with simultaneous increase in oxidative damage as a result of reactive oxygen species (ROS) accumulation. Increased activity of antioxidant enzymes, such as peroxidase and catalase, is related to increased production of ROS during ageing. Activity of biotransformation enzymes such as cytochrome P450 (CYP) and glutathion-S-transferase (GST) changes during ageing too. In general, activity of enzymes is lower in very young and very old individuals than in the period of adulthood. The aim of the present work was to study age-related changes in activities and expression of selected xenobiotic-metabolizing enzymes (GST, CYP 1A, CYP 2B, CYP 3A4) and antioxidant enzymes (catalase, peroxidase, superoxiddismutase) in rat liver. Male Wistar rats were sacrificed by decapitation under ether anesthesia at the age of 6 weeks and 21 months (6 individuals in each group). Subsequently, liver specimens were processed to obtain the subcellular...
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Formation of carbonyl compounds by non-enzymatic protein glycation
Krautová, Lenka ; Boušová, Iva (advisor) ; Dršata, Jaroslav (referee)
Author: Lenka Krautová Publishing place: Charles University in Prague, Faculty of Pharmacy in Hradec Králové Title: Formation of carbonyl compounds by non-enzymatic protein glycation Supervisor: PharmDr. Iva Boušová, Ph.D. This diploma thesis was aimed at the optimization of carbonyl groups determination in biological material. Carbonyl groups are formed by effect of protein oxidation or glycation. As a model protein, I used bovine serum albumin for spectrophotometric and aspartate aminotransferase for electrophoretic method of assessment, which were glycated by several glycating agents (glucose; glucose + copper; fructose; fructose + copper; methylglyoxal). I optimized different parameters of spectrophotometric determination: solvent, concentration of reagent, amount of protein, and duration of incubation. I managed to find suitable conditions of this determination, which I have used in subsequent experiments with different glycating agents. At electrophoretic determination, I performed SDS-PAGE with following silver staining or immunoblotting with chemiluminescent detection. The results of spectrophotometric determination showed that methylglyoxal has the highest glycating ability, followed by fructose with addition of cupric ions, fructose itself, glucose with addition of cupric ions, and the...
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Changes of lipid spectrum during body mass reduction in patients with diabetes mellitus
Šmídová, Barbora ; Dršata, Jaroslav (advisor) ; Boušová, Iva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Barbora Šmídová Supervisor: Prof. MUDr. Jaroslav Dršata, CSc. Supervisor - specialist: RNDr. Mgr. Alena Tichá, PhD. Title of diploma thesis: CHANGES OF LIPID SPECTRUM DURING BODY MASS REDUCTION IN PATIENTS WITH DIABETES MELLITUS The thesis deals with the determination of lipid parameters (plasma total fatty acids, plasma total cholesterol, lipoprotein cholesterol, plasma triglycerides, plasma cholesterol precursors (lathosterol and squalene) and markers of cholesterol absorption (β-sitosterol and campesterol)) in obese patiens with diabetes mellitus type 1 and type 2 who were tested with a seven-day fasting, followed by low-calorie diabetic diet. It is assumed that weight loss should improve insulin resistence. The aim of this work is to evaluate the lipid parameters during the body weight reduction in obese patients suffering with diabetes mellitus. Lipids were determined before and after a seven-day fasting and after one month from the beginning of fasting (in patiens with diabetes mellitus type 1 also after one year). Gas chromatography was used for the determination of fatty acids and non-cholesterol sterols and squalene. Cholesterol and triglycerides were determined by a routine...
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OCT activity and glycation damage in rat liver mitochondria
Merhautová, Lucie ; Dršata, Jaroslav (advisor) ; Boušová, Iva (referee)
OCT ACTIVITY AND GLYCATION DAMAGE IN RAT LIVER MITOCHONDRIA Lucie Merhutová Diploma thesis. Charles University in Prague, Faculty of Pharmacy in Hradec Králové. 2007 This work was created following investigations carried out in LBBCV (Laboratoire de Biologie et Biochimie Cellulaire du Vieillissement) at University Paris Diderot - Paris 7. The team of this laboratory studies the mechanisms of posttranslational non-enzymatic modifications of proteins, involved in the pathophysiology of aging. Glycation is one of the mechanisms responsible for the modification of intracellular macromolecules leading to the loss of their structure and function. In previous investigations, ornithine carbamoyltransferase (OCT), the second enzyme of urea cycle, has been identified as one of the markedly glycated proteins in liver mitochondria of senescent rats. The goal of this work was to investigate in vitro and ex vivo glycation of OCT and the effect of glycation on its function. Purified OCT and mitochondrial matrix extracts were incubated with methylglyoxal, one of the most reactive physiological glycating agents, and submitted to the enzymatic assay and immunochemical analysis. We demonstrated that methylglyoxal modifies OCT both in vitro and ex vivo and causes rapid and extensive decrease in its enzymatic activity....
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