This work is focused on the RTX (Repeats in ToXin) domains structure of selected RTX toxins and its impact on secretion and protein folding. The structural analysis included RTX domains of ApxI (Actinobacillus pleuropneumoniae-RTX-toxin I) from Actinobacillus pleuropneumoniae, HlyA (Alfa-hemolysin) from Escherichia coli and LtxA (Leukotoxin A) from Aggregatibacter actinomycetemcomitans and blocs 4 a 5 RTX domain CyaA (adenylate cyclase toxin) from Bordetella pertussis. The structures of LtxA RTX domain and CyaA RTX blocs 4 and 5 were obtained and characterized. Two models of CyaA RTX domain were built based on SAXS (Small Angle X-ray Scattering) model, previously solved RTX structures and RTX structures presented here.

The aim of this thesis is to analyze the phenomenon of female suicide terrorism in Chechnya, particularly with the focus on motivation of individual terrorists and organizations. Besides these two levels of analysis, it looks for facilitating and necessary conditions in the specific socio-political environment of Chechnya.

We study the dynamics of free test particles in a field of Schwarzschild black hole surrounded by an external exact thin axisymmetric solutions of Einstein's equations. Specifically, we use the Bach-Weyl ring and two member of the inverted Morgan-Morgan family of solutions as the additional sources. The fractal basin boundary and other meth- ods are used to detect and quantify chaos in time-like geodesic motion of the particles, primarily by computing box-counting dimension of said basin boundary. Our results mainly consist of the dependence of the chaoticity of these systems on mass and radius of the additional source as well as conserved energy and angular momentum of the test particles. We compare our results to literature and expand on them. 1

In this work we study properties of the higher-dimensional generally rotating black hole space-time so-called Kerr-NUT-(A)dS and the related spaces with the same explicit and hidden symetries as the Kerr-NUT-(A)dS spacetime. First, we search commuta- tivity conditions for classical (charged) observables and their operator analogues, then we investigate a fulfilment of these conditions in the metioned spaces. We calculate the curvature of these spaces and solve the charged Hamilton-Jacobi and Klein-Gordon equations by the separation of the variables for an electromagnetic field, which pre- serves integrability of motion of a charged particle and mutual commutativity of the corresponding operators.

1 Abstract Adenylate cyclase toxin (CyaA) is one of the major virulence factors of bacterium Bordetella pertussis, which is a causative agent of whooping cough. CyaA belongs to the family of RTX toxin-hemolysins. The toxin targets primarily cells expressing integrin receptor CD11b/CD18 but it can also penetrate cells lacking this receptor. CyaA acts on host cells by two independent activities. One is formation of small cation-selective channels, which can lead to colloid osmotic lysis of target cells. The second is disruption of cell signaling through the translocation of the adenylate cyclase (AC) domain to host cell cytosol, which leads to the conversion of ATP into cyclic AMP. It was recently shown that cholesterol affects endocytosis of CyaA. CyaA translocates it's AC domain after relocation of CyaA molecule to the cholesterol-rich lipid raft (Bumba et al. 2010). In this work I examined the effect of cholesterol on channel- forming activity and selectivity of ion channels created by CyaA. For measurements I used artificial membranes enriched with cholesterol. CyaA channels are voltage-dependent. The positive membrane potential on the side of toxin is rquired for incorporation of CyaA molecule into cell membrane. I tried to find out whether the value of voltage has effect on channels opening time....

Colicin U is a protein produced by strains of bacterium Shigella boydii. It exhibits antibacterial activity against some bacterial strains Shigella and Escherichia. Based on sequence homology with colicins A, B and N, the colicin U is classified as a pore-forming colicin. Interaction of colicin U with attacked bacteria is ensured by three-step mechanism: 1) First colicin U interacts with surface receptors OmpA, OmpF and core of LPS. 2) Thereafter the colicin is translocated to periplasm through interaction with Tol proteins. 3) Finally colicin U interacts with the inner membrane of the attacked bacteria causing its depolarization. In this thesis I demonstrated pore-forming features of colicin U and further observed characteristics and properties of these pores. Using methods of measuring on black lipid membranes I determined a single channel conductance (19 pS), ion selectivity, the influence of various conditions on the behaviour of the pores. These findings, in many cases, correspond to the findings on other related colicins. Furthermore, I successfully determined the pore diameter of colicin U ( ≈ 0,8 nm). The next section of the thesis focuses on creation of single cysteine mutations of colicin U. Subsequently I produced five mutant variants of colicin U and verified their functionality so that...

Thesis: The TV series Black Mirror as a substantial creation of British TV production Abstract: ENG The thesis deals with the analysis of the British TV anthology Black Mirror. It examines the program in terms of authorship, processing method, theme, genre, and impact on society. The aim is to determine the basic characteristics of the Black Mirror and to point out its uniqueness in the context of current trends in television content. In individual chapters, it focuses on the role of authorship, the definition of the genre and the definition of the main topic. The program is also subjected to a narrative and stylistic analysis that examines the way the story is constructed, the narrative structure, the work with the characters, the environment and the style. Furthermore this thesis also want to examine the the extent to which the topics of Black Mirror correspond to reality. The result of this research offers a complex picture of Black Mirror, including a comparison with selected British television programs.

The adenylate cyclase toxin-hemolysin (CyaA) plays a key role in virulence of the whooping cough agent Bordetella pertussis. It translocates an AC enzyme into cytosol of CD11b+ phagocytes and subverts their bactericidal functions by unregulated conversion of ATP to cAMP. In parallel, CyaA permeabilizes cellular membrane by forming cation-selective pores. The goal of my diploma thesis was an analysis of the mechanism of interaction of the segment linking the invasive adenylate cyclase domain and the RTX hemolysin moiety of CyaA with target membrane. Our data show that the segment linking the AC to the hydrophobic domain of CyaA is directly involved in the interaction of the toxin with the membrane and controls the formation of small cationt-selective pores. Our results generate new knowledge that will be of relevance to the entire field of toxin biology and will enable the design of improved CyaA- based vaccines. Keywords: Bordetella pertussis, adenylate cyclase toxin, membrane translocation, pore- forming activity, black lipid bilayers, liposomes

S vývojem informací, komunikací a technologií se moderní průmyslové řídicí systémy (ICS) potýkají čím dál více s otázkami automatického testováni pro zabezpečení stability a bezpečnosti systému. Z tohoto důvodu se testování stalo jednou z nejdůležitějších částí životního cyklu všech softwarů. V této diplomové práci budu zvažovat možnost využití stávajících zkušebních metod a nástrojů pro získání dostatečné jakosti a bezpečnosti softwaru v kontinuálních integračních systémech.

This thesis is focused on design of black coal combustion boiler of 160t/h output. It consists of several parts. First, stechiometric calculations and enthalpy of air and gas are made, then heat balance of boiler and the boiler losses are calculated, and also thermal efficiency is determined. In next step, the combustion chamber is designed and its thermal calculation is made. Then, each of heating surfaces are designed, and in the last step overall energy balance of the boiler is checked. The scheme of pressure system and design documentation added.