A contribuição de células B para a memória imunológica se dá por duas distintas populações: células B de memória e células produtoras de anticorpos de longa vida (ASC). A inter-relação entre estas células bem como os mecanismos envolvidos para a manutenção destas tem sido pouco entendida. O veneno de Thalassophryne nattereri tem se mostrado capaz de induzir uma intensa resposta imune de memória. Nós avaliamos o efeito das Natterinas, que são as toxinas majoritárias e inéditas, na indução e manutenção da resposta imune de memória de células B. Este estudo, além de permitir um maior esclarecimento da resposta humoral de memória induzida pelo veneno do peixe T. nattereri, permitiu o estudo da complexa organização do compartimento de células B de memória e ASCs. Também evidenciamos a importância da atividade proteásica para a manutenção da cronicidade de resposta de células B no peritônio, no baço e na medula, como verificamos que a ativação de receptores inatos como osTLRs é decisiva para a geração e manutenção de ASCs B220pos/neg em resposta às Natterinas, dependentes das vias de sinalização MyD88 ou TRIF. Estas sinalizações controlam a magnitude, a qualidade e a longa duração da resposta humoral de memória. / The contribution of B cells for the immunological memory feels for two different populations: memory B cells and long-lived antibodies secreting cells (ASC). The interrelation among these cells as well as the mechanisms involved for the maintenance of these it has been little understood. The venom of Thalassophryne nattereri possesses the ability to induce an intense memory immune response. We evaluated the effect of Natterins that are majority toxins in the venom, in the induction and maintenance of the immune memory response of cells B. The study, besides allowing a larger explanation of the humoral memory response induced by the venom of the fish, it allowed the understanding of the complex organization of the memory B cells compartment, mainly of the subtype of long-lived cells (ASC). Also, we showed the importance of the protease activity of Natterins in the maintenance of the chronic B cell responses in the three analyzed compartments. We verify that the activation of Toll like receptors is decisive for the generation and maintenance of ASCs B220pos/neg in response to Natterins, dependent on the MyD88 or TRIF signaling that control the quality and the duration of the humoral memory response.

A contribuição de células B para a memória imunológica se dá por duas distintas populações: células B de memória e células produtoras de anticorpos de longa vida (ASC). A inter-relação entre estas células bem como os mecanismos envolvidos para a manutenção destas tem sido pouco entendida. O veneno de Thalassophryne nattereri tem se mostrado capaz de induzir uma intensa resposta imune de memória. Nós avaliamos o efeito das Natterinas, que são as toxinas majoritárias e inéditas, na indução e manutenção da resposta imune de memória de células B. Este estudo, além de permitir um maior esclarecimento da resposta humoral de memória induzida pelo veneno do peixe T. nattereri, permitiu o estudo da complexa organização do compartimento de células B de memória e ASCs. Também evidenciamos a importância da atividade proteásica para a manutenção da cronicidade de resposta de células B no peritônio, no baço e na medula, como verificamos que a ativação de receptores inatos como osTLRs é decisiva para a geração e manutenção de ASCs B220pos/neg em resposta às Natterinas, dependentes das vias de sinalização MyD88 ou TRIF. Estas sinalizações controlam a magnitude, a qualidade e a longa duração da resposta humoral de memória. / The contribution of B cells for the immunological memory feels for two different populations: memory B cells and long-lived antibodies secreting cells (ASC). The interrelation among these cells as well as the mechanisms involved for the maintenance of these it has been little understood. The venom of Thalassophryne nattereri possesses the ability to induce an intense memory immune response. We evaluated the effect of Natterins that are majority toxins in the venom, in the induction and maintenance of the immune memory response of cells B. The study, besides allowing a larger explanation of the humoral memory response induced by the venom of the fish, it allowed the understanding of the complex organization of the memory B cells compartment, mainly of the subtype of long-lived cells (ASC). Also, we showed the importance of the protease activity of Natterins in the maintenance of the chronic B cell responses in the three analyzed compartments. We verify that the activation of Toll like receptors is decisive for the generation and maintenance of ASCs B220pos/neg in response to Natterins, dependent on the MyD88 or TRIF signaling that control the quality and the duration of the humoral memory response.

Today’s carbon constrained world with its increasing demand for cheap energy and a fossil fuel intensive fleet of power producers is making carbon capture and storage (CCS) desirable. Several CCS technologies are under investigation by various research and development groups globally. One of the more promising technologies is oxy-fuel combustion, since it produces a CO2 rich flue gas which requires minor processing to meet storage condition requirements. In this study the economics of an advanced super critical oxy-coal power plant burning lignite, simulated in-house was assessed. A robust and user-friendly financial tool box has been developed with commonly acceptable default parameter settings. Capital, operation and maintenance costs were estimated along with corresponding levelized cost of electricity and CO2 avoidance costs calculated using the detailed financial model developed. A levelized cost of electricity of 131 $/MWhrnet along with a levelized CO2 avoidance cost of 64 $/tonne was estimated for an ASC oxy-coal power plant with CO2 capture. Also a levelized cost of electricity of 83 $/MWhrnet was estimated for an ASC air-fired coal power plant without CO2 capture capabilities as the base plant. The price of electricity was observed to increase from 83 $/MWhrnet to 131 $/MWhrnet translating into a 57% increase. The sensitivity of the overall economics of the process was assessed to several parameters. The overall economics was found sensitive to the choice chemical engineering plant cost index (CEPCI), capacity factor, size of power plant, debt ratio, fuel price, interest rate, and construction duration.

Today???s carbon constrained world with its increasing demand for cheap energy and a fossil fuel intensive fleet of power producers is making carbon capture and storage (CCS) desirable. Several CCS technologies are under investigation by various research and development groups globally. One of the more promising technologies is oxy-fuel combustion, since it produces a CO2 rich flue gas which requires minor processing to meet storage condition requirements. In this study the economics of an advanced super critical oxy-coal power plant burning lignite, simulated in-house was assessed. A robust and user-friendly financial tool box has been developed with commonly acceptable default parameter settings. Capital, operation and maintenance costs were estimated along with corresponding levelized cost of electricity and CO2 avoidance costs calculated using the detailed financial model developed. A levelized cost of electricity of 131 $/MWhrnet along with a levelized CO2 avoidance cost of 64 $/tonne was estimated for an ASC oxy-coal power plant with CO2 capture. Also a levelized cost of electricity of 83 $/MWhrnet was estimated for an ASC air-fired coal power plant without CO2 capture capabilities as the base plant. The price of electricity was observed to increase from 83 $/MWhrnet to 131 $/MWhrnet translating into a 57% increase. The sensitivity of the overall economics of the process was assessed to several parameters. The overall economics was found sensitive to the choice chemical engineering plant cost index (CEPCI), capacity factor, size of power plant, debt ratio, fuel price, interest rate, and construction duration.

This diploma thesis deals with specifics for reporting of construction contracts in financial statements according to czech accounting regulations, IFRS and US GAAP. These are contracts that are custom-made, so they are not serial production and they are long-term, so most of their production goes into more accounting periods. In 2018, new Standards for Recognition of Revenue from contracts with customers IFRS 15 and ASC 606 will enter into force. The aim of the thesis is to characterize individual approaches to the reporting of revenues from construction contracts with a focus on the specifics in this sector and their comparison. To achieve this goal, a comparison method will be used. The thesis is divided into the theoretical and practical part. The theoretical part describes requirements and aspects related to building contracts, individual accounting standards including new revenue standards. In the practical part, an example is illustrated that shows a different approach to reporting revenue from these contracts in accordance with Czech accounting regulations and IFRS or US GAAP and comments on the changes that may occur through the application of new standards. At the end of the thesis, individual approaches are compared.

The genetic systems involved in the utilisation of aromatic β-glucosides in E. coli consist of the bgl, asc, and chb operons and the locus bglA encoding phospho-β-glucosidase A. The bgl and asc operons are known as cryptic or silent systems since their expression is not sufficient for utilisation of these sugars in wild type strains of E. coli. Their transcriptional activation by different classes of mutations confers a Bgl+ phenotype to the mutant. The maintenance of cryptic genes without accumulating deleterious mutation in spite of being silent is an evolutionary puzzle. Several observations have suggested the possibility that these genes may be expressed under specific physiological conditions conferring a fitness advantage to the organism. The main aim of this study was to investigate the possible role of aromatic β-glucoside catabolic systems of E. coli in combating nutrient stress and microaerobic growth conditions. The results presented in Chapter 2 address the evolution of aromatic β-glucoside catabolic systems when exposed to a novel β-glucoside as the sole substrate. The results indicate that the bgl opeon, the primary system involved in the utilisation of the aromatic β-glucosides arbutin and salicin, is also involved in esculin utilisation. In the absence of bglB encoding the enzyme phospho-β-glucosidase B, activation of the silent asc operon enables esculin utilisation. The bglA gene encoding phospho-β-glucosidase A specific for arbutin, can undergo successive mutations to evolve the ability to hydrolyse esculin and salicin sequentially when bglB and ascB are absent. The Esc+ and Sal+ mutants retain their arbutin+ phenotype, indicating that the mutations enhance the promiscuity of the enzyme. Sequencing data indicate that the first step Esc+ mutant carries a four base insertion within the promoter of the bglA gene that results in enhanced transcription of bglA. RT-PCR studies confirm that both the steady-state levels as well as the half-life of the bglA mRNA are enhanced in the mutant. This is further corroborated by the observation that overexpression of wild type bglA in the parent strain using a multicopy plasmid confers an Esc+ phenotype. The second step Sal+ mutant carries a point mutation within bglA ORF, a thymine to guanine transversion at position 583 (T583G) of the bglA gene, resulting in an amino acid change from cysteine to glycine at position 195 (C195G) of the BglA ORF close to the active site. Presence of a plasmid carrying the T583G mutation, introduced by site-directed mutagenesis, results in a Sal+ phenotype, confirming the role of the transversion in conferring the Sal+ phenotype. Based on docking studies, the positioning of salicin into the substrate binding site of the mutant BglA enzyme is different compared to wild type BglA due to the loss of stearic hindrance for the binding of salicin when C195 is replaced by the smaller amino acid glycine in the mutant protein. These observations indicate that under conditions of nutrient deprivation, exposure to novel substrates can result in the evolution of new metabolic capabilities by the sequential modification of a pre-existing genetic system. In the case of one novel substrate, the mutation results in the overexpression of the hydrolytic enzyme, while in the case of the second substrate, a mutation close to its active site increases its substrate specificity. Results presented in Chapter 3 specifically deal with the involvement of the bgl operon under low levels of oxygen. Earlier observations have shown that there is a 22 fold enhancement in the expression of the bgl operon under anaerobic condition. The present results provide evidence that bgl expression has a physiological role under low levels of oxygen and in addition suggest a possible mechanism for the overexpression of the bgl operon that involves the ArcAB two component system known to mediate regulation under microaerobic and static conditions. Transcription studies using a lacZ reporter fused to the wild type bgl promoter show that there is enhanced transcription from the bgl promoter under microaerobic and static conditions in the presence of arcA encoding the response regulator compared to that in its absence. The positive effect of arcA on the expression of the bgl operon is dispensable in the absence of H-NS since presence or absence of arcA does not change the expression of the bgl operon in an hns-null background, implying that the involvement of ArcA is via antagonizing H-NS. Competition experiments indicate that there is growth advantage associated with the activated allele of the bgl operon under low levels of oxygen since Bgl+ strains carrying the activated allele of the bgl operon as well as strains expressing BglG constitutively can out-compete wild-type strains. Presence of the wild type arcA allele results in a strong growth advantage compared to its absence under static conditions but not aerobic condition. The bgl operon seems to be one of the possible downstream targets of ArcA under static condition since absence of the bgl operon results in a modest reduction of the growth advantage (GASP) phenotype conferred by arcA. The up-regulation of the bgl operon is likely to enable the cells to scavenge available nutrients from their niche more efficiently. These experiments also show that the GASP phenotype associated with BglG constitutive strains under static conditions involves downstream genes that are different from oppA known to be one of the downstream targets during aerobic growth. It is possible that under low level of oxygen, the bgl operon is regulating a different set of downstream genes involving a different mechanism. In summary, the results of this investigation show that the aromatic β-glucoside catabolic systems in E. coli play a role in the generation of new metabolic capabilities via mutations in pre-existing genetic systems as well as through changes in gene expression patterns. The mechanisms outlined in this study are likely to be of broader significance applicable to microbial evolution under stress in general.

The majority of children with ASC are educated within a mainstream secondary setting. The challenges within the new environment may lead to some children experiencing a breakdown in provision resulting in temporary or permanent exclusions. A gap in the literature highlights a need for research which seeks to understand the views and experiences of year 7 children with ASCs about theirtransition to mainstream secondary school. The research involved four year 7 children with ASC, from across three settings, who had recently transitioned from a mainstream primary school to a mainstream secondary school (without resource based provision). The views of their parents and teachers were also sought. A multiple-embedded case study design was employed involving four cases from across three school settings. This involved utilising a transition Q-sort and a semi-structured interview with four children and semi-structured interviews with six parents and three teachers. The data was analysed using content analysis and thematic analysis (Braun & Clarke, 2006). The findings highlight a need for individualised transition planning for the child with ASC; consideration of co-occurring difficulties and the importance of including the views of the child and parents in transition planning. The findings are discussed in relation to psychological theories and existing literature. The implications for future research, local authority policy, school staff, parents and educational psychologists are considered.

This thesis presents a proposal of newly designed concept for travel hydraulic circuit of vibratory single drum roller ASC 110. The maximum machine gradeability was chosen as the main parameter for hydraulic circuit proposal. After selecting the new hydraulic travel concept, suitable hydraulic components have been selected. Length and local losses for two different machine speeds were calculated for sufficient hydraulic circuit design. Hydraulic losses have also been calculated for use in different climatic conditions. A new machine cooling concept has also been proposed. Heating and cooling curves of hydraulic circuit were calculated. For the purpose of comparing gradeability and energy balance of the newly designed travel concept compared to the existing one currently in mass production, both machines were fitted with measuring points. On both machines were measured for example, pressure, flow and temperature in each hydraulic circuit branch and others. From the time dependencies of the above-mentioned parameters it was possible to determine the energy efficiency of the newly designed concept. To assess the maximum gradeability, the slope of the hill was measured. Comparing the two machines, it has been found that the two hydraulic circuit solution has significantly improved traction properties in demanding machine applications, especially when driving uphill and downhill. This concept also affects less fuel consumption and places less demand on the hydraulic oil cooler due to its energy efficiency.

Transplantation of multipotent mesenchymal progenitor cells is a valuable option for treating tendon disease. Tenogenic differentiation leading to cell replacement and subsequent matrix modulation may contribute to the regenerative effects of these cells, but it is unclear whether this occurs in the inflammatory environment of acute tendon disease. Equine adipose-derived stromal cells (ASC) were cultured as monolayers or on decellularized tendon scaffolds in static or dynamic conditions, the latter represented by cyclic stretching. The impact of different inflammatory conditions, as represented by supplementation with interleukin-1β and/or tumor necrosis factor-α or by co-culture with allogeneic peripheral blood leukocytes, on ASC functional properties was investigated. High cytokine concentrations increased ASC proliferation and osteogenic differentiation, but decreased chondrogenic differentiation and ASC viability in scaffold culture, as well as tendon scaffold repopulation, and strongly influenced musculoskeletal gene expression. Effects regarding the latter differed between the monolayer and scaffold cultures. Leukocytes rather decreased ASC proliferation, but had similar effects on viability and musculoskeletal gene expression. This included decreased expression of the tenogenic transcription factor scleraxis by an inflammatory environment throughout culture conditions. The data demonstrate that ASC tenogenic properties are compromised in an inflammatory environment, with relevance to their possible mechanisms of action in acute tendon disease.

The aim of this thesis is to create a comprehensive view of financial reporting of lease contracts from lessee's and lessor's point of view in accordance with International Financial Reporting Standards (IFRS) and US Generally Accepted Accounting Principles (US GAAP). The thesis is divided into four parts. The first part is devoted to basic characteristics of the lease and its reporting according to the currently applicable standards - IAS 17 and ASC 840. Content of the next part is Convergence of Accounting Standards. Second half of the thesis focuses on the Exposure Draft and Comment Letters.