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Identification of antimicrobila peptides in spider venom
Benýšek, Jakub ; Tichá, Marie (advisor) ; Liberda, Jiří (referee)
Still increasing resistance to antibiotics leads to the need to find new active compounds with antimicrobial properties. This work is focused on the occurrence, chemical and physical description, mechanism of action and biological activity of such substances, found in spider venom. The second part is focused on isolation and identification of compounds with these properties from the venom of wild bees and a one spider. A novel peptide was isolated and identified from venom of bee Trachusa byssina. This novel peptide possess antimicrobial properties and low hemolytic activity. Molecular weight was estimated to 1749,9 ? 0,1contains 16 amino acids and is amidated on its C-terminus. Its primary structure GILSVLKNLLKKHMAS-NH2 was determined by using Edman degradation and ESI-QTOF mass spektrometry.
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Objective evaluation of the effect of therapy : Synapsys Posturography System
Markvartová, Anna ; Tichá, Marie (advisor) ; Jeníček, Jakub (referee)
Author's name: Anna Markvartová Bachelor thesis supervizor: MUDr. Marie Tichá Oponent: Title of bachelor thesis: Objective evaluation of the effect of therapy: Synapsys Posturography System Abstract: This bachelor thesis is based on objective evaluation. The author chose Synapsys Posturography System device for evaluating the effect of therapy. The theoretical part of this thesis defines basic concepts, especially brain damage, balance disorders and posturography. Practical part describes metodology and results of the research. 11 people participated in this research and two of them were patients after brain damage. Values measured in a group of patients and in reference group were compared in practical part. In the end of this thesis, there are some recomendations and final summary of results of the research. Key words: Objective evaluation, brain damage, balance disorders, posturography, Synapsys Posturography System
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Study of acid phosphatase in pea and tobacco
Šandová, Sandra ; Ryšlavá, Helena (advisor) ; Tichá, Marie (referee)
Phosphatases catalyze the hydrolytic fission of orthophosphoric acid ester. They are generally divided into acid and alkaline phosphatases according to their pH optimum. There was preparated the extract of leaves Nicotiana tabacum L. and Pissum sativum. Furthermore, there were determinated pea and tobacco phosphatases's rate constants. Reaction speed katalyzed by tobacco phosphatase is 10,0 mol/min.ml and by pea phosphatase 38,0 mol/min.ml using paraNP - phosphate as a substrate. Using a substrate phospho - L serine the reaction speed katalyzed by phosphatase of tobacco is 1,8 mol/min.ml and by phosphatase of pea is 0,4 mol/min.ml. Michaelis constant Km is 1,8 mM for tobacco phosphatase and 8,5 mM for pea phosphatase using paraNP - phosphate. Using a substrate phospho - L serine Michaelis constant Km of phosphatase of tobacco and pea is the same - 4,0 mM. The pH optimum of pea phosphatase is 5,0 using paraNP - phosphate as a substrate and 6,0 using phopho - L serine. pH optimum of tobacco phosphatase is 5,4 using paraNP - phosphate and 7,0 using phospho - L serine. Temperature optimum for pea phosphatase is 60 C and for tobacco phosphatase 55C.
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Determination of pepsinogens using IgY and IgG
Kulhavá, Lucie ; Pacáková, Věra (advisor) ; Tichá, Marie (referee)
A decreased concentration of pepsinogen A in serum was found to be a marker of gastric cancer, similarly as a low ratio of pepsinogen A to pepsinogen C. In the present study we have compared properties of immunoglobulin fraction isolated from the egg yolks after immunization of laying hens with pepsinogen isolated from porcine gastric mucosa with those of present in rabbit antiserum obtained after the animal immunization with the same antigen. The characteristics of chicken antibodies against porcine pepsinogen and the comparison with rabbit antibodies raised against the same antigen was carried out using the following methods: ELISA, affinity chromatography on immobilzed antigen and bovine serum albumin, SDS and native electrophoresis and MALDI-TOF MS/MS. While the rabbit specific antibodies interacted with the used antigen and only slightly with bovine serum albumin and there was a diference between pre-immune IgG and specific IgG, in the case of chicken antibodies IgY it did not work. No diference was observed between ELISA tests performed with pre-immune serum and the serum after immunization with porcine pepsinogen and a high interaction of IgY with bovine serum albumin in pre-immune serum and specific IgY after the immunization were detected. Similar results were obtained in experiments with...
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Immobilization of protease V8 on magnetic particles for application to proteolytic cleavage of pepsin A
Čepa, Adam ; Pacáková, Věra (advisor) ; Tichá, Marie (referee)
This thesis is part of a long-standing research in the field of diagnosis of the stomach diseases, which is based on the gastric enzyme pepsin A mapping. It was found that a phosphorylation in the primary structure of this enzyme may serve as a marker of incipient stage of carcinogenesis. This thesis is focused on the immobilization of protease V8 isolated from microorganism Stafylococcus aureus to magnetic agarose beads. Protease V8 is a promising candidate for producing peptide maps of pepsin A. The influence of pH, temperature and reaction time on the enzyme to activity has been studied and the optimal conditions for hydrolytic catalysis of formation of peptide fragments of pepsin A.
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Implementation of Challenging Tasks in Mathematics Education
Růžičková, Lucie ; Kopáčková, Alena (advisor) ; Harminc, Matúš (referee) ; Tichá, Marie (referee)
Title: Implementation of Challenging Tasks in Mathematics Education Author: PhDr. Lucie Růžičková Department: Department of Mathematics and Mathematics Education Supervisor: RNDr. Alena Kopáčková, Ph.D. ABSTRACT The thesis deals with practical implementation of challenging tasks in mathematics education. The methodology for examining "the fidelity of implementation of mathematical tasks" is being designed to describe whether the task is used according to the original intention of the teacher or according to the intention of its author. This methodology is employed in the analysis of four experiments to examine mathematical tasks and task environments. Then, based on the experience with the analysis, the methodology is adapted to the needs of both the teaching practice and educational research. The main result of the thesis is the creation of the methodology for examining the fidelity of implementation of mathematical tasks which is based on contrasting the descriptions of four general elements of the mathematical task at different phases of its implementation. KEY WORDS: mathematics education, mathematical task, challenging task, implementation of tasks
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Characterization of phosphatases in tobacco plants (Nicotiana tabacum L.)
Růžičková, Kateřina ; Ryšlavá, Helena (advisor) ; Tichá, Marie (referee)
Phosphateses (EC 3.1.3.x) are a group of enzymes that hydrolyze phosphoesters. That way they affect the energetic metabolism of a cell and its regulation. Phosphatases that dephosphorylate proteins are an integral part of signaling pathways, stress responses and they modulate enzymatic activity. This thesis deals with the study of phosphatases obtained from tobacco leaves (Nicotiana tabacum, L.). Solution of enzymes was prepared by extraction in both acidic and alkaline buffers. Through the use of the chromogenic substrate pNP-phosphate it was determined that there is a higher phosphatase activity in the glycosylated fraction than in the fraction that did not bind to Con A Sepharose. The research of the ions effect on the phosphatase activity has determined that Mg2+ and Ca2+ show positive effect on the phosphatase activity while the effect of Co2+ and Mn2+ is inhibitory. The Zn2+ ions have shown no effect whatsoever. The glycosylated phosphatases also dephosphorylated low-weight-molecular substrates phosphoserine, ATP and glucose-6-phosphate. The research of protein phosphatase activity discovered the affinity to the substrate phosvitin, although neither to casein nor its tryptic cleaves. Detailed experiments have shown that the pH optima for all the substrates lie from pH 5 to 6. Glycosylated phosphatases...
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Perception and Interpretation of Fractions by Pupils in the 5th Grade of Primary School
Vašíčková, Jana ; Tichá, Marie (advisor) ; Macháčková, Jana (referee)
The essential part of this diploma thesis is a classification of works obtained from an investigation aimed on determination of conceptions of fractions by pupils in the fifth form of the primary school. The experiment consists of four tasks, which were given to children in four different classes. I have classified produced works according to bases of division, which I had established. Mainly concerned classifications are the thematic classifying, classifying in accordance with the "type of children's works" and classifying based on occurred interpretations of fractions. I was trying to find such criterions that would state in evidence of what conceptions and knowledge of fractions do the pupils in the fifth form have. According to gained results I think that I fulfilled this goal. When I was sorting the works of children, I found out that there is a huge amount of large-scaled works that can become basis for another research.
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