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Management of the Construction Project
Hašek, Jiří ; Rusek, Jan (referee) ; Nováková, Jana (advisor)
The bachelor thesis deals with the issue of construction project management and explains the basic concepts that concern this area. The aim of this work is to get acquiainted with the basic methods of project management and their aplication on concrete practical project.
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Risk Analysis in Transport Infrastructure Projects
Hašek, Jiří ; Holá, Michaela (referee) ; Hromádka, Vít (advisor)
The subject of the master’s thesis is a risk analysis in transport infrastructure projects. In the theoretical part, I deal with public sector, life cycle of the project, evaluation of public projects, conception of risk, clasification of risk, risk analysis and valuation of the risk. In the practical section I process risk analysis of the project in transport infrastucture.
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Membrane microdomains in regulation of lipid metabolism
Veselá, Petra ; Malínský, Jan (advisor) ; Hašek, Jiří (referee) ; Zimmermannová, Olga (referee)
The fluid mosaic model described by Singer and Nicolson in 1972 was timeless and remains relevant to understanding the structure, function and dynamics of biological membranes more than fifty years later. From the outset, its authors acknowledged the existence of lateral membrane regions that differ in composition and biological function from their immediate surroundings. Therefore, even contemporary studies demonstrating the existence of many different membrane microdomains do not pose a fundamental challenge to the validity of this model. In particular, research over the last twenty years has shown that a number of cellular processes (nutrient transport, signaling, regulation of nucleic acid metabolism, lipophagy and many others) are linked to membrane microdomains. However, the molecular details of these links remain hidden in many cases. The aim of this work is to find specific links between membrane microdomains and the metabolism of selected lipids. Using the yeast model S. cerevisiae, we document the connection of a specialized plasma membrane microdomain, the membrane compartment of arginine permease Can1 (MCC), to the metabolism of sphingolipids and the mitochondrial anionic phospholipids, phosphatidylglycerol (PG) and cardiolipin (CL). The initial two chapters deal with the elucidation of...
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The characteristics of stress granules in yeast Saccharomyces cerevisiae
Slabá, Renata ; Hašek, Jiří (advisor) ; Binarová, Pavla (referee)
9 ABSTRACT For proper function proteins should have a native conformation. If their conformation is impaired due to environmental stress or genetic mutation, proteins become prone to aggregation. There exist various types of protein aggregates. Stable non-membraneous inclusions can form which can serve for clearance of aberrant proteins from place where they can interfere with essential cellular processes. Another type of aggregates can serve as transient deposits of proteins thus protecting them from stress conditions. Stress granules (SG) are a such example of transient granules. Their formation is induced by heat shock for example. SGs contain mRNA, components of translation machinery, and other proteins. One of these proteins is Mmi1, small highly conserved protein with unknown function. Association of Mmi1 with stress granules and partial co-localization with chaperon Cdc48 and proteasom indicates Mmi1 can mediate heat stress damaged protein degradation. We have uncovered that yeast prion protein Sup35 is a component of stress granules as well. With regard to its aggregation capability there existed an assumption that prion domain of Sup35 could serve as scaffold for SG assembly. However as we show deletion of prion domain of Sup35 protein does not affect stress granules formation dynamics. Yeast...
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Functional characterization of selected microtubule regulatory
Vinopal, Stanislav ; Dráber, Pavel (advisor) ; Binarová, Pavla (referee) ; Hašek, Jiří (referee)
Microtubules (MTs) play crucial roles in intracellular organization and transport, cell polarity, motility, signalling, division and differentiation. MTs form complex arrays, which are, due to their highly dynamic nature, capable of rapid reorganization in response to cellular requirements. Dynamics, stability and spatial organization of MTs are regulated by many factors including MT regulatory proteins. In the presented study we functionally characterized three selected MT regulatory proteins: Ca2+ -sensor STIM1, MT severing protein spastin and γ-tubulin that is essential for MT nucleation. We found out that activation of bone marrow mast cells (BMMCs) leads to the formation of plasma membrane protrusions containing MTs. Formation of these MT protrusions is dependent on an influx of extracellular Ca2+ regulated by protein STIM1, located in endoplasmic reticulum. STIM1 associates with MTs and its depletion prevents formation of MT protrusions. This indicates that Ca2+ ions might be involved in MT regulation. Since STIM1 depletion also causes defects in chemotaxis, we propose that MT protrusions might be involved in sensing of external signals recognized by BMMCs. Glioblastoma multiforme is the most common and most aggressive malignant primary brain tumor in humans. We demonstrated that MT severing...
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Conventional and Novel Functions of the Exocyst Complex in Plants
Kulich, Ivan ; Žárský, Viktor (advisor) ; Baluška, František (referee) ; Hašek, Jiří (referee)
Exocyst is an octameric protein complex, conserved across all Eukaryotes. Its role, originally described in yeast, resides in a tethering of the secretory vesicles to the plasma membrane prior to the membrane fusion of the two membranes. Subunits SEC3 and EXO70 are believed to be spatial landmarks for the vesicles delivery. While yeast genome encodes single EXO70, we find dozens of them in land plants (23 in Arabidopsis). This work is focused at a role of the exocyst complex in plant cells. Its first part documents, that exocyst is essential for delivery of the cell wall components, namely pectins, but also for pathogen induced secondary cell wall thickening. Second part reveals an unconventional role of EXO70B1 subunit harboring exocyst subcomplex at an autophagic pathway to the vacuole and raises many questions about plant secretory pathway.
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Multiple regulatory roles of the transmembrane adaptor protein NTAL in gene transcription and mast cell physiology
Polakovičová, Iva ; Dráber, Petr (advisor) ; Vyklický, Ladislav (referee) ; Hašek, Jiří (referee)
(EN) This thesis focuses mainly on understanding of the regulatory roles of the transmembrane adaptor proteins, non-T cell activation linker (NTAL) and phosphoprotein associated with glycosphingolipid-enriched microdomains (PAG), in murine mast cell signaling. There are conflicting reports on the role of NTAL in the high affinity immunoglobulin E receptor (FcεRI) activation pathways in mast cells. Studies carried out on mast cells prepared from NTAL knock-out mice have indicated that NTAL is a negative regulator of FcεRI signaling, whereas experiments performed on human mast cells and rat basophilic leukemia cells with silenced NTAL expression have suggested its positive regulatory role. To thoroughly examine the involvement of NTAL in FcεRI-mediated signaling events in mouse mast cells and to determine whether different methodologies of NTAL ablation have different physiological consequences, we utilized a broad range of assays. Using bone marrow-derived mast cells (BMMCs) as a model, we obtained cells from NTAL wild type and knock-out cells and using lentiviral delivery approach we transduced part of the wild type cells, with vector bearing NTAL shRNA or empty vector to generate NTAL knock-down cells and control cells, respectively. Comparison of all four groups of generated cells in our assays...
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Human 4E protein family in stress granules granules and their further characterization
Hrbková, Pavlína ; Frydrýšková, Klára (advisor) ; Hašek, Jiří (referee)
Eukaryotic initiation factor 4E (eIF4E) is a key part of initiation and regulation of translation in human cells. Three members of human eIF4E proteins have been characterized: eIF4E1, eIF4E2 and eIF4E3. Cellular stress causes translation initiation inhibition followed by disassembly of the polysomes, those processes are accompanied by the assembly of cytoplasmic RNA granules, called stress granules (SG). Stress granules are dynamic structures whose composition may vary depending on the cell type and the stress stimulus. In this study, human cells were subjected to the following stress conditions: high temperature (HS), sodium arsenite (AS) or hypoxia. Using fluorescence microscopy, pairs of human translational initiation factors from the 4E protein family were visualized and their localization to SG was assessed with one GFP- 4E incorporated in the stable cell line and the other one detected endogenously. Here we show eIF4E1 being a part of all the SGs, both in HS and AS conditions. Next, the eIF4E1 and eIF4E3 proteins together form more SGs than proteins eIF4E1, respectively eIF4E3, with eIF4E2. And last, that the presence of the particular 4E protein has no effect on the composition of SGs. Furthermore, selected groups of proteins were assessed for their potential to localize to the SGs under HS...
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