| |
|
The preparation of oligoester nanoparticles by emulsion method
Šeuerová, Andrea ; Dittrich, Milan (advisor) ; Šnejdrová, Eva (referee)
The deal of the work was focused on the study of efficiency of complex emulsifying agent on the nanoparticulate systems preparation by the o/w emulsion-solvent distribution method. Poly (DL-lactic acid) was used as the carrier, terbinafine as a model drug. Complex emulsifying agent was realized by the mixing of soy or egg lecithin with various proportions of nonionic surfactants. Four nonionic surfactants were tested - polysorbate 20, and poloxamers: 470, 6400, and 6800. Optimal compositions in the aspect of nanoparticles minimal size and zeta potential maximal value were stated.
|
|
Study of phosphorylation of inorganic pyrophosphatase from Streptococcus pneumoniae
Štechová, Michaela ; Svobodová, Jaroslava (referee) ; Doubravová, Linda (advisor)
The human patogen Streptococcus pneumoniae encodes a single copy of eukaryotic-like Ser/Thr protein kinase StkP. StkP regulates virulence, competence, stress resistence, gene expression and plays a role in the regulation of cell division cycle. Analysis of phosphoproteome maps of the wild type and stkP mutant strain of S. pneumoniae showed that in vivo StkP phosphorylates several putative substrates including Mn-dependent inorganic pyrophosphatase PpaC. Mass spectrometry analysis identified two phosphorylation sites in an active site of the protein. Pyrophosphatases are essential enzymes that catalyze hydrolysis of inorganic pyrophosphate produced during various biosynthetic reactions that utilize ATP. Changes in pyrophosphatase activity have been described to have global effects on cell metabolism, growth and division of bacteria. The aim of this thesis was to investigate the phosphorylation of inorganic pyrophosphatase PpaC in S. pneumoniae. Gene ppaC was cloned, expressed in E. coli and protein was purified via affinity chromatography. Phosphorylation of PpaC by StkP was examined in a kinase assay but we did not confirm that PpaC is a direct substrate of StkP in vitro. Further we prepared a set of mutants in ppaC gene. We replaced two presumable phosphoaminoacids identified by mass-spectrometry with...
|
|
Monitoring antidepressants consumption in pharmacy in 2003-2005
Borská, Anna ; Vopršalová, Marie (advisor) ; Trejtnar, František (referee)
7. SUMMARY 87 The subject of this thesis is the research of medicine consumption in the selected pharmacy. We focused on the quantification of medication consumption between years 2003 - 2005 in Pharmcentrum Budějovická, Prague 4. In theoretical part of our thesis we summarized the recent knowledge and the best recommendation for the curing of depression. Key attributes of the active principles used at pharmacotherapy of depressions are mentioned as well. In the following part we worked with the figures of single medicine annual consumptions gained from the pharmacy software. Using the DDD/ATC system we have computed the total consumption of each individual medicine. We also tried to learn whether there are any differences in prescriptions of various doctors according to their expertise or age. The consumption of antidepressants in Pharmcentrum Budějovická in the years 2003 - 2005 had been permanently increasing. It corresponds with the numbers published by the VZP (General Medical Insurance Company). There is the significantly growing consumption of so called 3rd generation antidepressants (SSRI) with the most preferred substance Citalopram. With only minor exceptions the consumption of substances of the other new generations has been also increasing. Such trend complies with the recommendations of...
|
| |
|
Study of the microRNAs in patients with head and neck squamous cell carcinoma
Hudcová, Kristýna ; Šulc, Miroslav (referee)
Searching for markers of disease is at the forefront of scientific interest. Biomarker of a specific disease should help in disease management - whether for diagnosing a disease or predicting its progress. We picked microRNAs - short RNAs that regulate mRNA translation, due to their stability and function in the regulation of many cellular processes. Therefore they can be used as a biomarker for many diseases. In our study we focused on neck squamous cell carcinoma (HNSCC), since this type of cancer is widespread and clinically important markers are missing. It is in our interest to seek new kinds of markers, suitable for early diagnosis of the disease and the most accurate assessment of prognosis. We selected 3 candidate miRNA (miR-29c-3p, miR-200b-5p, miR-375-3p) and used qRT-PCR method to profile their expression in tissues of patients with HNSCC. MiRNA expressions in 42 tumor samples were compared with the miRNA expression in adjacent tissue of the tumor. Adjacent tissue surrounds the tumor and has interesting properties, as it looks like a histological normal tissue, but on closer inspection it is not. It is affected by tumor tissue and may contribute to tumor progression. We managed to distinguish between these two tissues using miR-29c-3p and miR-375-3p. Additional analyzes were focused on...
|
|
Differentiation within yeast populations of Saccharomyces cerevisiae: influence of cultivation conditions
Šimek, Jan ; Palková, Zdena (advisor) ; Dostál, Jiří (referee)
Long-time research of chronologically aging yeast populations of Saccharomyces cerevisiae laboratory strains revealed that yeasts are able to differentiate into specialized cell types. Differentiation of liquid cultures growing in glucose rich medium and differentiation of colonies growing on solid glycerol medium has been previously studied. These populations create two fractions of cells with diverse morphology which adapt their metabolism and physiology to enable a long-term survival of the yeast population in environment with limited nutritional potential. In this study, yeast subpopulations isolated from colonies growing on solid glucose medium and liquid cultures cultivated in glycerol medium were characterized. Newly isolated cell types were compared with already known cell types isolated from colonies and liquid cultures. Selected metabolic processes and stress resistance were analysed in studied populations. Based on previous studies of yeast differentiation, a spectrum of GFP-labelled marker proteins was choosen and production and localization of these marker proteins was monitored within yeast populations. Results of the analyses showed that type of medium and cultivation influence development and metabolism of each yeast cell type. Key words: Saccharomyces cerevisiae, BY4742, cell...
|
|
Study of cytotoxicity of potential antituberculotics using selected methods on liver and kidney cell line
Katrnošková, Simona ; Ramos Mandíková, Jana (advisor) ; Bárta, Pavel (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Mgr. Simona Katrnošková Supervisor: PharmDr. Jana Ramos Mandíková, Ph.D. Title of Thesis: Study of cytotoxicity of potential antituberculotics using selected methods on liver and kidney cell line During pharmacologic therapy, tissues of liver and kidney are frequently exposed to high doses of xenobiotics. Via in vitro assays during preclinical testing, we are able to predict potential toxicity which helps to prevent the possible serious adverse drug reactions in clinical practice. The aim of this rigorous thesis was to state the cytotoxic profile of 4 potential antituberculotic drug candidates using appropriate cell models and in vitro assays. Another aim was to compare the cytotoxic effect of the tested compounds among themselves and to 4-aminosalicylic acid (PAS) which antituberculotic activity is known for many years. The tested substances were 3 salicylanilide diethyl phosphate-based derivatives and 4-(trifluormethyl)benzoic acid. For the cytotoxic potential assessment, we used the parameter half maximal inhibitory concentration IC50. We have used methods determining the cell metabolic activity, aminopeptidase activity, lactate dehydrogenase leakage and activity of 3/7 caspases in this...
|
|
Study of interspecies differences in plasma protein binding of 18F-labeled radiopharmaceuticals
Nováková, Dana ; Lázníček, Milan (advisor) ; Bárta, Pavel (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Mgr. Dana Nováková Consultant: Prof. PharmDr. Ing. Milan Lázníček, CSc. Title of Thesis: Study of interspecies differences in plasma protein binding of 18 F-labeled radiopharmaceuticals In this thesis, the binding ability of three radiopharmaceuticals containing radionuclide 18 F to the plasma proteins was examined in bovine, pig, human and rat plasma, and the interspecies comparison was performed. Examined radiopharmaceuticals included 18 F-FDG, 18 F-fluorocholine and 18 F-thymidine. The two main methods used are equilibrium dialysis and ultrafiltration. These methods are based on separation of free ligand from the linked component. During the separation, semi-permeable membrane and special filters (AMICON and VIVASPIN) were used for equilibrium dialysis and ultrafiltration, respectively. Free fraction of drugs is considered to be an important parameter for the availability of the drug distribution into the tissues and cells, interaction with receptors, excretion and metabolism. All experiments were carried out at the physiological temperature of 37řC. Obtained results indicate low plasma protein binding of the three radiopharmaceuticals in all studied species, which suggests...
|
|
Analysis of gene expression in maternal and fetal tissues exposed to cigarette smoke
Fejglová, Kamila ; Zikánová, Blanka (referee) ; Bruchová, Hana (advisor)
Despite widespread awareness of the adverse health effects of smoking, pregnant women expose themselves to this factor (firsthand and/or secondhand smoke). Cigarette smoke exposure during pregnancy increases the risk of preterm delivery, abortion or newborn sudden death syndrom. To study molecular mechanisms of cigarette smoke exposure in tissues of pregnant women and their newborns, we employed genomic approach and assessed gene expression profiles in leukocytes from maternal peripheral blood and umbilical cord blood and in placenta as well. Gene expression profiles of these tissues were determined by Illumina HumRef8 v3 expression chips that enable analysis of 23 811 coding, annotated transcripts. Microarray data were normalized using the quantile method. Differential gene expression was analyzed using moderated t-statistic. A linear model was fitted for each gene given a series of arrays using lmFit function. Multiple testing correction was performed using the Benjamini and Hochberg method. Comparative analysis of gene expression profiles between smokers (N=23) and non-smokers (N=62) revealed many deregulated genes, demonstrating a negative effect of cigarette smoke on transcriptome of the tested tissues. In peripheral blood of smokers, we detected overexpression of these genes; e.g. IFI27, IFIT1L,...
|
guest ::
