Colistin is a last-resort antibiotic used to treat serious infections caused by Enterobacteriaceae and other multidrug resistant gram-negative bacteria. Recently discovered plasmid-borne colistin resistance, mediated by the mcr genes, poses a serious risk to colistin therapy. The aim of this diploma thesis was to map the occurrence of Enterobacteriaceae carrying the mcr-1 to 8 genes in hospitalized patients, travellers, prospective colistin-resistant clinical isolates and in a retrospective collection of Enterobacteriaceae using a combination of selective cultivation and qPCR. Isolates with a detected mcr gene were characterized by Whole-Genome Sequencing. The localization of mcr genes was determined and other resistance genes and plasmids were identified. Furthermore, the physiological profile of selected colistin- resistant Escherichia coli isolates was characterized. In the presence of a subinhibitory amount of colistin, a strain carrying the mcr-1 gene may be favored. Later, the mcr-9 gene was described and its occurence was subsequently tested retrospectively. Enterobacter spp. isolates carrying the mcr-9 gene were mostly colistin-sensitive but, in some cases, resistance was induced after exposure to sublethal doses of colistin. The results of the study show that the incidence of plasmid-mediated...

Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lucie Čermáková Supervisor: RNDr. Eva Novotná, Ph.D. Title of diploma thesis: The effect of acalabrutinib and ibrutinib on the efficacy of daunorubicin in cancer cells Leukemia presents malignant diseases of hematopoiesis, which essence is the malignant transformation of a hematopoietic stem cell at various levels of maturation and increased proliferative activity. Chemotherapy is the gold standard in the treatment of leukemia. One of the many treatments is the use of anthracycline chemotherapeutics, especially daunorubicin (DAU). Anthracyclines are widely used in clinical practice but have high cardiotoxic effects that limit their dosage. One of the main causes of side effects is the reduction of an anthracycline chemotherapeutic to the appropriate toxic metabolite, which accumulates in the heart. Carbonyl, reducing enzymes from the superfamily aldo-ketoreductase (AKR), and short-chain dehydrogenase/reductase (SDR) are involved in this reduction. At the same time, carbonyl reducing enzymes, has been shown to be involved in the mechanisms that cause tumor cells to be resistant to anthracyclines, thereby reducing the inhibition of the growth of these cells. In the diploma thesis we found that...

Molecular-epidemiological analysis of Mycobacterium tuberculosis strains isolated in the West-Bohemian Region of the Czech Republic including detailed characterisation of anti-tuberculosis drugs - resistant strains MUDr. Jana Amlerová Abstract Tuberculosis is contagious infectious disease that embodies significant epidemiological and clinical problem worldwide. Tuberculosis incidence differs considerably in various regions of the world but even the countries with low incidence engage strongly in epidemiology of tuberculosis. Tuberculosis belongs to one of the priorities of WHO, cooperation in this matter takes place on a global scale. Tuberculosis is a social illness; accordingly, the countries with high occurrence of tuberculosis are classified as developing countries. Mainly in Africa, there is the situation being complicated by coexistence of HIV. Generally, Europe represents a region with low incidence of tuberculosis. The Czech Republic is a country with the lowest incidence in the world with less than five new cases per 100 000 inhabitants every year. This situation is among others result of high-level tuberculosis surveillance and effective application of epidemiological arrangements based in legislation. This dissertation thesis examines several fields of tuberculosis, mainly focused on modern...

This thesis deals with evaluation of analytical and organoleptic properties of new PIWI varieties. The aim of the thesis is to describe and evaluate the suitability of selected PIWI varieties for cultivation in the Czech Republic winery. In the thesis fifty-two bottles of varieties and cuvée wines from the Czech Republic, Germany, the Netherlands, Denmark, Poland and Sweden are analysed, tasted, evaluated and briefly described. All nineteen tested varieties should be recommended to certain growing conditions in the Czech Republic. The top rated varieties with three or more samples are PIWI varieties of Cabernet Blanc, Solaris and Hibernal. All varieties proved the ability to produce on average up to above average harmonious wines which can be the basis for winemaking in organic viticulture in the Czech Republic.

Resistence population of pollen beattle on selected locations of Moravia and Silesia were observed during the 2017. Beattles were taked off in the field during spring and their sensitivity to insecticides active substances indoxacarb, chlorpyrifos, thiacloprid, tau-fluvalinate, cypermethrin and lambda-cyhalothrin were tested in laboratory. Tested using methods of IRAC – 11, 21, 25 and 27. Mortality of beattles at various concentrations of active substances was evaluated after 24 hours. Levels of resistance were changed for individual location.

Obesity, one of the most serious health problems of the 21st century, often occurs as a result of an imbalance between energy intake and energy expenditure. Dietary lipids play an important role in the development of obesity, partly because they represent the richest source of energy amongst all macronutrients. It is, however, not only the amount of consumed lipids, but also the composition of fatty acids, which strongly influences health effects of a particular diet. Saturated fatty acids (SFA) are generally considered as unhealthy due to their pro-inflammatory and lipotoxic properties, while monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) represent a healthier alternative, as they are more readily oxidized and do not disrupt biochemical properties of cellular membranes. Amongst PUFA, PUFA of n-3 series (Omega-3) represent an utterly unique class of lipids that have been documented to protect against cardiovascular disease and dyslipidemia in men and improve insulin sensitivity and glucose tolerance primarily in animal models of obesity. Some molecular mechanisms of Omega-3 action have been already uncovered, such as the modification of biological membranes composition, activation of various transcription factors and membrane receptors, and their role as precursors for...

Insulin resistance (IR) is considered to be an important factor influencing the progression of atherosclerosis and is associated with higher morbidity and mortality. IR is a common feature of diabetes mellitus Type 2 and obesity. Many authors consider IR being the crucial abnormality of the metabolic syndrome which is characterized by the essential hypertension, hyperliproteinemia, visceral obesity, endothel dysfunction and many other abnormalities. Impaired insulin action (IR) is also described in diabetes mellitus Type 1, however this phenomenon has not been fully explained. The subjects of dissertation thesis was directed on the IR importance in diabetic Type 1 patients as well as on the renin angiotensin system inhibition in patients with IR and metabolic syndrome with impaired glucose homeostasis. Hyperinsulinemic euglycemic clamp is used in combination with indirect calorimetry to estimate the IR in vivo in humans. In our project we focused on a) the existence of the metabolic inflexibility phenomenon in type 1 diabetic patients b) the methodological evaluation of the hyperinsulinemic euglycemic clamp procedure in the same group c) the influence of renin angiotensin system inhibition with angiotensin II type 1 receptor inhibitor telmisartan in patients with metabolic syndrome and impaired glucose...

This thesis was compiled during the solution of the project MA NAZV QJ1230077 and it is focuses on monitoring of pollen beetle (Brassicogethes aeneus, Fab., 1775) populations resistence to selected insecticides, in particular lambda-cyhalotrin, cypermetrin, tau-fluvalinate, Biscaya 240 OD and chlorpyrifos-ethyl in South Moravia. Beetles were collected during the spring 2016. Their sensitivity to these insecticides active ingredients was tested in laboratory. Various concentrations of active substances and beetles´ mortality after 24 hours was evaluated according to methodology IRAC (adult-vial-test met. 11, 21, 25, www.irac-online.org). Beetles´ sensitivity depends on the dose of the active substance and the location where the beetles were taken from.

Methicillin-resistant Staphylococcus aureus (MRSA) is a major nosocomial pathogen that has spread into the community since the 90s. In general, the community-associated (CA- MRSA) strains are more virulent, but less resistant to antimicrobials than the hospital- acquired strains (HA-MRSA). Some lineages of MRSA such as sequence type 8 have been transmitted more successfully around the world than others and there are situations where a dominant lineage has been replaced by a new one. The factors that are crucial for the selection of dominant lineage are often not clear. As part of this thesis, a longitudinal study of MRSA epidemiology in the Motol University Hospital during the period of 2018-2020 was performed and the occurrence of MRSA clonal complexes was characterized. A multicenter study of the epidemiological situation of MRSA in Slovakia during the period of January - April 2020 was also performed concomitantly. Moreover, several isolates from dominant lineages were further characterized by their phenotype. In the Motol University Hospital study, it was found that the HA-MRSA of the clonal complex 5 dominates and is represented mainly by spa-type t586; a finding that concurs with recent studies from the Czech Republic. In Slovakia, similar to the Czech Republic, MRSA lineages from clonal...

Univerzita Karlova Farmaceutická fakulta v Hradci Králové Katedra Farmakologie a toxikologie Student: Kateřina Novotná Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Title of diploma thesis: Interaction of gilteritinib with OCT1 and OCT2 transporters; relation to conventional therapy of acute myeloid leukemia. Gilteritinib is one of the recently approved drugs which is primarily used in the treatment of relapsed/refractory acute myeloid leukemia (AML) with mutated FMS-like tyrosine kinase 3 (FLT3) receptor. In this project, gilteritinib was investigated in terms of its ability to interact with solute carrier (SLC) membrane transporters, namely with OCT1 and OCT2. These membrane proteins play a role in uptake of endogenous compounds and also drugs into the cells of main elimination organs (liver, kidney), but also to cancer cells. In particular, we wanted to examine potential interaction with daunorubicin and mitoxantrone, drugs traditionally used in AML therapy. First, we performed accumulation study and evaluated, whether gilteritinib is potential inhibitor of OCT1 and OCT2 studying differential uptake of daunorubicin and mitoxantrone into MDCKII-OCT1 and MDCKII-OCT2 cells based on OCT1 and OCT2 inhibition by gilteritinib. Secondly, the study evaluating the transfer of gilteritinib across the...