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Interakce rostlin s pathogeny: význam proteasomu a cytoskeletu
Zelená, Pavla
Modification of substrate proteins by ubiquitin is one of the major regulatory events in eukaryotic cells, and plants use this modification to react to exogenous and endogenous cues. Mounting evidence indicates that the ubiquitin-proteasome system is also a key component in response to biotic stressors. This thesis entitled "Plant-pathogen interactions: The role of proteasome and cytoskeleton" reviews the role of proteasome in plant signaling and briefly summarizes interactions between cytoskeleton and pathogen and outlines the role of cytokinin in plant-pathogen interactions. In the experimental part, proteasome-mediated signaling is probed using growth response assay, identification of proteasome substrates and confocal microscopy of cytoskeleton structures. The results present evidence that the employed methodology will be useful for plant-pathogen interaction analyses.
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Cell layer cultivation in the microfluidic system
Kachan, Ksenia ; Svoboda, Ondřej (referee) ; Chmelíková, Larisa (advisor)
The theoretical part of this thesis describes the principles of cell culture in vitro and modern in vitro models for endothelial cell layer cultivation. It also describes the principles of confocal and fluorescence microscopy. The practical part is dedicated to working with cell cultures and realization of experiment with cell cultivation in microfluidic system. Cell layer was photographed using laser confocal scanning microscope Leica TCS SP8 X during the entire experiment. For processing and analysis of the obtained images, an algorithm in the software MATLAB was proposed.
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Analysis of adherent cells confluency in 2D culture
Bracková, Michaela ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
First part of this semester work dealing with the theoretical description of adherens cells, particularly structure and functions of this cells. Subsequently work is devoted to description of cell culturing, with mention of conditions which are necessary for cell culturing, following by substances which promote cell growth. Last part of theoretical research is concern with microscopy technique that is suitable for studying of adherent cells. Subsequently it is about fluorescence and confocal microscopes. Practical part dealing with cell culturing of adherent cells and the evaluation of realized experiments.
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Actuoecology of testate amoebae in fresh water and soil environment in enteraction with fungi and their analysis with new microscopic techniques
Burdíková, Zuzana ; Holcová, Katarína (advisor) ; Jansa, Jan (referee) ; Hudáčková-Hlavatá, Natália (referee)
4 Abstract The present thesis focuses on testate amoebae (TA) and their relationship to their natural environment, as well as on relevant microscopic imaging methods. The bulk of the data has been published in original scientific papers and is compiled into three separate chapters (Pt I, Pt II and Pt III), each annotated by a brief introduction. (Pt I) The methods section is devoted to specialized microscopic techniques employed to broaden the scope of the ecological analyses. In particular, precise discrimination between live and dead individuals, biomass determination inside individual tests and a multi-modal visualization of the cytoplasm and organelles enhance the data. Laser scanning confocal microscopy and two-photon microscopy are the main imaging modalities employed to study TA morphology in detail. The data have implications for taxonomy and ecophysiology, including the use of TA as bioindicators of pollution. (Pt II) An actuoecological analysis focuses on the seasonal variability of TA species composition in a freshwater ecosystem, namely the Komo any ponds in Prague, during the course of the year. The species composition variation is correlated to simultaneously recorded limnological parameters such as temperature, pH, contamination by (heavy) metals (As, Cd, Mn, Ni, Fe, Pb), polycyclic aromatic...
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Application of carbon based electrodes for electrochemical detection in flow system
Mika, Jan ; Dejmková, Hana (advisor) ; Fischer, Jan (referee)
Carbon is due to its electrochemical properties a favourite material for construction of electrodes suitable for detection in flow system. The two most often used techniques (coulometric and amperometric) will be discussed in this Thesis. Carbon paste electrodes (CPE) represent the amperometric detectors. CPE are unstable in the medium with high content of organic solvents, which might complicate their application in HPLC, where the mobile phase contains organic component. It is interesting, that glassy carbon paste electrodes are more stable under such conditions. The change of surface and response of the electrode caused by the effect of methanol are described in this Thesis along with the possible reason of stability of GCPE. Roughing of the surface exposed to methanol is demonstrated by confocal microscopy. This effect leads to increase of height of the peak, background current and noise and to the shift of peak potential to less positive values. These changes are dependent on the time of exposure and the concentration of methanol. Coulometric detectors work with high conversion effectiveness and sensitivity, but in case of their passivation, reactivation is a relatively complicated problem, often leading to the replacement of electrode material. Answer to this problem can be given by using...
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Mechanisms of invasiveness and transcription regulation in cancer cells
Tolde, Ondřej ; Folk, Petr (advisor) ; Kovář, Jan (referee) ; Brdička, Tomáš (referee)
The mechanisms of invazivity and regulation of transcription of cancer cells Cancer originates in cells that overcome the control mechanisms of the organism. Cancer cells can be eventually released from the site of origin and spread through tissues. Cancer cells can acquire certain mechanisms that enable them to more effectively invade surrounding tissue or layers of other cells. The research on the migration of cancer cells is important for the understanding of the origin and spreading of metastases and consequently for anticancer therapy. In my Ph.D. work, I participated in the research of the properties of invasive metastatic cells. We compared non-invasive rat sarcoma cell line with a higly metastatic cell line derived from it. We showed that cells of the invasive cell line use amoeboid mode of migration, have upregulated Rho/ROCK signaling, and have accumulated actin and myosin at the leading edge. It is at the leading edge where the cells generate their traction forces. Cells of non-invasive cell line use mesenchymal mode of migration and generate forces mainly at their retracting end. We also compared two breast cancer cell lines derived from a single carcinoma. We showed that the more invasive cell line, derived from its parental line by neoplastic transformation, displayed elevated cytoskeletal...
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The effect of elevated CO2 concentration and irradiation on quantitative parameters of mesophyll cells of Norway spruce
Kubínová, Zuzana ; Albrechtová, Jana (advisor) ; Radochová, Barbora (referee)
KUBÍNOVÁ, Zuzana. The effect of elevated CO2 concentration and irradiation on quantitative parameters of mesophyll cells of Norway spruce. Prague, 2010. 74 p. Master's degree thesis. Faculty of Science, Charles University in Prague. Abstract The aim of the present thesis was to choose and adjust a suitable methodology for counting particles in 3D space, which would be suitable for unbiased estimation of chloroplast number in needle mesophyll cells. The disector method was used for the first time to determine the number of chloroplasts. This method enables unbiased estimation of chloroplast number in needle volume from optical sections captured from fresh free-hand sections by confocal microscope. The sections did not need any pre-processing. Another aim was to compare selected photosynthetic and anatomical characteristics of sun and shade Norway spruce needles, which were grown under different CO2 concentration. The trees were grown for eight years in ambient (during the experiment increasing from 357 up to 370 µmol CO2 ∙ mol-1 ) CO2 concentration or elevated (700 µmol ∙ mol-1 ) CO2 concentration in special glass domes on an experimental research site of the Institute of Systems Biology and Ecology, Academy of Sciences of the Czech Republic at Bílý Kříž in Moravskoslezské Beskydy mountains. The sun needles...
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Simulation of adhesion and transmigration imune-cells through capillary wall
Morgaenko, Katsiarina ; Čmiel, Vratislav (referee) ; Chmelíková, Larisa (advisor)
Teoretická část této práce obsahuje biofyzikální popis endoteliální vrstvy a transmigrace buněk přes tuto vrstvu. Dále popisuje charakteristiky všech důležitých komponent pro vytvoření modelu cévy in vitro (endoteliální buňky, kmenové buňky, leukocyty, Calcein AM, PKH26). Praktická část práce je věnována sestavení in-vitro modelů dvojrozměrných a trojrozměrných endoteliálních vrstev, jejich zobrazení pomoci mikroskopu a testování interakce leukocytů a kmenových buněk s těmito endoteliálními vrstvami. V závěru je navrhnut nejlepší tvar a velikost mikrofluidních systémů simulujících kapiláry.
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