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Analysis of protein cargo of extracellualr vesicles isolated from the yeast Candida albicans
Hlubučková, Lucie ; Konečná, Klára (advisor) ; Janďourek, Ondřej (referee)
Charles University Faculty of Pharmacy in Hradec Králové Study program: Pharmacy Candidate: Lucie Hlubučková Consultant: RNDr. Klára Konečná, Ph.D. Title of thesis: Analysis of protein cargo of extracellular vesicles isolated from the yeast Candida albicans Backgroung: The aim of this diploma thesis was to analyze the protein cargo carried in extracellular vesicles released from the yeast Candida albicans (C. albicans), which is one of the most important mycotic agens. Extracellular vesicles (EVs) are utilized as "transport vehicles", for the delivery of effector molecules into extracellular milieu. These molecules and primarilly proteins can play different roles in host-pathogen interactions. Proteins isolated from EVs and identified by proteomic approach were sorted into categories according to their molecular function and localization for the purpose of finding out, which proteins are predominantly distributed via extracellular vesicles into extracellular space after induction of nutrition starvation. Analysis of EVs protein cargo with focus on virulence factors could extend the knowledge about extracellular vesicles and their potential role in pathogenesis. Methods: Chosen C. albicans yeast strain was a clinical isolate strain isolated from a premenopausal women suffering from recurrent...
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Biofilms and human body
Chlupáčová, Adéla ; Jílek, Petr (advisor) ; Konečná, Klára (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Adéla Chlupáčová Supervisor: PharmDr. Petr Jílek, CSc. Supervisor specialist: doc. MUDr. Pavel Čermák, CSc. Title of Diploma thesis: Biofilms and human body Subtitle: The occurrence of biofilm producers on central venous catheters and the effect of xanthohumol on the biofilm of selected clinical isolates Background: The aim of the work was to show the frequency of appearance of biofilm on the catheters, which leads to serious health complications. In the second part of the thesis we investigated the biofilm resistance of common biofilm agents such as Staphylococcus aureus and Enterococcus faecalis to natural substances, namely xanthohumol. Methods: Part of the thesis which monitors the occurrence of biofilm producers on catheters was processed using tables and charts in Microsoft Excel. To testing xanthohumol were used laboratory techniques such biofilm staining with crystal violet in microtiter plate, the minimal inhibitory concentrations of xanthohumol were determined using the broth microdilution method, the minimal bactericidal concentrations were determined on Columbia blood agar, inhibition of bacterial release from biofilm was monitored in microtiter plates with...
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Hotel with sports background
Karásek, Petr ; Konečná, Klára (referee) ; Müller, Jan (advisor)
The main subject of the diploma thesis is a new construction of the hotel with sporting facility in Kuřim. The Thesis is done in a form of a project documentation with all requests by the law and regulations included. The project is Composed of two buildings, a hotel and the sporting hall. The building is Located in slope in direction to south direction. The hotel building have four Storeys. The whole disposition solution of both buildings is connected. In a first floor is located the hotel restaurant, staff background, wellness and fitness Secon and third floor is supposed to be Used Mainly for accomodating of Sportsmans, and the last fourth floor is for holding meetings and conferences. The main Supporting structure is done with as monolitic frame construction of beams, columns and continuous slabs. All roofs are done as mono-layer roofs. First floor of the sporting hall is used by people during the sporting activity. This area is composed of changing rooms with a social background and other spaces for the proper function of the hall. The main Supporting construction of the hall is designed as glued lamellar beams supported by columns.
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Ames test in the drug development
Klaučová, Martina ; Pávek, Petr (advisor) ; Konečná, Klára (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Martina Klaučová Supervisor: prof. PharmDr. Petr Pávek, Ph.D. Consultant: PharmDr. Ivona Pávková, Ph.D. Diploma thesis title: Ames test in the drug development Background: Thesis objective is the determination of potential genotoxicity of newly developed drugs within primary testing and the introduction of the Ames microfluctuation test which can be used in common laboratory conditions. Methods: I used commercially supplied kit based on the principles of Ames test which detects reverse mutation through colour changes of the samples using bacterial strains S. typhimurium. At first I had to study literary sources and then I could design the procedures of the Ames microfluctuation test, preparation of the chemicals and storage of the strains which are optimal for all laboratories. Results: The drug samples T6445 and T6447 with 30 µM concentration tested by metabolic activation S9 on bacterial strain ST TA 98 show genotoxicity. The sample UOCHB1 with 30 µM concentration tested without activation shows possible genotoxicity on both strains ST TA 98 and ST TA 100. Other samples do not show any toxicity. I used 3 different procedures during the designation of assay. The most suitable version of the...
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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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Determination the mechanism of entry F. tularensis into B lymphocytes
Hadámková, Barbora ; Konečná, Klára (advisor) ; Janďourek, Ondřej (referee)
Barbora Hadámková Determination the mechanism of entry F. tularensis into B lymphocytes Diploma thesis Charles University, Faculty Of Pharmacy in Hradec Králové Study program: Pharmacy Background: Besides processing the research with basics knowledge of the problem, the main aim of the study was the analysis of mechanism of entrance of intracellular bacteria Francisella tularensis into B cells. Methods: The B cells, which we obtained through peritoneal lavage from mice Balb/c, we blocked using antibodies individual complement receptors, B cell receptor and Fcƴ receptor. The population of the cells was infected by bacteria F. tularensis LVS/GFP opsonized by complement and/or by antibodies. Using flow cytometry we measured the percentage of infection of individual subpopulations of B cell B1a, B1b and B2 and we evaluated the influence of blocking and opsonization on the infection. Results: From the measured data, we can say that the percentage of infected B cells after infection by F. tularensis opsonized by complement is increased. This increase was more distinct in subtype of B cells B1b and B2. On the other hand, the opsonization F. tularensis by antibodies did not affect the infection. We also found out, that blocking of Fcƴ receptor has decrease the infection, if we used for infection of B cells...
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Meningococcal infection
Malá, Markéta ; Vejsová, Marcela (advisor) ; Konečná, Klára (referee)
7 2. ABSTRACT Title of Bachelor thesis: Meningococcal infections Student: Markéta Malá Supervisor: Mgr. Marcela Vejsová, Ph.D. Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Backround: The aim of this bachelor thesis was to summarize the findings about meningococcal infections and their originator - Neisseria meningitidis. Further this bachelor thesis should provide infomation about progression in meningococcal vaccines development, treatment and diagnostics. Main findings: N. meningitidis spreads into the human body from nasopharynx, through the epithelium it travels into the bloodstream. Severity of meningococcal infection depends on a strain of meningococcus and on strenght of immune system on the side of a host. Meningococcus has got efective tools for invading human body, they are called virulence factors. Quick recognition of clinical signs and rapid confirmation of infection originator are important when the infection breaks out. Nowadays we can prevent meningococcal infection by effective vaccination - combining meningococcal tetravalent conjugate vaccine and meningococcal serogroup B vaccine. Conclusions: Infections caused by N. meningitidis varies from ordinary nasopharyngitis to bacteremia and severe acute meningitis. N. meningitidis...
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Advanced methods of antimicrobial activity screening
Dolinská, Dagmar ; Janďourek, Ondřej (advisor) ; Konečná, Klára (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Dagmar Dolinská Supervisor: PharmDr. Ondřej Janďourek, Ph.D. Work title: Advanced methods for testing antimicrobial activity Goal: The aim of the work in the theoretical part was to summarize methods of antimicrobial screening focusing on antimycotics and in the practical part to evaluate the antifungal activity of 47 tested substances using the microdilution broth method. Methods: The substances were tested using quantitative microdilution broth method on eight strains of yeast and fibrous fungi. These were Candida albicans, Candida krusei, Candida parapsilosis, Candida tropicalis, Aspergillus fumigatus, Aspergillus flavus, Absidia corymbifera and Trichophyton interdigitale. Results: The highest antimycotic activity was demonstrated by compound Ac-0a, where we recorded after 24 hours of incubation activity against all eight strains. However, it was not able to stop the growth of three strains after 48 hours of incubation, the strains concerned were Candida parapsilosis, Aspergillus fumigatus and Aspergillus flavus. Other samples which have shown activity were Cl-2, where activity was observed for Candida krusei, Aspergillus fumigatus and Trichophyton interdigitale strains, Cl-3 for...
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