As enzimas diidroorotato desidrogenases (DHODHs) são flavo-enzimas que catalisam a oxidação do diidroorotato em orotato na quarta etapa da biossíntese de novo de nucleotídeos de pirimidina. Durante a rápida proliferação celular em mamíferos, a via de salvação de pirimidinas é insuficiente para suprir deficiências na síntese de nucleotídeos. Além disso, certos parasitas não possuem a via de salvação e contam somente com a biossíntese de novo para a produção de nucleotídeos. Por esta razão, DHODH se tornou um excelente alvo na busca por inibidores que interrompam a síntese de nucleotídeos. As enzimas DHODHs de E. coli (EcDHODH) e de X. fastidiosa (XfDHODH) são membros da classe 2 das DHODHs e encontram-se associadas à membrana citoplasmática através de uma extensão em seu N-terminal, enquanto que DHODH de T. cruzi (TcDHODH), membro da classe 1 de DHODHs, é uma proteína citosólica. Neste trabalho, usamos uma combinação de metodologias de biologia molecular e bioquímica com técnicas espectroscópicas para obter informações estruturais e funcionais acerca da enzima DHODH. Assim, Ressonância Paramagnética Eletrônica (RPE) associada à marcação de spin sítio dirigida (SDSL) e simulação espectral foram empregadas para estudar a interação da EcDHODH com modelos de membrana. Mudanças na dinâmica estrutural das vesículas induzidas pela enzima foram monitoradas via marcadores de spin localizados em diferentes posições ao longo da cadeia acil de fosfolipídios. Além disso, técnicas de DNA recombinante e mutações sítio dirigidas foram utilizadas para produzir mutantes de EcDHODH no qual um sondas paramagnéticas foram seletivamente ligadas em resíduos localizados na extensão N-terminal da proteína para experimentos subseqüentes de RPE-SDSL. Esses são os primeiros experimentos de marcação de spin sítio dirigida realizados no Brasil e com os quais monitoramos a dinâmica experimentada na região do N-terminal. Além disso, várias tentativas foram feitas para se expressar e purificar a enzima XfDHODH e a estabilidade estrutural da enzima TcDHODH na presença de um de seus inibidores naturais, o orotato, foi monitorada através de experimentos de Dicroísmo Circular (CD). / Dihydroorotate dehydrogenases (DHODHs) are flavin-containing enzymes which catalyse the conversion of (S)-dihydroorotate to orotate, in the fourth step of the de novo biosynthesis of pyrimidine nucleotides. In rapidly proliferating mammalian cells, pyrimidine salvage pathway is insufficient to overcome deficiencies for nucleotide synthesis. Moreover certain parasites lack salvage enzymes, relying solely on the de novo pathway to produce nucleotides. Thus, DHODH has turned out an excellent target to the development of inhibitors that block nucleotide biosynthesis. E. coli DHODH (EcDHODH) and X. fastidiosa DHODH (XfDHODH) are class 2 DHODHs found associated to cytosolic membranes through an N-terminal extension, whereas T. cruzi DHODH (TcDHODH) is a class 1 DHODH localizated in the cytoplasm. In the present work, we used a combination of molecular biology and biochemical methodologies with spectroscopic techniques to obtain structural and functional information on DHODH. On one hand, Electronic Paramagnetic Resonance (EPR) associated with Site-directed Spin Labeling (SDSL) and spectral simulation were employed to study the interaction of EcDHODH with vesicles. Changes in vesicle dynamic structure induced by the enzyme were monitored via spin labels located at different positions along the phospholipid acyl chain and via spin labels located at enzyme specific positions. On the other hand, DNA techniques and site-directed mutagenesis were used to produce mutants of EcDHODH where a nitroxide spin probe was selectively attached to some residues located at the protein N-terminal extension for subsequent EPR-SDSL experiments. These are the first site-directed spin labeling experiments performed in Brazil and the spectra allowed us to monitor dynamics experienced by those residues at the EcDHODH N-terminal domain. Furthermore, molecular biology and biochemical assays were employed with the objective of expressing and purifying XfDHODH and Circular Dichroism (CD) was utilized to probe the structural stability of TcDHODH in the presence of its natural inhibitor (orotate).

A esquizofrenia é um transtorno psiquiátrico incapacitante que afeta estimadamente 1% da população mundial. Delírios, alucinações, desorganização de pensamento e prejuízo cognitivo são as principais marcas da Esquizofrenia. Fisiologicamente, além de anormalidades funcionais e estruturais, alterações na atividade neuronal são reportadas. Como a atividade neuronal possui uma relação direta com o fluxo sanguíneo cerebral (CBF, Cerebral Blood Flow), a técnica de Imagens por Ressonância Magnética, denominada Marcação dos Spins Arteriais (ASL, Arterial Spin Labeling), que permite a obtenção de mapa quantitativo de CBF, é uma ferramenta útil na avaliação funcional cerebral. Além disso, a ASL pode ser usada na avaliação da conectividade funcional, que é eficiente na investigação de rupturas funcionais entre as regiões do cérebro. Comparando com um grupo de sujeitos saudáveis, os pacientes com esquizofrenia, recrutados no Hospital das Clínicas de Ribeirão Preto (HCFMRP), apresentaram redução de CBF em regiões bilaterais do polo frontal e giro frontal superior, giro frontal medial direito, partes triangular e opercular do giro frontal inferior direito, divisão posterior do giro supramarginal esquerdo, divisão superior e inferior do córtex occipital lateral esquerdo e polo occipital. A conectividade funcional, avaliada por três diferentes métodos (baseado em semente, análise de componentes independentes e teoria dos grafos), se apresentou prejudicada em regiões envolvendo funções motoras, sensoriais e cognitivas dos pacientes. Portanto, utilizando uma técnica de imagem completamente não invasiva, foi possível observar déficits de CBF e alterações na organização funcional do cérebro de pacientes com esquizofrenia, relacionados com os sintomas e características da psicopatologia. / Schizophrenia is a disabling psychiatric disorder that affects around 1% of the population worldwide. Delusions, hallucinations, disorganized thought, and cognitive deficits are the main features of schizophrenia. Physiologically, in addition to functional and structural abnormalities, changes in neuronal activity are reported. Since the Cerebral Blood Flow (CBF) is directly related with neuronal activity, the Magnetic Resonance Imaging (MRI) technique called Arterial Spin Labeling (ASL), which allows the quantification of CBF, is a useful tool in brain functional evaluation. In addition, ASL can be used to assess functional connectivity, which is efficient in investigating functional impairment between regions of the brain. Patients with Schizophrenia, recruited at the Clinical Hospital (HCFMRP), presented a reduction of CBF in bilateral regions of the frontal pole and superior frontal gyrus, right medial frontal gyrus, triangular and opercular parts of the right inferior frontal gyrus, posterior division of left supramarginal gyrus, superior and inferior division of left lateral occipital cortex and occipital pole. Functional connectivity, assessed by three different methods (seed-based, independent component analysis and graph theory), was impaired in regions involving patients\' motor, sensory and cognitive functions. Therefore, using a noninvasive imaging technique, it was possible to observe CBF deficits and alterations in the functional organization of the brain of schizophrenia patients, related to the symptoms and characteristics of the psychopathology.

O presente trabalho tem como objetivo desenvolver modelos de tabelas informativas contendo atributos de desempenho técnico e ambiental de produtos da construção civil para os produtos forro de lã de vidro e impermeabilização de coberturas planas. Foram estudadas as normas para Declaração Ambiental (normas ISO 14.000) e um levantamento dos requisitos que demandam atributos específicos para materiais de construção e que constam nos sistemas de certificação ambiental de edifícios LEED BD+C 2009, AQUA Residencial e norma de desempenho NBR 15.575: 2013 - Parte 1. A pesquisa foi desenvolvida com a participação de agentes do meio produtivo, articulando as demandas de projetistas e dos segmentos industriais dos produtos supracitados. Foram selecionados atributos que influenciam a qualidade ambiental do edifício: toxicidade; conteúdo de compostos orgânicos voláteis; presença de substâncias carcinogênicas; porcentagem na composição e tipo de conteúdo reciclado; descarte do produto e da embalagem; certificações do produto e do fabricante. Atributos que influenciam o desempenho técnico do material quanto à segurança, funcionalidade e durabilidade foram selecionados para cada tipologia de produto: reação ao fogo; propriedades que afetam a estanqueidade, manutenção, o desempenho térmico e acústico; prazos de garantia e durabilidade. A combinação desses atributos permite, desde as fases iniciais do processo de projeto, exercício de análise e interpretação dos dados para a especificação de materiais adequados às necessidades do edifício e seus usuários. O modelo apresentado, denominado Tabela Ambiental, favorece a informação dinâmica aos envolvidos da cadeia construtiva, evoluindo juntamente com o conhecimento do mercado sobre o tema e suas inovações tecnológicas. A Tabela Ambiental apresenta uma proposta de comunicação simplificada e, por meio da referência de comprovação, incentiva a especificação consciente e responsável, podendo incrementar a educação ambiental e direcionar o mercado da construção civil a novos rumos na sustentabilidade. / The present paper aims to develop models of informative tables containing attributes of technical and environmental performance of civil construction products for glass wool ceiling and flat roofs waterproofing. Standards were studied for Environmental Declaration (ISO 14000 standards) and a survey of requirements which demand specific attributes for building materials in the environmental certification systems for buildings LEED BD + C 2009, AQUA Residential and performance standard NBR 15.575: 2013 - Part 1. The research was developed with productive agents articulating the demands of designers and industrial segments of the mentioned products. Attributes which influence on building environmental quality were selected: content and type of recycled; packaging and product disposal; maker and product certifications. Attributes that influence the environmental quality of the building were selected: toxicity; content of volatile organic compounds; presence of carcinogenic substances; percentage in the composition and type of recycled content; disposal of product and packaging; certifications of the product and the manufacturer. Attributes that influence the technical performance of the equipment for safety, functionality and durability were selected for each type of product: reaction to fire; properties that affect the tightness, maintenance, thermal and acoustic performance; warranty terms and durability. The combination of these attributes allow, from the early stages of the design process, exercise of analysis and data interpretation for specifying suitable materials to the building needs and its users. The model presented, called as Environmental Table, favors to the dynamics involved in constructive chain information, progressing with the market knowledge about the subject and its technological innovations. The Environmental Table presents a simplified communication proposal and through reference encourages responsible and conscious specification, it can increase environmental education and direct the civil construction market to new ways in sustainability.

Traditional students enroll in post-secondary institutions during emerging adulthood. College enrollment is increasing and adult weight gain occurs most rapidly during the college-age years, with poor food decisions as a potential contributing factor. The present study examined the awareness, perception, and self-reported purchasing behaviors of college students regarding four front-of-package (FOP) nutrition labeling systems and symbols. Students were sent a 24-question web-based survey, with 908 completed surveys that met the research criteria. There were 888 (98.3%) respondents who recognized at least one of the four presented FOP nutrition labels. There were no significant differences between the groups that recognized one to four of the FOP nutrition labels in their stated likelihood to purchase foods with the specified labels. Students’ awareness of four commercial FOP nutrition labeling systems and symbols on product packaging did not have an impact on their food purchasing behaviors.

This study deals with "open code dating," the movement by grocery manufacturers and distributors toward dating perishable food packages in such a manner that consumers can readily determine product freshness or length of time on store shelves. The study explores the desirability and feasibility of open code dating, placing greatest importance upon the response of the consumer to the concept. It was found that consumers were aware of open code dating and generally strongly desired its universal adoption. Shoppers were also confused by open dating and failed to understand freshness dates properly. The strongest desire for open dating was found in shoppers at the upper end of the socio-economic scale. Grocery retailers expressed satisfaction with open coding, believing it an aid in stock rotation and customer satisfaction. Possible disadvantages, such as increased throwaway costs and large conversion costs, were not perceived as being significant. The businessmen favored widespread adoption of open code dating. On the basis of data from interviews with shoppers, it is concluded that consumers desire adoption of open code dating and do use this service. It is also concluded that adoption of open code dating would be an economically sound decision which would constitute a desirable marketing strategy.

No description available.

The high prevalence and mortality of cerebrovascular disease has led to the development of several methods to measure cerebral blood flow (CBF) in vivo. One of these, arterial spin labeling (ASL), is a quantitative magnetic resonance imaging (MRI) technique with the advantage that it is completely non-invasive. The quantification of CBF using ASL requires correction for a tissue specific parameter called the brain-blood partition coefficient (BBPC). Despite regional and inter-subject variability in BBPC, the current recommended implementation of ASL uses a constant assumed value of 0.9 mL/g for all regions of the brain, all subjects, and even all species. The purpose of this dissertation is 1) to apply ASL to a novel population to answer an important clinical question in the setting of Down syndrome, 2) to demonstrate proof of concept of a rapid technique to measure BBPC in mice to improve CBF quantification, and 3) to translate the correction method by applying it to a population of healthy canines using equipment and parameters suitable for use with humans. Chapter 2 reports the results of an ASL study of adults with Down syndrome (DS). This population is unique for their extremely high prevalence of Alzheimer’s disease (AD) and very low prevalence of systemic cardiovascular risk factors like atherosclerosis and hypertension. This prompted the hypothesis that AD pathology would lead to the development of perfusion deficits in people with DS despite their healthy cardiovascular profile. The results demonstrate that perfusion is not compromised in DS participants until the middle of the 6th decade of life after which measured global CBF was reduced by 31% (p=0.029). There was also significantly higher prevalence of residual arterial signal in older participants with DS (60%) than younger DS participants (7%, p = 0.005) or non-DS controls (0%, p < 0.001). This delayed pattern of perfusion deficits in people with DS differs from observations in studies of sporadic AD suggesting that adults with DS benefit from an improved cardiovascular risk profile early in life. Chapter 3 introduces calibrated short TR recovery (CaSTRR) imaging as a rapid method to measure BBPC and its development in mice. This was prompted by the inability to account for potential changes in BBPC due to age, brain atrophy, or the accumulation of hydrophobic A-β plaques in the ASL study of people with DS in Chapter 2. The CaSTRR method reduces acquisition time of BBPC maps by 87% and measures a significantly higher BBPC in cortical gray matter (0.99±0.04 mL/g,) than white matter in the corpus callosum (0.93±0.05 mL/g, p=0.03). Furthermore, when CBF maps are corrected for BBPC, the contrast between gray and white matter regions of interest is improved by 14%. This demonstrates proof of concept for the CaSTRR technique. Chapter 4 describes the application of CaSTRR on healthy canines (age 5-8 years) using a 3T human MRI scanner. This represents a translation of the technique to a setting suitable for use with a human subject. Both CaSTRR and pCASL acquisitions were performed and further optimization brought the acquisition time of CaSTRR down to 4 minutes which is comparable to pCASL. Results again show higher BBPC in gray matter (0.83 ± 0.05 mL/g) than white matter (0.78 ± 0.04 mL/g, p = 0.007) with both values unaffected by age over the range studied. Also, gray matter CBF is negatively correlated with age (p = 0.003) and BBPC correction improved the contrast to noise ratio by 3.6% (95% confidence interval = 0.6 – 6.5%). In summary, the quantification of ASL can be improved using BBPC maps derived from the novel, rapid CaSTRR technique.

L’objectif de cette thèse est d’attribuer à chaque sommet x d’un graphe G à n sommets une étiquette L(x) de taille compacte O(log n) bits afin de pouvoir :1. construire, à partir des étiquettes d’un ensemble de sommets en panne X C V (G), une structure de donnée S(X)2. décider, à partir de S(X) et des étiquettes L(u) et L(v), si les sommets u et v sont connectés dans le graphe G n X.Nous proposons une solution à ce problème pour la famille des graphes 3-connexes de genre g (via plusieurs résultats intermédiaires).— Les étiquettes sont de taille O(g log n) bits— Le temps de construction de la structure de donnée S(X) est O(Sort([X]; n)).— Le temps de décision est O(log log n). Ce temps est optimal.Nous étendons ce résultat à la famille des graphes excluant un mineur H fixé. Les étiquettes sont ici de taille O(polylog n) bits. / We aim at assigning each vertex x of a n-vertices graph G a compact O(log n)-bit label L(x) in order to :1. construct, from the labels of the vertices of a forbidden set X C V (G), a datastructure S(X)2. decide, from S(X), L(u) and L(v), whether two vertices u and v are connected in G n X.We give a solution to this problem for the family of 3-connected graphs whith bounded genus.— We obtain O(g log n)-bit labels.— S(X) is computed in O(Sort([X]; n)) time.— Connection between vertices is decided in O(log log n) optimal time.We finally extend this result to H-minor-free graphs. This scheme requires O(polylog n)-bit labels.

Cette thèse s‘inscrit dans le contexte de l‘étude de matériaux fibreux tissés et traite de l‘analyse morphologique de leur texture et de la simulation des arrangements des fibres dans un fil. Le volet « analyse » consiste à extraire des données caractérisant la géométrie des fibres et des fils composant le renfort tissé et s‘appuie sur des images tridimensionnelles issues de tomographies rayon X à haute résolution. Une chaine de traitement d‘images visant à séparer et identifier les fils et les fibres est proposée. Elle s‘appuie sur un algorithme de labellisation du fil et des fibres. Les caractéristiques telles que les diamètres et les orientations de fibres et la densité locale de fibres dans le fil sont ensuite calculées. Le volet « simulation » vise à générer un renfort fibreux « virtuel ». Un algorithme fondé sur la résolution de modèles dynamiques permet de placer au sein d‘un fil, des objets représentant les fibres tout en respectant des contraintes issues des paramètres issus du volet « analyse » ou choisis arbitrairement par l‘utilisateur. L‘ensemble des fibres composant le fil sont alors synthétisés en respect de ces contraintes. Ces deux volets sont appliqués avec succès à la caractérisation et à la synthèse de renforts fibreux de composites thermostructuraux. Les caractéristiques géométriques, estimées sur des tomographies à haute résolution, font ressortir des phénomènes tels que le cisaillement intra fils. La synthèse permet de simuler le placement de fibres à partir de paramètres géométriques obtenus à haute résolution, au sein d‘un fil dont l‘enveloppe est obtenue à basse résolution et donc sur une longueur plus représentative du tissage. / The aim of this work is to study fibrous woven materials and to develop morphological analysis of their texture enabling simulation of fibrous layout in yarns. The ―analysis‖ stage consists in data extraction to characterize the geometry of fibers and yarns constituting the woven material, based on tridimensional images generated by high resolution X-ray tomography. An image processing workflow to separate and identify fibers and yarn is proposed. Then, diameters, fiber orientations and local fiber density in yarns are computed. The ―simulation‖ stage targets to generate virtual fibrous materials. An algorithm based on the resolution of a dynamic model allows placing objects representing fibers into a yarn. It uses as input the characteristic parameters previously extracted in the analysis stage or arbitrarily chosen ones. Consequently, all fibers are synthesized according to these constraints. These two stages are successfully applied to characterize and synthesize woven ceramic matrix composites. Geometrical characteristics, extracted from the high-resolution tomographic images, highlight yarn phenomena like shearing planes. The synthesis simulates fibers placement, merging geometrical parameters extracted on high and low resolution. Indeed, using fiber parameters extracted from high resolution images and yarn envelopes from low resolution ones enable generating a result with a more representative woven length.

This thesis analyzes the strengths and weaknesses of various types of eco-labels, focusing primarily on differences between mandatory and voluntary eco-labeling programs. I argue that many of the problems with eco-labeling could be addressed by improving regulations. The current regulation of eco-labeling in the United States is discussed, especially the shortcomings of the FTC’s Green Guides. I recommend creating enforceable national legislation to regulate environmental claims that includes involvement of key stakeholders, a list of acceptable environmental claims, enforceable national definitions of environmental terms, an avenue for manufacturers and consumers to challenge environmental claims, consumer education, and periodic review and revision.