Daňová zátěž a vliv na rozhodování managementu, vliv případných změn daňového systému na konkurenceschopnost podniku a na rozhodovací procesy, v praktické části aplikace na podnikatelský subjekt. Optimalizace daňové povinnosti u fyzické osoby, přechod na podnikání právnické osoby, důsledky daňové reformy v roce 2008 pro podnikatele, srovnání všech modelových situací a následné doporučení.

The aim of this project is detection of gass leak based on the ultrasound sensing. Work introduces ultrasound, mechanism of its propagation and attenuation. From all of the available microphones, the piezoelectric microphone was chosen. Project continues with description of the device, which can be capable of detection of the ultrasound leakage in dangerous, explosive environment. This type of device must meet requirements of SIL 2 standard and ATEX directive for zone 2. Project discusses LNA (Low Noise Amplifier), filters and detection circuits design. The device is modular what means that each module can be freely replaced. The role of these modules is to convert analog signal to digital form suitable for microcontroller HERCULES TSM57012.

No description available.

Zu den Eigenschaften eukaryotischer Zellen gehört ihre Kompartimentierung, welche durch die Abtrennung verschiedener Reaktionsräume durch Lipiddoppelschichten erreicht wird. Verschiedene Vesikel-Transportwege verbinden diese Kompartimente miteinander, einer dieser Wege in der Hefe Saccharomyces cerevisiae ist der sogenannte ALP-Weg. Dieser gehört zu den biosynthetischen Wegen, über die neue Proteine an ihren Bestimmungsort gebracht werden, in diesem Falle die Vakuole. Ausgehend vom Golgi-Apparat werden die Vesikel dieses Weges mit Hilfe des Adaptorproteinkomplexes-3 (AP-3) gebildet. Ein weiteres Protein, das eine spezifische Funktion in diesem Weg übernimmt, ist Vps41. Ein aktuelles Modell beschreibt seine Funktion in der Aufnahme der Vesikel an der Vakuole. Es konnte gezeigt werden, das Vps41 mit der sogenannten ear-Domäne von Apl5, einer Untereinheit des AP-3- Komplexes, interagiert. In dieser Arbeit konnte ich nachweisen, dass die Interaktionsstelle im Vps41 innerhalb einer konservierten PEST-Domäne liegt. Eine Deletion dieser Domäne beeinflußte die Funktion des Proteins im ALP-Weg jedoch nicht die in der homotypischen Vakuolenfusion und im CPY-Weg. Eine weitere Eingrenzung des deletierten Bereiches zeigte, dass die PEST-Domäne eine Sequenz enthält, die einem Di-Leucin- Sortierungssignal ähnlich ist. Dieses konnte ich als minimal notwendigen Bereich für die Wechselwirkung mit der Apl5-ear-Domäne bestimmen. Meine Daten zeigen, dass dieser Bereich des Proteins notwendig ist für das Docking der AP-3-Vesikel an der Vakuole. Weiterhin konnte ich eine kompetitive Bindung von Liposomen und Apl5 an die N-terminale Hälfte von Vps41 zeigen. Zusammengefasst und mit aktuellen Veröffentlichungen in Zusammhang gebracht, ergänzen meine Daten das Modell der Funktion von Vps41 in der Vesikelaufnahme an der Vakuole: Vps41 wird durch die Rab-GTPase Ypt7, als deren Effektorprotein, an späte Endosomen gebunden. An dieser stark gekrümmten Membran taucht ein kürzlich identifiziertes ALPS (amphipathic lipid packing sensor)-Motiv im Vps41 in die Membran des Organells ein und zieht so den N-terminalen Bereich mit der Bindestelle für die AP-3-Vesikel an die Oberfläche des Organells wodurch eine verfrühte Fusion der AP-3-Vesikel mit dem Endosom verhindert wird. Erst nach der Reifung zur Vakuole wird die PEST-Domäne für die Bindung an Apl5 verfügbar, da sich die Membrankrümmung ändert. Zusätzlich wird das ALPS-Motiv phosphoryliert, so dass dieses nicht mehr in die Membran eintauchen kann. Erst jetzt ist eine Interaktion zwischen Apl5 und Vps41 und damit eine Fusion der AP-3-Vesikel mit der Vakuole möglich. Der zweite Teil dieser Arbeit beschäftigt sich mit der Protonentranslokation durch den Fo-Teil der ATP-Synthase aus Escherichia coli. Durch Mutagenese wurden ATP-Synthasen hergestellt, in denen die beiden für den Protonentransport essentiellen Aminosäurereste D61 in der Untereinheit c und R210 in der Untereinheit a in der α-Helix in der sie liegen, entweder einzeln oder beide zusammen, um je eine Helixwindung nach oben oder unten verschoben wurden. Dies führt zu einer Verlängerung bzw. Verkürzung der Protonenzu- und austrittskanäle. Durch die Untersuchung der Funktionalität dieser ATPasen auf sowohl aktives und passives Protonenpumpen, als auch ATP-Synthese konnte ich zeigen, daß die Position der beiden essentiellen Aminosäurereste cD61 und aR210 zueinander nicht entscheidend ist. Werden beide Reste in die gleiche Richtung verschoben, so daß ihre Position zueinander gleich bleibt, kommt es unabhängig von der Richtung immer zu einem kompletten Funktionsverlust. Weiterhin läßt sich aus meinen Daten folgern, daß die Position des Restes aR210 in der Mitte der Membran wichtig ist. Beim Verschieben des Restes auf die Position 206 (a-up) geht die gesamte Funktion des Fo-Teiles verloren, während das Verschieben auf die Position 214 (a-down) zu einem passiven Ausströmen der Protonen durch den Fo-Teil führt. Die Position des Restes cD61 in der Membran ist flexibler. Obwohl die Repositionierung des Aspartats auf die Position 57 (c-up) jegliche Funktionalität des Fo-Teiles beeinträchtigt, ermöglicht ein Verschieben auf die Position 65 (c-down) aktives und passives Protonenpumpen, sowie die Synthese von ATP.

Proton channels are essential for many of the processes of life. The influenza A viral protein M2 is responsible for sensing the conditions necessary for viral RNA release. The proton-translocating FoF1 ATPase (ATP synthase) uses a proton gradient to drive adenosine triphosphate (ATP) synthesis. We have directly measured proton uptake in vesicles containing reconstituted M2 or FO by monitoring external pH after addition of valinomycin to vesicles with 100-fold diluted external [K+]. This proton flux assay was utilized to quantify proton flux through single M2 and Fo channels. Contrary to previous reports, proton uptake by M2 was not significantly altered by acidification of the extravesicular pH. We conclude that pH only weakly affects proton flux through M2 in the pH range of 5.4 - 7.0. Theoretical analysis utilized for such vesicle uptake assays illuminates the appropriate time scale of the initial slope and an important limitation that must be placed on inferences about channel ion selectivity. The rise in pH over 10 seconds after ionophore addition yielded time-averaged single channel conductances of 0.35±0.2 aS and 0.72±0.4 aS at pH 5.4 and 7.0 respectively. Such a low time-average conductance implies that M2 is only conductive 10^-6 to 10^-4 of the time. M2 selectivity for hydrogen over potassium is ~10^7. Fo translocates protons across membranes, converting electrochemical energy to rotational inertia. Previous experiments have been partially confounded by a contaminating channel, CL, which co-purifies with Fo and leaks cations. CL activity is shown to not decrease following deletion of the previously uncharacterized yraM open reading frame of E. coli. Fo purified from a deletion strain lacking yraM is just as active as Fo purified from the wild-type strain. Using Fo from the deletion strain, the single-hit hypothesis of DCCD inhibition of passive proton flux through Fo was examined. A DCCD-induced reduction in ATP synthase activity correlates with a reduction in the total initial slope, the number of functional Fo per µg protein, and the single channel proton flux. At least 2 DCCD per Fo are required to totally inactivate passive proton flux. M2 and Fo have similar single channel conductances but different open probabilities.

Since 2002, NASA's GRACE Satellite mission has allowed scientists of various disciplines to analyze and map the changes in Earth's total water storage on a global scale. Although the raw data is available to the public, the process of viewing, manipulating, and analyzing the GRACE data can be difficult for those without strong technological backgrounds in programming or geospatial software. This is particularly true for water managers in developing countries, where GRACE data could be a valuable asset for sustainable water resource management. To address this problem, I have a developed a utility for subsetting GRACE data to particular regions of interest and I have packaged that utility in a web app that allows water managers to quickly and easily visualize GRACE data these regions. Using the GLDAS-Noah Land Surface Model, the total water storage for the regions derived from the raw GRACE data is decomposed into surface water, soil moisture, and groundwater components. The GRACE Groundwater Subsetting Tool is easily deployed, open-source, and provides access to all of the major signal processing solutions available for the total water storage data. The application has been successfully applied to both developed and developing countries in various parts of the world, including the Central Valley region in California, Bangladesh, the La Plata River Basin in South America, and the SERVIR Hindu Kush Himalaya region. The groundwater data in this application has proven capable of monitoring groundwater use based on drought trends as well as agricultural demand in a number of locations and can assist in uniting decision makers and water users in the mission of sustainably managing the world's groundwater resources.

University of Macau / Faculty of Social Sciences and Humanities / Department of Government and Public Administration

Desde a década de 1980 diversos autores apresentaram correlações entre provas de carga estática e ensaios de carregamento dinâmico em estacas. Para uma boa correlação é fundamental que os testes sejam bem executados e que atinjam a ruptura segundo algum critério, como o de Davisson, por exemplo, além de levar em conta o intervalo de tempo entre a execução da prova de carga estática e do ensaio dinâmico, face ao efeito \"set up\". Após a realização do ensaio dinâmico realiza-se a análise CAPWAP que permite a determinação da distribuição do atrito lateral em profundidade, a carga de ponta e outros parâmetros dos solos tais como quakes e damping. A análise CAPWAP é realizada por tentativas através do procedimento \"signal matching\", isto é, o melhor ajuste entre os sinais de força medido pelos sensores e o calculado. É relativamente fácil mostrar que a mesma solução pode ser obtida através de dados de entrada diferentes. Isso significa que apesar de apresentarem cargas mobilizadas próximas o formato da curva da simulação de prova de carga estática, obtida pelo CAPWAP, assim como a distribuição do atrito lateral, podem ser diferentes, mesmo que as análises apresentem \"match quality\" (MQWU) satisfatórios. Uma forma de corrigir o formato da curva simulada do CAPWAP, assim como a distribuição do atrito lateral, é através da comparação com provas de carga estática (PCE). A sobreposição das duas curvas, a simulada e a \"real\", permite a determinação do quake do fuste através do trecho inicial da curva carga-recalque da prova de carga estática, que por sua vez permite uma melhor definição da distribuição do atrito lateral e da reação de ponta. Neste contexto surge o conceito de \"match quality de recalques\" (MQR). Quando a PCE não está disponível, propõe-se efetuar um carregamento estático utilizando o peso próprio do martelo do bate-estaca (CEPM). Mostra-se, através de dois casos de obra, em que estavam disponíveis ensaios de carregamento dinâmico e PCEs, que esse procedimento permite obter uma melhor solução do ponto de vista físico, isto é consistente com as características do subsolo e com a curva carga-recalque da PCE, e não apenas matemático, através da avaliação do \"match quality\" (MQWU). / Since the 1980s a lot of authors showed correlations between static load tests and dynamic load tests on piles. For a good correlation it is necessary a good execution of the load test, also it is necessary to choose a capacity value from the results of the static load test, for example, the Davisson Offset limit load. The time of execution between the static load test and the dynamic load test should be considered because of the set up effect. Dynamic data may be further analyzed by CAPWAP Method to evaluate the soil resistance distribution, the toe resistance, quake and damping values. It is a signal matching method. Its results are based on the \"best possible match\" between computed pile top variable such as the pile top force and its measured equivalent. It is easy to demonstrate almost the same pile capacity on CAPWAP using different soil parameters. It means that even the pile capacity is almost the same, the shape of the pile top load-displacement of the CAPWAP Method and the shaft friction distribution can be different, although all results confirm good match quality. One way to correct the shape of the top loaddisplacement of the CAPWAP Method, as well as the shaft friction distribution, is by comparisson to a static load test. Overlaying both curves, the static load test and the CAPWAP Method, it is possible to determine the shaft quake value on the initial loads on the top load-displacement curve, allowing this way a improvement of the shaft resistance distribution and the toe resistance. In this context arises the concept of \"match quality of settlements\". When the static load test is not avaliabe, this thesis proposes a static load test using the hammer\'s weight of the pile driving machine. It is shown by two case studies that were available static load tests and dynamic load tests, this procedure allows a better solution on physics aspects, this is consistent with the subsoil conditions and the load-settlement curve of the static load test, not only a mathematical solution based on match quality.

Recombinant strategies are today dominating in thedevelopment of modern subunit vaccines. This thesis describesstrategies for the production and recovery of protein subunitimmunogens, and how genetic design of the expression vectorscan be used to adapt the immunogens for incorporation intoadjuvant systems. In addition, different strategies fordelivery of subunit vaccines by RNA or DNA immunization havebeen investigated. Attempts to create general production strategies forrecombinant protein immunogens in such a way that these areadapted for association with an adjuvant formulation wereevaluated. Different hydrophobic amino acid sequences, beingeither theoretically designed or representing transmembraneregions of bacterial or viral origin, were fused on gene leveleither N-terminally or C-terminally to allow association withiscoms. In addition, affinity tags derived fromStaphylococcus aureusprotein A (SpA) or streptococcalprotein G (SpG), were incorporated to allow efficient recoveryby means of affinity chromatography. A malaria peptide, M5,derived from the central repeat region of thePlasmodium falciparumblood-stage antigen Pf155/RESA,served as model immunogen in these studies. Furthermore,strategies forin vivoorin vitrolipidation of recombinant immunogens for iscomincorporation were also investigated, with a model immunogendeltaSAG1 derived fromToxoplasma gondii. Both strategies were found to befunctional in that the produced and affinity purified fusionproteins indeed associated with iscoms. The iscoms werefurthermore capable of inducing antigen-specific antibodyresponses upon immunization of mice, and we thus believe thatthe presented strategies offer convenient methods for adjuvantassociation. Recombinant production of a respiratory syncytial virus(RSV) candidate vaccine, BBG2Na, in baby hamster kidney(BHK-21) cells was investigated. Semliki Forest virus(SFV)-based expression vectors encoding both intracellular andsecreted forms of BBG2Na were constructed and found to befunctional. Efficient recovery of BBG2Na could be achieved bycombining serum-free production with a recovery strategy usinga product-specific affinity-column based on a combinatoriallyengineered SpA domain, with specific binding to the G proteinpart of the product. Plasmid vectors encoding cytoplasmic or secreted variants ofBBG2Na, and employing the SFV replicase for self-amplification,was constructed and evaluated for DNA immunization against RSV.Both plasmid vectors were found to be functional in terms ofBBG2Na expression and localization. Upon intramuscularimmunization of mice, the plasmid vector encoding the secretedvariant of the antigen elicited significant anti-BBG2Na titersand demonstrated lung protective efficacy in mice. This studyclearly demonstrate that protective immune responses to RSV canbe elicited in mice by DNA immunization, and that differentialtargeting of the antigens expressed by nucleic acid vaccinationcould significantly influence the immunogenicity and protectiveefficacy. We further evaluated DNA and RNA constructs based on the SFVreplicon in comparison with a conventional DNA plasmid forinduction of antibody responses against theP. falciparumPf332-derived antigen EB200. In general,the antibody responses induced were relatively low, the highestresponses surprisingly obtained with the conventional DNAplasmid. Also recombinant SFV suicide particles inducedEB200-reactive antibodies. Importantly, all immunogens inducedan immunological memory, which could be efficiently activatedby a booster injection with EB200 protein. <b>Keywords</b>: Affibody, Affinity chromatography, Affinitypurification, DNA immunization, Expression plasmid, Fusionprotein, Hydrophobic tag, Iscoms, Lipid tagging, Malaria,Mammalian cell expression, Recombinant immunogen, RespiratorySyncytial Virus, Semliki Forest virus, Serum albumin,Staphylococcus aureusprotein A, Subunit vaccine,Toxoplasma gondii

In this study, the effects of submergence on local scour in a uniform cohesionless sediment bed by a plane turbulent wall jet and the resulting flow field were investigated experimentally. Here, submergence is defined as the ratio of the tailwater depth to the thickness of the jet at its origin. The main focus was to determine scour dimensions at an asymptotic state, examine whether there was similarity in the velocity profiles for the flow in the scour hole, and to determine the growth of the length scales and decay of the maximum velocity of the jet. Also examined were the relationships between the scales for the velocity field in the scour hole and the scour hole size.<p>In the experiments, the range of submergence was varied from 3-17.5, whereas the range of densimetric Froude number and the ratio of the boundary roughness to the gate opening (relative boundary roughness) were varied from 4.4-6.9 and 0.085-0.137 respectively. The velocity field in the scour hole at asymptotic state was measured using a SonTek 16-MHz MicroADV. Time development of the characteristic dimensions of the scour hole was also measured.<p>The dimensions of the scour hole were found to increase with increasing submergence for all experiments with a bed-jet flow regime. In the bed-jet flow regime, the jet remains near the bed throughout the scouring process. Further, the time development of the scour hole dimensions were observed to increase approximately linearly with the logarithm of time up to a certain time before the beginning of asymptotic state for experiments with either the bed-jet or surface-jet flow regimes.<p> The flow field results showed that the velocity profiles in the region of forward flow and the recirculating region above the jet were similar in shape up to about the location of the maximum scour depth. Relationships describing this velocity profile, including its velocity and length scales, were formulated. The decay rate of the maximum velocity, the growth of the jet half-width, and the boundary layer thickness were also studied. The decay and the growth rate of the jet length scales were found to be influenced by the submergence ratio, densimetric Froude number, and the relative boundary roughness.<p>Two distinct stages in the decay of the maximum streamwise velocity, with distance along the direction of flow, were observed for the jet flows having a bed-jet flow regime. The first stage of velocity decay was characterized by a curvilinear decay of velocity, which followed that of a wall jet on a smooth, rigid bed for streamwise distance approximately equal to 2L. For the surface-jet flow regime, the decay of velocity was observed to be similar to that of a free-jump on a smooth, rigid bed for a streamwise distance approximately equal to L. Here, L is defined as the streamwise distance measured from the end of the rigid apron to where the maximum streamwise velocity in the jet is half the velocity of the jet at the end of apron. The streamwise maximum velocity of the jet was then seen to increase in what was called the recovery zone.<p>A relationship for the streamwise decay of the maximum velocity within the scour hole is proposed. Moreover, other scales representing the flow inside the scour hole such as the streamwise distance from the end of the apron to where the streamwise maximum velocity starts to deviate from curvilinear to linear decay and the streamwise distance to where maximum streamwise velocity starts to increase are suggested. Some new results on the velocity distribution for the reverse flow for a bed-jet flow regime are also presented. Finally, some dimensionless empirical equations describing the relationship between the jet scales for the jet flow in a scour hole and the scour hole size are given.