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Recombinant expression of RhoA protein with affinity tag.
Filipová, Barbora ; Novák, Petr (advisor) ; Šulc, Miroslav (referee)
The rate of intestinal infections associated with antibiotics has been rising steeply. Virulent toxins of C. difficile, Yersinia and many other bacteria target the RhoA protein. The protein is a GTPase involved in signalling pathways, its major function being the regulation of the actin cytoskeleton. The above mentioned toxins render this protein non- functional, which may lead up to a fatal destruction of the cell. The present diagnostic methods of these infections are insufficient. The use of protein chips and mass spectrometry to detect any RhoA protein modifications seems to provide a feasible method of determining these intestinal bacterial diseases. Therefore the purpose of this study is to use recombinant methods to prepare the desired RhoA protein with a bound affinity probe which will serve to its immobilisation on a biochip. The thus prepared protein will be subsequently used to diagnose the previously described diseases. In Czech.
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Depletion of Treg cells for potentiation of cancer treatment with HPMA copolymer-bound cytostatic drug conjugates"
Dvořáková, Barbora ; Kovář, Marek (advisor) ; Reiniš, Milan (referee)
Tumor diseases are severe problem worldwide with increasing number of patients suffering from various types of malignancies. Many of approved therapeutics cause serious side toxicities. Therefore, there are intensive efforts to improve cancer treatment protocols. The aim of this study was to deplete regulatory T (Treg) cells without affecting other immunocompetent cells playing a positive role in tumor eradication. Treg cells were reported to hamper anti-tumor immunity and promote tumor growth and survival. Thus, their selective elimination could lead to induction of anti-tumor responses and tumor rejection if combined with chemotherapy with selected N-(2- hydroxypropyl)methacrylamide (HPMA) copolymer-bound drug conjugates. Original approach was to deplete of Treg cells without the use of anti-CD25 mAb that has been widely exploited for Treg cell elimination; however, its long-term persistence in circulation together with inhibitory effect on activated effector cells (CD25+ ) are its main disadvantages. Thus, Treg cells were sensitized to cell cycle-specific cytostatic drugs via application of IL-2/anti-IL-2 JES6.1 mAb immunocomplexes that induce vigorous selective proliferation of this cell population. Subsequent application of cell cycle-specific cytostatics showed steep decrease of Treg cell...
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Recombinant expression of RhoA protein with affinity tag.
Filipová, Barbora ; Novák, Petr (advisor) ; Šulc, Miroslav (referee)
The rate of intestinal infections associated with antibiotics has been rising steeply. Virulent toxins of C. difficile, Yersinia and many other bacteria target the RhoA protein. The protein is a GTPase involved in signalling pathways, its major function being the regulation of the actin cytoskeleton. The above mentioned toxins render this protein non- functional, which may lead up to a fatal destruction of the cell. The present diagnostic methods of these infections are insufficient. The use of protein chips and mass spectrometry to detect any RhoA protein modifications seems to provide a feasible method of determining these intestinal bacterial diseases. Therefore the purpose of this study is to use recombinant methods to prepare the desired RhoA protein with a bound affinity probe which will serve to its immobilisation on a biochip. The thus prepared protein will be subsequently used to diagnose the previously described diseases. In Czech.
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Depletion of Treg cells for potentiation of cancer treatment with HPMA copolymer-bound cytostatic drug conjugates"
Dvořáková, Barbora ; Kovář, Marek (advisor) ; Reiniš, Milan (referee)
Tumor diseases are severe problem worldwide with increasing number of patients suffering from various types of malignancies. Many of approved therapeutics cause serious side toxicities. Therefore, there are intensive efforts to improve cancer treatment protocols. The aim of this study was to deplete regulatory T (Treg) cells without affecting other immunocompetent cells playing a positive role in tumor eradication. Treg cells were reported to hamper anti-tumor immunity and promote tumor growth and survival. Thus, their selective elimination could lead to induction of anti-tumor responses and tumor rejection if combined with chemotherapy with selected N-(2- hydroxypropyl)methacrylamide (HPMA) copolymer-bound drug conjugates. Original approach was to deplete of Treg cells without the use of anti-CD25 mAb that has been widely exploited for Treg cell elimination; however, its long-term persistence in circulation together with inhibitory effect on activated effector cells (CD25+ ) are its main disadvantages. Thus, Treg cells were sensitized to cell cycle-specific cytostatic drugs via application of IL-2/anti-IL-2 JES6.1 mAb immunocomplexes that induce vigorous selective proliferation of this cell population. Subsequent application of cell cycle-specific cytostatics showed steep decrease of Treg cell...
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The symbiotic bacteria of lice of genus \kur{Polyplax}: general phylogenetic and genomic characterisation
ŘÍHOVÁ, Jana
Blood-sucking lice of the genus Polyplax harbour two bacterial endosymbionts, one from family Legionellales and the other from family Neisseriales. Fylogenetic and genomic analyses reveal typical features of endosymbiotic bacteria and coevolution history with the host. In the genome of the endosymbiont from family Legionellales I found operon, which encodes biosynthesis of biotin, essential vitamin, especially for blood-sucking insects living on low-nutrient diet. This operon was probably horizontally transfer from Wolbachia, nutrient endosymbiont of bedbug Cimex lectularius.
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Modifikované peptidy ve spojení s kvantovými tečkami
Janů, Libor
In this study, water soluble CdTe QDs of different emission wavelengths were prepared. Microwave synthesis was used to prepare CdTe QDs coated with mercaptopropionic acid (MPA). MPA ensures biocompatibility and good water solubility of QDs. Therefore QDs can be used directly in bioanalytics. Further, QDs modified with biotinylated glutathione (GSH) were prepared. In this case mercapto group (--SH) ensures a bonding of GSH to QDs surface and biotin serves as high specific linker for streptavidin, avidin or neutravidin modified analyte. Finally, peptide mediated immuno-quantum dots were prepared. CdTe QDs were coated with heptamer HWRGWVC which is able to bind Fc region of human immunoglobulin G.
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The evaluation of horse nutrition with chosen preparats in sommer and winter season
HANETŠLÉGROVÁ, Petra
ABSTRACT The aim of this work was to provide a detailed breakdown of horse nutrition, to mention the importance of dietary supplements. The main part deals with the basics of horse nutrition, the bare necessities of nutrients, the proportion of maintenance rations and the technique of feeding. Afterwards, I outline the properties and effects of varied herbal plants such as garlic, hemp, hyssop, white horehound, coltsfoot and oats. Flaxseed, biotin, zinc, yeast and probiotics are dietary supplements improving the quality of fur, horsehair and hoofs. The feed will be analysed by standard methods. As far as horse excrement is concerned, we are going to observe its shape, colour, consistency, pH value and smell. The assessment of horse excrement will serve as a judgement about the activity of digestive system. The ethology observations will provide information on daily activity of a horse, on particular activities, to which the horse gives its attention during a day, and the amount of time it dedicates to them. As far as horses with respiratory disorders are concerned, we will observe the signs of their health problems and the rate of these problems occurring both at rest and in action. The information will be completed by the measurement of body proportions useful to body weight calculation. The statistical and economic methods will be selected in terms of the results accomplished during the research.
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