Recommendations for the detection of Leptospira in urine by PCR

In the present study PCR was applied to detect leptospires in human urine. Several approaches for sample processing were evaluated to optimize the detection of leptospires in urine mixed with this bacterium. Furthermore, some changes in the composition of the reaction mix were studied. No amplification was observed in acidic urine, therefore neutralization of the sample immediately after collection is strongly recommended. PBS gave better results than Tris or NaOH as neutralizing reagents. Freezing and thawing of samples before processing yielded negative results. Elimination of epithelial cells, leukocytes and crystals by centrifugation at 3,000 rpm at room temperature increased sensitivity. In addition, both the washing step after collecting leptospires by centrifugation and the inclusion of 0.1% bovine serum albumin in the reaction mix minimized the interference of other inhibitory compounds. These modifications were useful to improve the detection of Leptospira in urine by PCR.

Service-oriented infrastructure to support the control, monitoring and management of a shop floor system

Dissertação para obtenção do Grau de Mestre em Engenharia Electrotécnica e de Computadores

Barreiras e facilitadores à implementação de um modelo de gestão de cuidados em Saúde Mental

RESUMO: Enquadramento teórico - Os estudos epidemiológicos demonstram que apesar de todo o progresso científico, muitas pessoas continuam sem acesso aos Serviços de Saúde Mental (SSM) e que, em muitos casos, os cuidados não têm a qualidade suficiente. A experiência de vários países mostra que os processos de implementação de modelos de intervenção terapêutica, como é o da Gestão de Cuidados, são lentos e complexos, não dependendo somente do grau de efectividade ou da complexidade das práticas a implementar. O Modelo de Gestão de Cuidados (MGC), é definido como uma prática baseada na evidência, utilizada para ajudar os doentes nos seus processos de recuperação. As estratégias para implementar práticas baseadas na evidência são críticas para a melhoria dos serviços. Existem, apesar de toda a evidência, muitas barreiras à implementação. Ao constatarmos que as práticas validadas pela ciência estão longe de estar claramente disseminadas nos serviços de saúde mental, fundamentamos a necessidade de utilizar metodologia de implementação que, além da efectividade das práticas, permita uma efectividade da implementação. Para responder às necessidades de formação e no âmbito da implementação do Plano Nacional de Saúde Mental, foram formados, em Portugal, 170 profissionais de saúde mental provenientes de serviços públicos e do sector social, de todas as regiões de Portugal Continental. Considerando que estes profissionais adquiriram competências específicas no MGC, através de um programa de formação nacional idêntico para todos os serviços de saúde mental, investigámos o grau de implementação deste modelo, bem como os facilitadores e as barreiras à sua correcta implementação. Existem vários estudos internacionais sobre as barreiras e os facilitadores à implementação de práticas baseadas na evidência, embora a maior parte desses estudos seja baseado em entrevistas semi-estruturadas a profissionais. Por outro lado, não existem, em Portugal, estudos sobre as barreiras e os facilitadores à implementação de práticas de saúde mental. Objectivos 1. Estimar o grau da implementação do MGC nos serviços de saúde mental portugueses 2. Caracterizar as regiões onde a implementação do MGC tenha ocorrido em maior grau. 3. Identificar os factores facilitadores e as barreiras à implementação do MGC, entre as regiõesde saúde do país. 4. Explorar as relações entre a fidelidade da implementação, as barreiras e os facilitadores da implementação, a cultura organizacional e as características dos serviços de saúde mental. Metodologia Estudo observacional, transversal e descritivo, com características exploratórias. População: profissionais dos serviços de saúde mental públicos e do sector social que frequentaram o Programa Nacional de Formação em Saúde Mental Comunitária no curso “Cuidados Integrados e Recuperação”, da Coordenação Nacional para a Saúde Mental / Ministério da Saúde, entre Outubro de 2008 e Dezembro de 2009, (n=71). Avaliação Fidelidade de implementação do Modelo de Gestão de Cuidados - IMR-S (Illness Management and Recovery Scale); Qualidade das guidelines utilizadas na implementação do Modelo de Gestão de Cuidados - AGREE II-PT (Appraisal of Guidelines, for Research and Evaluation); Avaliação das Barreiras e Facilitadores à implementação do MGC - BaFAI (Barriers and Facilitators Assessment Instrument); Avaliação da Cultura Organizacional dos serviços de saúde mental - CVF-I (Competing Values Framework Instrument). Análise Estatística Para a descrição dos dados foram aplicados métodos de estatística descritiva. Para a comparação de subgrupos foram utilizados os testes de Mann Whitney e Kruskall-Wallis. Para a investigação de associações foram utilizados os métodos de correlação de Spearman e a Regressão Múltipla. O tratamento e análise dos dados foram realizados utilizando o programa estatístico IBM SPSS Statistics® para Mac/Apple® nas versões 19 e 20. Resultados Serviços: A articulação com os cuidados de saúde primários existe na maioria dos serviços (56.34%) e 77.46% dos serviços têm autonomia para definir os cuidados a prestar. A maioria dos serviços (63.38%) realiza duas ou mais reuniões clínicas por mês e a quase totalidade (95.77%) recebe estagiários e/ou internos. A área da investigação tem níveis considerados baixos, quando comparados com outros países da Europa, tanto para a globalidade das áreas de investigação (25.35%), como para as áreas psicossociais (22.54%). Considerando componentes fundamentais para a implementação de modelos de gestão de cuidados, os resultados nacionais indicam que 66.20% dos serviços fazem registos em processo clínico único. As percentagens de utilização de planos individuais de cuidados são globalmente baixas (46.48%). Por seu turno, a utilização de guidelines, nos serviços do país, tem uma percentagem média nacional de 57.75%. Profissionais: São, na sua maioria, do sexo feminino (69.01%), com idades entre os 25 e os 56 anos (média 38.9, ± 7.41). Pertencem, maioritariamente, aos grupos profissionais da enfermagem (23.94%) e da psicologia (49.30%). A formação dos profissionais é de nível superior em todos os grupos, com uma percentagem total de licenciados de 80.3%, tendo os restantes uma formação ao nível do mestrado. Apesar dos valores baixos (17%) de formação prévia em modelos de gestão de cuidados, 39% dos profissionais indicou utilizar algumas vertentes destes modelos na sua prática. Apesar de 97,18% dos profissionais ter participado em dois ou mais encontros científicos, num período de dois anos, apenas 38.03% apresentou alguma comunicação científica no mesmo intervalo. Guideline: Os resultados da avaliação da guideline do MGC indicaram percentagens mais altas, quanto à qualidade do seu desenvolvimento, nos Domínios 1 (Objectivo e finalidade, com 72.2%) e 4 (Clareza de Apresentação, 77.7%). O Domínio 5 (Aplicabilidade) foi pontuado no limite inferior do desenvolvimento com qualidade suficiente (54.1%), ao passo que a guideline obteve uma pontuação negativa nos Domínios 2 (Envolvimento das partes interessadas, com 41.6%) e 3 (Rigor do Desenvolvimento, com 28.1%). Adicionalmente não foi possível às avaliadoras cotar o Domínio 6 (Independência editorial), por ausência de referências neste contexto. A guideline teve uma avaliação global positiva (66%), com recomendação de aceitação com modificações. Cultura Organizacional: O perfil de liderança com maior frequência nos serviços de saúde mental portugueses foi o de Mentor (45.61%). As percentagens mais baixas pertenceram aos perfis Monitor e Inovador (3.51%). Na perspectiva da cultura organizacional dos serviços, apontuação mais alta foi a da Cultura das Relações Humanas (74.07%). A estratégia de liderança, com predomínio em todas as regiões, foi a estratégia de Flexibilidade (66.10%). Os resultados mostram que a única associação positivamente significativa com o grau da implementação do MGC é a do perfil Produtor, com um peso específico de 14.55% na prevalência dos perfis de liderança nos serviços de saúde mental portugueses. Barreiras: As barreiras à implementação da prática do MGC, identificadas pelos profissionais dos serviços de saúde mental, com percentagens mais altas nos totais do país, foram: o tempo (57.7%), o conhecimento sobre o modelo e a motivação (40.8%), a colaboração dos outros profissionais (33.7%), o número de contactos reduzidos com os doentes (35.2%), as insuficiências do ponto de vista dos espaços (70.4%) e dos instrumentos disponíveis (69%) para implementar o MGC. Existiu uma variação entre as regiões de saúde do país. Os resultados mostram que houve uma correlação negativa, de forma significativa, entre a implementação do MGC e as barreiras: da resistência à utilização de protocolos, do formato da prática, da necessidade de mais treino e da não cooperação dos profissionais. Foram encontradas diferenças estatisticamente significativas entre as barreiras à implementação e as características dos serviços, dos profissionais e da cultura organizacional. Implementação: A média nacional da fidelidade de implementação do MGC (41.48) teve valores aproximados aos de estudos similares. Na pontuação por regiões, a implementação com maior fidelidade ocorreu no Alentejo. Se considerarmos a implementação com fidelidade esta ocorreu em 57.75% dos serviços e uma boa implementação em 15.49%. Os métodos de regressão permitiram confirmar a capacidade preditiva das barreiras e da cultura organizacional quanto à fidelidade da implementação do MGC. Discussão: No universo das hipóteses inicialmente colocadas foi possível verificar a variação da implementação do MGC entre as regiões do país. O estudo permitiu, adicionalmente, concluir pela existência de denominadores comuns de maior sucesso da implementação do MGC. Foi ainda possível verificar uma relação significativa, existente entre o grau de implementação e as dimensões das barreiras, a cultura organizacional e os recursos dos SSM (aqui definidos pelas características dos serviços e dos profissionais). De uma forma mais conclusiva podemos afirmar que existem outros factores, que não estão relacionados com a avaliação restrita dos recursos financeiros ou humanos, associados à qualidade da implementação de práticas baseadas na evidência, como o MGC. Exemplo disso são os achados referentes à região de saúde do Alentejo, onde a distância dos grandes centros urbanos e as conhecidas dificuldades de acessibilidade, combinadas com os problemas conhecidos da falta de recursos, não impediram que fosse a região com os valores mais altos da fidelidade de implementação. Conclusões: Foram encontradas inúmeras barreiras à implementação do MGC. Existem barreiras diferentes entre regiões, que resultam das características dos serviços, dos profissionais e da cultura organizacional. Para existir implementação é necessária a consideração de metodologias próprias que vão para além dos tradicionais programas de formação. As práticas baseadas na evidência, amplamente defendidas, exigem implementações baseadas na evidência.-------------ABSTRACT: Introduction - Several epidemiological studies show that, despite all scientific progress, many people still continue to have no access to mental health services and in many situations the quality of care is poor. The experiences of several countries show that progress towards case management implementation is slow and complex, depending not only from the degree of effectiveness or the complexity of the practice. Case management is defined as an evidence-based practice used to help patients in the recovery process. Strategies to implement evidence-based practices are critical to services improvement. There are many barriers to their implementation, despite all available evidence. Realising that practices of proved scientific value are far from being clearly implemented, justifies the need to use implementation methodologies that, beyond practice effectiveness, allow implementation effectiveness. To answer training needs and in the framework of the National Mental Health Plan implementation, 170 mental health (MH) professionals from portuguese public and private sectors were trained. Considering that case management skills were acquired, as a result of this training programme, we decided to study the degree of implementation in the services.Barriers and facilitators to the implementation were studied as well. There are several studies related with barriers and facilitators to the implementation of evidence-based practices, but most of them use semi-structured interviews with professionals. Additionally, there are no studies in Portugal related with barriers and facilitators to the implementation of mental health practices. Objectives1. Estimate the degree of case management implementation in Portuguese MH Services. 2.Describe regions where implementation occurred with higher fidelity degree. 3. Identify barriers and facilitators to case management implementation across country regions. 4. Explore the relationships between implementation, barriers and facilitators, organisational culture and services characteristics. Methodology - Cross sectional, descriptive study. Assessments - Implementation fidelity - IMR-S (Illness Management and Recovery Scale); Guideline quality - AGREE II-PT (Appraisal of Guidelines, for Research and Evaluation); Barriers and facilitators assessment - BaFAI (Barriers and Facilitators Assessment Instrument); Organisational culture assessment - CVF-I (Competing Values Framework Instrument). Statistical analysis - Descriptives and cross-tabs. Subgroups comparison: Mann-Witney and Kruskall-Wallis. Associations between variables were calculated using Spearman correlation's and Multiple Regression. Results - Services: Liaison with primary care is done in most services (56.34%) and 77.46% have autonomy to determine care. Most services have regular clinical meetings and almost all give internship training (95.77%). Research activity is low compared with other European countries, for both general and psychosocial research. Considering key components for the case management implementation, 66.20% of all services use single clinical records. The use of individual care plans is globally low (46.48%) and there is a use of guidelines in 57.75% of services. Human Resources: most are women (69.01%), with age ranging from 25-56 (average 39.9, SD 7.41). The majority are psychologists (49.30%) and nurses (23.94%). All have a university degree, 19.7% have a masters degree and 83% didn’t have any case management training before the above mentioned national training. Despite the low levels of preceding case management training, 39% have used model components in day-to-day practice and although 97.18% of the workforce have attended scientific meetings in the last 2 years, only 38.03% presented communications in the same period. Guideline: Results show that higher scores were obtained in Domain 1. Scope and Purpose (72.2.%),and Domain 4. Clarity of presentation (77.7%). Domain 5. pplicability scored near low boundary (54.1%) and negative scores were found in Domain 2. Stakeholder Involvement (41.6%) and Domain 3. Rigour of Development (28.1%). Global score was 66% and the guideline was recommended with modifications. Organisational Culture: The most frequent leadership profile was the Mentor profile (45.61%). Lower scores belonged to Innovator and Monitor profiles (3.51%). On the organisational culture overall, higher scores were found in the Human Relations culture (74.07%). The higher leadership strategy was the strategy of flexibility (66.10%). The results additionally showed that the only leadership profile associated with case management implementation was the Producer profile, representing 14.55% of all leadership profiles in the country.Barriers: The barriers identified by MH professionals, with high percentages, were: lack of time (57.7%), knowledge and motivation (40.8%), other colleagues cooperation (33.7%), low number of contacts with patients (35.2%), lack of facilities (70.4%) and lack of instruments (69%) to implement case management, varying across regions. Results show that there was a negative correlation between implementation and the following barriers: using protocols, practice format, need for more training and lack of cooperation from colleagues. Additionally, statistical differences were found between barriers to implementation and: services characteristics, workforce characteristics, organisational culture. Implementation: The national average results of case management implementation fidelity was (41.48), close to values found in similar studies. In the regional scores South Region Alentejo had the highest implementation score. If we look at minimum scores to assume implementation fidelity, these occurred in 57.75% of services and a good implementation occurred in 15.49% of these. Regression methods allowed to confirm that implementation score prediction was possible using the combination of barriers and organisational culture scores. Discussion - Considering the initial study hypotheses, it was possible to confirm the variation of case management implementation across country regions. Additionally, we could conclude that common denominators exist when successful implementation occurred. It was possible to observe a significant relationship between implementation degree and the dimensions of barriers, organisational culture and services resources (defined as professionals and services characteristics). In a more conclusive way, we can say that there are factors, other than financial and human resources, that are associated with evidence based practices implementation like case management. An example is the Alentejo region, were the distance from urban centres, and the known difficulties associated with accessibility, plus the lack of financial and human resources, have not impeded the regional higher score on implementation. Conclusions: Case management implementation had several barriers to implementation. There are different barriers across country regions, resulting from organisational culture, services and professionals characteristics. To reach implementation it is necessary to consider specific methodologies that go beyond traditional training programs and evident practices, widely promoted. Evidence-based practices require evidence-based implementations.

Assessment of Notch1 ligands production - key protein targets in breast cancer

Cell-to-cell communication is required for many biological processes in development and adult life. One of the most common systems utilized by a wide range of eukaryotes is the Notch signalling pathway. Four Notch receptors and five ligands have been identified in mammals that interact via their extracellular domains leading to transcription activation. Studies have shown that the Notch ligands expression is undetectable in normal breast tissues, but moderate to high expression has been detected in breast cancer. Thus, any of the Notch1 ligands can be studied as possible therapeutic targets for breast cancer. To study Notch pathway proteins there is the need to obtain stable protein solutions. E. coli is the host of excellence for recombinant proteins for the ease of use, fast growth and high cell densities. However, the expression of mammalian proteins in such systems may overwhelm the bacterial cellular machinery, which does not possess the ability for post-translational modifications, or dedicated compartments for protein synthesis. Mammalian cells are therefore preferred, despite their technical and financial increased demands. We aim to determine the best expression and purification conditions for the different ligand protein constructs, to develop specific function-blocking antibodies using the Phage Display technology. Moreover, we propose to crystallize the Notch1 ligands alone and in complex with the phage display selected antibodies, unveiling molecular details. hJag2DE3 and hDll1DE6 proteins were purified from refolded inclusion bodies or mammalian cell culture supernatants, respectively, and purity was confirmed by SDS-PAGE (>95%). Protein produced in mammalian cells showed to be more stable, apparently with the physiological disulfide pattern, contrary to what was observed in the refolded protein. Several nano-scale crystallization experiments were set up in 96-well plates, but no positive result was obtained. We will continue to pursue for the best expression for the Notch ligand constructs in both expression systems.

Human serum antibody reactivity towards Paracoccidioides brasiliensis antigens treated with sodium metaperiodate

In this study, we evaluated the profile of anti-Paracoccidioides brasiliensis immunoglobulin isotypes in serum from patients with the acute and chronic forms of paracoccidioidomycosis, using the whole Paracoccidioides brasiliensis antigen and the antigen treated with sodium metaperiodate. All the immunoglobulin isotypes present in the serum from patients with the acute and chronic forms of paracoccidioidomycosis presented higher reactivity towards the whole antigen than to the antigen treated with metaperiodate (P < 0.05). The reactivity of IgG and IgM to the antigen treated with metaperiodate was greater in serum from patients with the acute form of the disease (P < 0.05), while IgA was more reactive in serum from patients with the chronic form (P < 0.05). There was greater reactivity of IgG1 and IgG2 to the whole antigen and the antigen treated with metaperiodate in the serum from patients with paracoccidioidomycosis than there was in serum from patients with other parasitic infections (P < 0.05). Furthermore, IgG1 from patients with the acute form recognized the 19kDa, 27kDa and 31kDa antigens in the western blot test. Thus, the results suggest that modifications to the epitopes of Paracoccidioides brasiliensis antigens may help to improve the immunodiagnosis of paracoccidioidomycosis.

Reactive hybrid knowledge bases

Hybrid knowledge bases are knowledge bases that combine ontologies with non-monotonic rules, allowing to join the best of both open world ontologies and close world rules. Ontologies shape a good mechanism to share knowledge on theWeb that can be understood by both humans and machines, on the other hand rules can be used, e.g., to encode legal laws or to do a mapping between sources of information. Taking into account the dynamics present today on the Web, it is important for these hybrid knowledge bases to capture all these dynamics and thus adapt themselves. To achieve that, it is necessary to create mechanisms capable of monitoring the information flow present on theWeb. Up to today, there are no such mechanisms that allow for monitoring events and performing modifications of hybrid knowledge bases autonomously. The goal of this thesis is then to create a system that combine these hybrid knowledge bases with reactive rules, aiming to monitor events and perform actions over a knowledge base. To achieve this goal, a reactive system for the SemanticWeb is be developed in a logic-programming based approach accompanied with a language for heterogeneous rule base evolution having as its basis RIF Production Rule Dialect, which is a standard for exchanging rules over theWeb.

Real-time collaborative editing of OutSystems DSL models

Real-time collaborative editing systems are common nowadays, and their advantages are widely recognized. Examples of such systems include Google Docs, ShareLaTeX, among others. This thesis aims to adopt this paradigm in a software development environment. The OutSystems visual language lends itself very appropriate to this kind of collaboration, since the visual code enables a natural flow of knowledge between developers regarding the developed code. Furthermore, communication and coordination are simplified. This proposal explores the field of collaboration on a very structured and rigid model, where collaboration is made through the copy-modify-merge paradigm, in which a developer gets its own private copy from the shared repository, modifies it in isolation and later uploads his changes to be merged with modifications concurrently produced by other developers. To this end, we designed and implemented an extension to the OutSystems Platform, in order to enable real-time collaborative editing. The solution guarantees consistency among the artefacts distributed across several developers working on the same project. We believe that it is possible to achieve a much more intense collaboration over the same models with a low negative impact on the individual productivity of each developer.

Characterization tests for insulation boards made from corn cob and natural glues

In recent years there has been a growing interest in developing news solutions for more ecologic and efficient construction, including natural, renewable and local materials, thus contributing in the search for more efficient, economic and environmentally friendly construction. Several authors have assessed the possibility of using various agricultural sub products or wastes, as part of the effort of the scientific community to find alternative and more ecologic construction materials. Corn cob is an agricultural waste from a very important worldwide crop. Natural glues are made from natural materials, non-mineral, that can be used as such or after some modifications to achieve the behaviour and performance required. Two examples of these natural glues are casein and wheat flour-based glues that were used in the present study. Boards with different compositions were manufactured, having as variables the type of glue, the dimension of the corn cob particles and the features of the pressing process. The tests boards were characterized with physical and mechanical tests, such as thermal conductivity (λ) with a ISOMET 2104 and 60 mm diameter contact probe, density (ρ) based on EN 1602:2013, surface hardness (SH) with a PCE Shore A durometer, surface resistance (SR) with a PROCEQ PT pendular sclerometer, bending behaviour (σ) based on EN 12089:2013, compression behaviour (σ10) based on EN 826:2013 and resilience (R) based on EN 1094-1:2008, with a Zwick Rowell bending equipment with 2 kN and 50 kN load cells (Fig. 1), dynamic modulus of elasticity (Ed) with a Zeus Resonance Meter equipment (Fig. 5) based on NP EN 14146:2006 and water vapour permeability (δ) based on EN 12086:2013. The various boards produced were characterized according to the tests and the ones with the best results were C8_c8 (casein glue, grain size 2,38-4,76 mm, cold pressing for 8 hours), C8_c4 (casein glue, grain size 2,38-4,76 mm, cold pressing for 4 hours), F8_h0.5 (wheat flour glue, grain size 2,38-4,76 mm, hot pressing for 0,5 hours), FEV8_h0.5 (wheat flour, egg white and vinegar glue, grain size 2,38-4,76 mm, hot pressing for 0,5 hours) and FEVH68_c4 (wheat flour, egg white, vinegar and 6 g of sodium hydroxide glue, grain size 2,38-4,76 mm, cold pressing for 4 hours). Taking into account the various boards produced and respective test results the type of glue and the pressure and pressing time are very important factors which strongly influence the final product. The results obtained confirmed the initial hypotheses that these boards have potential as a thermal and, eventually, acoustic insulation material, to use as coating or intermediate layer on walls, floors or false ceilings. This type of board has a high mechanical resistance when compared with traditional insulating materials.The integrity of these boards seems to be maintained even in higher humidity environments. However, due to biological susceptibility and sensitivity to water, they would be more adequate for application in dry interior conditions.

Caraterização do acesso ao Centro Hospitalar Lisboa Norte, EPE : análise do acesso e da utilização da consulta externa e da urgência do Centro Hospitalar Lisboa Norte, EPE

RESUMO - Introdução: As alterações verificadas no dimensionamento e na redistribuição da rede de prestação de cuidados hospitalares, nomeadamente no que confere à reorganização das áreas de influência, e, cumulativamente, à abertura do Hospital Beatriz Ângelo, tornam evidente a necessidade de se analisar o acesso e a utilização da consulta externa e da urgência, antes e depois destas alterações. Objetivo: Este trabalho de investigação tem como principal objetivo analisar o acesso e a utilização da consulta externa e da urgência do Centro Hospitalar Lisboa Norte e verificar a evolução desse mesmo acesso, no período de 2010 a 2013. Material e Métodos: Estudo descritivo que visa determinar a frequência e a distribuição dos episódios de Consulta Externa e de Urgência do Centro Hospitalar Lisboa Norte. Foram considerados todos os episódios de consulta externa (3785579 episódios) e de urgência (1140052 episódios) ocorridos entre 1 de janeiro de 2010 e 31 de dezembro de 2013. Resultados: Entre 2010 e 2013, verificou-se o aumento de +2,3% no número de primeiras consultas e de +5%, em consultas subsequentes. As especialidades médicas com maior afluência são Oftalmologia, Dermatologia I, Pediatria e Otorrinolaringologia I. Observou-se, no total de consultas, qual a percentagem que, em cada ano, obteve um tempo de espera inferior a 1 mês, entre a marcação e a realização da consulta: 56,7%, em 2010; 54,3%, em 2011; 55% em 2012; 56,2%, em 2013. Verificou-se o decréscimo do número de atendimentos de urgência, no período em análise, de -38,9%. Cerca de 90% dos atendimentos de urgência respeitam a utentes que acorrem diretamente à urgência sem virem referenciados de outras unidades de saúde. Os meses de janeiro e fevereiro são aqueles que registam maior afluência, 9,9% e 8,8%, respetivamente. O período de maior afluência regista-se entre as 9 e as 11 horas, representando 20,7% dos episódios, em 2010, 21,2%, em 2011, 21% e 20,5%, em 2012 e 2013, respetivamente. Conclusão: Face à alteração da área de influência do Centro Hospitalar Lisboa Norte, torna-se necessária a adaptação da capacidade instalada para haja um sobredimensionamento face à procura, bem como a adequação de recursos internos, financeiros, humanos e tecnológicos.

Analyzing Geospatial patterns of syrian refugee flows in southeastern Turkey by use of remote sensing and complementary data

Crisis-affected communities and global organizations for international aid are becoming increasingly digital as consequence geotechnology popularity. Humanitarian sector changed in profound ways by adopting new technical approach to obtain information from area with difficult geographical or political access. Since 2011, turkey is hosting a growing number of Syrian refugees along southeastern region. Turkish policy of hosting them in camps and the difficulty created by governors to international aid group expeditions to get information, made such international organizations to investigate and adopt other approach in order to obtain information needed. They intensified its remote sensing approach. However, the majority of studies used very high-resolution satellite imagery (VHRSI). The study area is extensive and the temporal resolution of VHRSI is low, besides it is infeasible only using these sensors as unique approach for the whole area. The focus of this research, aims to investigate the potentialities of mid-resolution imagery (here only Landsat) to obtain information from region in crisis (here, southeastern Turkey) through a new web-based platform called Google Earth Engine (GEE). Hereby it is also intended to verify GEE currently reliability once the Application Programming Interface (API) is still in beta version. The finds here shows that the basic functions are trustworthy. Results pointed out that Landsat can recognize change in the spectral resolution clearly only for the first settlement. The ongoing modifications vary for each case. Overall, Landsat demonstrated high limitations, but need more investigations and may be used, with restriction, as a support of VHRSI.

Functional genomic analysis of heat stress in Vitis vinifera

Grapevine (Vitis vinifera) is one of most agro-economically important fruit crops worldwide, with a special relevance in Portugal where over 300 varieties are used for wine production. Due to global warming, temperature stress is currently a serious issue affecting crop production especially in temperate climates. Mobile genetic elements such as retrotransposons have been shown to be involved in environmental stress induced genetic and epigenetic modifications. In this study, sequences related to Grapevine Retrotransposon 1 (Gret1) were utilized to determine heat induced genomic and transcriptomic modifications in Touriga Nacional, a traditional Portuguese grapevine variety. For this purpose, growing canes were treated to 42 oC for four hours and leaf genomic DNA and RNA was utilized for various techniques to observe possible genomic alterations and variation in transcription levels of coding and non-coding sequences between non-treated plants and treated plants immediately after heat stress (HS-0 h) or after a 24 hour recovery period (HS-24 h). Heat stress was found to induce a significant decrease in Gret1 related sequences in HS-24 h leaves, indicating an effect of heat stress on genomic structure. In order to identify putative heat induced DNA modifications, genome wide approaches such as Amplified Fragment Length Polymorphism were utilized. This resulted in the identification of a polymorphic DNA fragment in HS-0 h and HS-24 h leaves whose sequence mapped to a genomic region flanking a house keeping gene (NADH) that is represented in multiple copies in the Vitis vinifera genome. Heat stress was also found to affect the transcript levels of various non-coding and gene coding sequences. Accordingly, quantitative real time PCR results established that Gret1 related sequences are up regulated immediately after heat stress whereas the level of transcript of genes involved in identification and repair of double strand breaks are significantly down regulated in HS-0 h plants. Taken together, the results of this work demonstrated heat stress affects both genomic integrity and transcription levels.

Predicting market direction with hidden Markov models

This paper develops the model of Bicego, Grosso, and Otranto (2008) and applies Hidden Markov Models to predict market direction. The paper draws an analogy between financial markets and speech recognition, seeking inspiration from the latter to solve common issues in quantitative investing. Whereas previous works focus mostly on very complex modifications of the original hidden markov model algorithm, the current paper provides an innovative methodology by drawing inspiration from thoroughly tested, yet simple, speech recognition methodologies. By grouping returns into sequences, Hidden Markov Models can then predict market direction the same way they are used to identify phonemes in speech recognition. The model proves highly successful in identifying market direction but fails to consistently identify whether a trend is in place. All in all, the current paper seeks to bridge the gap between speech recognition and quantitative finance and, even though the model is not fully successful, several refinements are suggested and the room for improvement is significant.

Staying under the radar - the multifunctional LANA Protein

Many viruses have developed numerous strategies to recruit and take advantage of cellular protein degradation pathways to evade the cellular viral immune system. One such virus is the Kaposi´s Sarcoma associated herpesvirus (KSHV), first discovered in Kaposi´s Sarcoma lesions found in AIDS patients. Latency-Associated Nuclear Antigen (LANA) is a KSHV multifunctional protein responsible for tethering viral DNA to the chromosome ensuring maintenance and segregation of the viral genome during cell division. Besides its main role of viral maintenance, LANA also physically interacts with several host proteins to modulate cell functions. One such function is to recruit the EC5S ubiquitin-ligase complex by interacting with Elongin BC complex and Cullin 5 protein, which in turn ubiquitinate substrates such as NF-κB and p53 to allow persistent viral infection. Like any other post-translation modifications, ubiquitination is reversible through deubiquitination enzymes (DUBs). LANA also interacts with ubiquitin specific protease 7 (USP7), a deubiquitination enzyme involved in regulation of several proteins including p53. Interaction with USP7 is made through a conserved peptide motif, which is also present in LANA. This work addresses the role of LANA in the recruitment and modulation of the ubiquitination and deubiquitination pathways. Despite the continued efforts in uncovering new LANA interacting partners to form a functional EC5S ubiquitin-ligase complex, only MHV-68 LANA interacted directly with Elongin BC, other interactions were not direct and may require a linker protein. On the other hand, LANA interaction with USP7 was able to be analysed by X-ray structure determination. In addition to a conserved P/AxxS motif, a novel Glutamine (Gln) residue from KSHV LANA was shown to make a specific interaction with USP7. This Gln residue is also present in other herpesvirus protein and hence it might be a conserved motif within herpesviruses.

HIV-1: maternal prognosis

Profound modifications in the profile of patients are currently being observed within the epidemic context of AIDS, especially with respect to pauperization and feminization of the disease. The population most frequently affected is in the reproductive age, and among adults aged 18 to 24 years, the ratio is 1 man to 1 woman, a phenomenon occurring uniformly all over the world. One of the main challenges for HIV-1-infected pregnant women and their doctors is the effect of the interaction between HIV infection and pregnancy. The present article is a review of the literature; and its objective is to assess the influence of HIV-1 infection seen from the maternal perspective, with a discussion of immunologic function, maternal prognosis, and the HIV-abortion interface. At present, we cannot conclude that pregnancy has a short-term effect on the evolution of HIV infection, but the concomitance of HIV and pregnancy may adversely affect the prognosis of gestation, especially in view of its frequent association with increased abortion and puerperal morbidity rates.