963 resultados para Task-Oriented Methodology


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This dissertation primarily describes chemical-scale studies of G protein-coupled receptors and Cys-loop ligand-gated ion channels to better understand ligand binding interactions and the mechanism of channel activation using recently published crystal structures as a guide. These studies employ the use of unnatural amino acid mutagenesis and electrophysiology to measure subtle changes in receptor function.

In chapter 2, the role of a conserved aromatic microdomain predicted in the D3 dopamine receptor is probed in the closely related D2 and D4 dopamine receptors. This domain was found to act as a structural unit near the ligand binding site that is important for receptor function. The domain consists of several functionally important noncovalent interactions including hydrogen bond, aromatic-aromatic, and sulfur-π interactions that show strong couplings by mutant cycle analysis. We also assign an alternate interpretation for the linear fluorination plot observed at W6.48, a residue previously thought to participate in a cation-π interaction with dopamine.

Chapter 3 outlines attempts to incorporate chemically synthesized and in vitro acylated unnatural amino acids into mammalian cells. While our attempts were not successful, method optimizations and data for nonsense suppression with an in vivo acylated tRNA are included. This chapter is aimed to aid future researchers attempting unnatural amino acid mutagenesis in mammalian cells.

Chapter 4 identifies a cation-π interaction between glutamate and a tyrosine residue on loop C in the GluClβ receptor. Using the recently published crystal structure of the homologous GluClα receptor, other ligand-binding and protein-protein interactions are probed to determine the similarity between this invertebrate receptor and other more distantly related vertebrate Cys-loop receptors. We find that many of the interactions previously observed are conserved in the GluCl receptors, however care must be taken when extrapolating structural data.

Chapter 5 examines inherent properties of the GluClα receptor that are responsible for the observed glutamate insensitivity of the receptor. Chimera synthesis and mutagenesis reveal the C-terminal portion of the M4 helix and the C-terminus as contributing to formation of the decoupled state, where ligand binding is incapable of triggering channel gating. Receptor mutagenesis was unable to identify single residue mismatches or impaired protein-protein interactions within this domain. We conclude that M4 helix structure and/or membrane dynamics are likely the cause of ligand insensitivity in this receptor and that the M4 helix has an role important in the activation process.

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O estudo situa-se no âmbito das investigações voltadas para documentos que sistematizam o trabalho do professor, dentre eles, o manual do professor que organiza a atividade docente junto ao livro didático. A dissertação analisa manuais do professor dos livros de espanhol selecionados pelo MEC para serem distribuídos a professores, em 2005, em função da lei 11161 da obrigatoriedade do ensino da língua espanhola para o ensino médio em todo o território nacional. O objetivo foi identificar imagens discursivas de docente e de ensino de espanhol como língua estrangeira neles construídas. Os fundamentos teóricos adotados advêm da Análise do Discurso de base enunciativa, além de recorrermos aos conceitos de dialogismo (BAKHTIN, 1979) e de polifonia (BAKHTIN, 1979; DUCROT, 1987). Os resultados nesses manuais apontam para a construção de imagens de professor como: aquele que necessita ser guiado em sua tarefa, incapaz de realizar suas escolhas em sala de aula, um professor recebedor de ordens; desatualizado com as metodologias de ensino atuais, necessitando, portanto, de atualização profissional; há ainda um professor que busca instruções facilitadoras para seu trabalho. Já no que se refere à visão de língua, deparamo-nos com um manual que dá ênfase ao trabalho com a leitura, voltado para uma concepção que valoriza aspectos discursivos; outros que afirmam seguir a abordagem comunicativa, com um olhar para a língua em uso, porém adotam procedimentos pautados numa concepção de língua como estrutura e/ou misturam ambas perspectivas

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Eutrophication of fresh waters through anthropogenic enrichment by phosphorus is a global problem. The role of phosphorus enrichment in the formation of blooms of toxic blue-green algae (Cyanobacteria) in fresh waters is well established and of considerable concern in terms of human and animal health, loss of water resources and amenities, threats to fish stocks, and aesthetic considerations. Cultural eutrophication also poses threats to the ecosystem balance in fresh waters, with implications for wildlife. This article examines phosphorus enrichment in fresh waters from a systems perspective, and explores systems solutions that may be helpful in the development of more sustainable policies.

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The evaluation and comparison of internal cluster validity indices is a critical problem in the clustering area. The methodology used in most of the evaluations assumes that the clustering algorithms work correctly. We propose an alternative methodology that does not make this often false assumption. We compared 7 internal cluster validity indices with both methodologies and concluded that the results obtained with the proposed methodology are more representative of the actual capabilities of the compared indices.

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In this book section, the theoretical background to the methodology is outlined, questionnaire development described, sample selection outlined and biases and shortcomings to the survey noted.

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These minutes report on colloquium on the methodology of radiation measurement under water. The meeting was held on 3-5 January 1957, at the Biological Station, Lunz, Austria. The participants of the colloquium discussed various methodologies of radiation measurements, basic methods such as Secchi Disc and underwater photometer as well as specialist equipment like the absolute radiation apparatus.