Understanding protein function highly benefits from the knowledge of its three-dimensional structure, especially in the case of protein-ligand complexes. Structural biology methods such as X-ray crystallography, SAXS and NMR are therefore widely used for structural studies of protein-ligand interaction. In this work, these methods were used to understand two biological processes involving protein interactions: X-ray structural analysis was used to study binding of effector molecule to a prokaryotic transcription factor. NMR and SAXS techniques were used to study interaction of a monoclonal antibody with its protein antigen. Transcriptional regulator DeoR negatively regulates the expression of catabolic genes for the utilization of deoxyribonucleosides and deoxyribose in Bacillus subtilis. DeoR comprises an N-terminal DNA-binding domain and a C-terminal effector-binding domain (C-DeoR), and its function is regulated by binding of a small-molecular effector deoxyribose-5-phosphate. We determined crystal structures of C-DeoR both in the free form and in complex with deoxyribose-5-phosphate. Structural analysis revealed unique covalent binding of effector molecule through a reversible Schiff-base double bond with an effector-binding-site lysine residue. The physiological nature of this binding mode was...

Cardiovascular diseases represent the most important health risk factors because they are responsible for more than 50% of total mortality. Among them, the ischemic heart disease is leading cause of mortality. From the whole spectrum of different cardioprotective phenomena we have selected: 1) adaptation to chronic normobaric hypoxia (CNH) as the traditional experimental model in our laboratory area and 2) protective effect of exercise which in recent years represents promising and clinically relevant protective mechanism. The whole thesis is based on two studies. Aim of the first study was to characterize the expression of the main pro-inflammatory cytokine, TNF-α, in hearts of rats adapted to CNH. Chronic TNF-α inhibition by infliximab was used for discovering of certain role of TNF-α in CNH. We showed that increased myocardial level of TNF-α during adaptation to CNH was contributed via its receptor TNFR2 and nuclear factor κB-dependent activation of protective redox signalling with increased antioxidant defence. This adaptive pathway participates on the infarct size-limiting effect of CNH. Aim of the second study was find out whether exercise training and CNH could play synergy in cardiac protection in rats model. We reported that CNH and exercise reduced infarct size but their combination...

Fragestellung: Wir gingen der Frage nach, inwieweit genetische Variationen im Gen für den Tumor Nekrose Faktor-alpha einen Einfluß auf die Krankheitsprogression oder die Entstehung bzw. Ausprägung HIV-assoziierter Erkrankungen haben. Methoden: Die Promotorregion sowie die kodierenden Sequenzen des TNF-alpha-Gens wurden mittels SSCP-Analyse auf genetische Variationen untersucht. Anschließend erfolgte die Charakterisierung der häufigsten bekannten Promotorpolymorphismen mittels Restriktionsfragment-Längenpolymorphismus-Analyse (RFLP). Die Bestätigung der Polymorphismen der RFLP-Analyse erfolgte an ausgewählten Proben durch DNA-Fluoreszenzsequenzierung. In sämtlichen Fällen handelte es sich um singuläre Basentransitionen von Guanin zu Adenin. Ergebnisse: Unter Einbeziehung verschiedener Progressionsparameter wie der CD4-Zellzahl, des Zeitraumes vom ARC-Stadium zum AIDS-Stadium und AIDS-assoziierter Krankheiten wie dem Wasting Syndrom und der HIV-Enzephalopathie, erfolgte anschließend die statistische Analyse in Korrelation mit den ermittelten Genotypen. Es zeigte sich bei keiner der statistischen Analysen eine signifikante Assoziation mit einem bestimmten TNF-alpha-Genotyp. Schlußfolgerung: Es ist kritisch anzumerken, daß für einige Subanalysen die Größe der untersuchten Patientengruppe zu gering war, um eine statistische Aussagekraft für seltene Allele zu erreichen. Anhand der hier vorgelegten Ergebnisse hat der TNF-alpha-Genotyp weder einen Einfluß auf die Progression der HIV-Erkrankung noch auf die Ausbildung HIV-assoziierter Erkrankungen wie dem Wasting Syndrom oder der HIV-Enzephalopathie. / Objective: We determined whether variation of the tumor necrosis factor-alpha gene had an impact on HIV disease progression or the prevalence of hiv-associated diseases. Methods: The promotor region of the TNF-alpha gene were examined with SSCP analysis for polymorphisms in the promotor region. The most common promotor polymorphisms were characterized with restriction fragment length polymorphism analysis (RFLP). To confirm RFLP results DNA fluorescence sequenzing analyses were performed with selected samples. In all cases with diagnosis of promotor polymorphisms single base transitions from guanine to adenine were confirmed. Results: Statistical analyses correlated the genotypes with different markers for disease progression e.g. CD4-count, the period from ARC to AIDS and the occurance of HIV associated diseases (wasting syndrome, hiv encephalopathy). In none of the statistical analyses significant association with a certain TNF-alpha genotyp could be demonstrated. Conclusion: For some subanalysis the sample sizes were too small in order to be able to make safe statistical statements concerning rare allels. Regarding our results, none of the examined tumor necrosis factor-alpha promotor polymorphisms had an impact on HIV disease progression or the prevalence of hiv-associated diseases.

Es wurden die Effekte von Plättchenaktivierendem Faktor (PAF, 1-O-Alkyl-2-azetyl-sn-glyzero-3-phosphocholin) auf intrazelluläre Kalziumkonzentration und Zelllänge isolierter und feldstimulierter Kardiomyozyten der Ratte untersucht. Intrazelluläre Kalziumkonzentration und Zelllänge der feldstimulierten Zellen wurden mittels Laser-Raster-Mikroskopie simultan unter Verwendung des Kalzium-Fluoreszenzfarbstoffes Fluo-3 bestimmt. PAF (0.001nM - 10nM) inhibierte den systolischen Anstieg der intrazellulären Kalziumkonzentration zeit- und konzentrationsabhängig. Die maximalen Effekte wurden nach einer Inkubationszeit von 6-8 min beobachtet. Es kam zu 17% (0.001nM), 41% (0.1nM) und 52% (10nM PAF) Reduktion des systolischen Kalziumanstiegs. Zusätzlich konnte eine zeit- und konzentrationsabhängige Verringerung der simultan gemessenen Zellverkürzung nachgewiesen werden. Die Zellverkürzung war nach einer Inkubationszeit von 8 min um 10% (0.001nM), 32% (0.1nM) und 50% (10nM PAF) reduziert. Die Wirkungen von PAF konnten durch den Einsatz des spezifischen PAF-Rezeptorantagonisten WEB 2170 inhibiert werden. Diese Ergebnisse zeigen einen rezeptorvermittelten negativ inotropen Effekt von PAF, hervorgerufen durch eine Verringerung der systolischen intrazellulären Kalziumkonzentration ohne Desensibilisierung der Myofilamente. / We investigated the effects of platelet-activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) on intracellular calciumconcentration and cell length in isolated and field-stimulated rat cardiomyocytes. Intracellular calciumconcentration and cell length of field-stimulated cells were determined simultaneously by confocal laser scan microscopy by using the fluorescent calcium dye Fluo-3. PAF (0.001nM - 10nM) inhibited systolic intracellular calciumconcentration increase in a time- and concentration-dependent manner. Maximal effects were observed after an incubation time of 6-8 min, resulting in a 17% (0.001nM), 41% (0.1nM) and 52% (10nM PAF) inhibiton of systolic calcium increase. A time- and concentration-dependent decrease in simultaneously measured cell shortening also was demonstrated. Cell shortening was inhibited by 10% (0.001nM), 32% (0.1nM) and 50% (10nM PAF) after an incubation time of 8 min. The effects of PAF could be antagonized by the PAF-receptor antagonist WEB 2170. These data demonstrate that PAF receptor-dependently induces a negativ inotropic effect, which is correlated with a decrease in systolic intracellular calciumconcentration and is most likely not due to a decrease in myofilament sensitivity.

Terrain Breakline Detection Using Sky-view factor Abstract The subject of this diploma thesis is to propose the procedure of detecting of terrain breaklines using Sky-view factor. The theoretical part of the diploma thesis first introduces the issue of terrain breaklines detection, which presents the methods known to date and Sky- view factor, which is the focal point of the proposed method. Implementation over real data takes place at four locations in the Czechia (Vítkovice, Hradiště, Strnady and Velenka). Data in the form of raster format is tested. At the end, the results of the proposed method and its verification using two selected methods of field edge detection are presented and evaluated. Achieved results point to Sky-view factor for terrain edge detection. Keywords: terrain breakline, digital terrain model, Sky-view factor, detection terrain breaklines methods

<p>Predložen je i testiran model za energetsko vrednovanje kogenerativnih postrojenja pri čemu se proračun koeficijenta gubitka električne snage obavlja pomoću postupka koji je posebno naveden i predložen.<br />Sam model može i treba da predstavlja snažan alat prilikom kreiranja energetske politike, u delu koji se odnosi na kogeneraciju, s obzirom na to da pruža mogućnost jasnog energetskog vrednovanja postrojenja u smislu efikasnosti i uštede primarne energije kao i definisanja koji deo i koliko kogeneracija treba da bude stimulisana, odnosno potencijalno zastupljena u okviru nekog energetskog sistema.<br />Predloženi model je testiran na složenom kogenerativnom postrojenju snage 150 MW.</p> / <p>A model for energy evaluation of a cogeneration plant is proposed<br />and tested with the calculation of electric power loss done by means<br />of a procedure which is particularly specified and proposed.<br />The model itself can and should be used as a powerful tool for the<br />creation of energy policies in the part regarding cogeneration since it<br />gives opportunities for a clear energy evaluation of a plant in terms of<br />efficiency and primary energy saving, as well as for defining which<br />part and to what extent cogeneration should be stimulated, that is,<br />potentially represented within a specified energy system.<br />The proposed model has been tested on a cogeneration plant with<br />the capacity of 150 MW.</p>

Terrain Breakline Detection Using Sky-view factor Abstract The subject of this diploma thesis is to propose the procedure of detecting of terrain breaklines using Sky-view factor. The theoretical part of the diploma thesis first introduces the issue of terrain breaklines detection, which presents the methods known to date and Sky- view factor, which is the focal point of the proposed method. Implementation over real data takes place at four locations in the Czechia (Vítkovice, Hradiště, Strnady and Velenka). Data in the form of raster format is tested. At the end, the results of the proposed method and its verification using two selected methods of field edge detection are presented and evaluated. Achieved results point to Sky-view factor for terrain edge detection. Keywords: terrain breakline, digital terrain model, Sky-view factor, detection terrain breaklines methods

Automatiska flygningssystem och kontrollpresentationsenheter utvecklas hela tiden inom flygteknik. Avionik företag skapar datoriserade system som ska kunna automatisera ytterligare aspekter av de operationer som sker i cockpit. Att flyga ska inte bedömas vara något riskfyllt och för att säkerställa säkerhetssynen är det betydelsefullt att ta reda på om människor uppfattar teknikens framgång som positiv. Syftet är att ta reda på människans uppfattning och inställning till automatiserade flygplan. Frågeställningen besvaras genom en empirisk studie där först en noggrann bakgrundsundersökning gjorts för att få fram information om var automatiserade flygplan befinner sig gällande dess utveckling och framgång. Med hjälp av en gedigen bakgrund har en kvantitativ enkät undersökning genomförts som på bästa sätt ger svar på frågeställningen om automatiserade flygplan anses vara ett säkert transportmedel. Majoriteten av respondenterna, 86,9%, anser det vara viktigt eller väldigt viktigt att det finns en pilot som styr flygplanet. Automatiserade fraktflygplan är det som ligger närmast i framtiden jämfört med civila flygplan. Det har framförts att ökad förståelse om teknik ger en ökad tillit till AI och ju mer tillit till AI desto större chans till att skapa en bättre trygghetskänsla till automatiserade flygplan. Det kan även konkluderas att pilotlösa flygplan som styrs från kontrollrum på marken inte bör agera som en övergång till helt automatiserade flygplan. Istället föredras en stegvis automatisering i form av en digital andrepilot som hjälpmedel. Det måste dessutom skapas en större medvetenhet i hur flygplanen opererar för att få passagerare mottagbara till vidare utveckling. Slutligen måste inte automatiserade flygplan vara helt perfekta och eftersträva en omöjlig standard. / Automatic flight systems and control presentation units are constantly being developed in aviation technology. Avionic companies create computerized systems that will be able to automate additional aspects of the operations that take place in the cockpit. Flying should not be judged to be somewhat risky and to ensure safety, it is important to find out whether people perceive the technology's success as positive. The purpose is to find out about the people’s perception and attitude towards automated aircraft. The question is answered through an empirical study in which a thorough background study was first made to obtain information about where automated aircraft are located in relation to its development and success. With the help of a solid background, a quantitative questionnaire survey has been conducted that best answers the question of whether automated aircraft are considered a safe means of transport. The majority of respondents, 86.9%, consider it important or very important that there is a pilot who controls the aircraft. Automated cargo aircraft are the closest in the future compared to civil aircraft. It has been argued that increased understanding of technology gives increased confidence in AI and the more confidence in AI the greater the chance of creating a better sense of security for automated aircraft. It can also be concluded that pilotless aircraft controlled from control rooms on the ground should not act as a transition to fully automated aircraft. Instead, a step-by-step automation in the form of a digital second pilot is preferred. In addition, a greater awareness must be created in how the aircraft operate to get passengers susceptible to further development. Finally, automated aircraft need not be perfectly perfect and strive for an impossible standard.

<p>The purpose of this thesis have been to evaluate to what extend the ongoing development of Nato Network Enabling Capabilities Maturity levels, ( NNEC ML) have incorporated the human factor, or if the development is focusing to much on the technological aspects. As a background I have described the NNEC Maturity levels and to central elements; Defence lines of development and NNEC Value chain. In my thesis I point out different areas where the human factor is not taken enough into consideration, and I argue for a shift in focus from technology to the human factor. My conclusion is that it is necessary to shift focus from technology to the human factor in the following development if we shall succeed in bringing NATO and the different nations to the highest maturity level, Coherent. There is no doubt that technology is important, but it seems like the word networked is interpreted to be only a matter of technology. Concerning aspects like cultural understanding and the western linear way of thinking, it is necessary to broaden the meaning of the word networked to include the human factor in a greater sense. NNEC ML 5, Coherent means a maturity level where it is unnatural to operate in any other way than networked. A prerequisite to achieve this maturity level is that the people in the network feel familiar with the system they are operating, and trust their coalition partners. If we shall succeed in bringing the NATO coalition and the different nations to the highest maturity level it is essential that the human factor is prioritized in the following development. For the time being, the human factor is not emphasized enough.</p>

The purpose of this thesis have been to evaluate to what extend the ongoing development of Nato Network Enabling Capabilities Maturity levels, ( NNEC ML) have incorporated the human factor, or if the development is focusing to much on the technological aspects. As a background I have described the NNEC Maturity levels and to central elements; Defence lines of development and NNEC Value chain. In my thesis I point out different areas where the human factor is not taken enough into consideration, and I argue for a shift in focus from technology to the human factor. My conclusion is that it is necessary to shift focus from technology to the human factor in the following development if we shall succeed in bringing NATO and the different nations to the highest maturity level, Coherent. There is no doubt that technology is important, but it seems like the word networked is interpreted to be only a matter of technology. Concerning aspects like cultural understanding and the western linear way of thinking, it is necessary to broaden the meaning of the word networked to include the human factor in a greater sense. NNEC ML 5, Coherent means a maturity level where it is unnatural to operate in any other way than networked. A prerequisite to achieve this maturity level is that the people in the network feel familiar with the system they are operating, and trust their coalition partners. If we shall succeed in bringing the NATO coalition and the different nations to the highest maturity level it is essential that the human factor is prioritized in the following development. For the time being, the human factor is not emphasized enough.