Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Author: Marcela Vaisová Supervisor: Lucie Křivčíková Title of diploma thesis: Effect of tyrosol on minimal inhibitory concentration of antimycotics Previous studies concerning quorum sensing in Candida albicans discovered two quorum sensing molecules, farnesol and tyrosol. Tyrosol has been formerly confirmed as a substance which shortens the lag phase of Candida albicans cell cycle and stimulates its conversion to the hyphal form, farnesol stimulates blastospores. This work covers the effect of tyrosol combined with two antimycotics, fungistatic fluconazole and fungicidal amphotericine B. Candida albicans strains used in the experiment were mostly isolated from samples taken from patients suffering from variously located Candida albicans infections and four laboratory standard strains were also used. The method used for the experiment was a modified broth dilution method using combinations of both tyrosol and antimycotics various concentrations, which were prepared by two-fold dilution. Using amphotericine B, 28 strains did not show any difference in MIC compared with the control sample, 6 strains showed mildly higher MIC and 3 strains shower mildly lower MIC. Using fluconazole, 24...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Study program: Specialist in Laboratory Methods Candidate: Bc. Lucie Davidová Supervisor: PharmDr. Ondřej Janďourek, Ph.D. Title of diploma thesis: In vitro screening of novel potentially active antibacterial compounds. The diploma thesis deals with the antibiotics policy and is focused on resistance and mechanism of action of the antibiotics. A great attention is also paid to novel substances testing in terms of their impact on the selected bacteria. Theoretical part of thesis describes the system of antibiotic policy including its structure and organization in the Czech Republic. It also briefly summarizes this issue globally as well as individual organizations involved in the organization of the antibiotic policy. The theoretical part is also concerned with the history, classification, effects and significance of the antibiotics. A particular attention is paid to the chapter describing the macromolecular mechanism of action of 2 selected groups of substances (antibiotics) on bacteria, including physiological functions of some bacterial structures. And, last but not least, the theoretical part summarizes principles of bacterial resistance including its origin and development. The diploma thesis...

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...

Charles University, Faculty of Pharmacy, Hradec Králové Field of study: Specialist in laboratory methods Author: Bc. Tomáš Hampl Supervisor: PharmDr. Ondřej Janďourek, Ph.D. Title of diploma thesis: Mycobacterium tuberculosis as causative agent of grave illness The aim of this diploma thesis is to deal with tuberculosis, describe the pathogen, taxonomic classification, describe the origin, course and clinical manifestations of the disease, diagnosis, including laboratory diagnostics, epidemiology of tuberculosis, treatment and prevention. Tuberculosis (TBC) is one of the deadliest infectious diseases in the world caused by bacterium M. tuberculosis in humans. TBC is sometimes referred as a disease of the past, but the recent rise in cases of the disease, and especially the spread of forms of drug-resistant TBC, makes the disease very dangerous. Currently, a major problem is the alarming rise in resistant and multi-resistant tuberculosis, which is not counteracted by commonly used first-line drugs. Therefore, there is a constant emphasis around the world on discovering and testing new potential antituberculotics that could help patients with resistant and multidrug-resistant TBC. In the experimental part, the thesis deals with screening of antimycobacterial activity of potential antituberculotics...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Study program: Specialist on Laboratory Methods Autor: Bc. Marcela Vízková Supervisor: PharmDr. Ondřej Janďourek, Ph.D. Title: In vitro screening of novel potentially active antibacterial compounds III Since the discovery of penicillin, antibiotics have become part of modern therapeutic approaches. But the global spread of resistance makes their future uncertain. It is necessary to find new antibacterial substances useful in practice. As resistance is a global threat, the theoretical part deals with this issue. In addition to resistance, the theoretical part also briefly describes the antibiotics used, including new antibiotics, which were recently registered or likely to be registered. For selected groups, their mechanism of action is described in detail. Sensitivity should be determined to correctly indicate the antibiotic treatment. For this reason, the theoretical part also describes methods by which the sensitivity of a microbe to an antibiotic can be determined qualitatively or quantitatively. The theoretical part does not neglect the history of the development of antibacterial substances and familiarization with antibiotic policy in the Czech Republic. This diploma thesis is based on...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Study program: Specialist in Laboratory Methods Candidate: Bc. Aneta Janků Supervisor: PharmDr. Ondřej Janďourek, Ph.D. Title of diploma thesis: In vitro screening of novel potentially active antimycotic compounds III Key words: yeasts, moulds, antimycotics, mechanism of action of polyenes, microdilution broth method, MIC - minimal inhibition concentration. The main topic of this diploma thesis is in vitro screening of new, potentially active antifungal compounds. The introductory chapter in the theoretical part of the work is devoted to the epidemiology of fungal infections. The classification of micromycetes and the characteristics of the fungal eukaryotic cell are given below, focusing on composition of the plasma membrane and the cell wall. A separate chapter also deals with specific species of fungi and the diseases they cause. The main topic is drugs from the group of antifungals, where more attention is focused on polyene antifungals, their mechanism of action in detail and the emergence of resistance. An integral part are also the stages of development of new drugs and a list of novels developing antifungals. The final chapter describes the types of methods designed for in vitro evaluation...

Charles University Faculty of Pharmacy in Hradec Kralove Department of Biological and Medical Sciences Study program: Specialist on Laboratory Methods Author: Bc. Tereza Pchálková Supervisor: PharmDr. Ondřej Janďourek, Ph.D. Title of diploma thesis: In vitro screening of potential antimycobacterial compounds active against fast growing strains of Mycobacterium genus II Key words: Mycobacteria, Tuberculosis, Antituberculotics, Microdilution broth method, Minimum inhibition concentration Background: The aim of this diploma thesis has been screening of in vitro antimycobacterial activity of novel compounds against fast growing strains of the genus Mycobacterium (Mycobacterium smegmatis and Mycobacterium aurum). Another aim has been predicting the structure-activity relationships for tested compounds. Methods: The technique used for activity determination was microdilution broth method. The value of minimum inhibition concentration for each compound was determined. The evaluation was performed visually and spectrophotometrically using the Alamar blue indicator. Results: A total of 79 compounds were tested. 22 tested compounds showed significant activity against mycobacteria. The substances were sorted according to the similarity in chemical structure into 10 groups. From a chemical point of view, these...

The problem of high proportion of zeroes has long been an interest in data analysis and modeling, however, there are no unique solutions to this problem. The solution to the individual problem really depends on its particular situation and the design of the experiment. For example, different biological, chemical, or physical processes may follow different distributions and behave differently. Different mechanisms may generate the zeroes and require different modeling approaches. So it would be quite impossible and inflexible to come up with a unique or a general solution. In this dissertation, I focus on cases where zeroes are produced by mechanisms that create distinct sub-populations of zeroes. The dissertation is motivated from problems of chronic toxicity testing which has a data set that contains a high proportion of zeroes. The analysis of chronic test data is complicated because there are two different sources of zeroes: mortality and non-reproduction in the data. So researchers have to separate zeroes from mortality and fecundity. The use of mixture model approach which combines the two mechanisms to model the data here is appropriate because it can incorporate the mortality kind of extra zeroes. A zero inflated Poisson (ZIP) model is used for modeling the fecundity in <i> Ceriodaphnia dubia</i> toxicity test. A generalized estimating equation (GEE) based ZIP model is developed to handle longitudinal data with zeroes due to mortality. A joint estimate of inhibition concentration (ICx) is also developed as potency estimation based on the mixture model approach. It is found that the ZIP model would perform better than the regular Poisson model if the mortality is high. This kind of toxicity testing also involves longitudinal data where the same subject is measured for a period of seven days. The GEE model allows the flexibility to incorporate the extra zeroes and a correlation structure among the repeated measures. The problem of zero-heavy data also exists in environmental studies in which the growth or reproduction rates of multi-species are measured. This gives rise to multivariate data. Since the inter-relationships between different species are imbedded in the correlation structure, the study of the information in the correlation of the variables, which is often accessed through principal component analysis, is one of the major interests in multi-variate data. In the case where mortality influences the variables of interests, but mortality is not the subject of interests, the use of the mixture approach can be applied to recover the information of the correlation structure. In order to investigate the effect of zeroes on multi-variate data, simulation studies on principal component analysis are performed. A method that recovers the information of the correlation structure is also presented. / Ph. D.

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...

Charles University Faculty of Pharmacy in Hradec Kralove Department of Biological and Medical Sciences Study program: Specialist on Laboratory Methods Author: Bc. Lenka Králíková Supervisor: PharmDr. Ondřej Janďourek, Ph.D. Title of diploma: Atypical mycobacteria as causative agents of human diseases The aim of this diploma thesis is to summarize the history of mycobacteria, their classification, epidemiology of nontuberculous infections, clinical significance, diagnosis, resistance, and treatment options. Within the epidemiology, the way of managing tuberculosis and non-tuberculous mycobacterioses is compared. Mycobacterioses are rare diseases, which incidence is gradually increasing with the increasing number of immunocompromised patients. Based on the spot of infection, we divide them into lymphadenitis, skin lesions, lung disease and disseminated infections. The most frequently isolated pathogens are Mycobacterium avium complex, Mycobacterium ulcerans, Mycobacterium abscessus, Mycobacterium marinum and Mycobacterium kansasii. The experimental part is devoted to the antimycobacterial activity of newly synthesized compounds, which could find application in the future in the treatment of tuberculosis and other mycobacterioses. Substance testing was performed on strains of the genus Mycobacterium...