Development of triplet repeat primed PCR (TP-PCR) technique as a support of molecular test in Machado-Joseph disease

Dissertação de Mestrado, Ciências Biomédicas, 3 de Fevereiro de 2016, Universidade dos Açores.

Amplificação em imunohistoquímica: estudo comparativo de sistemas de polímeros

Desde o início da utilização da imunohistoquímica em anatomia patológica, um dos objetivos tem sido detetar as quantidades mais ínfimas de antigénio, tornando-o visível ao microscópio ótico. Vários sistemas de amplificação têm sido aplicados de forma a concretizar este objetivo, tendo surgido um grupo genérico de métodos simples e que apresentam uma amplificação superior: são os denominados métodos do polímero indireto. Tendo em conta a variedade de métodos disponíveis, o autor propõe-se a comparar a qualidade de quatro sistemas de amplificação, que recorrem ao método do polímero indireto com horseradish peroxidase (HRP). Foram utilizadas lâminas de diferentes tecidos, fixados em formol e incluídos em parafina, nos quais se procedeu à identificação de 15 antigénios distintos. Na amplificação recorreu-se a quatro sistemas de polímero indireto (Dako EnVision+ System – K4006; LabVision UltraVision LP Detection System – TL-004-HD; Leica NovoLink – RE7140-k; Vector ImmPRESS Reagent Kit – MP-7402). A observação microscópica e classificação da imunomarcação obtida foram feitas com base num algoritmo que enquadra intensidade, marcação específica, marcação inespecífica e contraste, num score global que pode tomar valores entre 0 e 25. No tratamento dos dados, para além da estatística descritiva, foi utilizado o teste one-way ANOVA com posthoc de tukey (alfa=0.05). O melhor resultado obtido, em termos de par média/desvio-padrão, dos scores globais foi o do NovoLink (22,4/2,37) e o pior foi o do EnVision+ (17,43/3,86). Verificou-se ainda que existe diferença estatística entre os resultados obtidos pelo sistema NovoLink e os sistemas UltraVision (p=.004), ImmPRESS (p=.000) e EnVision+ (p=.000). Concluiu-se que o sistema que permitiu a obtenção de melhores resultados, neste estudo, foi o Leica NovoLink.

Using test infrastructures for (remote) online evaluation of the sensitivity to SEUs of FPGAs

This paper proposes an online mechanism that can evaluate the sensitivity of single event upsets (SEUs) of field programmable gate arrays (FPGAs). The online detection mechanism cyclically reads and compares the values form the external and internal configuration memories, taking into account the mask information. This remote detection method also signals any mismatch as a result of a SEU that affects both used and not-used FPGA parts, which maximizes the monitored area. By utilizing an external, Web-accessible controller that is connected to the test infrastructure, the possibility of running the same operation in a remote manner is enabled. Moreover, the need for a local memory to store the mask values is also eliminated.

From design-for-test to design-for-debug-and-test: analysis of requirements and limitations for 1149.1

The increasing complexity of VLSI circuits and the reduced accessibility of modern packaging and mounting technologies restrict the usefulness of conventional in-circuit debugging tools, such as in-circuit emulators for microprocessors and microcontrollers. However, this same trend enables the development of more complex products, which in turn require more powerful debugging tools. These conflicting demands could be met if the standard scan test infrastructures now common in most complex components were able to match the debugging requirements of design verification and prototype validation. This paper analyses the main debug requirements in the design of microprocessor-based applications and the feasibility of their implementation using the mandatory, optional and additional operating modes of the standard IEEE 1149.1 test infrastructure.

Mode III interlaminar fracture of carbon/epoxy laminates using the edge crack torsion (ECT) test

The mode III interlaminar fracture of carbon/epoxy laminates was evaluated with the edge crack torsion (ECT) test. Three-dimensional finite element analyses were performed in order to select two specimen geometries and an experimental data reduction scheme. Test results showed considerable non-linearity before the maximum load point and a significant R-curve effect. These features prevented an accurate definition of the initiation point. Nevertheless, analyses of non-linearity zones showed two likely initiation points corresponding to GIIIc values between 850 and 1100 J/m2 for both specimen geometries. Although any of these values is realistic, the range is too broad, thus showing the limitations of the ECT test and the need for further research.

Development of paper-based color test-strip for drug detection in aquatic environment: Application to oxytetracycline

The wide use of antibiotics in aquaculture has led to the emergence of resistant microbial species. It should be avoided/minimized by controlling the amount of drug employed in fish farming. For this purpose, the present work proposes test-strip papers aiming at the detection/semi-quantitative determination of organic drugs by visual comparison of color changes, in a similar analytical procedure to that of pH monitoring by universal pH paper. This is done by establishing suitable chemical changes upon cellulose, attributing the paper the ability to react with the organic drug and to produce a color change. Quantitative data is also enabled by taking a picture and applying a suitable mathematical treatment to the color coordinates given by the HSL system used by windows. As proof of concept, this approach was applied to oxytetracycline (OXY), one of the antibiotics frequently used in aquaculture. A bottom-up modification of paper was established, starting by the reaction of the glucose moieties on the paper with 3-triethoxysilylpropylamine (APTES). The so-formed amine layer allowed binding to a metal ion by coordination chemistry, while the metal ion reacted after with the drug to produce a colored compound. The most suitable metals to carry out such modification were selected by bulk studies, and the several stages of the paper modification were optimized to produce an intense color change against the concentration of the drug. The paper strips were applied to the analysis of spiked environmental water, allowing a quantitative determination for OXY concentrations as low as 30 ng/mL. In general, this work provided a simple, method to screen and discriminate tetracycline drugs, in aquaculture, being a promising tool for local, quick and cheap monitoring of drugs.

TEC4SEA - A Modular Platform for Research, Test and Validation of Technologies Supporting a Sustainable Blue Economy

This paper presents the TEC4SEA research infrastructure created in Portugal to support research, development, and validation of marine technologies. It is a multidisciplinary open platform, capable of supporting research, development, and test of marine robotics, telecommunications, and sensing technologies for monitoring and operating in the ocean environment. Due to the installed research facilities and its privileged geographic location, it allows fast access to deep sea, and can support multidisciplinary research, enabling full validation and evaluation of technological solutions designed for the ocean environment. It is a vertically integrated infrastructure, in the sense that it possesses a set of skills and resources which range from pure conceptual research to field deployment missions, with strong industrial and logistic capacities in the middle tier of prototype production. TEC4SEA is open to the entire scientific and enterprise community, with a free access policy for researchers affiliated with the research units that ensure its maintenance and sustainability. The paper describes the infrastructure in detail, and discusses associated research programs, providing a strategic vision for deep sea research initiatives, within the context of both the Portuguese National Ocean Strategy and European Strategy frameworks.

An Exact Schedulability Test for Global FP Using State Space Pruning

Presented at 23rd International Conference on Real-Time Networks and Systems (RTNS 2015). 4 to 6, Nov, 2015, Main Track. Lille, France.

Adaptação Cultural e Linguística do Trunk Control Test: avaliação das propriedades psicométricas

Introdução: O controlo postural do tronco é um fator preditivo de autonomia, sendo fundamental a existência de instrumentos válidos e fiáveis a fim da sua avaliação na população portuguesa. Objetivo: Traduzir e adaptar o Trunk Control Test (TCT) para a população portuguesa em indivíduos após AVE e avaliar as suas propriedades psicométricas. Métodos: O TCT foi sujeito aos processos de tradução e retroversão para a população portuguesa por dois tradutores bilingues e realizadas duas reuniões com painel de peritos na área. Avaliou-se a validade, a fiabilidade, a sensibilidade, a especificidade e o poder de resposta em 19 indivíduos com AVE. Para avaliar a validade de critério os indivíduos foram adicionalmente submetidos à Escala de Equilíbrio de Berg (EEB), à Avaliação Motora de Rivermead (AMR) e à Escala de Comprometimento do Tronco (ECT). A fiabilidade inter-observadores foi garantida por uma segunda amostra de 25 fisioterapeutas, através da avaliação do desempenho de um participante no TCT. Os dados foram analisados no programa SPSS 22.0. Resultados: O TCT apresentou baixa consistência interna ( =0,523) e fiabilidade inter-observadores substancial (k=0,662). Obteve-se forte correlação do TCT com a ECT (r=0,885) e AMR (r=0,864), e correlação moderada com a EEB (r=0,700). A validade de construção aponta para uma moderada correlação entre itens (KMO=0,755; Bartlett=0,001). Não foi possível obter os valores de sensibilidade, especificidade e poder de resposta do TCT. Conclusão: O estudo demonstrou que o TCT é um instrumento válido e fiável na avaliação da população portuguesa após AVE.

Effects of quantitative easing on the USA economy: A test for policy effectiveness

The catastrophic disruption in the USA financial system in the wake of the financial crisis prompted the Federal Reserve to launch a Quantitative Easing (QE) programme in late 2008. In line with Pesaran and Smith (2014), I use a policy effectiveness test to assess whether this massive asset purchase programme was effective in stimulating the economic activity in the USA. Specifically, I employ an Autoregressive Distributed Lag Model (ARDL), in order to obtain a counterfactual for the USA real GDP growth rate. Using data from 1983Q1 to 2009Q4, the results show that the beneficial effects of QE appear to be weak and rather short-lived. The null hypothesis of policy ineffectiveness is not rejected, which suggests that QE did not have a meaningful impact on output growth.

A practical approach to execute and test a lean startup

This paper practically applies the “Lean Startup Approach” by identifying, analyzing and executing a newly developed web-based business idea. Hypotheses were designed and tested with the construction of a minimum viable product – i.e. a landing page. In-depth interviews allowed deciding either to pivot or persevere the initial launch strategy. Overall, the aim was to collect as much valuable response as possible from customers and ultimately decide for a superior strategy while devoting the smallest amount of time and money.

A Wireless Mesh Network NS-3 Simulation Model: Implementation and Performance Comparison With a Real Test-Bed

Wireless mesh networks present an attractive communication solution for various research and industrial projects. However, in many cases, the appropriate preliminary calculations which allow predicting the network behavior have to be made before the actual deployment. For such purposes, network simulation environments emulating the real network operation are often used. Within this paper, a behavior comparison of real wireless mesh network (based on 802.11s amendment) and the simulated one has been performed. The main objective of this work is to measure performance parameters of a real 802.11s wireless mesh network (average UDP throughput and average one-way delay) and compare the derived results with characteristics of a simulated wireless mesh network created with the NS-3 network simulation tool. Then, the results from both networks are compared and the corresponding conclusion is made. The corresponding results were derived from simulation model and real-worldtest-bed, showing that the behavior of both networks is similar. It confirms that the NS-3 simulation model is accurate and can be used in further research studies.

Octopus Standard Automated Perimetry, Pulsar Perimetry, Moorfields Motion Displacement Test and Heidelberg Retinal Tomography in Glaucoma Detection - A Comparison of Diagnostic Accuracy

Purpose: To investigate the accuracy of 4 clinical instruments in the detection of glaucomatous damage. Methods: 102 eyes of 55 test subjects (Age mean = 66.5yrs, range = [39; 89]) underwent Heidelberg Retinal Tomography (HRTIII), (disc area<2.43); and standard automated perimetry (SAP) using Octopus (Dynamic); Pulsar (TOP); and Moorfields Motion Displacement Test (MDT) (ESTA strategy). Eyes were separated into three groups 1) Healthy (H): IOP<21mmHg and healthy discs (clinical examination), 39 subjects, 78 eyes; 2) Glaucoma suspect (GS): Suspicious discs (clinical examination), 12 subjects, 15 eyes; 3) Glaucoma (G): progressive structural or functional loss, 14 subjects, 20 eyes. Clinical diagnostic precision was examined using the cut-off associated with the p<5% normative limit of MD (Octopus/Pulsar), PTD (MDT) and MRA (HRT) analysis. The sensitivity, specificity and accuracy were calculated for each instrument. Results: See table Conclusions: Despite the advantage of defining glaucoma suspects using clinical optic disc examination, the HRT did not yield significantly higher accuracy than functional measures. HRT, MDT and Octopus SAP yielded higher accuracy than Pulsar perimetry, although results did not reach statistical significance. Further studies are required to investigate the structure-function correlations between these instruments.

Effects of selection and drift on G matrix evolution in a heterogeneous environment: a multivariate Qst-Fst Test with the freshwater snail Galba truncatula.

Unraveling the effect of selection vs. drift on the evolution of quantitative traits is commonly achieved by one of two methods. Either one contrasts population differentiation estimates for genetic markers and quantitative traits (the Q(st)-F(st) contrast) or multivariate methods are used to study the covariance between sets of traits. In particular, many studies have focused on the genetic variance-covariance matrix (the G matrix). However, both drift and selection can cause changes in G. To understand their joint effects, we recently combined the two methods into a single test (accompanying article by Martin et al.), which we apply here to a network of 16 natural populations of the freshwater snail Galba truncatula. Using this new neutrality test, extended to hierarchical population structures, we studied the multivariate equivalent of the Q(st)-F(st) contrast for several life-history traits of G. truncatula. We found strong evidence of selection acting on multivariate phenotypes. Selection was homogeneous among populations within each habitat and heterogeneous between habitats. We found that the G matrices were relatively stable within each habitat, with proportionality between the among-populations (D) and the within-populations (G) covariance matrices. The effect of habitat heterogeneity is to break this proportionality because of selection for habitat-dependent optima. Individual-based simulations mimicking our empirical system confirmed that these patterns are expected under the selective regime inferred. We show that homogenizing selection can mimic some effect of drift on the G matrix (G and D almost proportional), but that incorporating information from molecular markers (multivariate Q(st)-F(st)) allows disentangling the two effects.