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Effect of parasite on activity of biotransformation and antioxidant enzymes in host intestine
Mládková, Zuzana ; Szotáková, Barbora (advisor) ; Skálová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Zuzana Mládková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Effect of parasite on activity of biotransformation and antioxidant enzymes in host intestine Hymenolepis diminuta, known as a rat tapeworm, is commonly used in science as a model of Cestoda for studying physiology, biochemistry and drug metabolism in tapeworms. Recently, Hymenolepis diminuta has been studied for helminth-based therapy for a potential treatment of inflammatory bowel disease. The aim of this study was to determine how H. diminuta influences the activity of detoxification enzymes of the host intestine. At first 6 male rats (Wistar breed) were infected by cysticercoids of Hymenolepis diminuta previously isolated from the beetles Tenebrio molitor (intermediate host). At the same time the physiological saline solution was administered to the control group of 6 male rats. All rats were housed for 2 months in animal quarters with 12 h light/dark cycle. Thereafter, the tapeworms were removed from their intestines. Intestinal mucosa containing metabolic active enzymes was isolated. Subsequently, the subcelullar fractions were prepared and used for in vitro experiment. The activity of enzymes was...
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Metabolomic analysis of bile acids in various biological samples.
Hadravská, Pavlína ; Skálová, Lenka (advisor) ; Raisová Stuchlíková, Lucie (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences University of Eastern Finland in Kuopio, School of Pharmacy Department of Pharmaceutical Chemistry Candidate: Pavlína Hadravská Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Consultant: prof. Seppo Auriola Title of Diploma thesis: Metabolomic analysis of bile acids in various biological samples This thesis aimed to establish a suitable qualitative and quantitative analysis of selected bile acids in mouse liver, human plasma and especially in human hair. Nine selected bile acids were analyzed (cholic acid, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, taurocholic acid, taurodeoxycholic acid, glycocholic acid and glycochenodeoxycholic acid). An identification and a quantification of the bile acids have been performed by UHPLC with two types of hybrid mass spectrometers (Quadrupole connected to Time of Flight and Quadrupole connected to Orbitrap). We tried to develop a method that allows a detection of both unconjugated bile acids and their glycine- and taurine-conjugates in biological samples. To develop and optimize this method, mouse liver and human plasma were used. Afterwards, the method was used to detect bile acids in human hair, because the bile acids could serve as...
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The effect of flavonoids on selected biotransformation enzymes
Procházková, Eliška ; Szotáková, Barbora (advisor) ; Skálová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Eliška Procházková Supervisor: Prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: The effect of flavonoids on selected biotransformation enzymes In recent years, catechins which belong to flavonoids have been studied polyphenolic constituents of green tea. Especially their beneficial effects on the human health are mostly studied. The popularity of dietary supplements with high levels of flavonoids is growing and thus they can affect a wide range of biotransformation enzymes and also the metabolism of many drugs. The aim of this diploma thesis was to find out how the selected catechins - catechin (C), epicatechin (EC), epicatechin 3-gallate (ECG), epigallocatechin (EGC), epigallocatechin 3-gallate (EGCG) and the green tea extract - polyphenon 60 (P) influence the viability of the Caco-2 cells and the activity and expression of the selected biotransformation enzymes in these cells - cytochromes P450 (CYP) isoforms 1A1, 1A2, and 3A, aldehyde reductase (AKR1A1), aldo-keto reductase 1C (AKR1C), carbonyl reductase 1 (CBR1), UDP-glucuronosyltransferase (UGT), sulfotransferase (SULT) and glutathione S-transferase (GST). The viability tests (NRU test and MTT test) proved that catechins (EGC,...
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The effect of flubendazole on the efficacy of paclitaxel in brest cancer cells
Vítovcová, Barbora ; Skálová, Lenka (advisor) ; Ambrož, Martin (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Barbora Vítovcová Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The effect of flubendazole on the efficacy of paclitaxel in breast cancer cells Flubendazole (FLU) is a common anthelmintic drug for veterinary and human use. Its molecular mechanism of action is based on binding to β-tubulin subunit of microtubules. FLU was previously identified to have antitumor activity. The aim of this study was to evaluate the effect of FLU on antiproliferative efficacy of paclitaxel (PTX) in breast cancer cells. For this study, three breast cancer cell lines were used - non-metastatic line MCF-7 and two metastatic cell lines, MDA-MB-231 and BT-474. The effects of FLU, PTX and FLU + PTX combination on viability of the cells (determined using WST-1 analysis) and on the expression of molecules involved in cell adhesion, migration and proliferation on mRNA (RT-PCR analysis) and protein levels (western blotting) were studied and compared. The activity of caspases after PTX and FLU treatment was evaluated using luminescence measurement. FLU has significant antiproliferative effect in all tested cell lines. MCF-7 and MDA-MB-231 have been shown as the most sensitive to PTX and FLU treatment....
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Metabolomic analysis of bile acids in various biological samples.
Hadravská, Pavlína ; Skálová, Lenka (advisor) ; Raisová Stuchlíková, Lucie (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences University of Eastern Finland in Kuopio, School of Pharmacy Department of Pharmaceutical Chemistry Candidate: Pavlína Hadravská Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Consultant: prof. Seppo Auriola Title of Diploma thesis: Metabolomic analysis of bile acids in various biological samples This thesis aimed to establish a suitable qualitative and quantitative analysis of selected bile acids in mouse liver, human plasma and especially in human hair. Nine selected bile acids were analyzed (cholic acid, deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid, lithocholic acid, taurocholic acid, taurodeoxycholic acid, glycocholic acid and glycochenodeoxycholic acid). An identification and a quantification of the bile acids have been performed by UHPLC with two types of hybrid mass spectrometers (Quadrupole connected to Time of Flight and Quadrupole connected to Orbitrap). We tried to develop a method that allows a detection of both unconjugated bile acids and their glycine- and taurine-conjugates in biological samples. To develop and optimize this method, mouse liver and human plasma were used. Afterwards, the method was used to detect bile acids in human hair, because the bile acids could serve as...
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Effect of mebendazole on the activity of selected enzymes in tapeworm Hymenolepis diminuta
Lukačiková, Karolína ; Vokřál, Ivan (advisor) ; Skálová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Karolína Lukačiková Supervisor: PharmDr. Ivan Vokřál, Ph.D. Title of diploma thesis: Effect of mebendazole on the activity of selected enzymes in tapeworm Hymenolepis diminuta The resistance of parasitic helminths to anthelmintic drugs is a growing worldwide phenomenon and a concerning issue. Xenobiotic metabolizing enzymes play an important role in drug resistance development as they can lower the concentration of the anthelmintics in the parasite's body and therefore protect the parasite from the anthelmintic effect. The role of drug metabolizing enzymes in drug resistance development has been already described in the group of roundworms and flukes. Limited information is available about this topic in tapeworms. In our study we decided to test the possibility of the anthelmintic mebendazole to affect the activity of these enzymes and possibly to influence the drug resistance development in rat tapeworm (Hymenolepis diminuta). Our first goal was the isolation of adult tapeworms from the definitive host (rat, Rattus norvegicus). We used mealworm beetle (Tenebrio molitor) as an intermediate host. After the successful isolation, adult tapeworms were incubated with the mebendazole (1 and 10µM) in...
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Sesquiterpenes in cancer therapy
Ambrož, Martin ; Skálová, Lenka (advisor) ; Hodek, Petr (referee) ; Souček, Pavel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Martin Ambrož Supervisor: Prof. RNDr. Lenka Skálová, Ph.D. Title of Doctoral Thesis: Sesquiterpenes in cancer therapy. Chemotherapy is an important part of treatment of cancer. Currently, the use of several anticancer drugs in combination seem to be more advantageous. This combination therapy prevents drug resistance development and make it possible to lower the doses of single drugs, which prevents the side effects in non-cancerous tissues. Large pool of potential drugs for combination therapy can be found among natural compounds, for example among sesquiterpenes. Our project was designed to evaluate the effect of Myrica rubra essential oil (MEO) and selected sesquiterpenes in combination cancer therapy. We focused on the potential ability of MEO and sesquiterpenes to increase the antiproliferative efficacy of classical cytostatics (doxorubicin, oxaliplatin, 5-fluorouracil). In addition, we also studied the possible interaction of sesquiterpenes with other drugs in combination therapy, which could be caused by sesquiterpene-mediated modulation of drug-metabolizing enzymes. For these purposes, a panel of various cell lines (both cancerous and non-cancerous) and primary culture of hepatocytes...
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Induction and inhibition of acenaphthenol dehydrogenase
Štěrbová, Klára ; Skálová, Lenka (advisor) ; Szotáková, Barbora (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Title, Name, Surname of candidate: Klára Štěrbová Title, Name, Surname of tutor: Doc. RNDr. Lenka Skálová, Ph.D. Title of a diploma work: Induction and inhibition of acenaphthenol dehydrogenase Breast cancer belongs to hormone-dependent tumours. Apart from lifetime exposure to estrogens, the volume of female active sex hormones in breast tissue also plays a big part in the development of this disease. One of mechanisms regulating concentration of estrogens in the target tissue at the pre-receptor level is their interconversion from inactive estron to its active form, 17β-estradiol enzymes from the aldo-ketoreduktas 1C (AKR1C) subfamily. On the basis of specific substrates, inductive and inhibitive studies, enzymes involved in the conversion of individual hormones can be characterized. Relatively specific substrate for AKR1C is acenaphthenol (AcNOH). However, due to the fact that in a multi-enzyme model system the involvement of other enzymes in the reaction cannot be excluded, the enzymes which take part in conversion of acenaphthenol labeled "acenaphtenol dehydrogenase" (AND). The Aim of my diploma paper is to spectrofluorimetrically determine the activity of AND in cytosole taken from breast...
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Biological activity of antioxidant compounds in L-6 myoblasts from rat skeletal muscles
Staňková, Veronika ; Boušová, Iva (advisor) ; Skálová, Lenka (referee)
Charles University in Prague University Roma Tre Faculty of Pharmacy in Hradec Králové Department of Biology Department of Biochemical Sciences Candidate: Veronika Staňková Supervisor: PharmDr. Iva Boušová, Ph.D. Prof. Sandra Incerpi, Prof. Jens Z. Pedersen, Prof. Luciano Saso Title of diploma thesis: Biological activity of antioxidant compounds in L-6 myoblasts from rat skeletal muscles. This work was carried out in the laboratory of physiology at University Roma Tre in Rome, under the supervision of Prof. Sandra Incerpi. The team of this laboratory studies the antioxidants of different structures and evaluates their ability to prevent the production of reactive oxygen species and oxidative stress. The reactive oxygen and nitrogen species play an important role as regulatory mediators of physiological responses. However, they can be also very harmful because they can damage DNA and proteins, cause peroxidation of lipids and injure other biomoleculs. In my work, I tested the antioxidant effect of different polyphenols (baicalein, 5,6-dihydroxyflavone, negletein, mosloflavone, gallic acid and its derivative) in the cell culture (L-6 myoblasts from rat skeletal muscles) and in the in vitro test. The state of oxidative stress was induced by cumene hydroperoxide (generator of cumene hydroperoxide...
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The role of Wnt pathway in nociception
Pospíšilová, Blanka ; Skálová, Lenka (advisor) ; Pávek, Petr (referee)
Charles University in Prague University Roma Tre Faculty of Pharmacy in Hradec Králové Department of Biology Department of Biochemical Sciences Candidate: Veronika Staňková Supervisor: PharmDr. Iva Boušová, Ph.D. Prof. Sandra Incerpi, Prof. Jens Z. Pedersen, Prof. Luciano Saso Title of diploma thesis: Biological activity of antioxidant compounds in L-6 myoblasts from rat skeletal muscles. This work was carried out in the laboratory of physiology at University Roma Tre in Rome, under the supervision of Prof. Sandra Incerpi. The team of this laboratory studies the antioxidants of different structures and evaluates their ability to prevent the production of reactive oxygen species and oxidative stress. The reactive oxygen and nitrogen species play an important role as regulatory mediators of physiological responses. However, they can be also very harmful because they can damage DNA and proteins, cause peroxidation of lipids and injure other biomoleculs. In my work, I tested the antioxidant effect of different polyphenols (baicalein, 5,6-dihydroxyflavone, negletein, mosloflavone, gallic acid and its derivative) in the cell culture (L-6 myoblasts from rat skeletal muscles) and in the in vitro test. The state of oxidative stress was induced by cumene hydroperoxide (generator of cumene hydroperoxide...
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