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Entry of bacteria Mycobacterium bovis BCG into B lymphocytes
Šamajová, Marianna ; Konečná, Klára (advisor) ; Kubelková, Klára (referee)
Charles University Faculty of Pharmacy in Hradec Králové Study program: Pharmacy Author: Marianna Šamajová Supervisor: RNDr. Klára Konečná, Ph.D. Consultant: plk. gšt. doc. RNDr. Zuzana Kročová, Ph.D. Title of diploma thesis: Entry of bacteria Mycobacterium bovis BCG into B lymphocytes Background: The objective of this work was to evaluate the entry of bacterium Mycobacterium bovis BCG into B lymphocytes and the role of selected receptors in this process. Methods: Peritoneal cell suspensions with unblocked and/or blocked receptors on BALB/c mouse B lymphocytes we infected by bacterium M. bovis BCG-GFP unopsonized and/or opsonized by fresh murine serum ("complement") or immune serum ("antibodies"). Using flow cytometry we evaluated the entry of bacterium M. bovis BCG-GFP into B lymphocytes and their subpopulations B1a, B1b and B2. Results: M. bovis BCG-GFP actively enters into B lymphocytes. Depending on the subpopulation, it most infects B1a, less B1b and at least B2 lymphocytes. Only the subpopulation B2 responds significantly to the opsonization by complement. Opsonization by antibodies had no significant effect on the infection. Entry into CD19+ cells is mediated through the BCR receptor, especially in subpopulations B1a and B1b. Under the opsonized conditions, the CR1/2 complement receptor is...
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Nanoparticle encapsulation of bacterial antigen for vaccination purposes
Nemeškalová, Alžběta ; Jílek, Petr (advisor) ; Konečná, Klára (referee)
Author: Alžběta Nemeškalová Title: Nanoparticle encapsulation of bacterial antigen for vaccination use Charles University in Prague, Faculty of Pharmacy in Hradec Králové Study program: Pharmacy Shigellosis, an infectious diarrheal disease caused by Shigella spp., is currently a major health concern in developing countries and there is still no vaccine available. Non-living vaccines seem to be the safest option: our approach is therefore based on using Outer membrane vesicles (OMVs) as a subunit vaccine. However, mucosally administered OMVs are not capable of inducing as appropriate protection and a suitable adjuvant must be added to modify the immune response. Polymeric nanoparticles (NP) were studied in this work as mucosal adjuvats. The aim of this project was to prepare particles based on a foodborne protein (Protein P, under patent). Apart from simple proteic particles, two different ligands (Compound A, Compound B) were used to modify the particle surface, which resulted in four different formulations (P, P-A, P-B, P-A-B). We loaded these NPs with OMVs and described their properties, such as size, polydispersity and Z-potential. Their capability to encapsulate and carry the bacterial antigens was also determined. Furthermore, a method of dissolving proteic NP was developed in order to...
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Study of interspecies differences in plasma protein binding of 18F-labeled radiopharmaceuticals
Pašková, Barbora ; Konečná, Klára (advisor) ; Jílek, Petr (referee)
Mgr. Barbora Pašková The study of antimycotic activity of newly synthetized substances Rigorous thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Study program: Pharmacy Background: The aim of this rigorous thesis was to evaluace the antimycotic activity of 77 substances prepared at the Department of Inorganic and Organic Chemistry, Faculty of Pharmacy in Hradec Králové of Charles University in Prague. Method: The substances were tested by the microdilution broth method and all substances were tested at eight strains of yeasts and filamentous fungi. The tested substances were divided into five groups according to their chemical structure to salicylanilide carbamates, sulfoneamide ureas - derivates of sulphamethoxazole, sulphonamide imidazolidinetriones - derivates of sulphamethoxazole, derivates of isoniazide and substances with disunited structure. Results: The highest antimycotic activity was discovered in group of salicylanilide carbamates. There were tested 28 derivates of this group, from which 18 of them showed antimycotic activity. On the other hand, substances from group of derivates of isoniazide showed no antimycotic activity. The most susceptible strain was Trichophyton mentagrophytes and the most resistant was strain Aspergillus fumigatus. Conclusions: To...
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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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Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants
Kollárová, Nikola ; Janďourek, Ondřej (advisor) ; Konečná, Klára (referee)
Candidate: Nikola Kollárová Title of diploma thesis: Detection of Sap2 in the secretome of Candida albicans cell wall and secretory mutants Charles University, Faculty of Pharmacy in Hradec Králové, Department of Biological and Medicinal Sciences Complutense University of Madrid, Faculty of Pharmacy, Department of Microbiology II Study program: Pharmacy Backgound: The aim of this diploma thesis was to search for C. albicans proteins involved in the secretion of the secreted aspartyl proteinase 2 enzyme (Sap2) evaluating the ability to degrade BSA (bovine serum albumin) as a source of nitrogen in several cell wall and secretory mutants of C. albicans. The work was carried out at the Department of Microbiology II, Faculty of Pharmacy, Complutense University of Madrid. Methods: The supernatant samples of several Candida albicans mutants were tested by SDS-PAGE electrophoresis and stained. Bands corresponding to BSA were observed and compared to controls. The other method was counted with 96-well plate. Results: The correlation between optical density and degradation of BSA was observed. Some mutants with disability to degrade BSA were found in a pilot screening of the ability to degrade BSA using 96-well plate method. That fact was confirmed by SDS-PAGE electrophoresis. C. albicans mutants showing...
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Microbiological diagnostics of causative agents of infectious diseases associated with central venous catheter insertion
Hošková, Miroslava ; Konečná, Klára (advisor) ; Janďourek, Ondřej (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Subject of study: Specialist in laboratory methods Student: Bc. Miroslava Hošková Supervisor: RNDr. Klára Konečná, Ph.D. Title: Microbiological diagnostics of causative agents of infectious diseases associated with central venous catheter insertion Background: The purpose of the thesis is to provide a comprehensive insight into the issue of microbiological diagnosis of agents that cause central venous catheter related infectious processes. The primary objective of the thesis is to evaluate the frequency of occurrence of individual microbiological agents and draw a comparison between published reports and empirical findings. The secondary objective of the thesis is to consider the extent of central venous catheter microbial colonization, i.e., in how many cases of positive culture cultivation, their contamination or significant colonization have been determined, and eventually whether system catheter infection has been proved in correlation with the results of blood culture. Methods: One hundred and seventy-one central venous catheters from different inpatient departments were delivered to the microbiological section of hospital Nemocnice Boskovice s. r. o. for culture examination have been...
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Effectiveness of disinfectants for hygienic hand disinfection against clinically important strains of enterococci
Malíková, Martina ; Janďourek, Ondřej (advisor) ; Konečná, Klára (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Study program: Bioanalytical laboratory diagnostics in health care Candidate: Bc. Martina Malíková Thesis supervisor: PharmDr. Ondřej Janďourek, Ph.D. Consultant: RNDr. Irena Hanovcová, CSc. Title of diploma thesis: Effectiveness of disinfectants for hygienic hand disinfection against clinically important strains of enterococci Alcohol-based hand disinfectants for hygienic hand disinfection are a key tool for the control of nosocomial infections worldwide. The aim of this study was to verify the effectiveness of these products, which are used in healthcare facilities, against vancomycin-sensitive and vancomycin-resistant enterococci isolated from clinical materials. Testing of the efficacy of disinfectants was performed according to the Czech technical standard ČSN EN 1040 - Chemical disinfectants and antiseptics - Quantitative test using suspension to determine the basic bactericidal effect of chemical disinfectants and antiseptics - Test method and requirements (phase 1). We used the dilution method with a neutraliser for our testing. There were used a total of 35 strains of enterococci to determine efficacy, 12 of which were vancomycin sensitive (four E. faecium strains and eight E. faecalis...
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Microbiological diagnostics of causative agents of infectious diseases associated with central venous catheter insertion
Hošková, Miroslava ; Konečná, Klára (advisor) ; Janďourek, Ondřej (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Subject of study: Specialist in laboratory methods Student: Bc. Miroslava Hošková Supervisor: RNDr. Klára Konečná, Ph.D. Title: Microbiological diagnostics of causative agents of infectious diseases associated with central venous catheter insertion Background: The purpose of the thesis is to provide a comprehensive insight into the issue of microbiological diagnosis of agents that cause central venous catheter related infectious processes. The primary objective of the thesis is to evaluate the frequency of occurrence of individual microbiological agents and draw a comparison between published reports and empirical findings. The secondary objective of the thesis is to consider the extent of central venous catheter microbial colonization, i.e., in how many cases of positive culture cultivation, their contamination or significant colonization have been determined, and eventually whether system catheter infection has been proved in correlation with the results of blood culture. Methods: One hundred and seventy-one central venous catheters from different inpatient departments were delivered to the microbiological section of hospital Nemocnice Boskovice s. r. o. for culture examination have been...
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