Surface plasmon resonance as a tool in the functional analisys of an immunodominant site in foot-and-mouth disease virus

A fast and direct surface plasmon resonance (SPR) method for the kinetic analysis of the interactions between peptide antigens and immobilised monoclonal antibodies (mAb) has been established. Protocols have been developed to overcome the problems posed by the small size of the analytes (< 1600 Da). The interactions were well described by a simple 1:1 bimolecular interaction and the rate constants were self-consistent and reproducible. The key features for the accuracy of the kinetic constants measured were high buffer flow rates, medium antibody surface densities and high peptide concentrations. The method was applied to an extensive analysis of over 40 peptide analogues towards two distinct anti-FMDV antibodies, providing data in total agreement with previous competition ELISA experiments. Eleven linear 15-residue synthetic peptides, reproducing all possible combinations of the four replacements found in foot-and-mouth disease virus (FMDV) field isolate C-S30, were evaluated. The direct kinetic SPR analysis of the interactions between these peptides and three anti-site A mAbs suggested additivity in all combinations of the four relevant mutations, which was confirmed by parallel ELISA analysis. The four-point mutant peptide (A15S30) reproducing site A from the C-S30 strain was the least antigenic of the set, in disagreement with previously reported studies with the virus isolate. Increasing peptide size from 15 to 21 residues did not significantly improve antigenicity. Overnight incubation of A15S30 with mAb 4C4 in solution showed a marked increase in peptide antigenicity not observed for other peptide analogues, suggesting that conformational rearrangement could lead to a stable peptide-antibody complex. In fact, peptide cyclization clearly improved antigenicity, confirming an antigenic reversion in a multiply substituted peptide. Solution NMR studies of both linear and cyclic versions of the antigenic loop of FMDV C-S30 showed that structural features previously correlated with antigenicity were more pronounced in the cyclic peptide. Twenty-six synthetic peptides, corresponding to all possible combinations of five single-point antigenicity-enhancing replacements in the GH loop of FMDV C-S8c1, were also studied. SPR kinetic screening of these peptides was not possible due to problems mainly related to the high mAb affinities displayed by these synthetic antigens. Solution affinity SPR analysis was employed and affinities displayed were generally comparable to or even higher than those corresponding to the C-S8c1 reference peptide A15. The NMR characterisation of one of these multiple mutants in solution showed that it had a conformational behaviour quite similar to that of the native sequence A15 and the X-ray diffraction crystallographic analysis of the peptide ? mAb 4C4 complex showed paratope ? epitope interactions identical to all FMDV peptide ? mAb complexes studied so far. Key residues for these interactions are those directly involved in epitope ? paratope contacts (141Arg, 143Asp, 146His) as well as residues able to stabilise a particular peptide global folding. A quasi-cyclic conformation is held up by a hydrophobic cavity defined by residues 138, 144 and 147 and by other key intrapeptide hydrogen bonds, delineating an open turn at positions 141, 142 and 143 (corresponding to the Arg-Gly-Asp motif).

Interaction between information systems and work in the Brazilian banking sector

This article presents the results of a research to understand the conditions of interaction between work and three specific information systems (ISs) used in the Brazilian banking sector. We sought to understand how systems are redesigned in work practices, and how work is modified by the insertion of new systems. Data gathering included 46 semi-structured interviews, together with an analysis of system-related documents. We tried to identify what is behind the practices that modify the ISs and work. The data analysis revealed an operating structure: a combination of different practices ensuring that the interaction between agents and systems will take place. We discovered a structure of reciprocal conversion caused by the increased technical skills of the agent and the humanization of the systems. It is through ongoing adjustment between work and ISs that technology is tailored to the context and people become more prepared to handle with technology.

Robosonic: Randomness-Based Manipulation of Sounds Assisted by Robots

In this text, we intend to explore the possibilities of sound manipulation in a context of augmented reality (AR) through the use of robots. We use the random behaviour of robots in a limited space for the real-time modulation of two sound characteristics: amplitude and frequency. We add the possibility of interaction with these robots, providing the user the opportunity to manipulate the physical interface by placing markers in the action space, which alter the behaviour of the robots and, consequently, the audible result produced. We intend to demonstrate through the agents, programming of random processes and direct manipulation of this application, that it is possible to generate empathy in interaction and obtain specific audible results, which would be difficult to otherwise reproduce due to the infinite loops that the interaction promotes.

Imagem percebida de Lisboa como destino turístico por parte do mercado finlandês

RESUMO: O presente estudo pretende analisar a imagem percebida de Lisboa como destino turístico por parte do mercado finlandês, considerando as seguintes perguntas de partida: a) quais os elementos que caracterizam a imagem percebida de Lisboa como destino turístico por parte dos turistas finlandeses; b) quais os factores com maior predominância na imagem percebida e c) qual o perfil do turista finlandês que visita Lisboa. Para atingir os objectivos deste estudo, foi realizado um inquérito por questionário a turistas finlandeses em Lisboa, de forma autoministrada, nos meses de Maio, Junho, Setembro e Outubro de 2008, no último dia da sua estada. Para além de questões que visaram compreender a imagem percebida de Lisboa como destino turístico, por parte dos inquiridos, o questionário incluiu perguntas sobre as suas características sócio-demográficas. Os resultados confirmaram as três componentes da imagem propostas no modelo de Echtner e Ritchie: psicológica-funcional, atributos-holística e comum-única. Os mesmos permitem também concluir que os inquiridos valorizam principalmente as características históricas e estéticas da cidade, como também o bom clima e a simpatia da população local. Por sua vez, os aspectos relativos à programação, eventos, diversão nocturna, interacção com a população local e preços, são os factores menos associados à cidade pelos inquiridos. As qualidades e as características únicas de Lisboa foram destacadas, tendo-se também verificado um elevado grau de satisfação com a visita à cidade e um vasto leque de sugestões dos inquiridos para melhorar Lisboa como destino turístico. ABSTRACT: This study aims to analyze the perceived image of Lisbon as a tourist destination by the Finnish market, considering the following questions: a) what are the elements that characterize the perceived image of Lisbon as a tourist destination by Finnish tourists b ) what features/attributes are most predominant in the perceived image c) what is the sociodemographic profile of the Finnish tourist who visits Lisbon. Data was collected in May, June September and October 2008. The questions were used seeking to identify the image of Lisbon of the respondents as a tourist destination, as well as the socio-demographic characteristics of the tourists. The results identified the three components of the image model proposed by Ritchie and Echtner: psychological-functional, attributes-holistic and commonunique. The results also allowed to conclude that respondents identify and value particularly historical and aesthetic characteristics, as well as the good weather and the friendliness of the local people whereas factors of the city such as the events, the nightlife, the interaction with the local population and the price level, didn’t particularly convinced the respondents. The qualities and the unique characteristics of Lisbon were highlighted, having also noticed a high degree of satisfaction on the visit to the city and a wide range of suggestions obtained from respondents to improve Lisbon as a tourist destination. TIIVISTELMÄ: Perehdyin tutkielmassani suomalaisten kokemusperäiseen mielikuvaan Lissabonista matkakohteena. Tutkimuksen tavoitteena oli selvittää, millaisia kokemuksia ja mielikuvia suomalaisilla matkailijoilla on Lissabonista matkakohteena. Tutkimus pyrki vastaamaan kolmeen kysymykseen: a) mitkä tekijät luonnehtivat suomalaisten matkailijoiden kokemusperäistä mielikuvaa Lissabonista matkakohteena, b) mitkä ominaisuudet hallitsevat kyseistä mielikuvaa sekä c) mihin sukupuoli-, ikä-, koulutus- ja yhteiskuntaluokkaan Lissabonissa vierailevat suomalaiset matkailijat kuuluivat. Tiedot kerättiin touko-, kesä-, syys- ja lokakuussa 2008. Kyselyssä keskityttiin pääosin matkailijoiden kokemuksiin ja mielikuviin Lissabonista sekä matkan ominaisuuksiin ja matkailijoiden matkakohdetta koskeviin tietoihin. Lisäksi matkailijoita pyydettiin kirjoittamaan, miten Lissabonin asemaa matkakohteena voitaisiin parantaa. Alkuperäisen olettamuksen mukaan tulokset osoittivat Echtnerin ja Richien ehdottaman mallin mukaisesti mielikuvien muodostuvan kolmesta osatekijästä: toiminnallinen-psykologinen, attribuutti-kokonaisvaltainen ja yleinenainutlaatuinen. Tulokset osoittivat vastaajien yhtyvän mielikuvissaan ensisijaisesti kaupungin toiminnallisiin osatekijöihin. Sen sijaan kaupungin tapahtumiin ja yöelämään tai vuorovaikutukseen paikallisten kanssa taikka hintatasoon liittyviä ominaisuuksia ei koettu erityisen merkittäviksi. Vastauksista tulivat selvästi esille Lissabonin vetovoimatekijät ja ainutlaatuiset ominaisuudet matkakohteena. Lisäksi vastaukset osoittivat matkailijoiden olleen suurimmaksi osaksi erittäin tyytyväisiä vierailuunsa kaupungissa. Vastauksiin sisältyi myös useita ehdotuksia Lissabonin aseman parantamiseksi matkailukohteena.

Studies in search of a suitable experimental insect model for xenodiagnosis of hosts with Chagas' disease: 3 - On the interaction of vector species and parasite strain in the reaction of bugs to infection by Trypanosoma cruzi

The reaction of nine vector species of Chagas' disease to infection by seven different Trypanosoma cruzi strains; Berenice, Y, FL, CL, S. Felipe, Colombiana and Gávea, are examined and compared. On the basis of the insects' ability to establish and maintain the infection, vector species could be divided into two distinct groups which differ in their reaction to an acute infection by T. cruzi. While the proportion of positive bugs was found to be low in Triatoma infestans and Triatoma dimidiata it was high, ranging from 96.9% to 100% in the group of wild (Rhodnius neglectus, Triatoma rubrovaria)and essentially sylvatic vectors in process of adaptation to human dwellings, maintained under control following successful insecticidal elimination of Triatoma infestans (Panstrongylus megistus, Triatoma sordida and Triatoma pseudomaculata). An intermediate position is held by Triatoma brasiliensis and Rhodnius prolixus. This latter has been found to interchange between domestic and sylvatic environments. The most important finding is the strikingly good reaction between each species of the sylvatic bugs and practically all T. cruzi strains herein studied, thus indicating that the factors responsible for the excellent reaction of P.megistus to infection by Y strain, as previously reported also come into operation in the reaction of the same vector species to acute infections by five of the remaining T.cruzi strains. Comparison or data reported by other investigators with those herein described form the basis of the discussion of Dipetalogaster maximus as regards its superiority as a xenodiagnostic agent.

Equilibrium self-assembly of colloids with distinct interaction sites: Thermodynamics, percolation, and cluster distribution functions

We calculate the equilibrium thermodynamic properties, percolation threshold, and cluster distribution functions for a model of associating colloids, which consists of hard spherical particles having on their surfaces three short-ranged attractive sites (sticky spots) of two different types, A and B. The thermodynamic properties are calculated using Wertheim's perturbation theory of associating fluids. This also allows us to find the onset of self-assembly, which can be quantified by the maxima of the specific heat at constant volume. The percolation threshold is derived, under the no-loop assumption, for the correlated bond model: In all cases it is two percolated phases that become identical at a critical point, when one exists. Finally, the cluster size distributions are calculated by mapping the model onto an effective model, characterized by a-state-dependent-functionality (f) over bar and unique bonding probability (p) over bar. The mapping is based on the asymptotic limit of the cluster distributions functions of the generic model and the effective parameters are defined through the requirement that the equilibrium cluster distributions of the true and effective models have the same number-averaged and weight-averaged sizes at all densities and temperatures. We also study the model numerically in the case where BB interactions are missing. In this limit, AB bonds either provide branching between A-chains (Y-junctions) if epsilon(AB)/epsilon(AA) is small, or drive the formation of a hyperbranched polymer if epsilon(AB)/epsilon(AA) is large. We find that the theoretical predictions describe quite accurately the numerical data, especially in the region where Y-junctions are present. There is fairly good agreement between theoretical and numerical results both for the thermodynamic (number of bonds and phase coexistence) and the connectivity properties of the model (cluster size distributions and percolation locus).

Interaction of formaldehyde and tobacco smoking in the frequency of micronucleus and the XRCC3 Thr241Met polymorphism

Formaldehyde is classified by IARC as carcinogenic to humans (nasopharyngeal cancer). Tobacco smoke has been epidemiologically associated to a higher risk of development of cancer, especially in the oral cavity, larynx and lungs, as these are places of direct contact with many carcinogenic tobacco’s compounds. XRCC3 is involved in homologous recombination repair of cross-links and chromosomal double-strand breaks (Thr241Met polymorphism). The aim of the study is to determine whether there is an in vivo association between genetic polymorphism of the gene XRCC3 and the frequency of genotoxicity biomarkers in subjects exposed or not to formaldehyde and with or without tobacco consumption.

Percolation of colloids with distinct interaction sites

We generalize the Flory-Stockmayer theory of percolation to a model of associating (patchy) colloids, which consists of hard spherical particles, having on their surfaces f short-ranged-attractive sites of m different types. These sites can form bonds between particles and thus promote self-assembly. It is shown that the percolation threshold is given in terms of the eigenvalues of a m x m matrix, which describes the recursive relations for the number of bonded particles on the ith level of a cluster with no loops; percolation occurs when the largest of these eigenvalues equals unity. Expressions for the probability that a particle is not bonded to the giant cluster, for the average cluster size and the average size of a cluster to which a randomly chosen particle belongs, are also derived. Explicit results for these quantities are computed for the case f = 3 and m = 2. We show how these structural properties are related to the thermodynamics of the associating system by regarding bond formation as a (equilibrium) chemical reaction. This solution of the percolation problem, combined with Wertheim's thermodynamic first-order perturbation theory, allows the investigation of the interplay between phase behavior and cluster formation for general models of patchy colloids.

Effects of the interaction of tobacco smoke and alcohol consumption on buccal micronucleus in workers exposed occupationally to formaldehyde

Occupational exposure to formaldehyde (FA) has been shown to induce nasopharyngeal cancer and has been classified as carcinogenic to humans (group 1) on the basis of sufficient evidence in humans. Tobacco smoke has been associated to a higher risk of development of cancer, especially in the oral cavity, larynx and lungs, as these are places of direct contact with many carcinogenic tobacco’s compounds. Alcohol is a recognized agent that influence cells in a genotoxic form, been citied as a strong agent with potential in the development of carcinogenic lesions. Epidemiological evidence points to a strong synergistic effect between cigarette smoking and alcohol consumption in the induction of cancers in the oral cavity. Approximately 90% of human cancers originate from epithelial cells. Therefore, it could be argued that oral epithelial cells represent a preferred target site for early genotoxic events induced by carcinogenic agents entering the body via inhalation and ingestion. The MN assay in buccal cells was also used to study cancerous and precancerous lesions and to monitor the effects of a number of chemopreventive agents.

Projecto timecloud: software de gestão de tempo laboral numa plataforma cloud

O presente projecto tem como objectivo a disponibilização de uma plataforma de serviços para gestão e contabilização de tempo remunerável, através da marcação de horas de trabalho, férias e faltas (com ou sem justificação). Pretende-se a disponibilização de relatórios com base nesta informação e a possibilidade de análise automática dos dados, como por exemplo excesso de faltas e férias sobrepostas de trabalhadores. A ênfase do projecto está na disponibilização de uma arquitectura que facilite a inclusão destas funcionalidades. O projecto está implementado sobre a plataforma Google App Engine (i.e. GAE), de forma a disponibilizar uma solução sob o paradigma de Software as a Service, com garantia de disponibilidade e replicação de dados. A plataforma foi escolhida a partir da análise das principais plataformas cloud existentes: Google App Engine, Windows Azure e Amazon Web Services. Foram analisadas as características de cada plataforma, nomeadamente os modelos de programação, os modelos de dados disponibilizados, os serviços existentes e respectivos custos. A escolha da plataforma foi realizada com base nas suas características à data de iniciação do presente projecto. A solução está estruturada em camadas, com as seguintes componentes: interface da plataforma, lógica de negócio e lógica de acesso a dados. A interface disponibilizada está concebida com observação dos princípios arquitecturais REST, suportando dados nos formatos JSON e XML. A esta arquitectura base foi acrescentada uma componente de autorização, suportada em Spring-Security, sendo a autenticação delegada para os serviços Google Acounts. De forma a permitir o desacoplamento entre as várias camadas foi utilizado o padrão Dependency Injection. A utilização deste padrão reduz a dependência das tecnologias utilizadas nas diversas camadas. Foi implementado um protótipo, para a demonstração do trabalho realizado, que permite interagir com as funcionalidades do serviço implementadas, via pedidos AJAX. Neste protótipo tirou-se partido de várias bibliotecas javascript e padrões que simplificaram a sua realização, tal como o model-view-viewmodel através de data binding. Para dar suporte ao desenvolvimento do projecto foi adoptada uma abordagem de desenvolvimento ágil, baseada em Scrum, de forma a implementar os requisitos do sistema, expressos em user stories. De forma a garantir a qualidade da implementação do serviço foram realizados testes unitários, sendo também feita previamente a análise da funcionalidade e posteriormente produzida a documentação recorrendo a diagramas UML.

SIAT:sistema de informações e alertas de trânsito

A evolução tecnológica e das sociedades permitiu que, hoje em dia, uma boa parte da população tenha acesso a dispositivos móveis com funcionalidades avançadas. Com este tipo de dispositivos, temos acesso a inúmeras fontes de informação em tempo-real, mas esta característica ainda não é, hoje em dia, aproveitada na sua totalidade. Este projecto tenta tirar partido desta realidade para, utilizando os diversos dispositivos móveis, criar uma rede de troca de informações de trânsito. O utilizador apenas necessita de servir-se do seu dispositivo móvel para, automaticamente, obter as mais recentes informações de trânsito enquanto, paralelamente, partilha com os outros utilizadores a sua informação. Apesar de existirem outras alternativas no mercado, com soluções que permitem usufruir do mesmo tipo de funcionalidades, nenhuma utiliza este tipo de dispositivos (GPS’s convencionais, por exemplo). Um dos requisitos necessário na implementação deste projecto é uma solução de geocoding. Após terem sido testadas várias soluções, nenhuma cumpria, na totalidade, os requisitos deste projecto, o que originou o desenvolvimento de uma nova solução que cumpre esses requisitos. A solução é, toda ela, muito modular, formada por vários componentes, cada um com responsabilidades bem identificadas. A arquitectura desta solução baseia-se nos padrões de desenvolvimento de uma Service Oriented Architecture. Todos os componentes disponibilizam as suas operações através de web services, e a sua descoberta recorre ao protocolo WS-Discovery. Estes vários componentes podem ser divididos em duas categorias: os do núcleo, responsáveis por criar e oferecer as funcionalidades requisitadas neste projecto e os módulos externos, nos quais se incluem as aplicações que apresentam as funcionalidades ao utilizador. Foram criadas duas formas de consumir a informação oferecida pelo serviço SIAT: a aplicação móvel e um website. No âmbito dos dispositivos móveis, foi desenvolvida uma aplicação para o sistema operativo Windows Phone 7.

Sensor de glicose polarimétrico

Esta tese tem como principal objectivo a investigação teórica e experimental do desempenho de um sensor polarimétrico baseado num cristal líquido para medição da concentração de glicose. Recentemente uma série de sensores polarimétricos baseados em cristais líquidos foram propostos na literatura e receberam considerável interesse devido as suas características únicas. De facto, em comparação com outros moduladores electro-ópticos, o cristal líquido funciona com tensões mais baixas, tem baixo consumo de energia e maior ângulo de rotação. Além disso, este tipo de polarímetro pode ter pequenas dimensões que é uma característica interessante para dispositivos portáteis e compactos. Existem por outro lado algumas desvantagens, nomeadamente o facto do desempenho do polarímetro ser fortemente dependente do tipo de cristal líquido e da tensão a ele aplicada o que coloca desafios na escolha dos parâmetros óptimos de operação. Esta tese descreve o desenvolvimento do sensor polarimétrico, incluindo a integração dos componentes de óptica e electrónica, os algoritmos de processamento de sinal e um interface gráfico que facilita a programação de diversos parâmetros de operação e a calibração do sensor. Após a optimização dos parâmetros de operação verificou-se que o dispositivo mede a concentração da glicose em amostras com uma concentração de 8 mg/ml, com uma percentagem de erro inferior a 6% e um desvio padrão de 0,008o. Os resultados foram obtidos para uma amostra com percurso óptico de apenas 1 cm.

Substrate interaction with recombinant amidase from Pseudomonas aeruginosa during biocatalysis

The interaction of a variety of substrates with Pseudomonas aeruginosa native amidase (E.C. 3.5.1.4), overproduced in an Escherichia coli strain, was investigated using difference FTIR spectroscopy. The amides used as substrates showed an increase in hydrogen bonding upon association in multimers, which was not seen with esters. Evidence for an overall reduction or weakening of hydrogen bonding while amide and ester substrates are interacting with the enzyme is presented. The results describe a spectroscopic approach for analysis of substrate-amidase interaction and in situ monitoring of the hydrolysis and transferase reaction when amides or esters are used as substrates.

Placas de OSB: investigação e estudo laboratorial das suas características de desempenho

A utilização de placas de OSB – Oriented Strand Board está em franca expansão no território nacional e parece ser uma alternativa ecológica e económica aos painéis de derivados de madeira normalmente usados. O OSB é obtido num processo de fabrico resultante da interacção de diversas variáveis, que conferem aos painéis uma vasta gama de aplicações estruturais ou não estruturais (paredes exteriores e interiores, pavimentos, coberturas, componentes de vigas, cofragens, embalagens, móveis, elementos de decoração, etc.). Neste trabalho pretendeu-se investigar, através de pesquisa bibliográfica as características das placas OSB assim como seu processo de fabrico e condicionantes existentes no seu decurso. Em seguida estas foram comparadas com os seus principais concorrentes, o contraplacado e o aglomerado de madeira e cimento. O trabalho experimental realizado teve como objectivo analisar as propriedades de OSB, proveniente do mercado nacional, de acordo com a EN 300:2006 e com os dados recolhidos na pesquisa bibliográfica. Foram executados ensaios de flexão (EN 310:2002), inchamento em espessura (EN 317:2002) e ensaios de flexão após envelhecimento (EN 321:2010). Concluiu-se que o OSB é um material com algumas características de destaque, no entanto a substituição de outros derivados de madeira por este deve ser acompanhada por estudos, principalmente em ambientes húmidos. É importante realçar que este possui modelos com características específicas para determinadas utilizações e ambientes que possibilitam uma optimização do seu comportamento, no entanto, nos ensaios realizados apenas foi utilizado um modelo base de OSB.

Developmental and evolutionary assumptions in a study about the impact of premature birth and low income on mother–infant interaction

In order to study the impact of premature birth and low income on mother–infant interaction, four Portuguese samples were gathered: full-term, middle-class (n=99); premature, middle-class (n=63); full-term, low income (n=22); and premature, low income (n=21). Infants were filmed in a free play situation with their mothers, and the results were scored using the CARE Index. By means of multinomial regression analysis, social economic status (SES) was found to be the best predictor of maternal sensitivity and infant cooperative behavior within a set of medical and social factors. Contrary to the expectations of the cumulative risk perspective, two factors of risk (premature birth together with low SES) were as negative for mother–infant interaction as low SES solely. In this study, as previous studies have shown, maternal sensitivity and infant cooperative behavior were highly correlated, as was maternal control with infant compliance. Our results further indicate that, when maternal lack of responsiveness is high, the infant displays passive behavior, whereas when the maternal lack of responsiveness is medium, the infant displays difficult behavior. Indeed, our findings suggest that, in these cases, the link between types of maternal and infant interactive behavior is more dependent on the degree of maternal lack of responsiveness than it is on birth status or SES. The results will be discussed under a developmental and evolutionary reasoning