Movement of NH3 through the human urea transporter B: a new gas channel

Aquaporins and Rh proteins can function as gas (CO2 and NH3) channels. The present study explores the urea, H2O, CO2, and NH3 permeability of the human urea transporter B (UT-B) (SLC14A1), expressed in Xenopus oocytes. We monitored urea uptake using [14C]urea and measured osmotic water permeability (Pf) using video microscopy. To obtain a semiquantitative measure of gas permeability, we used microelectrodes to record the maximum transient change in surface pH (∆pHS) caused by exposing oocytes to 5% CO2/33 mM HCO3- (pHS increase) or 0.5 mM NH3/NH4+ (pHS decrease). UT-B expression increased oocyte permeability to urea by >20-fold, and Pf by 8-fold vs. H2O-injected control oocytes. UT-B expression had no effect on the CO2-induced ∆pHS but doubled the NH3-induced ∆pHS. Phloretin reduced UT-B-dependent urea uptake (Jurea * ) by 45%, Pf * by 50%, and (- ∆pHS * )NH3 by 70%. p-Chloromercuribenzene sulfonate reduced Jurea * by 25%, Pf * by 30%, and (∆pHS * )NH3 by 100%. Molecular dynamics (MD) simulations of membrane-embedded models of UT-B identified the monomeric UT-B pores as the main conduction pathway for both H2O and NH3 and characterized the energetics associated with permeation of these species through the channel. Mutating each of two conserved threonines lining the monomeric urea pores reduced H2O and NH3 permeability. Our data confirm that UT-B has significant H2O permeability and for the first time demonstrate significant NH3 permeability. Thus the UTs become the third family of gas channels. Inhibitor and mutagenesis studies and results of MD simulations suggest that NH3 and H2O pass through the three monomeric urea channels in UT-B.

Eliciting nature and landscape writing through outdoor experiences

Nature and landscape writing includes creative writing about wild places. However, most authors have a literary background and are not outdoor ‘educators’. Using a hermeneutic phenomenological approach, the reasons suggested are a lack of framing of outdoor experiences for this intent, the need for learning the skills of interpretation and lexicon and the offer of prolonged, powerful experiences and time for creative thinking and responses, such as an extended solo. It is suggested that outdoor educators may be too busy ‘experiencing’ to write, that they do not go ‘slow’ enough or that they are encapsulated in the ‘edginess of existence’ through adventure and just pass through their surroundings rather than connect with them. Outdoor educators have much to offer as they experience metaphorical or literal journeys comprising ‘flow’ rather than episodic encounter through lived experience to create rich embodied stories with ideological and social aspects so often overlooked in narrative.

Indicadores de saude em mulheres acima de 50 anos de idade participantes de um programa de atividades fisicas no complexo da Mare-RJ

Univesidade Estadual de Campinas. Faculdade de Educação Física

The azo dye Disperse Orange 1 induces DNA damage and cytotoxic effects but does not cause ecotoxic effects in Daphnia similis and Vibrio fischeri

Azo dyes constitute the largest group of colorants used in industry and can pass through municipal waste water plants nearly unchanged due to their resistance to aerobic treatment, which potentially exposes humans and local biota to adverse effects. Unfortunately, little is known about their environmental fate. Under anaerobic conditions, some azo dyes are cleaved by microorganisms forming potentially carcinogenic aromatic amines. In the present study, the azo dye Disperse Orange 1, widely used in textile dyeing, was tested using the comet, Salmonella/microsome mutagenicity, cell viability, Daphnia similis and Microtox (R) assays. The human hepatoma cell line (HepG2) was used in the comet assay and for cell viability. In the mutagenicity assay. Salmonella typhimurium strains with different levels of nitroreductase and o-acetyltransferase were used. The dye showed genotoxic effects with respect to HepG2 cells at concentrations of 0.2, 0.4, 1.0, 2.0 and 4.0 mu g/mL. In the mutagenicity assay, greater responses were obtained with the strains TA98 and YG1041, suggesting that this compound mainly induces frameshift mutations. Moreover, the mutagenicity was greatly enhanced with the strains overproducing nitroreductase and o-acetyltransferase, showing the importance of these enzymes in the mutagenicity of this dye. In addition, the compound induced apoptosis after 72 h in contact with the HepG2 cells. No toxic effects were observed for either D. similis or Vibrio fischeri. (C) 2011 Elsevier B.V. All rights reserved.

Ameloblast apoptosis and IGF-1 receptor expression in the continuously erupting rat incisor model

Enamel-producing cells (ameloblasts) pass through several phenotypic and functional stages during enamel formation. In the transition between secretory and maturation stages, about one quarter of the ameloblasts suddenly undergo apoptosis. We have studied this phenomenon using the continuously erupting rat incisor model. A special feature of this model is that all stages of ameloblast differentiation are presented within a single longitudinal section of the developing tooth. This permits investigation of the temporal sequence of gene and growth factor receptor expression during ameloblast differentiation and apoptosis. We describe the light and electron microscopic morphology of ameloblast apoptosis and the pattern of insulin-like growth factor-1 receptor expression by ameloblasts in the continuously erupting rat incisor model. In the developing rat incisor, ameloblast apoptosis is associated with downregulated expression of the insulin-like growth factor-1 receptor. These data are consistent with the hypothesis that ameloblasts are hard wired for apoptosis and that insulin-like growth factor-1 receptor expression is required to block the default apoptotic pathway. Possible mechanisms of insulin-like growth factor-1 inhibition of ameloblast apoptosis are presented. The rat incisor model may be useful in studies of physiological apoptosis as it presents apoptosis in a predictable pattern in adult tissues.

Multi-fluorescent silica colloids for encoding large combinatorial libraries

Large chemical libraries can be synthesized on solid-support beads by the combinatorial split-and-mix method. A major challenge associated with this type of library synthesis is distinguishing between the beads and their attached compounds. A new method of encoding these solid-support beads, 'colloidal bar-coding', involves attaching fluorescent silica colloids ('reporters') to the beads as they pass through the compound synthesis, thereby creating a fluorescent bar code on each bead. In order to obtain sufficient reporter varieties to bar code extremely large libraries, many of the reporters must contain multiple fluorescent dyes. We describe here the synthesis and spectroscopic analysis of various mono- and multi-fluorescent silica particles for this purpose. It was found that by increasing the amount of a single dye introduced into the particle reaction mixture, mono- fluorescent silica particles of increasing intensities could be prepared. This increase was highly reproducible and was observed for six different fluorescent dyes. Multi-fluorescent silica particles containing up to six fluorescent dyes were also prepared. The resultant emission intensity of each dye in the multi-fluorescent particles was found to be dependent upon a number of factors; the hydrolysis rate of each silane-dye conjugate, the magnitude of the inherent emission intensity of each dye within the silica matrix, and energy transfer effects between dyes. We show that by varying the relative concentration of each silane-dye conjugate in the synthesis of multi-fluorescent particles, it is possible to change and optimize the resultant emission intensity of each dye to enable viewing in a fluorescence detection instrument.

The entrapment of particles by windbreaks

A theory is developed for calculating the entrapment of particles by a windbreak, with four results. (1) The fraction of particles in the oncoming flow which pass through the windbreak, or transmittance of the windbreak for particles (sigma), is related to the optical porosity (tau). The very simple approximation sigma=tau works well for most applications involving the interception of spray droplets by windbreaks. Results from a field experiment agree with the theoretical predictions. (2) A new equation for the bulk drag coefficient of a windbreak is tested against numerical, wind tunnel and field experiments. This enables the bleed velocity for the flow through the windbreak to be predicted in terms of the screen pressure coefficient (k) of the barrier. (3) The relationship between k and tau is different for a vegetative barrier than for a screen across a confined duct, implying a lower Fc for given tau. (4) The total deposition of particles to a windbreak is determined by a trade-off between particle absorption and throughflow, implying an optimum value of tau for maximum total deposition. For particles larger than 30 mum and vegetation elements smaller than 30 mm, this occurs near tau = 0.2. (C) 2001 Elsevier Science Ltd. All rights reserved.

Organização parlamentar, processo decisório e produção legislativa no cenário político capixaba : uma análise da Assembléia do Estado do Espírito Santo na 16ª legislatura (2007-2010)

Esta pesquisa é um estudo sobre o processo legislativo e a produção de leis na ALES, durante a 16ª Legislatura. O escopo do estudo foi o de analisar o impacto das regras, contidas no regimento interno e na Constituição estadual, na produção de leis entre 2007 e 2010. O pressuposto geral da abordagem advém do Novo Intitucionalismo como movimento teórico da Ciência Política contemporânea, que nos permite inferir que as regras institucionais, para além de serem simples regras de organização das Casas Legislativas, influenciam no processo decisório e, consequentemente, na produção de leis. Importando as análises dos modelos distributivo, informacional e partidário de organização dos Legislativos, a pesquisa permitiu concluir que, a ALES, de 2007 a 2010, manteve seus trabalhos alinhados à versão distributiva. O parlamentar capixaba atuou, individualmente, a fim de manter suas bases eleitorais. Com relação à atuação do sistema comissional, foi identificada a inaptidão das comissões da ALES para influenciar o processo decisório. As comissões permanentes não possuem capacidade de moldar os projetos que por elas tramitam. O regime de tramitação influencia diretamente a produção de leis da ALES, notadamente, nas leis de autoria do Executivo, já que estas, tramitaram, quase que na totalidade, em regime de urgência. No que tange aos partidos, a fragmentação partidária identificada na ALES acaba por refletir nos dados obtidos sobre a produção de leis, na medida em que não existe uma verdadeira articulação em torno dos partidos como atores determinantes no processo legislativo da ALES. Enfim, em apertada síntese, foi possível constatar a imposição da agenda de trabalhos pelo Executivo. Os projetos aprovados, de temas importantes e abrangência estadual, foram, em sua maioria, propostos pelo Executivo; a urgência impacta frontalmente os projetos do Executivo, que são aprovadas em pequeno espaço de tempo; as comissões não possuem poder mínimo de influência nos projetos de autoria do Executivo; e os partidos, altamente fragmentados, não constituem instância com capacidade de articulação.

Higienização de utensílios alimentares em estabelecimentos de ensino pré-escolar: um estudo em duas instituições de ensino

A transmissão de microrganismos patogénicos pode ocorrer através da sua presença em resíduos de alimentos ou água depositados sobre os utensílios que se encontram em deficientes condições de higienização. Para que seja assegurada a higiene alimentar torna-se essencial a conservação e higienização dos utensílios, desempenhando os manipuladores de alimentos um papel determinante no que diz respeito à execução de um eficaz protocolo de higienização, uma correcta manipulação e a adopção de normas higiénicas que evitem a contaminação dos utensílios. Da totalidade das amostras realizadas aos utensílios alimentares provenientes das duas instituições de ensino pré-escolar analisados neste estudo, 27% encontravam-se contaminadas e num insuficiente estado de higienização devido à presença de microrganismos mesófilos aeróbios a 30ºC em número superior a 100 UFC/peça. Foi verificado a presença de fungos e a pesquisa positiva de enterobactérias e bactérias presumíveis de E. coli. Em ambos os estabelecimentos de ensino, os utensílios constituídos por material plástico apresentaram níveis de contaminação na generalidade superiores em relação aos constituídos por aço inoxidável. O tipo de material e o estado dos utensílios são também aspectos relevantes que podem influenciar o processo de adesão microbiana e consequente formação de biofilme, pelo que na aquisição dos utensílios devem ser consideradas as características inerentes aos diferentes materiais, devendo os mesmos ser substituídos periodicamente devido ao desgaste causado pelo seu frequente uso. Para além disso, os utensílios devem passar constantemente por uma avaliação microbiológica para controlo da eficiência do processo de higienização, contribuindo para a garantia da higiene alimentar e segurança do alimento. A garantia da higiene alimentar constitui uma base importante em idades de maior vulnerabilidade.

À descoberta das emoções sociais: vergonha e culpa : implementação e avaliação de duas sessões de um programa para a promoção de competências emocionais : um enfoque comunitário

Dissertação de Mestrado em Psicologia da Educação, especialidade em Contextos Comunitários.

Simulação da aplicação da metodologia Balanced Scorecard numa PME: O estudo do caso PME-T.H.

Mestrado em Contabilidade

Simulação e controlo de veículos autónomos terrestres

A implementação e venda de robôs autónomos tem sido um sector que nos últimos anos tem adquirido cada vez mais quota no mercado, nomeadamente no sector militar, agrícola e da vigilância. Como tal, tem sido também de grande importância a capacidade de implementar e testar robôs por parte das entidades que os fabricam. Uma das formas que tem garantido o sucesso do desenvolvimento de robôs é a simulação prévia dos mesmos antes que estes passem a fase de produção. Sendo assim, o LSA como entidade de desenvolvimento de robôs autónomos, tem necessidade de adquirir um sistema que simule os robôs em desenvolvimento. O trabalho desta tese consiste na realização de um sistema que simule robôs autónomos terrestres de forma que se possa observar o comportamento da cinemática, dinânica e hardware dos robôs em ambiente 3D. Esta aplicação de simulação pode mais tarde ser utilizada pelo laboratório para testar missões, validar alterações de estrutura, sensores, etc. Para além disso, com recurso ao simulador Player/Stage/Gazebo testar o robô LINCE e implementar algoritmos de controlo para o mesmo. Os algoritmos de controlo implementados baseiam-se em primitivas de controlo básico para serem utilizadas pelo sistema de navegação e gerar trajectórias complexas. Os algoritmos desenvolvidos nesta tese baseiam-se nas equações cinemáticas do veículo estudado. Estes algoritmos depois de testados no simulador, poderão ser colocados no Hardware do robô. Desta forma consegue-se desenvolver algoritmos para determinado robô sem que este esteja operacional.

People counting system using existing surveillance video camera

The Casa da Música Foundation, responsible for the management of Casa da Música do Porto building, has the need to obtain statistical data related to the number of building’s visitors. This information is a valuable tool for the elaboration of periodical reports concerning the success of this cultural institution. For this reason it was necessary to develop a system capable of returning the number of visitors for a requested period of time. This represents a complex task due to the building’s unique architectural design, characterized by very large doors and halls, and the sudden large number of people that pass through them in moments preceding and proceeding the different activities occurring in the building. To achieve the technical solution for this challenge, several image processing methods, for people detection with still cameras, were first studied. The next step was the development of a real time algorithm, using OpenCV libraries and computer vision concepts,to count individuals with the desired accuracy. This algorithm includes the scientific and technical knowledge acquired in the study of the previous methods. The themes developed in this thesis comprise the fields of background maintenance, shadow and highlight detection, and blob detection and tracking. A graphical interface was also built, to help on the development, test and tunning of the proposed system, as a complement to the work. Furthermore, tests to the system were also performed, to certify the proposed techniques against a set of limited circumstances. The results obtained revealed that the algorithm was successfully applied to count the number of people in complex environments with reliable accuracy.

As duas deusas: a partir da obra de vários autores: trabalho de projecto

Trabalho de Projeto submetido à Escola Superior de Teatro e Cinema para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Teatro, Artes Performativas – Especialização em Interpretação.

The transport of carboxylic acids and important role of the Jen1p transporter during the development of yeast colonies

On solid substrates, yeast colonies pass through distinct developmental phases characterized by the changes in pH of their surroundings from acidic to nearly alkaline and vice versa. At the beginning of the alkali phase colonies start to produce ammonia, which functions as a quorum-sensing molecule inducing the reprogramming of cell metabolism. Such reprogramming includes, among others, the activation of several plasma membrane transporters and is connected with colony differentiation. In the present study, we show that colony cells can use two transport mechanisms to import lactic acid: a ‘saturable’ component of the transport, which requires the presence of a functional Jen1p transporter, and a ‘non-saturable’ component (diffusion) that is independent of Jen1p. During colony development, the efficiency of both transport components changes similarly in central and outer colonial cells. Although the lactate uptake capacity of central cells gradually decreases during colony development, the lactate uptake capacity of outer cells peaks during the alkali phase and is also kept relatively high in the second acidic phase. This lactate uptake profile correlates with the localization of the Jen1p transporter to the plasma membrane of colony cells. Both lactic acid uptake mechanisms are diminished in sok2 colonies where JEN1 expression is decreased. The Sok2p transcription factor may therefore be involved in the regulation of non-saturable lactic acid uptake in yeast colonies.