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Pistacia lenstiscus essential oil: in vitro testing of cytotoxicity and anti-inflammatory activity
Elstnerová, Marie ; Szotáková, Barbora (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Marie Elstnerová Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Title of diploma thesis: Pistacia lentiscus essential oil: in vitro testing of cytotoxicity and anti-inflammatory activity This project, which has been initiated in cooperation with University of Sassari (Italy), is aimed to find out more details about biological activity of essential oil from Pistacia lentiscus. We want to contribute to the decision, if this essential oil can be used as a part of oral health products. The essential oil was prepared from leaves of wild occurring plants growing in North Sardinia. The cytotoxicity and anti-inflammatory activity of not only the pure essential oil but also of the lecithin nanoemulsion with P. lentiscus oil and Tween 80 nanoemulsion with P. lentiscus oil and moreover, the two most abundant terpenes in the essential oil, (−)-α-pinene and (−)-terpinen-4-ol, were tested. The cytotoxicity was measured by the colorimetric assay using tetrazolium salt WST-1 on four oral cell lines (gingival fibroblasts, periodontal ligament fibroblasts, dysplastic oral keratinocytes and primary gingival keratinocytes). The viability of human oral cells was the most diminished by lecithin nanoemulsion, pure...
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Degradation of hyaluronic acid in the skin and percutaneous absorption of exogenous hyaluronic acid through the skin
Exnerová, Andrea ; Skálová, Lenka (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Andrea Exnerová Supervisor: Mgr. Petra Žádníková, Ph.D. prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Degradation of hyaluronic acid in the skin and percutaneous absorption of exogenous hyaluronic acid through the skin Up to half of the body's total hyaluronic acid (HA) content is present in the skin. HA acts as an organizer of the extracellular matrix and is involved in maintaining skin hydration and elasticity. In the last decade, several important findings have been added to the field of HA degradation in the skin, and one of the aims of this thesis was to create a comprehensive overview. Degradation of HA in the dermis was elucidated by the discovery of the essential role in the degradation of HA in dermal fibroblasts played by the KIAA1199 protein. In the epidermis, the lysosomal enzyme hyaluronidase 1 is overexpressed in the stratum granulosum just below the stratum corneum (SC). It is uncertain how HA is internalized into the keratinocytes or whether only intracellular HA is degraded in this way. Epidermal degradation of HA remains elusive. HA is part of a range of topically applied products on the skin, mainly due to its unique moisturizing properties, but it is not yet...
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Effect of monoterpene citral on the expression of detoxification enzymes in HepaRG cells
Švábová, Tereza ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Tereza Švábová Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Consultant: Mgr. Michaela Šadibolová Title of diploma thesis: Effect of monoterpene citral on the expression of detoxification enzymes in HepaRG cells Citral is an acyclic monoterpene, which is mainly produced by higher plants. It is one of the many secondary metabolites, which serve the plant as detoxifying factors, morphoregulators and attractants for pollinators or defense substances. Besides other things, it has numerou positive effects such as antibacterial, anti-inflammatory or antitumor, but it may be toxic, especially for the liver, if used incorrectly. The main goal of this thesis was to find out the effect of citral on selected biotransformation and antioxidant enzymes in HepaRG cells. Differentiated HepaRG cells were incubated with 10 µM rifampicin, 10 µM β-naphthoflavone, 30 µM oltipraz, 10 µM citral, 30 µM citral and 100 µM citral for mRNA expression detection for 12 and 24 hours and for protein expression evaluation for 24 and 48 hours. At the mRNA level, rifampicin induced CYP3A4 at 12- and 24-h incubation, β-naphthoflavone induced CYP1A2 at 12- and 24-h incubation and aldo-ketoreductase (AKR1C) at 24-h incubation,...
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Identification of new virulence factors in intracellular pathogen Francisella tularensis
Daňková, Věra ; Szotáková, Barbora (advisor) ; Krejsek, Jan (referee) ; Prokešová, Ludmila (referee)
ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Věra Daňková Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Title of Doctoral Thesis: IDENTIFICATION OF NEW VIRULENCE FACTORS IN INTRACELULLAR PATHOGEN FRANCISELLA TULARENSIS This dissertation thesis is focused on the role of bacterial tetratricopeptide repeat-like (TPR-like) proteins in the pathogenesis of infection. As a model organism we have chosen highly virulent intracellular bacterium Francisella tularensis (F. tularensis), whose mechanism of pathogenesis is not completely described. In the first part of dissertation thesis we took advantage of bioinformatic methods and identified three genes (FTS_0201, FTS_1680, and FTS_0778) with predicted TPR-like domains. Mutants defective in protein expression were prepared by TargeTron insertion mutagenesis. Prepared mutant strains were used for studying the role of selected proteins in pathogenicity and immunogenicity of F. tularensis subsp. holarctica strain employing in vivo and in vitro models and further for studying the involvement of these proteins in stress tolerance. Our results showed that the FTS_1680 protein is required for intracellular replication and full virulence of bacterium. We also described impaired ability...
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Effect of albendazol on the expression of selected enzymes in Haemonchus contortus
Božíková, Martina ; Matoušková, Petra (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Martina Božíková Supervisor: doc. Ing. Petra Matoušková, Ph.D. Title of diploma thesis: Effect of albendazol on the expression of selected enzymes in Haemonchus contortus Haemonchus contortus is the dominant gastrointestinal pathogen of small ruminants that can cause weakening or even death of the host organism. Losses are mainly recorded in young animals, which subsequently reflects on the farm economy itself. Therefore, prevention and therapy still play the most important role. On the other hand, the increased ability of worms to survive this treatment poses a serious problem. One of the possibilities that might be causing this problem is the increased activity of enzymes responsible for the metabolism of xenobiotics. The aim of this master's thesis was to investigate the effect of albendazole (ABZ) on the gene expression of selected biotransformation enzymes; UDP-glycosyltransferases (UGTs). For this purpose, two strains of H. contortus - anthelmintics-susceptible ISE (Inbred Susceptible Edinburgh) and anthelmintics-resistant WR (White River) were selected. One group of the parasites was cultivated for 12 hours in a media containing albendazole, the other group, which served as control, was...
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Anthelmintics in plants - uptake, biotransformation and transcriptional response
Syslová, Eliška ; Szotáková, Barbora (advisor) ; Petrová, Šárka (referee) ; Tůmová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate Mgr. Eliška SYSLOVÁ Supervisor prof. Ing. Barbora SZOTÁKOVÁ, Ph.D. Consultant RNDr. Radka PODLIPNÁ, Ph.D. Title of Doctoral Thesis: Anthelmintics in Plants - Uptake, Biotransformation and Transcriptional Response The use of anthelmintics, drugs against parasitic worms, is almost necessary in veterinary medicine today. However, anthelmintics may enter the environment with the excrements of treated animals, and influence non-target organisms there. Plants which can receive, biotransform or accumulate these substances, are an integral part of the environment. Biotransformation of anthelmintics may affect the capacity of the antioxidant system, interfere with endogenous plant metabolism or polyphenol biosynthesis where plants use the same detoxifying enzymes. The aim of this thesis is to study the influence of the most commonly used anthelmintics albendazole (ABZ), fenbendazole (FBZ), flubendazole (FLU) and ivermectin (IVM) on plants. To study their uptake by plants, their biotransformation in plants and transcriptional response of plant cells to their presence. The results of the dissertation include the finding that model systems of cell suspensions and in vitro regenerants of Plantago lanceolata...
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Anthelmintics in plants - uptake, biotransformation and transcriptional response
Syslová, Eliška ; Szotáková, Barbora (advisor) ; Petrová, Šárka (referee) ; Tůmová, Lenka (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate Mgr. Eliška SYSLOVÁ Supervisor prof. Ing. Barbora SZOTÁKOVÁ, Ph.D. Consultant RNDr. Radka PODLIPNÁ, Ph.D. Title of Doctoral Thesis: Anthelmintics in Plants - Uptake, Biotransformation and Transcriptional Response The use of anthelmintics, drugs against parasitic worms, is almost necessary in veterinary medicine today. However, anthelmintics may enter the environment with the excrements of treated animals, and influence non-target organisms there. Plants which can receive, biotransform or accumulate these substances, are an integral part of the environment. Biotransformation of anthelmintics may affect the capacity of the antioxidant system, interfere with endogenous plant metabolism or polyphenol biosynthesis where plants use the same detoxifying enzymes. The aim of this thesis is to study the influence of the most commonly used anthelmintics albendazole (ABZ), fenbendazole (FBZ), flubendazole (FLU) and ivermectin (IVM) on plants. To study their uptake by plants, their biotransformation in plants and transcriptional response of plant cells to their presence. The results of the dissertation include the finding that model systems of cell suspensions and in vitro regenerants of Plantago lanceolata...
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Toxicity of pulegone and menthofuran and their effect on miRNA expression in human liver slices
Lancošová, Bibiána ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bibiána Lancošová Supervisor: doc. PharmDr. Boušová Iva, Ph.D Title of diploma thesis: Toxicity of pulegone and menthofuran and their effect on miRNA expression in human liver slices Monoterpenes are secondary plant metabolites, characterised by their odour, which have a positive impact on human body and are used not only in cosmetic and food industry, but also in medication. R-pulegone (PUL) and its metabolite R-menthofuran (MF) are the most represented in plants of genus Mentha (Mint), with higher doses of these substances exerting certain risk of hepatotoxicity. The aim of this thesis was to assess the concentration, at which the viability of liver cells decreases and also to observe the changes in individual microRNA expression as a result of influence of PUL, MF and paracetamol, as the reference compound. Five samples of human liver obtained from patients of both genders and at age of 57 to 73 years represented biological material for hepatotoxicity testing. The samples were stored in Euro-Collins solution and subsequently cut into precision-cut liver slices with thickness of 150- 170 µm, placed to 12-well plate and put into incubation medium with the tested substances for 24 hours. The...
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Effect of selected sesquiterpenes on carbonyl reductase 1 activity in vitro
Mazurková, Veronika ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Veronika Mazurková Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of rigorous thesis: Effect of selected sesquiterpenes on carbonyl reductase 1 activity in vitro Sesquiterpenes, the main components of plant essential oils, are integral parts of spices, traditional food and beverages. They are used in folk medicines, pharmaceutical industry as well as in cosmetics. Several sesquiterpenes possess interesting biological activities but they could interact with concurrently administered drugs via the modulation of activity and/or expression of drug-metabolizing enzymes. Enzyme carbonyl reductase 1 (CBR1) is one of the enzymes participating in the metabolism of a number of endogenous and xenobiotic compounds. One of the CBR1 substrates is chemotherapeutic agent doxorubicin, which is metabolized by this enzyme to the less effective metabolite doxorubicinol, which is responsible for the doxorubicin cardiotoxicity. The aim of this rigorous thesis was to find out the ability of five selected sesquiterpenes (i.e. valencene, β-caryophyllene oxide, α-humulene, cis-nerolidol and trans-nerolidol) to affect the activity of CBR1 in vitro. The effect of studied substances on the activity of this enzyme was...
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