The development of chromatography for the characterization of protein interactions: a personal perspective

This article reviews the progress of a personal endeavour to develop chromatography as a quantitative procedure for the determination of reaction stoichiometries and equilibrium constants governing protein interactions. As well as affording insight into an aspect of chromatography with which many protein chemists are unfamiliar, it shows the way in which minor adaptations of conventional chromatographic practices have rendered the technique one of the most powerful methods available for the characterization of interactions. That pathway towards quantification is followed from the introduction of frontal gel filtration for the study of protein self-association to the characterization of ligand binding by the biosensor variant of quantitative affinity chromatography.

Use of confocal microscopy to follow the development of penetrative hyphae during growth of Rhizopus oligosporus in an artificial solid-state fermentation system

Two methods were compared for determining the concentration of penetrative biomass during growth of Rhizopus oligosporus on an artificial solid substrate consisting of an inert gel and starch as the sole source of carbon and energy. The first method was based on the use of a hand microtome to make sections of approximately 0.2- to 0.4-mm thickness parallel to the substrate surface and the determination of the glucosamine content in each slice. Use of glucosamine measurements to estimate biomass concentrations was shown to be problematic due to the large variations in glucosamine content with mycelial age. The second method was a novel method based on the use of confocal scanning laser microscopy to estimate the fractional volume occupied by the biomass. Although it is not simple to translate fractional volumes into dry weights of hyphae due to the lack of experimentally determined conversion factors, measurement of the fractional volumes in themselves is useful for characterizing fungal penetration into the substrate. Growth of penetrative biomass in the artificial model substrate showed two forms of growth with an indistinct mass in the region close to the substrate surface and a few hyphae penetrating perpendicularly to the surface in regions further away from the substrate surface. The biomass profiles against depth obtained from the confocal microscopy showed two linear regions on log-linear plots, which are possibly related to different oxygen availability at different depths within the substrate. Confocal microscopy has the potential to be a powerful tool in the investigation of fungal growth mechanisms in solid-state fermentation. (C) 2003 Wiley Periodicals, Inc.

Development of a library for clients of ONVIF video cameras: challenges and solutions

The interoperability of IP video equipment is a critical problem for surveillance systems and other video application developers. ONVIF is one of the two specifications addressing the standardization of networked devices interface, and it is based on SOAP. This paper addresses the development of an ONVIF library to develop clients of video cameras. We address the choice of a web services toolkit, and how to use the selected toolkit to develop a basic library. From that, we discuss the implementation of features that ...

DEVELOPMENT OF A MECHATRONIC SYSTEM FOR BEDRIDDEN PEOPLE SUPPORT

This work is developed in the context of Ambient Assisted Living (AAL) and has, as main purpose, the development of a mechatronic system that allows caring of bedridden patients with ongoing medical care terminal (MCT), by a single person. This system allows higher autonomy in domiciliary care, safety, comfort and hygiene of bedridden patients. It contributes to a large increase in their quality of life as well as the ease of monitoring by providers of continuous care, which, in many cases, may be the family itself. The product includes an embedded processing interface for acquiring physiological data to support online monitoring. The development of this project was focused on improving the quality of life, autonomy, participation in social life and reducing healthcare costs in the area AAL. The developed societies currently face severe demographic changes: the world is aging at an unprecedented rate. In 2000, about 420 million people, or about 7 percent of the world population were over 65 years old. In 2050, that number will be near 1500 million people, about 16 percent of the world population. This demographic trend will be accompanied by the increase of people with physical limitations. This will impose new challenges for traditional health systems, not only for Portugal but also for all European countries. There is an urgent need to find solutions to improve the lives of people in their preferred environment by increasing their autonomy, self-confidence and mobility. Therefore, in the case of household scenarios, the provision of effective health services is of fundamental importance to the welfare and economic development of each country. This ongoing project aims to develop a mechatronic system to meet the diverse needs, namely: improving life, health care, safety, comfort, and remote monitoring of bedridden person.

Territorial Identity and Sustainable Development: from Concept to Analysis

A necessidade de (re)afirmar a identidade dos lugares e regiões com o objectivo de aumentar a sua competitividade no mercado global de bens, serviços e ideias tem sido geralmente adoptado nas políticas de desenvolvimento regional e local, mais de um modo retórico do que operacional. Faltam, de facto, instrumentos analíticos adequados para a avaliação da identidade territorial no contexto do nexo local-global. Neste artigo é discutido o enquadramento conceptual e metodológico necessário para o estudo das mudanças nas identidades territoriais e é apresentada evidência empírica dos conhecimentos, atitudes e práticas dos agentes de desenvolvimento local em Portugal.

Competitiveness of tourist destinations: the study of 65 key destinations for the development of regional tourism

Tourism has reached world importance in the economy. The competition in tourism has intensified between destinations, whether the destinations are cities, regions or countries. In this sense, the evaluation of the tourism competitiveness of these destinations may be helpful in planning and prioritizing actions that will benefit the industry. This article discusses the concept of competitiveness by the multidimensional view of performance, efficiency and unit analysis. Using the theoretical framework lifted, this article shows the 'Study on the competitiveness of the 65 destinations inducers of regional tourism development' prepared by the Tourism Ministry, the Brazilian Support Service to Micro and Small Enterprises (Sebrae) and Getulio Vargas Foundation (FGV), its assumptions that guided this study as well its methodological aspects. Based on this methodology, it was carried out a diagnosis of these 65 destinations selected by the Brazilian Ministry of Tourism to be inducers of tourism in their respective regions. The result of competitiveness reached by these 65 inductors destinations is presented in this article, providing a map of the level of competitiveness of tourism in Brazil.

What factors of satisfaction and motivation are affecting the development of the academic career in Portuguese higher education institutions?

An important constituent group and a key resource of higher education institutions (HEIs) is the faculty or academic staff. The centrality of the faculty role makes it a primary sculptor of institutional culture and has implications for the quality of the institution and therefore has a major role in achieving the objectives of the institution. Demand for academic staff in higher education has been increasing and may be expected to continue to increase. Moreover the performance of academic staff as teachers and researchers determines much of the student satisfaction and has an impact on student learning. There are many factors that serve to undermine the commitment of academics to their institutions and careers. Job satisfaction is important in revitalizing staff motivation and in keeping their enthusiasm alive. Well motivated academic staff can, with appropriate support, build a national and international reputation for themselves and the institution in the professional areas, in research and in publishing. This paper aims to identify the issues and their impacts on academic staff job satisfaction and motivation within Portuguese higher education institutions reporting an ongoing study financed by the European Union through the Portuguese Foundation for Science and Technology.

Policies for decentralization and development of health education

How the policy of action and professional standards have influenced the development and decentralization of health education is discussed. It is concluded that a review of policies both of developing as well as of developed countries could help to put some observations into a perspective that is closer to the Brazilian reality.

Investigation on the development of bidding strategies for a wind farm owner

In this paper, the development of bidding strategies is investigated for a wind farm owner. The optimization model is characterized by making the analysis of scenarios. The proposed approach allows evaluating alternative production strategies in order to submit bids to the electricity market with the goal of maximizing profits. The problem is formulated as a linear programming problem. An application to a case study is presented

Development of fire-resistant (B1 & B2) winter 1 KPU foams applied by gun or straw adapter and optimization of a gun applied winter 1 KPU foam

A realização do presente trabalho teve como principais objectivos o desenvolvimento de espumas de poliuretano de um componente com propriedades de resistência à chama superiores (B1 & B2), aplicadas por pistola ou por adaptador/tubo e a optimização de uma espuma de poliuretano de um componente de inverno aplicada por pistola. Todo o trabalho desenvolvido está dividido em dois projectos distintos: i. O primeiro projecto consistiu em desenvolver espumas de um componente com propriedades de resistência à chama (classificadas como B1 e B2 de acordo com a norma alemã DIN 4102), aplicadas por pistola (GWB1 e GWB2) ou por adaptador/tubo (AWB), utilizando polióis poliésteres aromáticos modificados e aditivos retardantes de chama halogenados. Estas espumas deveriam apresentar também propriedades aceitáveis a baixas temperaturas. Após realizar várias formulações foi possível desenvolver uma espuma AWB2 com apenas 3,3% de poliol poliéster no pré-polímero e com propriedades equivalentes às da melhor espuma comercial mesmo a 5/-10 (temperatura da lata/cura da espuma em °C) e também com uma altura de chama de apenas 11 cm. A partir de duas formulações (AWB2) que passaram o Teste B2, foram obtidas também, uma espuma GWB2 e outra GWB1 com propriedades equivalentes às da melhor espuma da concorrência a -10/-10 e a 23/5, respectivamente, embora não tenham sido submetidas ao teste B2 e B1 após as modificações efectuadas. ii. O segundo projecto consistiu em optimizar uma espuma de poliuretano de um componente de inverno aplicada por pistola (GWB3). A espuma inicial tinha problemas de glass bubbles quando esta era dispensada a partir de uma lata cheia, sendo necessário ultrapassar este problema. Este problema foi resolvido diminuindo a razão de GPL/DME através do aumento da percentagem em volume de DME no pré-polímero para 14% no entanto, a estabilidade dimensional piorou um pouco. O reagente FCA 400 foi removido da formulação anterior (6925) numa tentativa de diminuir o custo da espuma, obtendo-se uma espuma aceitável a 23/23 e a 5/5, com uma redução de 4% no custo da produção e com uma redução de 5,5% no custo por litro de espuma dispensada, quando comparada com a sua antecessora. Por último, foi avaliada a influência da concentração de diferentes surfactantes na formulação 6925, verificando-se o melhoramento da estrutura celular da espuma para concentrções mais elevadas de surfactante, sendo este efeito mais notório a temperaturas mais baixas (5/5). Dos surfactantes estudados, o B 8871 mostrou o melhor desempenho a 5/5 com a concentração mais baixa, sendo portanto o melhor surfactante, enquanto o Struksilon 8003 demonstrou ser o menos adequado para esta formulação específica, apresentando piores resultados globais. Pode-se ainda acrescentar que os surfactantes L-5351, L-5352 e B 8526 também não são adequados para esta formulação uma vez que as espumas resultantes apresentam cell collapse, especialmente a 5/5. No caso dos surfactantes L-5351 e L-5352, esta propriedade piora com concentrações mais elevadas. Em cada projecto foram também efectuados testes de benchmark em determinadas espumas comerciais com o principal objectivo de comparar todos os resultados das espumas desenvolvidas, em ambos os projectos, com espumas da concorrência.

The development of a scale to measure concepts of schizophrenia: experience among Brazilian psychiatrists

INTRODUCTION: Among psychiatric disorders schizophrenia is often said to be the condition with the most disputed definition.The Bleulerian and Schneiderian approaches have given rise to diagnostic formulations that have varied with time and place. Controversies over the concept of schizophrenia were examined within European/North American settings in the early 1970s but little has since been reported on the views of psychiatrists in developing countries. In Brazil both concepts are referred to in the literature. A scale was developed to measure adherence to Bleulerian and Schneiderian concepts among psychiatrists working in S. Paulo. METHODOLOGY: A self-reported questionnaire comprising seventeen visual analogue-scale statements related to Bleulerian and Schneiderian definitions of Shizophrenia, plus sociodemographic and training characteristics, was distributed to a non-randomised sample of 150 psychiatrists. The two sub-scales were assessed by psychometric methods for internal consistency, sub-scale structure and test-retest reliability. Items selected according to internal consistency were examined by a two-factor model exploratory factor analysis. Intraclass correlation coefficients described the stability of the scale. RESULTS: Replies were received from 117 psychiatrists (mean age 36 (SD 7.9)), 74% of whom were made and 26% female. The Schneiderian scale showed better overall internal consistency than the Bleulerian scale. Intra-class correlation coefficients for test-retest comparisons were between 0.5 and 0.7 for Schneiderian items and 0.2 and 0.7 for Bleulerian items. There was no negative association between Bleulerian and Schneiderian scale scores, suggesting that respondents may hold both concepts. Place of training was significantly associated with the respondent's opinion; disagreement with a Bleulerian standpoint predominated for those trained at the University of S. Paulo. CONCLUSIONS: The less satisfactory reliability for the Bleulerian sub-scale limits confidence in the whole scale but on the other hand this questionnaire contributes to the understanding of the controversy over Bleulerian and Schneiderian models for conceptualisation of schizophrenia, the former requiring more inference and therefore being prone to unreliability.

Development of a biosensor for urea assay based on amidase inhibition, using an ion-selective electrode

A biosensor for urea has been developed based on the observation that urea is a powerful active-site inhibitor of amidase, which catalyzes the hydrolysis of amides such as acetamide to produce ammonia and the corresponding organic acid. Cell-free extract from Pseudomonas aeruginosa was the source of amidase (acylamide hydrolase, EC 3.5.1.4) which was immobilized on a polyethersulfone membrane in the presence of glutaraldehyde; anion-selective electrode for ammonium ions was used for biosensor development. Analysis of variance was used for optimization of the biosensorresponse and showed that 30 mu L of cell-free extract containing 7.47 mg protein mL(-1), 2 mu L of glutaraldehyde (5%, v/v) and 10 mu L of gelatin (15%, w/v) exhibited the highest response. Optimization of other parameters showed that pH 7.2 and 30 min incubation time were optimum for incubation ofmembranes in urea. The biosensor exhibited a linear response in the range of 4.0-10.0 mu M urea, a detection limit of 2.0 mu M for urea, a response timeof 20 s, a sensitivity of 58.245 % per mu M urea and a storage stability of over 4 months. It was successfully used for quantification of urea in samples such as wine and milk; recovery experiments were carried out which revealed an average substrate recovery of 94.9%. The urea analogs hydroxyurea, methylurea and thiourea inhibited amidase activity by about 90%, 10% and 0%, respectively, compared with urea inhibition.