Sequence analysis of part of a large plasmid carried by Australian environmental isolate of Listeria monocytogenes strain DRDC8 has lead to identification of an islet of genes that encode proteins similar to copper binding and transport genes found in other Gram positive bacteria. Comparative sequence analysis showed that there are at least four genes (pCT0017, pCT0018, pCT0019 and ctpA) on this islet predicted to be involved in copper homeostasis. One of these, ctpA, is predicted to encode a P-type ATPase with a function analogous to CopA, a copper transporting gene in Enterococcus hirae. ORF pCT0017 is likely to be a CopY-like regulatory protein which could control the expression of ctpA. ORF pCT0019 is predicted to be a Cu²⁺ binding protein. In addition, two genes located downstream of the ctpA are predicted to encode a two component regulatory system region. The predicted function of ORF pCT0018 is not clear. A related chromosomal gene (cutR) is predicted to also encode a copper transporting P-type ATPase. To investigate the role of the protein encoded by pCT0018, the growth behavior of L. monocytogenes strain DRDC8, other strains carrying mutations within pCT0018, pCT0019, cutR and ctpA, as well as strains cured of the large plasmid, were grown under conditions of copper stress and starvation. The growth data showed that with the exception of strain DRDC8 and other strains carrying ctpA, most were unable to grow at higher copper concentration (>15 mM CuSO₄) and suggested that the copper homeostasis genes located on the large plasmid are associated with tolerance to high levels of copper. Strain DSE955PL, which carries a cutR mutation and is cured of the large plasmid, was the most sensitive (<5 mM CuSO₄). This indicated that proteins encoded by plasmid genes work synergistically to confer tolerance to copper. Of most interest was the fact that a pCT0018 mutant was more sensitive (<15 mM CuSO₄) to high levels of copper than the wild type parent DRDC8 (<20 mM CuSO₄). This suggested that ORF pCT0018 was necessary for copper tolerance. To investigate the effects of insertion mutations in pCT0017, pCT0018 and ctpA on copper uptake and export, the levels of copper accumulated by these strains was assessed using atomic absorption spectroscopy. A significant difference in copper accumulation among the bacteria strains was observed when either LEB or BHI media were used to culture the bacteria. This data suggested that the growth medium chemicals influence the levels of copper accumulated by cells. However, the effect of these media on bacteria growth rates during copper stress was not significant. Atomic absorption analysis of intracellular copper accumulation suggested that DSE955PL and DSE955 (a chromosome mutant) were able to accumulate copper (80 - 110 mg.gˉ¹ dry weight of cells), whereas DRDC8 and strains carrying mutations in pCT0018, ctpA, and strains cured of the large plasmid, were less able to accumulate copper (30 - 70 mg.gˉ¹ dry weight of cells). This data suggested that cutR may encode a copper export system and that ctpA is involved in copper uptake. To investigate the gene expression profile for pCT0018 under elevated copper, reverse transcriptase PCR was used to detect transcripts encoding pCT0017, pCT0018, pCT0019 and pCT0020 from RNA extracted from L. monocytogenes strain DRDC8 following culture at elevated levels of copper. Although transcripts for each of the target genes were detected, transcription was not responsive to copper, nor was the pattern of transcription consistent with that expected for a single operon. To directly determine whether the protein encoded by the pCT0018 open reading frame was able to bind copper, this gene was cloned in pET15b in frame with an N-terminal Histag and expressed in E. coli. The expressed protein was purified with a Ni-NTA column and shown to contain copper. Attempts to directly show that protein pCT0018 could bind copper by Cu-IMAC were unable to unequivocally show that the protein was immobilized on the column. Purified protein was used to raise a polyclonal antiserum in rabbit and the antiserum was used for Western analysis to test expression of pCT0018 by wild type L. monocytogenes DRDC8 and specific gene mutants. Although the antiserum bound to purified protein, it was not possible to demonstrate binding to native pCT0018 in cell lysates prepared from L. monocytogenes DRDC8. SDS-PAGE of cytoplasmic and cell envelope proteins isolated from L. monocytogenes strains was used to identify proteins expressed in response to copper stress and starvation. No significant differences in protein profiles for cytoplasmic protein were observed. However, copper-immobilized metal affinity chromatography (Cu-IMAC) showed that expression of a number of copper binding proteins were differentially expressed by DRDC8 following growth in copper stress and starvation conditions. Three of these proteins were selected for amino sequence analysis by MALDI-TOFF MS. Two were confirmed to be L. monocytogenes non-heme iron-binding ferritin and a thiol peroxidase, both of which bind copper. The other protein was similar to an unknown protein from L. monocytogenes. Interestingly, no proteins directly implicated with the copper homeostasis islet were identified. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1374407 / Thesis (M.Sc.) - University of Adelaide, School of Molecular and Biomedical Science, 2009

Thesis (Ph.D.). / Written for the Dept. of Biochemistry. Title from title page of PDF (viewed 2009/06/09). Includes bibliographical references.

Thesis (M.Sc.). / Written for the Dept. of Human Genetics. Title from title page of PDF (viewed 2009/06/18). Includes bibliographical references.

Submitted by Gisele Mara Hadlich (gisele@ufba.br) on 2018-06-26T14:04:32Z No. of bitstreams: 1 DISSERTA_Rodolfo GASSER_revGisele_24nov_OK.pdf: 5953775 bytes, checksum: f1ccd70ec56e9d5d7feb19a1796fdb6a (MD5) / Made available in DSpace on 2018-06-26T14:04:33Z (GMT). No. of bitstreams: 1 DISSERTA_Rodolfo GASSER_revGisele_24nov_OK.pdf: 5953775 bytes, checksum: f1ccd70ec56e9d5d7feb19a1796fdb6a (MD5) / O aquífero cárstico Salitre situa-se na Bacia Carbonática de Irecê, na região centro-norte da Bahia, no contexto geológico da Formação Salitre, no domínio do Grupo Una. É constituído por espessas sequências de calcarenitos, calcilutitos, calcissiltitos e dolomíticos intercalados, por vezes, com sequências terrígenas subordinadas, constituídas por silexitos, arenitos, siltitos, laminitos e margas. O objetivo principal do estudo é avaliar a vulnerabilidade intrínseca à contaminação natural do aquífero cárstico Salitre, no município de Cafarnaum, para isso usou-se a metodologia COP. Além da vulnerabilidade, buscou-se a caracterização hidroquímica, identificando os principais constituintes químicos presentes e identificar os possíveis fatores naturais e antropogênicos influentes na qualidade das águas subterrâneas. A metodologia deste estudo utiliza métodos hidroquímicos, hidrodinâmicos e estatísticos/geoestatísticos para compreender as condições hidrogeoquímicas, além do uso de imagens de satélites para a individualização das feições cársticas. Foram coletadasamostras de água retiradas de 31 poços tubulares distribuídos no município, com o uso da sondamultiparâmetros, foram medidos in situ os parâmetros físico-químicos: pH, EH, condutividadeelétrica, sólidos totais dissolvidos, temperatura, turbidez, oxigênio dissolvido. No laboratório, foram determinados os íonsNO₃¯, F¯, Cl¯, SO₄²¯, PO₄³¯, HCO₃¯, Ba⁺, Ca²⁺, Sr²⁺, Mg²⁺, Na⁺, Zn⁺², e K⁺, utilizando-se de técnicas fluorimétricos, titrimétricos e espectrofométricos. Para a distribuição espacial dos resultados hidroquímicos foram confeccionados mapas de pontos, utilizando o método de krigagem ordinária.Para a classificação do tipo deágua, foi utilizado o diagrama triangular de Piper, para o diagnóstico da água para irrigação, foiusado o diagrama SAR.Os aquíferos cársticos são especialmente vulneráveis à contaminação devido a sua pouca profundidade, a alta velocidade do fluxo de água, a sua conexão direta com a superfície e a sua recarga através de formas de absorção abertas. Para a avaliação da vulnerabilidade natural do aquífero cárstico à contaminação através da metodologia COP, levam-se em consideração três parâmetros hidrogeológicos: concentração do fluxo, camadas sobrepostas e regime de precipitação. A análise do mapa de vulnerabilidade intrínseca a contaminação, permitiu a delimitação de 4 classes de vulnerabilidade na área: muito alta (12,5% - 70,20 km²), alta (0,5% - 2,0 km²), moderada (32,7%, 183,30 km²) e baixa (54,4%, 305,70 km). A individualização das feições cársticase de sua área de influência é o fator que teve um papel significativo para a delimitação da classe de vulnerabilidade muito alta.O aquífero é livre e anisotrópico, com direção preferencial de fluxo potenciométrico SW – NE. Foram obtidos valores mais elevados que os valores máximos permitidos para os seguintes elementos: cálcio, nitrato, magnésio, sólidos totais dissolvidos, sulfato, cloreto, fluoreto e sódio. O mecanismo de dissolução das rochas e o processo evaporação controlam a mineralização da água. Os resultados obtidos mostraram que as águas estudadas classificam-se de maneira geral comosulfatas ou cloretadas e águas cálcicas ou magnesianas, consideradas de risco alto a muito alto de salinidade e risco baixo a médio de sódio. A detecção de elevadas concentrações de NO₃¯, em regiões com alta densidade populacional, sugere a contaminação por efluentes domésticos, justamente pela falta de esgotamento sanitário em grande parte do município.

Ever since the governments began to meet for the Conferences of the Parties (COP) to try to come to agreement over an international climate agreement, civil society organizations involved in the climate issue have also gathered for their own conferences in relation to the COP. Still the role of these re-occurring “COP counter summits” in global climate governance have not been researched in depth. In order to contribute to a better understanding of the role the COP counter summit in climate governance in general, the aim of this study is to conduct a qualitative single-case study of one of these meetings. This is carried out by a field study to the COP counter summit in Paris 2015 with methods of participatory observation and semi-structured interviews with informants. An analytical framework was developed based on theory within related research fields, like parallel summits, social movements and globalization studies. The study contributes with research about what functions of the COP counter summit in Paris had for civil society and how these functions were performed.

This diploma thesis focuses on the phenomenon of global climate change from the perspective of climate change law. The thesis defines the phenomenon in legal terms and deals with problems it brings to the environment and the world population. Its major part is dedicated to the international law of climate change, including all past Conferences of the Parties (COP) and their outcomes together with the most important treaties - United Nations Framework Convention in terms of Kyoto Protocol and the unsuccessful Doha Amendment. The Paris Agreement, which is the most recent international treaty, already ratified by 160 countries, could be a game changer in terms of effectivity in both climate change mitigation and adaptation legislation as well as much needed international solution. This thesis also focuses on the EU law of climate change, especially on the progress and future of Emissions Trading System and other legislation. Not only is the EU now more focused on the adaptation legislation, this is an important topic also for the Czech Republic. Especially when many Czech laws stem from EU regulations, there is still a great potential in the adaptation legislation and major progress is expected in the near future. Key words Climate change, Paris Agreement, emissions allowances, COP

The diploma thesis deals with design of a suitable heat source for heating and hot water. There are dealt with operational costs of different types of heat sources, which are compared with one another. The next part is the implementation of specific proposal of the heat source - heat pump ground/water with deep wells and measuring system. The last part describes measuring several operating modes of the heat pump.

This diploma work aims to evaluate the functional connection of the cogeneration unit with an internal combustion engine and an absorption heat pump to increase the heating effect of the cogeneration. The aim of this work is to design a functional system and evaluate it from the technical - economic terms. In the introduction of the theoretical work the applied cogeneration technologies used in combined heat and power production are presented. Then there is described the principle of heat pumps, their basic components and theoretical comparison of compression and absorption heat pumps. In the practical part of this work is for a specific application in the area Technická 2 proposed merger of the cogeneration unit with an internal combustion engine and an absorption heat pump, while detailed design parameters used in absorption heat circulation are set. At the end of this paper there are designed various heat exchangers of applied absorption heat pump. The final part of the work is devoted to technical - economic assessment of applied design. In this assestment three different types of heat pumps are compared: Absorption heat pump with working solution H2O-LiBR, absorption heat pump with working fluids NH3-H2O and compressor heat pump.

This thesis deals with the formulation and application of reduced order models based on extraction of dominant structures from a system utilizing the method of proper orthogonal decomposition. Time evolution of computed modes is described by a system of ordinary differential equations, which is gained by means of Galerkin projection of these modes onto the Navier-Stokes equations. This methodology was applied on two test cases Kármán vortex street and vortex rope. In both cases, a CFD simulation of one refference point was carried out and by utilizing gained modes, the corresponding reduced order models were formulated. Their results were assessed by comparing to the refference simulation.

Title: Neurophysiologic intervention induced changes of the functional foot characteristics in school children. Objectives: The aim of the research was to identify the functional characteristics of the foot in school children selected on the basis of the activity of physical activity in the highest intensity (sporting children) and in the lowest intensity (non-sporting children) and to find out possibilities of influencing the functional characteristics of the foot by targeted neurophysiological intervention. The whole research study was divided into 3 related parts, which were examined separately. The first study aim was an identification of group specific functional characteristics of the foot in 2 groups of school children - actively sporting children and non-sporting children. The second study aim was the influence determination of long-term duration high performance sport activity on the functional characteristics of the school children foot in adolescent elite soccer players. And the aim of the third study was to verify the effect of the targeted neurophysiologic active intervention on the functional foot characteristics in actively sporting children with foot dysfunction of pronated foot and/or flatfoot type. Methods: The research study was divided into three study parts consisted of...