Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Anna Giannitsi Supervisor: RNDr. Jakub Hofman, Ph.D Title of diploma thesis: The role of biotransformation enzymes in the resistance of cancer cells against standard cytostatics Drug resistance is currently one of the major problems of chemotherapy. Tumor cells are able to defend themselves against the effect of cytostatic drugs due to various mechanisms which leads to a failure of anticancer therapy. The effort to describe new mechanisms of resistance and to develop new therapeutic methods, which would limit this therapeutic obstacle, is logically the subject of many studies. The activity of drug metabolizing enzymes and the subsequent decrease of intercellular concentration of anticancer drugs belongs to one of the possible mechanisms of pharmacokinetic resistance. Enzymes of I. and II. phase of biotransformation participate in this phenomenon. Cytochromes P450, main enzymes of the I. phase, play a major role in the metabolism of many cytostatic agents producing either pharmacologically active or inactive metabolites. Increased expression in tumors and the involvement of individual isoforms into the overall metabolism of cytostatic, which is deactivated by their activity, seems to be one of the...

No description available.

Damages, which weeds caused annually are difficult to quantify and its negative influence manifests annually in all crops and all fields.Chemical weed control is presently considered an important method for controlling weeds. The essence of chemical pest weeds consists in the use of herbicides to reduce weed species population under the economic threshold of harmfulness. In an effort to control weed populations using three methods: mechanical, chemical and biological. In the thesis was mapped weeds in potatoes on private land in Plasná, district of Jindřichův Hradec in region of South Bohemia. In the practical part of this thesis, I conducted an experiment that was aimed at monitoring the effects of selected herbicides in crops of potatoes. The aim of the experiment was to verify the effects of herbicides on the population most frequently occurring weeds in the root crops. The results showed that the herbicide is more effective for preemergence weed monitored. Assessment of weed infestation was carried out numerical method (ANOVA, Tukey = 0,05) in the ARM 8 (GyllingDataManage ment, Inc.) Spring 2013 - Fall 2013.

This diploma thesis deals with resistance of microorganisms to antibiotics and public awareness of this topic. The aim of the thesis was to find out what the public awareness of the given topic is and what knowledge the public has about the possibilities of prevention of creation and spreading of resistant microorganisms. Three main hypotheses have been set: Hypothesis 1: The public has no deeper knowledge of antibiotic treatment. Hypothesis 2: The public will in most cases get information on antibiotic treatment from a doctor. Hypothesis 3: Most people are unaware that they can contribute by rational use of antibiotics to prevent antibiotic resistance. In order to achieve the stated goals of the diploma thesis, a quantitative method of research was used. Data collection was conducted through anonymous questionnaire survey of the public population using the Click4Survey.cz web server and distribution of paper questionnaires in FN Plzeň. Results of the questionnaire survey were evaluated based on statistical data processing in Microsoft Office Excel 2007, and a Chi-squared test of independence in the pivot table and a relative frequency test were used to verify the hypotheses. Based on the evaluation of the results, it was found that among the groups of respondents, depending on whether they filled in a questionnaire on the Internet or a printed questionnaire at the hospital, there are fundamental statistical differences in knowledge and awareness of the problem. That is why it was necessary to further evaluate these two groups separately. Antibiotic resistance nowadays is a very important medical issue. The main cause is inconsistent use or overuse of antibiotics in both human and veterinary medicine, as well as insufficient awareness of the extent of the problem and the consequences of antibiotic resistance. For this reason, I have chosen this issue as a topic for my diploma thesis to highlight the risks of antibiotic resistance and to provide information primarily to the general public.

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...

1 Abstract The cluster of obesity, insulin resistance and other associated comorbidities represents a significant health risk for the affected individuals as well as the whole population. Chronic low-grade inflammation of adipose tissue is considered one of the main mechanisms respon- sible for the progression from simple obesity to a fully developed metabolic syndrome. The aim of our study was to explore two different approaches that could potentially ameliorate adipose tissue inflammation - therapeutic hypothermia and the adipocytokine clusterin. In the first part, we showed that a period of deep hypothermia associated with the an- oxic phase of cardiac surgery significantly delayed the onset of systemic inflammatory re- sponse induced by surgery. The relative gene expression of the studied genes was not altered during the hypothermic period, but was significantly increased in five out of ten studied genes (IL-6, MCP-1, TNF-α, HIF1-α, GLUT1) and decreased in two genes (IRS1, GPX1) at the end of surgery. We conclude that deep hypothermia delays the onset of local adipose tissue hy- poxia and inflammation. These results could partially explain the positive effects of therapeu- tic deep hypothermia on postoperative morbidity and mortality in cardiac surgery patients. In the second part, we examined plasma...

It is known that entomopathogenic nematodes rarely live in fields where pesticides and fertilizers are used. Because they are organisms with a high potential for biological and integrated pest control the thesis tried to determine the influence of pesticides on the infectivity and mortality of selected nematodes: Steinernema feltiae, S. arenarium and S. kraussei in water solution of some pesticides. The results are that preparations Vydate (a. i. oxamyl) and Sulka (a. i. sulphur) have the highest negative influence on both characteristics. These pesticides cause a mortality of 94.3 - 99.9 % and reduce infectivity by about 50 {--} 75 % (level 3). The following preparations may be identified as having higher potential for infectivity reduction: above all, Starane 250 EC (fluroxypyr), Sumithion super (fenithrothion), and Novozir MN 80 (mancozeb); on the other hand, Treflan 48 EC (trifluralin) significantly increases mortality. Another important observation is a fact that infectivity is significantly influenced only by concentration of pesticides, not by exposure time. During an experiment with enhacement of resistance to oxamyl (Vydate) I falling mortality and increasing infectivity of entomopathogenic nematodes were observed after five rounds.

This master‘s thesis is focused on the optimization of sample preparation and analytical method for the determination of fluoroquinolone. The theoretical part describes the principles of analysis in soil samples, environmental aspects of antibiotic resistance, characteristics of UPLC-MS / MS methods, description of quinolone. The experimental part deals with the identification and determination of fluoroquinolone substance (ciprofloxacin, enrofloxacin) in soil in which manure from poultry litter was incorporated using liquid chromatography with mass spectrometry (MS / MS, triple quadrupole). Internal standard and standard addition methods were used to quantify ciprofloxacin and enrofloxacin. The amount of CIP in the samples ranged from 0.2030 to 12.7088 µg·g-1 and for ENR from 0.2537 to 10.0224 µg·g-1.

Obesity leads to many severe metabolic disorders, e.g. dyslipidemia, insulin resistance, ectopic fat accumulation in the liver and skeletal muscles, non-alcoholic fatty liver disease and finally diabetes mellitus type 2. Metformin (1,1-dimethylbiguanide) is the most favored medicament for the treatment and prevention of these disorders. It stimulates cellular glucose uptake and normalizes blood levels of lipid metabolites without triggering insulin secretion. Research on insulin resistance and diabetes is often realized through developing diet- induced obesity in laboratory animals. The aim of this project is to compare metabolic effects of two different high-fat diets named HFD and HSD. The HFD diet consists chiefly of n-6 polyunsaturated fatty acids (corn oil) and starch (100% glucose). The HSD diet contains mainly saturated fatty acids (lard) and sucrose (50% glucose and 50% fructose). I also studied metabolic effects of metformin by adding it continuously to the drinking water given to obese mice fed with the HFD or the HSD diet. Methods: Intraperitoneal glucose tolerance test (IPGTT), blood and tissue levels of lipid metabolites assessment, radio-immunological assessment of blood levels of insulin, assessment of AMPK activity in liver by western blotting. Results: Increased consumption of the...

Trichomonas vaginalis is a human pathogen that affects annually approximately 258 million people worldwide. This parasite possesses organelles of mitochondrial origin called hydrogenosomes, which generate ATP under anaerobic conditions. The identification of the protein content at the subcellular level may provide new targets for antiparasitic drugs developments as well as it contributes for our understanding of the organelles function and evolution. The availability of protocols for organelles purification and the complete genome sequence allow the study of the organellar proteomes using mass spectrometry and bioinformatics, providing a powerful strategy that combine cell biology and proteomics. In our research, we used several approaches to identify the protein composition in hydrogenosomes and mitosomes. We performed transcriptomic and proteomic analysis to investigate the molecular responses of Trichomonas vaginalis upon iron availability. Furthermore, the changes in the proteome during the development of metronidazole resistance were also studied. The organelles separated by differential and Optiprep-sucrose gradient centrifugation were analyzed with nano- RP-HPLC/MALDI-TOF/TOF. We also used Triton X-114 phase partitioning to separate membrane proteins and iTRAQ technique to label the peptides...