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Produtos de glicação avançada, carbamilação e estresse oxidativo contribuem como fatores de risco não tradicionais para a aterosclerose na doença renal crônica (DRC), em parte, por prejudicarem o metabolismo lipídico e por representarem um mecanismo de injúria memorizado ao longo do desenvolvimento da doença renal. A albumina sérica, isolada de animais com DRC, reduz a remoção de colesterol mediado por apoA-I e subfrações de HDL, prejudicando o fluxo de colesterol de macrófagos arteriais ao fígado por meio do transporte reverso de colesterol. Objetivo: Avaliou-se a influência do tratamento com N-acetilcisteína (NAC) em ratos com DRC sobre a concentração plasmática de produtos de oxidação e glicação avançada e o reflexo sobre os efeitos da albumina sérica sobre o efluxo de colesterol e o estresse de retículo endoplasmático em macrófagos. Métodos: Ratos Wistar com 2 meses de idade, pesando aproximadamente 200-250g foram submetidos à nefrectomia 5/6 e mantidos por 60 dias (grupo DRC) com ou sem tratamento com N-acetilcisteína na água (600mg/L), após o 7° dia de indução da DRC (grupo DRC + NAC). Animais controles foram falso-operados (grupo C) e um subgrupo submetido ao tratamento com NAC (C + NAC). No início e no final do estudo foram determinadas as concentrações plasmáticas de glicose, colesterol (CT), triglicérides (TG), ureia, creatinina e na urina, excreção urinária de proteína de 24 h. AGE totais, pentosidina, TBARS (marcador de peroxidação lipídica) e pressão arterial sistólica (PAS) foram determinados no final do estudo. A albumina sérica foi isolada por cromatografia rápida para separação de proteínas e purificada por extração alcoólica. Macrófagos J774 foram incubados por 18 h com as albuminas dos diferentes grupos experimentais para determinação do conteúdo dos receptores de HDL (ABCA-1 e ABCG-1) e de marcadores de estresse de retículo endoplasmático (chaperonas Grp 78, Grp94 e proteína dissulfeto isomerase, PDI) por imunolbot e efluxo de colesterol, mediado por apo A-I e HDL2. Para isto, as células foram previamente enriquecidas com LDL-acetilada e 14C-colesterol. Macrófagos foram também incubados isoladamente com concentrações crescentes de NAC para avaliação do conteúdo dos receptores de HDL. Resultados: Ao final do estudo, o peso corporal foi 10% menor no grupo DRC em comparação ao C (p=0,006). Esta alteração foi prevenida pelo tratamento com NAC. A PAS (mmHg) foi maior no grupo DRC (130 ± 3) em comparação ao grupo DRC+NAC (109±3; p=0,0004). Ureia, creatinina, CT, TG (mg/dL), proteinúria (mg/24 h), AGE total, pentosidina (unidades arbitrárias de fluorescência) e TBARS (nmol/mL) foram maiores nos grupos DRC em comparação ao grupo C (122 ± 8 vs. 41 ± 0,9 ; 0,9 ± 0,07 vs. 0,4 ± 0,03; 151 ± 6 vs. 76 ± 2,7; 83 ± 4 vs. 51,5 ± 3; 46 ± 2,5 vs. 14 ± 0,9; 32620 ± 673 vs. 21750 ± 960; 16700 ± 1370 vs. 5314 ± 129; 6,6 ± 0,5 vs. 2 ± 0,2, respectivamente) (p < 0,0001) e nos grupos DRC+NAC em comparação ao grupo C+NAC (91,4 ± 5 vs. 40 ± 0,9 ; 0,6 ± 0,02 vs. 0,3 ± 0,02; 126 ± 7,5 vs. 76 ± 2,6; 73 ± 6 vs. 68 ± 4; 51 ± 3,5 vs. 18,4 ± 1,5; 24720 ± 1114 vs. 20040 ± 700; 10080 ± 748 vs. 5050 ± 267; 4,5 ± 0,5 vs. 1,8 ± 0,2, respectivamente) (p < 0,0001). No grupo DRC + NAC, PAS, CT, ureia, creatinina, AGE total, pentosidina e TBARS foram, respectivamente, 17% (p=0,0004), 17% (p=0,02), 25% (p=0,02), 33% (p=0,06), 24% (p < 0,0001), 40% (p=0,0008), 28% (p=0,009) menores do que no grupo DRC. A glicemia foi maior nos grupos C + NAC (107+-4,6) e DRC + NAC (107+-2,6) em comparação ao C (96+-1,8) e DRC (98+-1,6), respectivamente. Macrófagos tratados com albumina-DRC apresentaram maior conteúdo de PDI (5 vezes; p=0,02 e 7 vezes p=0,02) e Grp94 (66 %; p =0,02 e 20 %; p=0,02) quando comparados aos tratados com albumina-C ou albumina-DRC + NAC, respectivamente. O conteúdo do receptor ABCA-1 foi menor 87% e 70% (p < 0,01) nos macrófagos tratados com albumina-C+NAC e albumina-DRC, respectivamente em comparação com albumina-C. O conteúdo de ABCG-1 foi, respectivamente, 4 e 7,5 vezes maior nos macrófagos tratados com albumina-C+NAC e albumina-DRC+NAC em comparação as respectivas situações sem tratamento. O efluxo de colesterol mediado por apo A-I foi 59 % e 70 % (p < 0,0001) menor nos macrófagos tratados com albumina-C+NAC e albumina-DRC, respectivamente em comparação a albumina-C. O efluxo de colesterol mediado pela HDL2 foi 52 % maior nos macrófagos tratados com albumina-C+NAC em comparação as células tratadas com albumina-C. Não houve diferença no conteúdo do receptor ABCA-1 nos macrófagos tratados com concentrações crescentes NAC por 8 h. No entanto, após 18 h, o ABCA-1 diminuiu 50 %, 69 % e 72 % nos macrófagos tratados respectivamente com 10 mM, 20 mM e 30 mM de NAC isoladamente em comparação aos macrófagos controles. O conteúdo de ABCG-1 nos macrófagos tratados com NAC, em 8 h e 18 h não sofreu alteração. Conclusão: A N-acetilcisteína reduz produtos de oxidação e glicação avançada no plasma de animais com DRC e previne o estresse de RE em macrófagos, induzido pela albumina isolada destes animais. Apesar de diminuir o conteúdo de ABCA-1 e o efluxo de colesterol mediado por apo A-I, a NAC aumenta o conteúdo de ABCG-1. Desta forma, a NAC pode contribuir para atenuar os efeitos deletérios da albumina modificada na DRC sobre o acúmulo lipídico em macrófagos, contribuindo para a prevenção da aterosclerose / Advanced glycation, carbamylation and oxidative stress c contribute to atherosclerosis in chronic kidney disease (CKD) as nontraditional risk factors. They impair lipid metabolism and promote a long last injury during the development of CKD. Serum albumin isolated from CKD-animals reduces cholesterol efflux mediated by apoa A-I and HDl subfractions, impairing the cholesterol flux from arterial wall macrophage to the the liver by the reverse cholesterol transport (RCT).Objective: In the present study it was analyzed the influence of N-acetylcysteine treatment in CKD-rats in plasma concentration of lipid peroxides and advanced glycation end products and the effect of serum albumin in macrophage cholesterol efflux and endoplasmic reticulum stress development. Methods: Two months male Wistar weighting 200-250g were submitted to a 5/6 nephrectomized maintained for 60 days (CKD group) treated or not with N-acetylcysteine in water (600 mg/L), after the seventh day of CKD induction (CKD+NAC group). Sham animals were false-operated (SHAM group) and a subgroup was treated with NAC (SHAM+NAC group). In the basal and final periods it was determined plasma concentration of glucose, total cholesterol (TC), triglycerides (TG), urea, creatinine and 24h-urinary protein excretion (UPE). Total AGE, pentosidine, thiobarbituric reactive substances (TBARS) levels and systolic blood pressure (SBP) were measured at the final period only. Serum albumin was isolated by fast protein liquid chromatography and purified by alcoholic extraction. J774 macrophage were incubated for 18 h with albumin isolated from the experimental groups in order to determine the content of HDL receptors and markers of endoplasmic reticulum stress (Grp78, Grp94 and protein dissulfide isomerase, PDI) by immunioblot and cholesterol efflux mediated by apo A-I and HDL2. For this, cells were previously overloaded with acetylated LDL and 14C-cholesterol. Macrophage were also incubated with different concentrations of NAC alone in order to measure HDL-receptors and cholesterole efflux. Results: In the end of the protocol, body weight was 10% lower in CKD group in comparison to SHAM group (p=0.006). This change was preserved by treatment with NAC. SBP (mmHg) was higher in CKD group (130±3) in comparison to CKD+NAC (109±3; p=0.0004). Urea, creatinine, TC, TG (mg/dL), UPE (mg/24 h), total AGE, pentosidine (arbitrary units of fluorescence) and TBARS (nmol/mL) were higher in CKD group in comparison to SHAM (122±8 vs. 41 ± 0.9; 0.9 ± 0.07 vs. 0.4 ± 0.03; 151 ± 6 vs. 76±2.7; 83 ± 4 vs. 51.5 ± 3; 46 ± 2.5 vs. 14 ± 0.9; 32620 ± 673 vs. 21750 ± 960; 16700 ± 1370 vs. 5314 ± 129; 6.6 ± 0.5 vs. 2 ± 0.2, respectively) (p < 0.0001) and in CKD+NAC in comparison to C+NAC (91.4±5 vs. 40±0.9 ; 0.6±0.02 vs. 0.3 ± 0.02; 126±7.5 vs. 76 ± 2.6; 73±6 vs. 68±4; 51 ± 3.5 vs. 18.4±1.5; 24720 ± 1114 vs. 20040±700; 10080±748 vs. 5050 ± 267; 4.5±0.5 vs. 1.8±0.2, respectively) (p < 0.0001). In CKD+NAC group, SBP, TC, urea, creatinine, total AGE, pentosidine and TBARS were, respectively, 17 % (p=0.0004), 17 % (p=0.02), 25 % (p=0.02), 33 % (p=0.06), 24 % (p<0.0001), 40 % (p=0.0008), 28 % (p=0.009) lower than CKD group. Glycemia was higher in SHAM+NAC (107+-4.6) and CKD+NAC (107+-2.6) in comparison to SHAM (96+-1.8) and CKD group (98+-1.6), respectively. Macrophages treat with CKD-albumin presented higher content of PDI (5 times; p=0.02 e 7 times p=0.02) and Grp94 (66 %; p=0.02 e 20 %; p=0.02) when compared to SHAM-albumin and CKD+NAC-albumin- treated cells, respectively. ABCA-1 protein content was 87 % and 70 % (p < 0.01) lower in macrophages treated with SHAM+NAC-albumin and CKD-albumin, respectively compared with SHAM-albumin. ABCG-1 protein level was respectively 4 and 7.5 times higher in macrophages treated with SHAM+NAC-albumin and CKD+NAC-albumin in comparison to their respective controls without treatment. The cholesterol efflux mediated by apo A-I was 59 % and 70 % (p < 0.0001) lower in macrophages treated with SHAM+NAC-albumin and CKD-albumin, respectively, when compared to SHAM-albumin. The HDL2-mediated cholesterol efflux was 52 % higher in macrophages treated with SHAM+NAC-albumin compared to macrophages treated with SHAM-albumin. No difference was observed in the ABCA-1 protein level in macrophages treated with crescent concentrations of NAC alone for 8 h. Nonetheless, after 18 h, ABCA-1 was 50 %, 69 % and 72 % reduced in macrophages treated, respectively, with 10 mM, 20 mM and 30 mM NAC in comparison to control cells. ABCG-1 content in macrophages treated with NAC for 8 h and 18 h was not changed. Conclusion: NAC reduces plasma lipid peroxidation and AGE in CKD animals and prevents the endoplasmic reticulum stress induced by CKD-albumin in macrophages. Despite diminishing ABCA-1 and apo A-I-mediated cholesterol efflux, NAC increases ABCG-1. Then, NAC may contribute to attenuate the deleterious effects of the in vivo modified albumin on lipid accumulation in macrophages helping to prevent atherosclerosis in CKD

Habitar ambientes dimensionados a partir de estudos antropométricos e Avaliações Pós-Ocupação (APOs) dos aspectos funcionais é conseqüência de processos de projeto arquitetônico a serem consolidados. A tecnologia tem permitido obter um maior controle sobre todas as fases da produção de edificações, sendo a fase de pré-projeto decisiva para o efetivo sucesso de um empreendimento. A presente pesquisa objetiva o desenvolvimento de um roteiro para a elaboração de um programa computacional de Projeto e Desenho Arquitetônico Auxiliado por Computador (CAADD - Computer Aided Architectural Design and Drafting) que contribua para o trabalho do arquiteto nas análises geométricas de dimensionamento funcional de edificações habitacionais sociais multi-familiares, no momento do pré-projeto, considerando as especificidades do conjunto de variáveis envolvidas em cada situação de projeto. Ao escolher as alternativas de composição de Programa de Necessidades no sistema, este geraria maquetes eletrônicas (modelo digital) a partir dos parâmetros definidos no próprio programa. Os volumes abstraídos em meio digital são tratados genericamente no sistema como objetos volumétricos (mobiliário, eletrodomésticos, peças sanitárias, esquadrias, estrutura, vedação). A tecnologia de representação gráfica digital escolhida foi o padrão aberto para a distribuição de conteúdo tridimensional denominado X3D. A montagem do roteiro contempla conceitos de Computação Gráfica e de Geometria Computacional, aplicando critérios para arranjo físico de ambientes da habitação, relação entre ambientes, distribuição de unidades habitacionais (apartamentos) e análises de locação da edificação no sítio urbano. Utilizam-se como referências dimensionais resultados de APOs (classificadas como dados dinâmicos, os quais se alteram com maior freqüência no tempo) e legislação e normas vigentes (classificados como dados estáveis, os quais se alteram com menor freqüência no tempo), visando melhorias espaciais e manutenção contínua da abordagem do projeto e desenho arquitetônico funcional nos novos projetos habitacionais. / The possibility of living in environments that have been sized on the basis of anthropometric studies and post-occupancy evaluations (POEs) of the functionality of buildings is the consequence of architectural design processes yet to be consolidated. This technology has enabled greater control over all stages in the construction of buildings, the pre-design stage being decisive for the success of an enterprise. The objective of the present study is to develop guidelines for designing a computer program for architectural designing and drafting (CAADD-Computer Aided Architectural Design and Drafting) that will serve as an aid to architects in terms of geometrical analyses of the functional dimensions of low-income housing during the pre-design stage, considering the specific aspects of the variables involved in each design. By choosing alternatives for programming the software, the system is set up to enable designers to generate digital models based on standards defined in the program itself. The digitally extracted volumes are treated generically in the system as volumetric objects (furniture, appliances, plumbing fixtures, door and window frames, structures, waterproofing and others). The open standard for distributing tridimensional contents, known as X3D, was chosen as the technology for digital graphic representation. The guidelines take into account concepts for graphic computing and computational geometry and apply criteria for the physical arrangement of living environments, relationships among environments, distribution of the apartments and analyses of the location of the buildings in their urban context. Dimensional references are used to establish these guidelines, including the results of POEs (classifying the data that changes most often over time as \"dynamic\"). Other factors that must be taken into consideration are the local laws, norms and regulations (classified as \"stable\" data, which change less often over time). The objective is to develop spatial improvements and continuous feedback of the approach of the design and functional architectural drafting in new housing designs.

Esta tese trata dos ambientes coletivos dos condomínios horizontais e dos loteamentos fechados construídos a partir de 1990 na cidade de Londrina Pr. Busca descobrir quais as necessidades dos usuários (moradores) e, a partir da avaliação de desempenho físico e de satisfação do usuário voltada aos ambientes construídos e de uso coletivo, com ênfase nos aspectos social, funcional, segurança, durabilidade e manutenabilidade, formula procedimentos metodológicos de desempenho de avaliação para dar suporte na definição de programas de necessidades para futuros empreendimentos similares. Propõe um modelo metodológico que encontra suporte no Analytic Hierarchy Process (AHP) para hierarquizar os atributos e as alternativas que influenciam nas decisões de projeto (critérios e subcritérios) e cuja calibragem do modelo se dá por meio da Avaliação Pós-Ocupação (APO), envolvendo os pontos de vista dos moradores, dos administradores ou síndicos, dos autores dos projetos destes empreendimentos, além dos levantamentos in loco realizado pelo pesquisador. / ABSTRACT This thesis deals with collective environments of gated communities and plots which started being built in 1990 in the city of Londrina-PR. It searches for finding out the needs of the users (residents) and, from the evaluation of their physical performance and satisfaction regarding the built and collective environments, with special emphasis on social, functional, safety, durability and maintenance aspects, conceives the methodological procedures of performance evaluation to support the definition of programs of needs for future similar developments. It proposes a methodological model supported by the Analytic Hierarchy Process (AHP) to form as a hierarchy the attributes and the alternatives which have influence on the design decisions (criteria and sub-criteria), whose calibration of the proposed methodological model took place through the Post-Occupancy Evaluation (POE), involving the viewpoints from the residents, the administrators or syndics and architects, besides the surveys in loco carried out by this researcher.

Esta tese trata dos ambientes coletivos dos condomínios horizontais e dos loteamentos fechados construídos a partir de 1990 na cidade de Londrina Pr. Busca descobrir quais as necessidades dos usuários (moradores) e, a partir da avaliação de desempenho físico e de satisfação do usuário voltada aos ambientes construídos e de uso coletivo, com ênfase nos aspectos social, funcional, segurança, durabilidade e manutenabilidade, formula procedimentos metodológicos de desempenho de avaliação para dar suporte na definição de programas de necessidades para futuros empreendimentos similares. Propõe um modelo metodológico que encontra suporte no Analytic Hierarchy Process (AHP) para hierarquizar os atributos e as alternativas que influenciam nas decisões de projeto (critérios e subcritérios) e cuja calibragem do modelo se dá por meio da Avaliação Pós-Ocupação (APO), envolvendo os pontos de vista dos moradores, dos administradores ou síndicos, dos autores dos projetos destes empreendimentos, além dos levantamentos in loco realizado pelo pesquisador. / ABSTRACT This thesis deals with collective environments of gated communities and plots which started being built in 1990 in the city of Londrina-PR. It searches for finding out the needs of the users (residents) and, from the evaluation of their physical performance and satisfaction regarding the built and collective environments, with special emphasis on social, functional, safety, durability and maintenance aspects, conceives the methodological procedures of performance evaluation to support the definition of programs of needs for future similar developments. It proposes a methodological model supported by the Analytic Hierarchy Process (AHP) to form as a hierarchy the attributes and the alternatives which have influence on the design decisions (criteria and sub-criteria), whose calibration of the proposed methodological model took place through the Post-Occupancy Evaluation (POE), involving the viewpoints from the residents, the administrators or syndics and architects, besides the surveys in loco carried out by this researcher.

Habitar ambientes dimensionados a partir de estudos antropométricos e Avaliações Pós-Ocupação (APOs) dos aspectos funcionais é conseqüência de processos de projeto arquitetônico a serem consolidados. A tecnologia tem permitido obter um maior controle sobre todas as fases da produção de edificações, sendo a fase de pré-projeto decisiva para o efetivo sucesso de um empreendimento. A presente pesquisa objetiva o desenvolvimento de um roteiro para a elaboração de um programa computacional de Projeto e Desenho Arquitetônico Auxiliado por Computador (CAADD - Computer Aided Architectural Design and Drafting) que contribua para o trabalho do arquiteto nas análises geométricas de dimensionamento funcional de edificações habitacionais sociais multi-familiares, no momento do pré-projeto, considerando as especificidades do conjunto de variáveis envolvidas em cada situação de projeto. Ao escolher as alternativas de composição de Programa de Necessidades no sistema, este geraria maquetes eletrônicas (modelo digital) a partir dos parâmetros definidos no próprio programa. Os volumes abstraídos em meio digital são tratados genericamente no sistema como objetos volumétricos (mobiliário, eletrodomésticos, peças sanitárias, esquadrias, estrutura, vedação). A tecnologia de representação gráfica digital escolhida foi o padrão aberto para a distribuição de conteúdo tridimensional denominado X3D. A montagem do roteiro contempla conceitos de Computação Gráfica e de Geometria Computacional, aplicando critérios para arranjo físico de ambientes da habitação, relação entre ambientes, distribuição de unidades habitacionais (apartamentos) e análises de locação da edificação no sítio urbano. Utilizam-se como referências dimensionais resultados de APOs (classificadas como dados dinâmicos, os quais se alteram com maior freqüência no tempo) e legislação e normas vigentes (classificados como dados estáveis, os quais se alteram com menor freqüência no tempo), visando melhorias espaciais e manutenção contínua da abordagem do projeto e desenho arquitetônico funcional nos novos projetos habitacionais. / The possibility of living in environments that have been sized on the basis of anthropometric studies and post-occupancy evaluations (POEs) of the functionality of buildings is the consequence of architectural design processes yet to be consolidated. This technology has enabled greater control over all stages in the construction of buildings, the pre-design stage being decisive for the success of an enterprise. The objective of the present study is to develop guidelines for designing a computer program for architectural designing and drafting (CAADD-Computer Aided Architectural Design and Drafting) that will serve as an aid to architects in terms of geometrical analyses of the functional dimensions of low-income housing during the pre-design stage, considering the specific aspects of the variables involved in each design. By choosing alternatives for programming the software, the system is set up to enable designers to generate digital models based on standards defined in the program itself. The digitally extracted volumes are treated generically in the system as volumetric objects (furniture, appliances, plumbing fixtures, door and window frames, structures, waterproofing and others). The open standard for distributing tridimensional contents, known as X3D, was chosen as the technology for digital graphic representation. The guidelines take into account concepts for graphic computing and computational geometry and apply criteria for the physical arrangement of living environments, relationships among environments, distribution of the apartments and analyses of the location of the buildings in their urban context. Dimensional references are used to establish these guidelines, including the results of POEs (classifying the data that changes most often over time as \"dynamic\"). Other factors that must be taken into consideration are the local laws, norms and regulations (classified as \"stable\" data, which change less often over time). The objective is to develop spatial improvements and continuous feedback of the approach of the design and functional architectural drafting in new housing designs.

Pantothenate synthetase (PS), which catalyzes the last step in the pantothenate (vitamin B5) biosynthesis, is a dimeric enzyme present in bacteria, fungi and plants. The enzymatic properties of PS from Escherichia Coli, Mycobacterium tuberculosi, Fusarium Oxysporum, Lotus japonicus, Oryza sativum, Brassica napus and Arabidopsis thaliana have been characterized. The chemical reaction and the proposed mechanism of reaction are identical for PS, irrespective of the source. However, from an enzyme mechanistic point of view, plant PS’s are dissimilar to their bacterial counterparts, in that they exhibit “allosteric behavior”, a property that has not been observed in the bacterial enzyme. The behavior is quite remarkable when one takes into consideration the fact that plant PS’s share a high degree of sequence identity (~ 40%) with the bacterial enzymes. Even more intriguing is the structural mechanism proposed to explain the observed differences in structure between the PS’s from E.Coli and M.tb, which share a 42% sequence identity. Till date there is no structural information available on the plant PS’s and of the substrate bound conformation of E.coli PS. This thesis aims to provide an understanding on some aspects of the structure – function relationship of this physiologically important enzyme. Specifically, the solution properties of E. coli PS have been examined using high-resolution multinuclear, multidimensional NMR methods. Given the large size of the full-length protein (~ 63 KDa), the structurally distinct N and C-terminal domains were cloned and expressed as individual proteins and their properties investigated. Towards this end, the tertiary fold of the 40 kDa dimeric amino-terminal domain of E. coli pantothenate synthetase has been determined using multidimensional multinuclear nuclear magnetic resonance (NMR) methods (PDB entry 2k6c). Sequence specific resonance assignments for backbone HN, 15N, 13Cα, 13C', sidechain 13Cβ and aliphatic 13CH3 (of isoleucine, leucine and valine residues) were obtained using perdeuterated ILV-methyl protonated samples (BMRB entry 6940). Secondary structure of nPS was determined from 13C secondary chemical shifts and from short and medium range NOEs. Global fold of the 40 kDa homo-dimeric nPS has been determined using a total of 1012 NOEs, 696 dihedral angles, 260 RDCs, 155 hydrogen bonds, radius of gyration potential, non-crystallographic symmetry potential and database derived potential based upon the Ramachandran map. The calculated structures, which show that the N-terminal domain forms a homo-dimer in solution, is of high stereochemical quality as judged by the Ramachandran statistics (70% of the residues have backbone dihedral angles in the allowed region, 25.5% in the additionally allowed region, 4.0% in generously allowed region, and only 0.5% in disallowed region). Dynamics of nPS, which has rotational correlation time τc of 17.3 ns, was investigated by 15N relaxometry measurements. Results of these studies indicate that the E. coli protein exhibits dynamic nature at the dimer interface. These structural and dynamic features of the protein were found to be of interest when correlated with NMR based substrate binding studies. Interaction of homo-dimeric amino-terminal domain (nPS) of E. coli pantothenate synthetase (PS; encoded by the gene panC; E.C. 6.3.2.1) with the substrates pantoate, β-alanine, ATP and the product pantothenate has been studied using isotopically edited solution NMR methods. Addition of pantoate prior to ATP has led to the interesting observation that pantoate binds each monomer of nPS at two sites. ATP displaces a molecule of pantoate from the ATP binding site. β-alanine and pantothenate do not bind the protein under the condition studied. Binding of pantoate and ATP also manifests as changes in residual dipolar couplings (RDCs) of backbone 1H-15N pairs in nPS when compared to the free form of the protein. Structures of homo-dimeric nPS bound to two molecules of pantoate (PDB entry 2k6e) as well as to pantoate + ATP (PDB entry 2k6f) were calculated by inclusion of hydrogen bonds between the ligands and backbone 1H-15N pairs of nPS in addition to other NMR derived restraints. The ligand bound structures have been compared to the similar forms of the M. tb PS. Structure of each monomer of nPS bound to pantoate and ATP shows the substrates in a favorable orientation for the intermediate pantoyl adenylate to form. Moreover, at all stages of substrate binding the symmetry of the dimer was preserved. A single set of resonances was observed for all protein-ligand complexes implying symmetric binding with full-occupancy of the ligands bound to the protein. In an effort to understand the structural basis of the observed enzymatic properties of plant PS’s, a structural model of the Arabidopsis PS was constructed. The results of these structural and substrate binding studies strongly suggest that 1 Substrate binding to PS occurs only at the active site. 2 There are no additional substrate binding sites which could potentially participate as regulatory sites. 3 Pantoate does not bind at the dimer interface to induce the observed homotropic effects. 4 The structural results presented on the substrate bound forms of nPS have direct implication for the development of novel antibacterial and herbicidal agents. Recently a great deal of interest has been evinced on the effects of molecular crowding on protein folding / unfolding pathways. Nuclear magnetic resonance is the only method by which high resolution structural information can be obtained on partially denatured states of a protein under equilibrium condition. Recent methodological advances have enabled the determination of high resolution structures using information embedded in the residual dipolar couplings. Molecular crowding using confinement may thus reveal important details about chaperone mediated protein folding. We have attempted to develop a protocol to study the effects of molecular confinement by sequestering proteins in poly-acrylamide gels and then subjecting these protein molecules to denaturation and then characterize these states by nuclear magnetic resonance. The preliminary results of these studies are described here.

Made available in DSpace on 2014-12-17T13:57:16Z (GMT). No. of bitstreams: 1 MatheusJCRD.pdf: 8164370 bytes, checksum: 41f511c06d1cf35f3a26af93806d4f33 (MD5) Previous issue date: 2007-02-27 / Confirming the Brazilian tendency in the field, the multifamily vertical condominium habitats in Natal are defined as buildings with three or more floors which have been an increasingly used solution. In the mentioned project, the connection between the projectarchitects and the user/ buyers are spread out, by which the first people conceive the realestate property as a creation for the market and not the individual client in specific. Such situation along with technical and financial limitations of the project lead to the adoption of standard solutions to be utilized by clients with different profiles. Besides that, there are various legal and urban parameters by the City Director Plan showing elements of great influence in the final solution adopted by the mentioned edifices. Moving to this subject in general, this project is focused on the case study of the Ed. Ville de Montpellier, having as a base of Post Occupancy Evaluation (POE), considered an efficient tool to analyze and keep up with the progress of the construction of the building, including technical approvals, the application of surveys with the local residents and the creation of informal interviews. The data shows that with time some items that initially motivated the acquisition of the realestate property (with a social common area) move to being less valued, and that the residents quickly alter the pre constructed space, thus seeking to alter the property in a more personal and conforming manner. The possibility of a new emphasis for projects on the mentioned space should also be in discussion, the created project calls for the attention of projected conceptual aspects and interdependence within project and construction which permits the indication of some recommendations for the projection of multifamily residential buildings within the studied realm / Confirmando tend?ncia brasileira na ?rea, em Natal, a habita??o em condom?nios verticais multifamiliares, constitu?dos por edif?cios com mais de tr?s pavimentos, tem sido uma solu??o crescentemente utilizada. Nesse tipo de projeto, no entanto, a liga??o entre os arquitetosprojetistas e os usu?rios/compradores ? difusa, pois os primeiros concebem o im?vel tendo em vista uma demanda de mercado e n?o um cliente espec?fico. Tal situa??o, aliada ?s limita??es t?cnicas e financeiras do empreendimento, conduz ? ado??o de solu??es standard a serem utilizadas por clientes de perfis diferentes. Al?m disso, os v?rios par?metros urban?sticos e legais definidos pelo Plano Diretor da cidade mostramse elementos de grande influ?ncia na solu??o final adotada para essas edifica??es. Partindo desse quadro geral, esse trabalho realiza o estudo de caso do Ed. Ville de Montpellier, tendo como base a avalia??o p?socupa??o (APO), considerada eficiente ferramenta para an?lise e acompanhamento do desempenho edil?cio. No trabalho, de cunho qualitativo descritivo, foram utilizados multim?todos envolvendo: an?lise de documentos relacionados ao pr?dio em quest?o, vistoria t?cnica, aplica??o de question?rios com moradores e realiza??o de entrevistas informais. Os dados mostram que, com o tempo, alguns itens que inicialmente motivaram a aquisi??o do im?vel (como a ?rea social comum) passam a ser menos valorizados, e que rapidamente os moradores alteram o espa?o padronizado, buscando adequ?lo aos seus desejos e necessidades. Discutindo a possibilidade de novo enfoque para projetos arquitet?nicos nesta ?rea, o trabalho chama a aten??o para aspectos de concep??o projetual e a interdepend?ncia entre projeto e obra, e permite a indica??o de algumas recomenda??es para a projeta??o de edif?cios residenciais multifamiliares na realidade estudada

Many developing countries, rich with natural resources, have turned to tourism as a source of national growth and subsequently an increasing amount of local communities are being "developed". Despite its importance to developing countries, tourism has been covered scarcely in the literature on development studies. Local communities that find themselves in a situation having to deal with tourism related development, face many challenges. Contrary to previous work on development which considered poor local communities a threat to the natural environment, current views emphasise the role of the community in bringing about participation, conservation and consequently Sustainable Development. Therefore, if tourism can be seen as a possible path to Sustainable Development, the role of the local community in its development is of vital importance. This research investigates the role of the local community in integrating conservation in tourism related development by means of a case study on Apo Island. / DEVELOPMENT STUDIES / MA(SS)(DEV. STUD)

Many developing countries, rich with natural resources, have turned to tourism as a source of national growth and subsequently an increasing amount of local communities are being "developed". Despite its importance to developing countries, tourism has been covered scarcely in the literature on development studies. Local communities that find themselves in a situation having to deal with tourism related development, face many challenges. Contrary to previous work on development which considered poor local communities a threat to the natural environment, current views emphasise the role of the community in bringing about participation, conservation and consequently Sustainable Development. Therefore, if tourism can be seen as a possible path to Sustainable Development, the role of the local community in its development is of vital importance. This research investigates the role of the local community in integrating conservation in tourism related development by means of a case study on Apo Island. / DEVELOPMENT STUDIES / MA(SS)(DEV. STUD)