Optimization of the Sybr Green real time PCR for the detection of Human Herpes Virus type 6 (HHV-6)

HHV-6 is the etiological agent of Exanthem subitum which is considered the sixth most frequent disease in infancy. In immuno-compromised hosts, reactivation of latent HHV-6 infection may cause severe acute disease. We developed a Sybr Green Real Time PCR for HHV-6 and compared the results with nested conventional PCR. A 214 pb PCR derived fragment was cloned using pGEM-T easy from Promega system. Subsequently, serial dilutions were made in a pool of negative leucocytes from 10-6 ng/µL (equivalent to 2465.8 molecules/µL) to 10-9 (equivalent to 2.46 molecules/µL). Dilutions of the plasmid were amplified by Sybr Green Real Time PCR, using primers HHV3 (5' TTG TGC GGG TCC GTT CCC ATC ATA 3)'and HHV4 (5' TCG GGA TAG AAA AAC CTA ATC CCT 3') and by conventional nested PCR using primers HHV1 (outer): 5'CAA TGC TTT TCT AGC CGC CTC TTC 3'; HHV2 (outer): 5' ACA TCT ATA ATT TTA GAC GAT CCC 3'; HHV3 (inner) and HHV4 (inner) 3'. The detection threshold was determined by plasmid serial dilutions. Threshold for Sybr Green real time PCR was 24.6 molecules/µL and for the nested PCR was 2.46 molecules/µL. We chose the Real Time PCR for diagnosing and quantifying HHV-6 DNA from samples using the new Sybr Green chemistry due to its sensitivity and lower risk of contamination.

A multi-objective optimization of the active and reactive resource scheduling at a distribution level in a smart grid context

In the traditional paradigm, the large power plants supply the reactive power required at a transmission level and the capacitors and transformer tap changer were also used at a distribution level. However, in a near future will be necessary to schedule both active and reactive power at a distribution level, due to the high number of resources connected in distribution levels. This paper proposes a new multi-objective methodology to deal with the optimal resource scheduling considering the distributed generation, electric vehicles and capacitor banks for the joint active and reactive power scheduling. The proposed methodology considers the minimization of the cost (economic perspective) of all distributed resources, and the minimization of the voltage magnitude difference (technical perspective) in all buses. The Pareto front is determined and a fuzzy-based mechanism is applied to present the best compromise solution. The proposed methodology has been tested in the 33-bus distribution network. The case study shows the results of three different scenarios for the economic, technical, and multi-objective perspectives, and the results demonstrated the importance of incorporating the reactive scheduling in the distribution network using the multi-objective perspective to obtain the best compromise solution for the economic and technical perspectives.

Zero-Time Kidney Histology Predicts Early Graft Function

Our purposes are to determine the impact of histological factors observed in zero-time biopsies on early post transplant kidney allograft function. We specifically want to compare the semi-quantitative Banff Classification of zero time biopsies with quantification of % cortical area fibrosis. Sixty three zero-time deceased donor allograft biopsies were retrospectively semiquantitatively scored using Banff classification. By adding the individual chronic parameters a Banff Chronic Sum (BCS) Score was generated. Percentage of cortical area Picro Sirius Red (%PSR) staining was assessed and calculated with a computer program. A negative linear regression between %PSR/ GFR at 3 year post-transplantation was established (Y=62.08 +-4.6412X; p=0.022). A significant negative correlation between arteriolar hyalinosis (rho=-0.375; p=0.005), chronic interstitial (rho=0.296; p=0.02) , chronic tubular ( rho=0.276; p=0.04) , chronic vascular (rho= -0.360;P=0.007), BCS (rho=-0.413; p=0.002) and GFR at 3 years were found. However, no correlation was found between % PSR, Ci, Ct or BCS. In multivariate linear regression the negative predictive factors of 3 years GFR were: BCS in histological model; donor kidney age, recipient age and black race in clinical model. The BCS seems a good and easy to perform tool, available to every pathologist, with significant predictive short-term value. The %PSR predicts short term kidney function in univariate study and involves extra-routine and expensive-time work. We think that %PSR must be regarded as a research instrument.

On the analysis of data emerging in non-linear and complex systemscomparison of two X-Ray prony spectra

New arguments proving that successive (repeated) measurements have a memory and actually remember each other are presented. The recognition of this peculiarity can change essentially the existing paradigm associated with conventional observation in behavior of different complex systems and lead towards the application of an intermediate model (IM). This IM can provide a very accurate fit of the measured data in terms of the Prony's decomposition. This decomposition, in turn, contains a small set of the fitting parameters relatively to the number of initial data points and allows comparing the measured data in cases where the “best fit” model based on some specific physical principles is absent. As an example, we consider two X-ray diffractometers (defined in paper as A- (“cheap”) and B- (“expensive”) that are used after their proper calibration for the measuring of the same substance (corundum a-Al2O3). The amplitude-frequency response (AFR) obtained in the frame of the Prony's decomposition can be used for comparison of the spectra recorded from (A) and (B) - X-ray diffractometers (XRDs) for calibration and other practical purposes. We prove also that the Fourier decomposition can be adapted to “ideal” experiment without memory while the Prony's decomposition corresponds to real measurement and can be fitted in the frame of the IM in this case. New statistical parameters describing the properties of experimental equipment (irrespective to their internal “filling”) are found. The suggested approach is rather general and can be used for calibration and comparison of different complex dynamical systems in practical purposes.

Contributo para o estudo do motor linear síncrono supercondutor

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

Sistemas de protecção sísmica semi-activos aplicados em estruturas de engenharia civil caso de estudo: amortecedores com fluído magneto-reológico

Dissertação para obtenção do Grau de Mestre em Engenharia Civil – Perfil Estruturas

Non-linear time series models

The last three decades have seen quite dramatic changes the way we modeled time dependent data. Linear processes have been in the center stage in modeling time series. As far as the second order properties are concerned, the theory and the methodology are very adequate.However, there are more and more evidences that linear models are not sufficiently flexible and rich enough for modeling purposes and that failure to account for non-linearities can be very misleading and have undesired consequences.

Towards Human-like Bimanual Movements in Anthropomorphic Robots: A Nonlinear Optimization Approach

Previously we have presented a model for generating human-like arm and hand movements on an unimanual anthropomorphic robot involved in human-robot collaboration tasks. The present paper aims to extend our model in order to address the generation of human-like bimanual movement sequences which are challenged by scenarios cluttered with obstacles. Movement planning involves large scale nonlinear constrained optimization problems which are solved using the IPOPT solver. Simulation studies show that the model generates feasible and realistic hand trajectories for action sequences involving the two hands. The computational costs involved in the planning allow for real-time human robot-interaction. A qualitative analysis reveals that the movements of the robot exhibit basic characteristics of human movements.

Analyzing Multiple Outcomes: Is it Really Worth the use of Multivariate Linear Regression?

In health related research it is common to have multiple outcomes of interest in a single study. These outcomes are often analysed separately, ignoring the correlation between them. One would expect that a multivariate approach would be a more efficient alternative to individual analyses of each outcome. Surprisingly, this is not always the case. In this article we discuss different settings of linear models and compare the multivariate and univariate approaches. We show that for linear regression models, the estimates of the regression parameters associated with covariates that are shared across the outcomes are the same for the multivariate and univariate models while for outcome-specific covariates the multivariate model performs better in terms of efficiency.

Gestão de Componentes em Dinâmico

A crescente evolução das tecnologias de informação e comunicação, aliadas ao desenvolvimento industrial, proporcionou um aumento de competitividade entre as indústrias, originando uma subida do nível da relação qualidade preço. Esta premissa causou uma maior preocupação com a procura contínua pela melhoria dos processos, de forma a aumentar as actividades de valor acrescentado, eliminando todo o tipo de desperdícios. Nesta conjuntura, a Grohe Portugal Componentes Sanitários, Lda propôs uma melhoria no âmbito da gestão de stocks de componentes existentes em dinâmico. Esta acção de melhoria passa pela definição e implementação de um método de gestão destes componentes, acompanhado por um conjunto de regras de identificação de actividades e respectivos intervenientes, por forma a optimizar os meios existentes e evitar a ocorrência de falhas de componentes nas linhas. Trata-se de um método baseado no cálculo das necessidades das linhas, que através da procura média semanal e constituição dos produtos finais define um nível de prioridade entre os componentes, identificando quais os mais requisitados pelas linhas e possibilitando a gestão do dinâmico. Na contínua tentativa de combater possíveis falhas, desenvolveu-se um sistema de gestão do tipo Kanban com a capacidade de gerir o produto semi-acabado para consumo interno. Foram, ainda, criadas melhorias que permitem um acréscimo de eficiência na gestão dos componentes em estante dinâmica, diminuindo o capital imobilizado investido em stocks, levando a um rearranjo de layouts, proporcionando melhores condições de trabalho e optimizando percursos e recursos. Descreve-se detalhadamente o processo de (i) actualização, definição e implementação do método de gestão de componentes em dinâmico, acompanhado pelo respectivo conjunto de regras, (ii) a implementação de um sistema do tipo Kanban orientado às reais preocupações da empresa, (iii) a redefinição de layouts em conformidade com a actualização dos dinâmicos e (iv) a identificação e execução de um conjunto de melhorias. Todas estas actividades acompanhadas pelo impacto financeiro na organização. Por fim, efectua-se o balanço deste projecto e sugerem-se oportunidades de melhoria.

Kalman tracking linear predictor for vowel intelligibility enhancement on european portuguese HMM based speech synthesis

The recent developments on Hidden Markov Models (HMM) based speech synthesis showed that this is a promising technology fully capable of competing with other established techniques. However some issues still lack a solution. Several authors report an over-smoothing phenomenon on both time and frequencies which decreases naturalness and sometimes intelligibility. In this work we present a new vowel intelligibility enhancement algorithm that uses a discrete Kalman filter (DKF) for tracking frame based parameters. The inter-frame correlations are modelled by an autoregressive structure which provides an underlying time frame dependency and can improve time-frequency resolution. The system’s performance has been evaluated using objective and subjective tests and the proposed methodology has led to improved results.

Modelling and optimization of the ion exchange membrane bioreactor for removal of anionic pollutants from drinking water streams

Dissertação para obtenção do Grau de Doutor em Engenharia Química, especialidade de Engenharia Bioquímica

Correlação entre a Extensão Linear do Tronco e o Alinhamento Segmentar do Membro Inferior Contralesional na Posição de Pé em Indivíduos Pós-AVE

O controlo postural (CP) do tronco é um pré-requisito para o movimento tendo como objetivo prevenir e minimizar as perturbações. Um CP adequado pressupõe estabilidade e orientação dos segmentos, sendo necessária uma orientação do tronco com componente de extensão linear para as atividades sentado e pé. Num acidente vascular encefálico (AVE), pode existir comprometimento do CP do tronco quer contralesional (CONTRA) como ipsilesional (IPSI). Neurofisiologicamente, parece haver uma projecção predominantemente ipsilateral para garantir um CP do tronco contralateralmente ao movimento, tornando-se importante perceber a correlação entre componentes do CP do tronco e o alinhamento do membro CONTRA. Contudo, atualmente são poucos os estudos direcionados para a o membro inferior (MI). Assim, uma vez que nesta população, assumir a posição de pé e realizar marcha constitui muitas vezes o seu principal objetivo, considerou-se relevante avaliar a correlação entre a extensão linear do tronco e o alinhamento segmentar do MI CONTRA na posição de pé. Participantes e Métodos: Estudo observacional, transversal, analítico com 11 indivíduos (idade média 70+10 anos, 54,5% homens). Como critérios de inclusão definiram-se AVE isquémico da artéria cerebral média, único e unilateral, em fase crónica. Recorrendo a um software de avaliação postural (SAPo) avaliou-se a extensão linear do tronco (alinhamento dos acrómios, alinhamento das espinhas ilíacas ântero-superiores (EIAS), ângulo formado entre os dois e o alinhamento vertical do tronco) e o alinhamento segmentar do MI (ângulo do joelho e ângulo entre o tronco e MI). Para a análise estatística inferencial utilizou-se o coeficiente de correlação de Pearson entre o ângulo dos acrómios e as EIAS, o alinhamento vertical do tronco e o alinhamento horizontal da pélvis IPSI com as variáveis de alinhamento do MI, para uma significância de 0,05. Resultados: Algumas correlações mostraram ser quase nulas (com valores entre -0,02 a 0,02) ou fracas como o alinhamento horizontal da pélvis IPSI com o ângulo do tronco e MI CONTRA (0,29 com p=0,39). O alinhamento do tronco e o ângulo do joelho apresentou moderada correlação (0,642), estatisticamente significativa (p=0,03). Conclusão: Os resultados deste estudo sugerem a existência de uma correlação positiva entre o alinhamento vertical do tronco e a extensão do joelho do MI CONTRA na posição de pé em indivíduos pós-AVE.

Estudo de avaliação de emissões de nanopartículas na soldadura por fricção linear de ligas de alumínio

Dissertação para obtenção do Grau de Mestre em Engenharia Mecânica