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In vitro effect of flavonoids on activity of selected biotransformation enzymes in intestinal cell lines
Pokorná, Petra ; Szotáková, Barbora (advisor) ; Matoušková, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Petra Pokorná Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: In vitro effect of flavonoids on activity of selected biotransformation enzymes in intestinal cell lines Anthocyanins belong to a class of flavonoids which are part of major natural pigments in plants. They are considered to be very beneficial for the human organism. Anthocyanins are widely consumed as fruits, vegetables and red wine and they represent significant constituents of dietary supplements. The popularity of dietary supplements with high concentrations of flavonoids is still increasing, but in addition to positive effects on human health it can also have side effects influence drug metabolizing enzymes and thus affect either desirable or undesirable effects of many drugs. In this thesis in vitro effect of some anthocyanidins (cyanidin, delphinidin, malvidin, pelargonidin) on the proliferation of the intestinal cell line Caco-2 was investigated using the assay of metabolic activity (MTT) and determination of the integrity of the cytoplasmic membrane of cells (neutral red). Then was studied their effect on the activity of selected biotransformation enzymes: glutathione S-transferase (GST),...
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Crime on the open drug scene in Prague
Matoušková, Petra ; Fidesová, Hana (advisor) ; Janíková, Barbara (referee)
The main topic of this theses is the relationship between crime and open drug scene in Prague. The use of illegal drugs alone and crime related to drug use, fundamentally threatens, not only individuals, but the entire society. Therefore it is necessary to address this problem as best as possible and consequently seek the most effective ways of problem solving. The theoretical part of the study is trying to explain the character of the drug scene during the previous political regime as well as the present. It deals with the general relationship between the crime and the drug use. It shows an overview of the model connections between those phenomena. The qualitive part of the research has the goal to describe the lifestyle of proble drug users, a way of raising money for drugs and livelihood. It focuses on detection of criminal offenses for which respondents were prosecuted, convicted and sentenced. It is also asking for so-called tertiary crime. Statement here counts whether respondents were also sometimes the victims of crime. The aim of this theses is to verify the following hypotheses and try to define what kind of crime associates with the open drug scene. The research results were obtained by semistructured interview between clients outreach program. The study involved 33 clients from the...
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Role of disulphide bonds in hA2A subtype adenosine receptor
Zappe, Lukáš ; Matoušková, Petra (advisor) ; Bárta, Pavel (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Lukáš Zappe Supervisor: Ing. Petra Matoušková Ph.D., PD Dr. Anke C. Schiedel Title of diploma thesis: Role of disulphide bonds in hA2A subtype adenosine receptor The adenosine A2A receptor belongs to the G protein coupled receptor family (GPCR). GPCRs are the targets of almost 40% of the drugs on the market. GPCRs are characterized by seven transmembrane helices, which are linked by three extracellular and three intracellular loops (ECL and ICL). The structure of the receptor has been revealed by crystallography, hence we know that ECL1 and ECL2 are connected by several disulphide bonds. The ECL2 is believed to be involved in ligand binding and recognition. In order to understand the relevance of those disulphide bonds involved in this process, four adenosine A2A receptor mutants were generated by one-site direct mutagenesis, in which the cysteine residues were replaced with serine residues (C146S, C159S, C166S and C146S- C159S). These receptor mutants were expressed in the mammalian cell line, CHO K1(Chinese Hamster Ovary) and the receptor expression was tested with ELISA (Enzyme- linked immunosorbent assay). The determination of ligand binding has been carried out by radioligand...
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Determination of mRNA of DHRS3 enzyme in human tissues by real-time SYBR green qPCR
Rusnáková, Pavlína ; Zemanová, Lucie (advisor) ; Matoušková, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Pavlína Rusnáková Supervisor: RNDr. Lucie Zemanová, PhD. Title of diploma thesis: Determination of mRNA of DHRS3 enzyme in human tissues by real-time SYBR green qPCR Dehydrogenase/reductase (SDR family) member 3 (DHRS3, also known as SDR16C1, retSDR1) is NADPH dependent carbonyl reductase, member of the short-chain dehydrogenases/reductases superfamily. DHRS3 was characterized for the first time by Haeseleer et al. as an enzyme that reduces all-trans-retinal to all-trans-retinol in the visual cycle that is localised in the outer segment of the cone photoreceptors in bovine retina. However, expression of DHRS3 was shown in some other tissues. It was proven that DHRS3 plays an important role in embryogenesis and tumour development. It was also established that DHRS3 participates in the reduction of some endogenous compounds (androstendione, estrone, DL-glyceraldehyde) and xenobiotics (acetohexamide, 4-nitroacetophenone, benzil) in vitro. DHRS3 seems to be an important enzyme with participation in significant metabolic pathways. Nevertheless, knowledge of some DHRS3 properties, including expression in human tissues, is quite poor. The aim of this study was to determine expression of DHRS3...
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Influence of glutamate-induced obezity on antioxidant enzymes in mice
Duchačová, Kateřina ; Boušová, Iva (advisor) ; Matoušková, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Kateřina Duchačová Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: Influence of glutamate-induced obezity on antioxidant enzymes in mice. Obesity is currently one of the most widespread chronic diseases in the world. Many scientific teams are trying to clarify the causes of this disease and also examine changes in the body that may be related to this disease. The aim of this study was to determine the activity and expression of the enzymes superoxide dismutase (SOD) and catalase (CAT) and to assess activity of peroxidase (Px) in liver cytosol of experimental NMRI mice with obesity induced by monosodium glutamate (MSG) and healthy controls. Activities and expressions of selected enzymes were compared with respect to the pathological conditions (MSG mice, osmolality controls and unaffected mice), age (8 weeks x 20 weeks) and sex of the experimental mice. Activities were determined using spectrophotometric methods and polyacrylamide gel electrophoresis followed by immunoblotting was employed to monitor protein expressions. Differences in specific activities of SOD and CAT among the groups were only small. The most significant changes related to the induction of obesity...
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Effect of alpha-humulene on adhesion of breast cancer cell line MDA-MB-231
Marešová, Markéta ; Matoušková, Petra (advisor) ; Jirkovská, Anna (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Markéta Marešová Supervisor: Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of alpha-humulene on adhesion of breast cancer cell line MDA-MB-231 α-humulene is a sesquiterpene contained in the essential oil of Chinese Bayberry (Myrica rubra), which has various biological effects. The aim of this thesis was to study the cell adhesion of tumor cell line MDA-MB-231, and the effects of α-humulene on expression of cell adhesion molecules. Furthermore, the cytotoxic effect of α-humulene on this cell line was verified. Effect of α-humulene on cell proliferation was evaluated by neutral red uptake test (NRU test) and by xCelligence system. Cell adhesion was also continuously monitored by xCelligence. Expression ganges of cell adhesion molecules upon α-humulene treatment were determined by Western blotting and by quantitative polymerase chain reaction on the protein and mRNA levels, respectively. The proliferation of the cells was significantly affected by α-humulene after 24 hour treatment (IC50 13.6 µg/ml). α-humulene alone caused slightly increased cell adhesion. Adhesion molecules EpCAM and β-cathenin were almost unaffected. Level of ICAM1 mRNA was increased after 12 hours and...
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Effect of valencene on adhesion of breast cancer cell line MDA-MB-231
Mottlová, Petra ; Matoušková, Petra (advisor) ; Schröterová, Ladislava (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Petra Mottlová Supervisor: Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of valencene on adhesion of breast cancer cell line MDA-MB-231 Valencene belongs to sesquiterpenes and is constituent of essential oils of many plants. It is the main citrus flavoring component. Valencene is mainly used in the food and cosmetic industries, but recent studies have confirmed its biological activity. Its antitumor, antiinflammatory, antioxidant and antiallergic effects have been already proven. The objective of this study was to determine the cytotoxic effect and influence on the adhesion of sesquiterpene valencene on breast cancer cell line MDA-MB-231. Another objective was to study the mechanism of its effect from the perspective of the selected adhesion molecules, which have an important role in tumorigenesis and metastasis. Valencene's cytotoxic effect was tested with use of neutral red (NRU test). Valencene influence on cell adhesion was continuously monitored by means of the X-Celligence device and expression of selected adhesion molecules was studied by Western blot and qPCR methods. The results showed a slight cytotoxic effect of valencene. Cell viability was over 70% at a...
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Inhibitory effect of anthocyanidins on hepatic carbonyl reductase
Hlaváčová, Jitka ; Skálová, Lenka (advisor) ; Matoušková, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Jitka Hlaváčová Supervisor: Prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Inhibitory effect of anthocyanidins on hepatic carbonyl reductase Carbonyl reductase (CBR) is a reducing enzyme that plays a significant role in the first phase of biotransformation of both endogenous and exogenous substrates. It participates in detoxification of many xenobiotics, among them important drugs. Differences in expression and activity of hepatic biotransforming enzymes may affect efficiency and safety of treatment. The aim of this work was to compare the activity and expression of CBR in old and young individuals and to define an inhibitory effect of selected anthocyanidins on the activity of CBR in humans and rats and to determine a value of IC50 and a type of inhibition for the strongest inhibitors. Meaningly higher activity of CBR was detected in the senescent rats compared to the young ones and the old rats also showed larger interindividual variability. No significant age- related differences have been identified by measurement of activity CBR in human samples. Anthocyanidins showed a mild to moderate inhibitory effect on CBR. The enzyme was most strongly inhibited by delphinidin (IC50...
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