Mechanism of interleukin-8 induction by human cytomegalovirus UL76 protein

Dissertation presented to obtain the Ph.D degree in Biology

Spatiotemporal mechanisms for actomyosin ring assembly and contraction in budding yeast cell division

Dissertation presented to obtain the Ph.D degree in Molecular Medicine

Gestão de infra-estruturas e avaliação de equipamentos laboratoriais

RESUMO: A confiabilidade e qualidade dos resultados clínicos de um laboratório, estão dependentes da qualidade do equipamento onde estes são processados. A qualidade de um equipamento consiste em apresentar resultados com uma confiabilidade metrológica de acordo com os processos de medição realizados. Desta forma, os equipamentos de medição devem conter características metrológicas de acordo com os requisitos dos processos que estão inseridos, tais como: erro máximo, incerteza, resolução… Todos os equipamentos por muito bons que sejam, apresentam perdas das suas características metrológicas ao longo do tempo, o que se torna importante a determinação periódica das suas calibrações, manutenções e o seu respectivo ajuste. A periodicidade das calibrações, devem ser baseadas, em critérios bem estabelecidos de modo a evitar a ocorrência da não conformidade, devido às medições com os equipamentos fora das suas especificações, garantindo-se assim, a confiabilidade metrológica e o aumento da produtividade devido ao controlo dos processos, evitando-se desta forma qualquer tipo de avaria por parte destes. Os normativos de Qualidade e Certificação estabelecem como requisito a calibração dos equipamentos de medição em intervalos adequados, mas não definem claramente qual o intervalo adequado. O importante é gerir as etapas de calibração e manutenção de acordo com os equipamentos de modo a garantir que os resultados emitidos por estes sejam mantidos em condições confiáveis, dentro do período entre calibrações e manutenções, de modo a que nãoocorra uma condição de falha, e por conseguinte, medições com grande quantidade de erros.------------ ABSTRACT: The reliability and quality of the results of a clinical laboratory, are dependent on the quality of the equipment where they are processed. The quality of a product is to present results with a metrological reliability according to the measuring processes carried out. Thus, measuring equipment must contain metrological characteristics according to the requirements of the processes that are included, such as: maximum error, uncertainty, resolution ... All equipment for very good they are, show losses of its metrological characteristics over time, it becomes important to determine its periodic calibrations, maintenance and its adjustment. The frequency of calibrations should be based on well-established criteria in order to avoid the occurrence of non-compliance due to the measurements with the equipment beyond its specifications, thus ensuring the metrological reliability and increased productivity due to the control of processes, thus avoiding any kind of damage on the part thereof. The normative and Quality Certification as a requirement to establish calibration of measuring equipment at appropriate intervals, but not clearly stipulate the proper range. It is important to manage the steps of calibration and maintenance equipment according to ensure that the results emitted by these conditions are maintained in trusted within the period between calibrations and maintenance, sothat there occurs a fault condition, and therefore, large amount of measurements with errors.

Plux: Designing a solution for growth in the sports sector

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

Qualidade e satisfação do sistema de seguimento à distância de cardioversores-desfibrilhadores implantáveis

RESUMO - Com o aumento da esperança de vida e das doenças crónicas, cada vez se tem implantado mais cardioversores-desfibrilhadores (CDI) para prevenção de morte súbita. O aumento exponencial no número de implantes aumenta o número de seguimentos dos dispositivos, sobrecarregando os profissionais de saúde e comprometendo a qualidade dos serviços prestados. Segundo as recomendações internacionais, um CDI deverá ser vigiado a cada 3 meses, o que perfaz 4 consultas por ano/doente no mínimo, se não existirem episódios de “choques” ou “descompensações” clínicas, e mensalmente quando é atingido o indicador de substituição electiva do gerador. A evolução da tecnologia, de algoritmos, visualização de episódios e terapias requer recursos técnicos e humanos diferenciados e um gasto de tempo considerável no seguimento. Em países como os Estados Unidos da América, em que os doentes têm de percorrer distâncias muito grandes para aceder aos cuidados de saúde, tornou-se preemente a necessidade de um sistema de vigilância alternativo. Nesse sentido, e usando o conceito da telemedicina, foi criado o seguimento/monitorização à distância de dispositivos cardíacos. Este reduz os custos em consultas, deslocações e recursos humanos, uma vez que contempla apenas uma consulta presencial por ano. Por outro lado, aumenta a segurança do doente com a monitorização periódica e a criação de alarmes, permitindo uma assistência de qualidade e intervenção adequada imediata. Aproveitando as vantagens que este tipo de sistema de transmissão remota oferece, procedeu-se no meu serviço, à distribuição inicial de 62 comunicadores a doentes portadores de CDI´s ou com Ressincronizadores Cardíacos (TRC´s1). Apesar de ser considerada uma melhoria na qualidade dos serviços prestados, é também uma mudança importante na metodologia da consulta feita até aqui. Segundo vários autores, a avaliação da qualidade dos cuidados em saúde está intrinsecamente ligada ao grau de satisfação dos doentes com esses serviços, ou seja, à relação entre as suas expectativas e os resultados percebidos por eles, sendo considerado um importante indicador de qualidade dos serviços. Com este trabalho, pretende-se avaliar a percepção dos doentes face ao novo seguimento em termos de aceitação, satisfação, validade, segurança e confiança no novo sistema. Se este mantém os mesmos padrões de qualidade que o seguimento presencial. Trata-se de um estudo transversal com uma componente retrospectiva de avaliação da nova metodologia de consulta à distância. Para tal, foi elaborado um questionário, que foi aplicado a 40 doentes (17,5% do género feminino e 82,5% do género masculino; média de idades de 65 anos) que constituíram a amostra do estudo. Verificou-se uma média de 5 anos de tempo de implante do CDI. Dos dados obtidos, é de realçar que 70% dos inquiridos estão satisfeitos e 30% estão muito satisfeitos com esta nova metodologia de consulta e cerca 67,4% prefere a consulta à distância. Quando solicitados para comparar a qualidade do serviço prestado entre as duas consultas, 65% respondeu igual e 27,5% melhor. Todos os inquiridos responderam ter confiança e segurança com o sistema de consulta à distância. Cerca de 87,5% dos inquiridos vê-se mesmo a continuar com este tipo de consulta. Os resultados obtidos são bastante satisfatórios no que diz respeito à transição do modo como a consulta de CDI´s é feita. Reflectem também que a tecnologia não é necessariamente uma barreira no acesso aos profissionais de saúde, desde que suportada por algum contacto directo (telefone e através de uma consulta presencial por ano). 1 TRC – Terapia de Ressincronização Cardíaca

Inventory management for the health sector: ABC analysis approach

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

Extracção automática de tópicos de documentos

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para a obtenção do grau de Mestre em Engenharia Informática

Amorphous silicon e 3D sensors applied to object detection

Nowadays, existing 3D scanning cameras and microscopes in the market use digital or discrete sensors, such as CCDs or CMOS for object detection applications. However, these combined systems are not fast enough for some application scenarios since they require large data processing resources and can be cumbersome. Thereby, there is a clear interest in exploring the possibilities and performances of analogue sensors such as arrays of position sensitive detectors with the final goal of integrating them in 3D scanning cameras or microscopes for object detection purposes. The work performed in this thesis deals with the implementation of prototype systems in order to explore the application of object detection using amorphous silicon position sensors of 32 and 128 lines which were produced in the clean room at CENIMAT-CEMOP. During the first phase of this work, the fabrication and the study of the static and dynamic specifications of the sensors as well as their conditioning in relation to the existing scientific and technological knowledge became a starting point. Subsequently, relevant data acquisition and suitable signal processing electronics were assembled. Various prototypes were developed for the 32 and 128 array PSD sensors. Appropriate optical solutions were integrated to work together with the constructed prototypes, allowing the required experiments to be carried out and allowing the achievement of the results presented in this thesis. All control, data acquisition and 3D rendering platform software was implemented for the existing systems. All these components were combined together to form several integrated systems for the 32 and 128 line PSD 3D sensors. The performance of the 32 PSD array sensor and system was evaluated for machine vision applications such as for example 3D object rendering as well as for microscopy applications such as for example micro object movement detection. Trials were also performed involving the 128 array PSD sensor systems. Sensor channel non-linearities of approximately 4 to 7% were obtained. Overall results obtained show the possibility of using a linear array of 32/128 1D line sensors based on the amorphous silicon technology to render 3D profiles of objects. The system and setup presented allows 3D rendering at high speeds and at high frame rates. The minimum detail or gap that can be detected by the sensor system is approximately 350 μm when using this current setup. It is also possible to render an object in 3D within a scanning angle range of 15º to 85º and identify its real height as a function of the scanning angle and the image displacement distance on the sensor. Simple and not so simple objects, such as a rubber and a plastic fork, can be rendered in 3D properly and accurately also at high resolution, using this sensor and system platform. The nip structure sensor system can detect primary and even derived colors of objects by a proper adjustment of the integration time of the system and by combining white, red, green and blue (RGB) light sources. A mean colorimetric error of 25.7 was obtained. It is also possible to detect the movement of micrometer objects using the 32 PSD sensor system. This kind of setup offers the possibility to detect if a micro object is moving, what are its dimensions and what is its position in two dimensions, even at high speeds. Results show a non-linearity of about 3% and a spatial resolution of < 2µm.

Creation of a web application for smart park system and parking prediction study for system integration

In the last few years, we have observed an exponential increasing of the information systems, and parking information is one more example of them. The needs of obtaining reliable and updated information of parking slots availability are very important in the goal of traffic reduction. Also parking slot prediction is a new topic that has already started to be applied. San Francisco in America and Santander in Spain are examples of such projects carried out to obtain this kind of information. The aim of this thesis is the study and evaluation of methodologies for parking slot prediction and the integration in a web application, where all kind of users will be able to know the current parking status and also future status according to parking model predictions. The source of the data is ancillary in this work but it needs to be understood anyway to understand the parking behaviour. Actually, there are many modelling techniques used for this purpose such as time series analysis, decision trees, neural networks and clustering. In this work, the author explains the best techniques at this work, analyzes the result and points out the advantages and disadvantages of each one. The model will learn the periodic and seasonal patterns of the parking status behaviour, and with this knowledge it can predict future status values given a date. The data used comes from the Smart Park Ontinyent and it is about parking occupancy status together with timestamps and it is stored in a database. After data acquisition, data analysis and pre-processing was needed for model implementations. The first test done was with the boosting ensemble classifier, employed over a set of decision trees, created with C5.0 algorithm from a set of training samples, to assign a prediction value to each object. In addition to the predictions, this work has got measurements error that indicates the reliability of the outcome predictions being correct. The second test was done using the function fitting seasonal exponential smoothing tbats model. Finally as the last test, it has been tried a model that is actually a combination of the previous two models, just to see the result of this combination. The results were quite good for all of them, having error averages of 6.2, 6.6 and 5.4 in vacancies predictions for the three models respectively. This means from a parking of 47 places a 10% average error in parking slot predictions. This result could be even better with longer data available. In order to make this kind of information visible and reachable from everyone having a device with internet connection, a web application was made for this purpose. Beside the data displaying, this application also offers different functions to improve the task of searching for parking. The new functions, apart from parking prediction, were: - Park distances from user location. It provides all the distances to user current location to the different parks in the city. - Geocoding. The service for matching a literal description or an address to a concrete location. - Geolocation. The service for positioning the user. - Parking list panel. This is not a service neither a function, is just a better visualization and better handling of the information.

Métodos de avaliação do impacto económico dos eventos adversos em meio hospitalar

RESUMO - O sistema de saúde é constantemente sujeito a pressões sendo as mais relevantes a pressão para o aumento da qualidade e a necessidade de contenção de custos. Os Eventos Adversos (EAs) ocorridos em meio hospitalar constituem um sério problema de qualidade na prestação de cuidados de saúde, com consequências clinicas, sociais, económicas e de imagem, que afectam pacientes, profissionais, organizações e o próprio sistema de saúde. Os custos associados à ocorrência de EAs em meio hospitalar, incrementam significativamente os custos hospitalares, representando cerca de um em cada sete dólares gastos no atendimento dos doentes. Só na última década surgiram estudos com o objectivo principal de avaliar esse impacto em meio hospitalar, subsistindo ainda uma grande indefinição quanto às variáveis e métodos a utilizar. O objectivo principal deste trabalho de projecto foi conhecer e caracterizar as diferentes metodologias utilizadas para avaliação dos custos económicos, nomeadamente dos custos directos, relacionados com a ocorrência de eventos adversos em meio hospitalar. Tendo em atenção as dificuldades referidas, utilizou-se como metodologia a revisão narrativa da literatura, complementada com a realização de uma técnica de grupo nominal. Os resultados obtidos foram os seguintes: i) a metodologia utilizada na maioria dos estudos para determinar a frequência, natureza e consequências dos EAs ocorridos em meio hospitalar, utiliza matrizes de base observacional, analítica, com base em estudos de coorte retrospectivo recorrendo aos critérios definidos pelo Harvard Medical Practice Study; ii) a generalidade dos estudos realizados avaliam os custos directos dos EAs em meio hospitalar, iii) verificou-se a existência de uma grande diversidade de métodos para a determinação dos custos associados aos EAs. A generalidade dos estudos determina esse valor com base na contabilização do número de dias adicionais de internamento, resultantes do EA, valorizados com base em custos médios; iv) o grupo de peritos, propôs como metodologia para a determinação do custo associado a cada EA, a utilização de sistemas de custeio por doente; v) propõe-se o desenvolvimento de uma plataforma informática, que permita o cruzamento da informação disponível no registo clinico electrónico do doente com um sistema automático de identificação de EAs, a desenvolver, e com sistemas de custeio por doente, de modo a valorizar os custos por doente e por tipo de EA. A avaliação dos custos directos associados à ocorrência de EAs em contexto hospitalar, pelo impacto económico e social que tem nos doentes e organizações, será seguramente uma das áreas de estudo e investigação futuras, no sentido de melhorar a eficiência do sistema de saúde e a qualidade e segurança dos cuidados prestados aos doentes.

Nanostructuring silicon probes via electrodeposition: characterization of electrode coatings for acute in vivo neural recordings

Understanding how the brain works will require tools capable of measuring neuron elec-trical activity at a network scale. However, considerable progress is still necessary to reliably increase the number of neurons that are recorded and identified simultaneously with existing mi-croelectrode arrays. This project aims to evaluate how different materials can modify the effi-ciency of signal transfer from the neural tissue to the electrode. Therefore, various coating materials (gold, PEDOT, tungsten oxide and carbon nano-tubes) are characterized in terms of their underlying electrochemical processes and recording ef-ficacy. Iridium electrodes (177-706 μm2) are coated using galvanostatic deposition under different charge densities. By performing electrochemical impedance spectroscopy in phosphate buffered saline it is determined that the impedance modulus at 1 kHz depends on the coating material and decreased up to a maximum of two orders of magnitude for PEDOT (from 1 MΩ to 25 kΩ). The electrodes are furthermore characterized by cyclic voltammetry showing that charge storage capacity is im-proved by one order of magnitude reaching a maximum of 84.1 mC/cm2 for the PEDOT: gold nanoparticles composite (38 times the capacity of the pristine). Neural recording of spontaneous activity within the cortex was performed in anesthetized rodents to evaluate electrode coating performance.

Rehabilitation of renders of old buildings in Portugal: survey, supporting methodology proposal and case study

The main purpose of the research is to present a proposal for a methodology to support the rehabilitation project of renders of old buildings in Portugal. To achieve the objective it was considered essential to define the main types of participants and aspects to integrate the proposal. The research methodology consists in an inquiry presented to professional participants in rehabilitation, a market study of materials and products available in Portugal, the design of a methodology proposal and its application to a case study. The inquiry sample totals 24 answers from the targeted professionals. A sequence of relevant supporting procedures consists in the proposal, which aims to provide a supporting methodology to decide and project in this context and also to be tested with its application to the building. This proposal was applied to an old building with load-bearing stone masonry walls and air-lime based renders. It was concluded that the assessment of the building and external renderings’ condition, its diagnosis and of the supporting walls, the definition of intervention, the specification of materials to be used and performance requirements to comply, and also plans for conservation and periodic maintenance, are crucial. From the inquiry, compatibility between materials and complementary roles and points of view of different types of participants in rehabilitation must be highlighted. A proposal for a methodology to support the project could provide useful guidance particularly for architects and construction engineers, and improve the understanding of direct participants on site, therefore contributing for the correct implementation of intervention.

Towards modification of Medicago truncatula epigenome: genome editing with engineered nucleases

Periodic drought is the primary limitation of plant growth and crop yield. The rise of water demand caused by the increase in world population and climate change, leads to one of the biggest challenges of modern agriculture: to increase food and feed production. De novo DNA methylation is a process regulated by small interfering RNA (siRNAs), which play a role in plant response and adaptation to abiotic stress. In the particular case of water deficit, growing evidences suggest a link between the siRNA pathways and drought response in the model legume Medicago truncatula. As a first step to understand the role of DNA methylation under water stress, we have set up several bioinformatics and molecular methodologies allowing the design of Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems and the assembly of TALENs (transcription activator-like effector nucleases), to target both dicer-like 3 (MtDCL3) and RNA-Dependent RNA polymerase (MtRDR2), enzymes of the RNA-directed DNA methylation pathway. TALENs efficiency was evaluated prior to plant transformation by a yeast-based assay using two different strategies to test TALENs activity: Polyacrylamide gel electrophoresis (PAGE) and Single strand conformation polymorphisms (SSCP). In this assay, yeast cells triple transformation emerged as good and rapid alternative to laborious yeast mating strategies. PAGE analysis might be a valuable tool to test TALENs efficacy in vivo if we could increase TALENs activity. SSCP-based approach proved to be ineffective due to the generation of several false positives. TALENs and CRISPR/Cas9 system constructed and designed in this work will in the future certainly enable the successful disruption of DCL3 and RDR2 genes and shed the light on the relationship between plant stress resistance and epigenetic regulation mediated by siRNAs in M.truncatula.

Improving silicon probe performance through layer-by-layer coating

Fully comprehending brain function, as the scale of neural networks, will only be possi-ble with the development of tools by micro and nanofabrication. Regarding specifically silicon microelectrodes arrays, a significant improvement in long-term performance of these implants is essential. This project aims to create a silicon microelectrode coating that provides high-quality electrical recordings, while limiting the inflammatory response of chronic implants. To this purpose, a combined chitosan and gold nanoparticles coating was produced allied with electrodes modification by electrodeposition with PEDOT/PSS in order to reduce the im-pedance at 1kHz. Using a dip-coating mechanism, the silicon probe was coated and then charac-terized both morphologically and electrochemically, with focus on the stability of post-surgery performance in anesthetized rodents. Since not only the inflammatory response analysis is vital, the electrodes recording degradation over time was also studied. The produced film presented a thickness of approximately 50 μm that led to an increase of impedance of less than 20 kΩ in average. On a 3 week chronic implant, the impedance in-crease on the coated probe was of 641 kΩ, compared with 2.4 MΩ obtained for the uncoated probe. The inflammatory response was also significantly reduced due to the biocompatible film as proved by histological tests.

Rehabilitation of renders of old buildings in Portugal: survey, supporting methodology proposal and case studies

The main purpose of the research is to present a proposal for a methodology to support the rehabilitation project of renders of old buildings. To achieve the objective it was considered essential to define the main types of participants and aspects to integrate the proposal. The research methodology consists in an inquiry presented to several professional participants in rehabilitation, a market study of materials and products available in Portugal, the design of a methodology proposal and its application to a case study. The inquiry sample totals 24 answers from the targeted professionals. A sequence of relevant supporting procedures consists in the proposal, which aims to provide a supporting methodology to decide and project in this context and also to be tested with its application to the building. This proposal was applied to an old building with load-bearing stone masonry walls and air-lime based renders. It was concluded that the assessment of the building and external renderings’ condition, its diagnosis and of the supporting walls, the definition of intervention, the specification of materials to be used and performance requirements to comply, and also plans for conservation and periodic maintenance, are crucial. From the inquiry, compatibility between materials and complementary roles and points of view of different types of participants in rehabilitation must be highlighted. A proposal for a methodology to support the project could provide useful guidance particularly for architects and construction engineers, and improve the understanding of direct participants on site, therefore contributing for the correct implementation of the intervention.