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Non-conventional bacterial signaling pathways
Krupička, Jiří ; Branny, Pavel (advisor) ; Beranová, Jana (referee)
Two component systems were traditionally considered as main phosphorylation systems of bacteria involved in cell signalling. Recently, attention focuses increasingly on bacterial eukaryote-like Ser/Thr protein kinases (eSTKs). These protein kinases are structurally similar to their eukaryotic counterparts. Some eSTKs possess additional domains such as extracellular PASTA domains that were discovered in a variety of gram-positive bacteria. It has been proved that these domains can act as sensors for unlinked peptidoglycan fragments. However, majority of environmental signal molecules still remains unknown. eSTKs phosphorylate a broad spectrum of substrates including proteins involved in various cell processes such as virulence, cell wall biosynthesis, cell division, and central and secondary metabolism. Cross talk between eSTKs and two component systems also occurs. In this thesis, the current knowledge about eSTKs and their significant substrates in different bacterial species is discussed.
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Klinicky významné druhy kvasinek a jejich interakce s hostitelem
Novosadová, Zuzana ; Palková, Zdena (advisor) ; Beranová, Jana (referee)
Pathogenic yeasts are nowadays a serious threat for mammalian host. They can cause dangerous diseases, which in many cases even result in death. Pathogens increase the chances of systemic infections by many virulence mechanisms. Experiments addressing the pathogenhost interactions are crucial for defeating these types of infections within the human body. Host-pathogen interactions are very complex and include all components of multicellular host organism. Recently, scientists have focused on the interaction of the mammalian immune system and pathogenic yeasts in more detail. This work summarises interactions of pathogen with selected host cells, especially with macrophages. Yeast pathogens, especially Candida albicans, are capable of influencing the gene expression in interacting cells. These pathogens are capable of modulating the expression while engulfed inside macrophages and other cells of the immune system. Pathogenic yeasts can also change the overall characteristics of their surrounding environment. C. albicans can sense pH and influence it. Therefore, it can increase its virulence by the changes of pH leading to autoinduction of morphological transitions. This work briefly reviews how selected yeast pathogens influence their surroundings while interacting within the host organism. Deeper...
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Růst Mycobacterium smegmatis na agarovém médiu a agarovém médiu pokrytém celofánovou folií - morfologická a proteomová studie
Ramaniuk, Volha ; Weiser, Jaroslav (advisor) ; Beranová, Jana (referee)
Biofilm formation is one of the most common bacterial survival strategies. Majority of bacterial species are able to form these three-dimensional structures, including pathogens like Mycobacterium tuberculosis. Representatives of Mycobacterium genus widely occur in the nature, although they can cause serious problems when they appear in medical equipment and artificial replacements of the human body. Non-pathogenic Mycobacterium smegmatis mc2 155 was used as a model organism in our experiments. We investigated morphology of the three- and six-day-old colonies (in fact biofilms) on agar and agar covered with cellophane using Stereo microscope and Scanning Electron Microscope. We found that a type of surface as well as a carbon source has a great influence on the morphology of the M. smegmatis colonies. We isolated proteomes from the agar and cellophane cultures and from planktonic culture. Two-dimensional electrophoresis was used as the main proteomic method. Proteomic data were analyzed using PDQuest software. Then the sets of proteins detected by qualitative and quantitative analyses were compared using Venn diagrams. As a result, we recognized 7 unique proteins that might be specific for recognition and adhesion of bacteria to the cellophane, no unique protein in agar proteome and 46 unique...
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Cytoplasmic membrane of Bacillus subtilis Regulation of the physical parameters
Beranová, Jana ; Konopásek, Ivo (advisor) ; Branny, Pavel (referee) ; Holoubek, Aleš (referee)
Bacillus subtilis, a model Gram-positive soil bacterium, employs two distinct mechanisms in its membrane adaptation to low temperature: 1) Long-term adaptation to suboptimal temperature is accomplished by increasing the ratio of anteiso- to iso-branched fatty acids in the membrane lipids. 2) After a sudden temperature decrease, the oxygen-dependent fatty acid desaturase (Des) is induced which desaturates fatty-acyl chains incorporated in membrane lipids. The transcription of the gene encoding desaturase, des, is activated by the decrease of the membrane order, via two- component system DesK-DesR. In this work, I studied the influence of cultivation conditions on the mechanisms of B. subtilis membrane adjustments for a low temperature employing fatty acid analysis, fluorescence spectroscopy, differential scanning calorimetry and methods of molecular biology. In the first part of this work, I examined the impact of the cultivation medium on the composition and biophysical features of the B. subtilis cytoplasmic membrane during growth under the optimal (40 řC) and suboptimal (20 řC) cultivation temperature. I compared the nutrient-rich complex medium containing glucose and the mineral medium supplemented with either glucose or glycerol. The results obtained showed the crucial importance of medium...
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Interaction of Borrelia sp. with HL-60 cells and monocytes and cultivation of Anaplasma phagocytophilum in HL-60 cell culture
Marková, Lucie ; Hulínská, Dagmar (advisor) ; Beranová, Jana (referee)
Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum are causative agents of Lyme disease and human granulocytic anaplasmosis. Their common vector in Europe are the ticks from the genus Ixodes. In our work, we focused on interaction of innate immune cells with the causative agent of Lyme diseases, that are insubstitutable in their function in the early phase of the disease. Anaplasma phagocytophilum is hard to cultivate, the only possibility is to cultivate it in cell cultures. Successful cultivation of Anaplasma phagocytophilum acquired from patients in our geographic area is crucial for following experiments and for diagnostics too. In our experiments, we used validated cell cultures of HL-60 cells, canine monocytes DH82 and murine monocytes P388D1. During our studies of interaction of the causative agent of Lyme diseases with cells, we used two strains of different species Borrelia. Borrelia garinii M192 and Borrelia burgdorferi sensu stricto B31. These strains vary in virulence. The strain M192 is virulent, but the strain B31 lost its virulence by passages. We specialised in study of morphological changes using light microscopy (observation of dyed and fixed preparates and observation in dark field), eventually by transmision electron microscopy. During our experiments, we concluded that HL-60...
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The delta subunit of RNA polymerase from gram positive bacteria
Matějčková, Jitka ; Beranová, Jana (referee) ; Krásný, Libor (advisor)
1 Abstrakt Aby bakteriální buňka přežila neustále se měnící podmínky, musí se na ně adaptovat. Tato adaptace je podmíněna změnou genové exprese. Klíčovým krokem genové exprese je transkripce. Hlavním enzymem bakteriální transkripce je RNA polymerasa (RNAP), což je esenciální vícepodjednotkový enzym. RNAP je nejvíce prostudována u Escherichia coli, modelového organismu gram negativních bakterií. Porovnala jsem E. coli a Bacillus subtilis (zástupce gram pozitivních bakterií) a shrnula jsem rozdíly v RNAP a transkripci. Jejich RNA polymerasy se liší přítomností podjednotky δ u gram pozitivních bakterií. Tato podjednotka zvyšuje promotorovou selektivitu, recykluje jádro RNAP a celkově stimuluje syntézu RNA. Podjednotka δ ovlivňuje sporulaci a virulenci některých bakterií. V této práci jsem shromáždila současné poznatky o jednotlivých částech genové exprese, zejména o regulaci iniciace transkripce a o podjednotce δ RNAP.
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Impact of the economic crisis on unemployment in terms of sex, age and education in Czech Republic
Beranová, Jana ; Miskolczi, Martina (advisor) ; Průša, Ladislav (referee)
The objective of diploma thesis is to define specific groups of population, defined by sex, age or education, which are represented disproportionately in comparison of whole population of Czech Republic. In the thesis is described the population structure in terms of demography and economy, as well as factors which influence the labor market from short-term and long-term point of view. To achieve all marked out objectives is made explanatory statistics for unemployment of specified groups of population and the index analysis that allows to examine the inter-annual development of unemployment rate in terms of age, sex and education. Based on the index analysis are defined critical groups, which are evaluated by proportionality among unemployed persons. The analysis showed that critical groups are considered young people with lower level of education and women with high school education or women 30-39 years old.
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