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Western blot analysis of ICAM-1 and P-selectin protein expression in aorta of mice with high soluble endoglin levels.
Uhlířová, Barbora ; Rathouská, Jana (advisor) ; Konečná, Klára (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Barbora Uhlířová Supervisor: PharmDr. Jana Rathouská, Ph.D. Title of Diploma Thesis: Western blot analysis of ICAM-1 and P-selectin protein expression in aorta of mice with high soluble endoglin levels Background: Detection and subsequent evaluation of the expression of adhesion molecules ICAM-1 and P-selectin in aorta of two groups of transgenic mice differing by levels of human serum endoglin that were fed a special high fat and cholesterol diet. Methods: We used six-month-old female transgenic mice capable of expressing of human endoglin, which was able to be determined after cleavage in the circulation (Soluble endoglin - sENG) by the use of an ELISA assay. Therefore, we were able to divide the mice into two groups - an experimental group (high levels of sENG) and a control group (very low sENG levels). For 3 months, both groups were given a special high fat and cholesterol diet. Biochemical analysis (total cholesterol levels), and Western blot analysis of molecules ICAM-1 and P-selectin in the homogenate of mouse aortas were carried out. Results: Biochemical analysis of total cholesterol in the blood revealed statistically no significant differences in the measured values...
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Current methods in diagnostics, treatment and prevention of infection caused by methicilin resistant Staphylococcus aureus (MRSA)
Korejsová, Erika ; Konečná, Klára (advisor) ; Vejsová, Marcela (referee)
Candidate: Erika Korejsová Supervisor: Mgr. Klára Konečná, Ph.D. Consultant: MUDr. Marie Smolíková Title: Current methods in diagnostics, treatment and prevention of infection caused by methicilin resistant Staphylococcus aureus (MRSA) Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Study field: Medical Bioanalytics Staphylococcus aureus is a bacterium that is found in the nasal mucosa of healthy people (20-40%) and skin without causing disease. Under certain circumstances, such as damage or injury of skin, mucous membranes and in immuno-incompetent individuals, the MRSA infection can originate. Therefore, hospitalized patients are much more susceptible to this infection. The biggest dangers represent MRSA (methicillin-resistant Staphylococcus aureus) strains. These strains of Staphylococcus aureus are mostly multi-resistant, where in addition to resistance to oxacillin (methicillin), resistence to other antibiotik occurs as well. Treatment of these patients is problematic, because MRSA strains are extensively resistant and persist in the organism and in the environment. Their eradication is not simple. The patient's blood reinfection often occurs due to low levels of antibiotics. It means a considerable financial...
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Identification of membrane localized native peptides of intracellular bacteria Francisella tularensis
Hrebíková, Hana ; Konečná, Klára (advisor) ; Pávková, Ivona (referee)
Bc. Hana Hrebíková Identification of membrane localized native peptides of intracellular bacteria Francisella tularensis Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Specialist in Laboratory Methods Background: The main goal of this diploma thesis was develop new methods suitable for assume of concentration of peptides in biological sample and identification of membrane- associated proteins of bacteria Francisella tularensis (F. tularensis). Methods: Experimental development was tested on standard proteins like alfa kasein and BSA. These proteins had known concentration and they were treated by trypsine. The concentration of reached peptides were measured by modified and classic method of micro BCA (bicinchoninic acid protein assay). Method was applied on real bacterial sample of F. tularensis species strain FSC200. By cultivation of whole bacteria bacterial peptides were prepared. The cultivation was provided in different time intervals in solution, which contained trypsine. This method named shaving was described in literature. Results: The concentration of peptides was measured by classic micro BCA method. Peptides were separated by HPLC and identified by mass spectrometry. Identification of two bacterial peptides from protein named 17 kDa major membrane...
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Antimicrobial Additives and their use in top Coating Paint
Čechová, Petra ; Jílek, Petr (advisor) ; Konečná, Klára (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Petra Čechová Consultant: PharmDr. Petr Jílek, Csc. Title of Thesis: Antimicrobial additives and their use in top coating paint. Background: Evaluation of efficacy in antimicrobial photocatalytic paint designed by Synpo a.s. Pardubice Company (AMAc-1, AMAc-2, AMAc-3, AMAc-4, AMAc-5); Composition of paint unknown (we only known that paint include photocatalytic additives); Pathogen bacterial strains: E. coli, S. aureus, P. aeruginosa. Methods: Contact of microbes with antimicrobial photocatalytic paint: pre-activated with UVA-light, activated with UVA-light at the same time, non-activated with UVA-light. Main findings: Graphs show how number of CFU microbes is changing in time; Definition of efficacy (-k): negative value of tangent of line linear regression. Conclusions: Confirmation of hypothesis: grow of efficacy of antimicrobial photocatalytic paint after exposition UVA-light; Influence on fall of number microbial CFU of strain E. coli; Significant after UVA irradiation (activation); S. aureus: fluctuating of results, all the same definite fall of bacterial CFU after activation UVA-light; Most resistant to efficacy of antimicrobial paint was P. aeruginosa; Influence of...
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Susceptibility profile of biofilms of non-albicans Candida spp. to echinocandins
Pešková, Pavla ; Křivčíková, Lucie (advisor) ; Konečná, Klára (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Kralové Department of Biological and Medical Sciences Candidate: Pavla Pešková Supervisor: Ing. Lucie Křivčíková Diploma Thesis: Susceptibility profile of biofilms of non-albicans Candida spp. to echinocandins Yeasts of the genus Candida are one of the most frequent human fungal pathogens. Infections caused by them are related to a specific form of growth - biofilm (BF), which has increased their resistance to antifungal treatment. Since bloodstream infections caused by non-albicans Candida species are increasing, it is important to focus on their susceptibility characteristics. The main aim of our experiment was to examine the susceptibility profiles of BF produced by rare non-albicans Candida species to echinocandins. We tested 3 species of the genus Candida - C. lusitaniae, C. guilliermondii and C. krusei and 3 different echinocandins - anidulafungin (AND), caspofungin (CAS) and micafungin (MFG). Echinocandins have unique mechanisms of action. They inhibit the function of the enzyme β-1,3-glucan synthase. Disruption of its function leads to inhibition of β-1,3-glucan production, damage of fungal cell wall and loss of viability of the cell. In experimental part we used YNB medium and RPMI 1640 medium to grow Candida species BF and...
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Application of electromigration methods in the analysis of Francisella tularensis proteins
Žáková, Jitka ; Konečná, Klára (advisor) ; Jílek, Petr (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Kralove Department of Biological and Medical Sciences Degree program: Medical bioanalysis Field of study: Medical Laboratory Technician Author: Jitka Žáková Title: Application of electromigration methods in the analysis of Francisella tularensis proteins Bachelor thesis Theoretical part describes pathogenic bacterium Francisella tularensis (taxonomy, morphology, cultivation, mode of disease, sources of infection, epidemiology) and basic characterisation of electromigration methods (electrophoresis, izoelectric focusing, izotachophoresis, capillary electrophoresis). Experimental part contains methods and results of selected electromigration experiments (1DE SDS-PAGE, 2 DE SDS-PAGE). We chose the methods that prove to be usefull in generating data suitable for current publication in international journals, thus ilustrating the significance in analysis of F. tularensis proteome.
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Examination of blood for the infectious diseases for the transfusive medicine. Legislature of different countries of the world.
Dohnalová, Eva ; Konečná, Klára (advisor) ; Řeháček, Vít (referee)
Author: Bc. Eva Dohnalová Title: Blood-bank testing for infectious diseases Legislation of different countries of the world Supervisor-specialist: MUDr. Martin Šumpík Supervisor: Mgr. Klára Konečná Diploma thesis Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Study specialisation: Specialist in Laboratory Methods A list of most frequently detected infectious agents in blood transfusion service is presented in the first part of this thesis. The history, taxonomy, structure, possible ways of transmission and the most frequently selected diagnostics of detected causative agents are also described in this thesis. In the second part of the thesis, there is described one of the areas of transfusion medicine legislation - blood and blood products testing for infectious diseases. The legislation of some countries of the European Union (the Czech Republic, the Slovak Republic, Poland, Germany, Austria, the United Kingdom of Great Britain and Northern Ireland and France) and the United States of America and their mutual comparison is presented here as well.
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Production of secreted aspartic proteinases by pathogenic yeasts.
Šlajchová, Andrea ; Buchta, Vladimír (advisor) ; Konečná, Klára (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Andrea Šlajchová Supervisor: Doc. RNDr. Vladimír Buchta, CSc. Title of diploma thesis: Production of secreted aspartic proteinases by pathogenic yeasts The objective of this thesis was to identify the effect of farnesol as a quorum sensing molecule, which facilitates intracellular communication of Candida albicans, on the production of secreted aspartic proteinases (Sap). The theoretical section summarizes the morphology of Candida, the virulence factors, the illnessess cause by Candida and its treatment. A separate chapter focuses on secreted aspartic proteinases and on farnesol. The strains tested came from clinical isolates belonging to the Clinical Microbiology Institute of FN Hradec Kralove; two further strains were collected as well (10231 and 25188). The ATCC 10231 strain, which did not show farnesol production but the production of farnesol acid, was included in the test. Special media contained a source of bovine hemoglobin as the only source of nitrogen and had differing pHs. The strains were first pre-incubated to exogenous farnesol and later transfered to test media at various different concentrations of farnesol. The results were read after seven days of...
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. Changes in gene expression of B cell line Ramos RA-1 after infection with Francisella tularensis.
Matyášová, Simona ; Konečná, Klára (advisor) ; Kubelková, Klára (referee)
Francisella tularensis is a facultative intracellular bacterium causing disease tularemia of different species of mammals, including humans. It is able to infect phagocytotic as well as non-phagocytotic cells. We focused on the study of B cells, that role in the infection of F. tularensis have been, until recently, regarded as not significant. Over the last 10 years several studies showed important role of B cells in innate immunity to F.tularensis infection. The aim of this thesis was to idenify changes in the expression of genes that encode molecules of cytokines produced by B cells, after F. tularensis infection, due to stimulation with bacterial recombinant protein IglC and lipopolysaccharide. These changes were examinated on B cell line Ramos RA-1. Using reverse transcriptase polymerase chain reaction, we detected these cytokines: IFN-γ, IL-10 a TGF-β. Key words: Francisella tularensis, B cells, cytokines, reverse transcriptase polymerase chain reaction
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