Dpws middleware to support agent-based manufacturing control and simulation

Disserta����o apresentada na Faculdade de Ci��ncias e Tecnologia da Universidade Nova de Lisboa para obten����o do grau de Mestre em Engenharia Electrot��cnica e de Computadores

Mediterranean spotted fever and identification of new agents of rickettsioses in Portugal : epidemiological determinants, host and microbial features in portuguese patients

Resumo: a febre botonosa, tamb��m conhecida por febre escaro-nodular (FEN) �� uma doen��a end��mica nos Pa��ses da bacia do Mediterr��neo, ��frica, M��dio Oriente, ��ndia e Paquist��o. O agente etiol��gico respons��vel por esta patologia �� a bact��ria Rickettsia conorii. Contudo, em alguns pa��ses, como Portugal e It��lia, esta patologia �� causada por duas estirpes diferentes: R conorii Malish e R conorii Israeli spotted fever strain. O principal vector e reservat��rio �� o ixod��deo Rhipicephalus sanguineus. Mesmo com uma elevada taxa de subnotifica����o detectada no nosso Pa��s, a taxa incid��ncia da FEN �� de 8.4/105 habitantes (1989-2005), uma das mais altas quando comparada coom a de outros pa��ses da bacia do Mediterr��neo. De todos os distritos portugueses, Bragan��a e Beja s��o aqueles que apresentam as taxas de incid��ncia mais elevadas, 56,8/105 habitantes e 47,4 / 105 habitantes respectivamente. Em Portugal, as altera����es clim��ticas verificadas na ��ltima d��cada, nomeadamente a subida das temperaturas m��dias anuais, parecem ter influenciado o ciclo de vida do vector e a sua din��mica sazonal, permitindo ao R. sanguineus completar mais de um ciclo de vida por ano. Este facto, e a possibilidade deste vector se manter activo noutros meses do ano, nomeadamente nos meses de inverno, tem influenciado consequentemente o padr��o de distribui����o anual dos casos de FEN. A febre escaro-nodular caracteriza-se clinicamente como uma doen��a exantem��tica, com um processo de vasculite generalizado. Apesar de na generalidade ser considerada uma doen��a benigna (quando tratada atempadamente e com terap��utica adequada e espec��fica)e de estarem descritos casos graves em cerca de 5-6% dos doentes, em Portugal essa percentagem aumentou e consequentemente levou a um aumento de casos fatais. Este facto tornou-se mais evidente em 1997, no Hospital Distrital de Beja e no Hospital Garcia de Orta, onde a taxa de letalidade atingiu os 32% e 18% respectivamente.Para al��m dos factores de co-morbilidade encontrados nos doentes mais graves, como diabetes mellitus, ou o atraso na institui����o da terap��utica espec��fica, foi colocada de que a estirpe R. conorii Israel spotted fever strain pudesse ser mais virulenta ou ent��o estivesse associada a diferentes manifesta����es cl��nicas que dificultassem o diagn��stico cl��nico e a institui����o atempada da terap��utica. Houve ainda a necessidade de avaliar alguns par��metros imunol��gicos dos doentes e tentar identificar que factores, nomeadamente que citoquinas, poderiam estar envolvidos na resposta a uma infec����o por R.conorii.Face a estas quest��es foi avaliada e comparada a epidemiologia, manifesta����es cl��nicas e laboratoriais de 140 doentes (71 infectados com R. conorii Malish e 69 infectados com R. conorii Israel spotted fever strain). Concluiu-se que existe uma sobreposi����o de manifesta����es clinicas entre os dois grupos de doentes, mas que a percentagem da escara de inocula����o �� significativamente inferior em doentes infectados com R. conorii Israel spotted fever strain. Dos resultados mais importantes encontrados neste estudo concluiu-se que a estirpe R. conorii Malish e �� demonstrado, pela primeira vez, estatisticamente que o alcoolismo �� um factor de risco para a morte de doentes com FEN. Associadas a factores de um mau progn��sitco da doen��a, est��o as manifesta����es gastrointestinais, que poder��o ser ou n��o reflexo de altera����es do sistema nervoso central, e ainda a altera����o de par��metros laboratoriais como a presen��a de hiperbilirubinemia e aumento dos valores da ureia.A maior parte dos estudos realizados sobre os mecanismos da resposta imunit��ria �� infec����o por R. conorii e as interac����es hospedeiro - agente etiol��gico t��m sido elucidados com base em modelos animais. Poucos estudos t��m sido efectuados em doentes e nenhum estudo pr��vio tinha sido realizado no sentido de avaliar localmente (escara/pele) quais os mediadores ou outras mol��culas envolvidas na resposta imunit��ria ��s rickettsioses. Foi avaliado o n��vel de express��o g��nica de RNA mensageiro (RNAm)de diferentes citoquinas em amostras de pele de doentes com FEN pela t��cnica de PCR em tempo real.Os resultados deste estudo mostraram que, quando comparado com o grupo controlo, os 23 doentes analisados apresentavam n��veis estatisticamente significativos, mais elevados de express��o g��nica de interfer��o (IFN-��, Tumor necrosis factor (TFN-��, interleucina 10 (IL-10, RANTES (regulated by activation, normal T-cell-expressed and secreted chemokine)e indolamina 2-3 desoxigenase (IDO),uma enzima envolvida no controlo e limita����o do crescimento intracelular das rickettsias, atrav��s da degrada����o do triptofano. Seis dos 23 doentes apresentaram ainda niveis de express��o elevados de ��xido n��trico indut��vel (iNOS)que actua como microbicida. Encontrou-se uma correla����o positiva entre a express��o de RNAm de TNF-��, ��, iNOS e IDO e os casos menos graves de FEN sugerindo um tipo de resposta imunit��ria tipo Th1, i.e. com papel protector na resposta �� infec����o.Verificou-se tamb��m que os valores de express��o gen��tica do RNAm de IL-10, estavam inversamente correlacionados com a express��o do RNAm de TNF-�� e IFN-��. Os casos menos graves de FEN parecem assim envolver um balan��o entre a resposta pr��-inflamat��ria e anti-inflamat��ria. J�� os n��veis de express��o g��nica do RNAm de IL-10 estavam inversamente correlacionados com a express��o RNAm de TNF-�� e IFN-��. Os casos menos graves de FEN parecem assim envolver um balan��o entre uma resposta pr��-inflamat��ria e anti-inflamat��ria. J�� os n��veis de express��o RNAm da quimoquina RANTES foram estatisticamente mais elevados em doentes graves.Nesta disserta����o �� ainda descrita uma nova rickettsiose presente em Portugal, causada pela bact��ria R. sibirica mongolitimonae, que foi identificada laboratorialmente por isolamento do agente, e por detec����o do DNA em biopsia de pele. A presen��a deste agente foi ainda corroborada pela detec����o em paralelo do mesmo agente no ixod��deos como R. africae like e em pulgas como R. felis e R.typhi alertam para a possibilidade de exist��ncia de outras rickettsioses que possam estar diagnosticadas em Portugal. Abstract: Mediterranean spotted fever (MSF), a tick-borne disease caused by Rickettsia conorii, is widley distributed in the Old World, being endemic in the southern Europe, Africa, Middle East, India and Pakistan. In Portugal two strains cause disease: R.conorii Malish and R.conorii Israeli spotted fever.Rhipicephalus sanguineus, the brown dog tick, is considered the main vector and reservoir. MSF is characterized by seasonality, and most of cases are encountered in late spring and summer, peaking in July and August. However, CEVDI/INSA laboratory has observed that the incidence of MSF cases has changed during winter season.The increasing annual averages of air temperatures and warmer and drier winters might have influenced the dynamics of the life cycle and activity of R. sanguineus, and indirectley the number MSF cases during the so called MSF off-season.In the period of 1989-2005, the incidence rate of MSF was 8.4/105 inhabitants, one of the highest rates compared with other endemic countries. In the Portugal during the same period, the highest incidence rates were reported in the districts of Bragan��a, with 56.8/105 inhabitants, and Beja, with 47.4/105 inhabitants. Severe cases of MSF are reported in 6% of the patients, but it seems that this pattern of disease in Portugal has been changing.This factor became more evident in 1997, with a reported case fatality rate of 32% and 18% in patients with MSF admited at Beja and Garcia Orta Hospitals, respectively. Although it was found that diabetes mellitus and delay in therapy have been implicated as a risk factor for death, the hypothesis was considered, that the new ISF strain isolated from Portugueses patients in the same year (1997)causes different or atypical clinical conorii Malish strain. The local (skin biopsies) immune response to R. conorii infection was also evaluated.A prospective study was performed to characterized epidemiological, clinical, laboratory features and determined risk factors for a fatal outcome. One hundred forty patients (51% patients were infected with Rickettsia conorii Malish stain and 49% with Israeli spotted fever strain)with diagnosis documented with identification of the causative rickettsial strain were admitted to 13 Portugueses Hospitals during 1994-2006.Comparison of the clinical manifestations of MSF caused by Malish and ISF strains revealed tremendous overlap that would not permit clinical recognition of the strain envolved, but an eschar was observed in a significantly higher percentage of patients with Malish than ISF strain.A fatal outcome was significantly more likely for patients with ISF strain infection meaning that ISF strain was more virulent than Malish strain, and also alcoholism was a host risk factor for a fatal outcome.The pathophysiology of a fatal outcome involved significantly greater incidence of petechial rash, gastrointestinal symptoms, confusion/obtundation, dehydration, tachypnea, hepatomegaly, leukocytosis, coagulopathy, azotemia, hyperbilirubinemia, and elevated hepatic enzymes and creatine kinase. Multivariate analysis revealed that acute renal failure and hyperbilirubinemia were most strong associated with a fatal oucome of infections with both strains.The immune response to R. conorii infection determined with both strains. The immune response to R. conorii infection determined by the expression levels of inflammatory and immune mediators in skin biopsies collected from untreated patients with Mediterranean spotted fever reveal that intralesional expression of mRNA of TNF-��, IFN-��, IL-10, RANTES, and indoleamine-2, 3-dioxygenase (IDO)an enzyme involved in limiting rickettsial growth by tryptophan degradation, were elevated in skin of MSF patients compared to controls. Six patients had elevated levels of inducible nitric oxide synthase (NOS2, a source microbicidal nitric oxide.Positive correlations among TNF-��, IFN-��, NOS2,IDO and mild-to-moderate disease suggested that type 1 polarization plays a protective role. Significantly high levels of intralesional IL-10 were inversely correlated with IFN-�� and TNF-��. The chemokine RANTES was significantly higher in patients with several MSF. It seems that MSF patients with mild-to-moderate disease have a strong and balanced intralesional pro-inflammatory and anti-inflammatory response, while severe disease is associated with higher chemokine expression.Whether these findings are simply a correlate of mild and severe disease or contribute to anti-rickettsial immunity and pathogenesis remains to be determined.In this dissertation is also described a new rickettsiois present in Portugal caused by R.sibirica mongolitimonae strain, identified based on agent isolation and DNA detection by PCR technique in a skin biopsy.The presence of this agent corroborated by its detection also in Rhipicephalus pusillus tick. Also, pathogenic tick and flea-borne rickettsial agents such as R. africae strain detected in Rhipicephalus bursa tick, and R.felis and R.typhi detected in different fleas species raise the alert for the possible existence of other rickettsioses in Portugal that might be underdiagnosed.

Intelligent decision support systems for collaboration in industrial plants

Disserta����o apresentada para obten����o do Grau de Doutor em Sistemas de Informa����o Industriais, Engenharia Electrot��cnica, pela Universidade Nova de Lisboa, Faculdade de Ci��ncias e Tecnologia

Evaluation of Opuntia spp. bioactive products as promising natural chemotherapeutical agents- an in vitro approach

Dissertation to obtain a Master Degree in Biotechnology

Human activity recognition for an intelligent knee orthosis

Disserta����o para obten����o do Grau de Mestre em Engenharia Biom��dica

Lightweight robust behavior industrial agent methodology

Disserta����o para obten����o do Grau de Mestre em Engenharia Electrot��cnica e de Computadores

Internal consultants as positive change agents

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

Opening Pousada da Serra da Estrela in Covilh��: A two-way impact analysis - interacting with private agents

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

Opening Pousada da Serra da Estrela in Covilh��: A two-way impact analysis - Interacting with public agents

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

Novel insights into the action of antimicrobial agents against human pathogens

Dissertation presented to obtain the Ph.D degree in Biochemistry

Applications of the indole scaffold in medicinal chemistry: development of new antioxidants, COX inhibitors and antitubercular agents

Disserta����o para obten����o do Grau de Doutor em Qu��mica, especialidade Qu��mica Org��nica

Phage display as a tool for generation of bio-therapeutic agents for breast cancer

Notch is a conserved signalling pathway, which plays a crucial role in a multiple cellular processes such as stem cell self-renewal, cell division, proliferation and apoptosis. In mammalian, four Notch receptors and five ligands are described, where interaction is achieved through their extracellular domains, leading to a transcription activation of different target genes. Increased expression of Notch ligands has been detected in several types of cancer, including breast cancer suggesting that these proteins represent possible therapeutic targets. The goal of this work was to generate quality protein targets and, by phage display technology, select function-blocking antibodies specific for Notch ligands. Phage display is a powerful technique that allows the generation of highly specific antibodies to be used for therapeutics, and it has also proved to be a reliable approach in identifying and validating new cancer-related targets. Also, we aimed at solving the tri-dimensional structure of the Notch ligands alone and in complex with selected antibodies. In this work, the initial phase focused on the optimization of the expression and purification of a human Delta-like 1 ligand mutant construct (hDLL1-DE3), by refolding from E. coli inclusion bodies. To confirm the biological activity of the produced recombinant protein cellular functional studies were performed, revealing that treatment with hDLL1-DE3 protein led to a modulation of Notch target genes. In a second stage of this study, Antibody fragments (Fabs) specific for hDLL1-DE3 were generated by phage display, using the produced protein as target, in which one good Fab candidate was selected to determine the best expression conditions. In parallel, multiple crystallization conditions were tested with hDLL1-DE3, but so far none led to positive results.

A new credit monitoring framework for agents' networks

The purpose of this work is to develop a practicable approach for Telecom firms to manage the credit risk exposition to their commercial agents��� network. Particularly it will try to approach the problem of credit concession to clients��� from a corporation perspective and explore the particular scenario of agents that are part of the commercial chain of the corporation and therefore are not end-users. The agents��� network that served as a model for the presented study is composed by companies that, at the same time, are both clients and suppliers of the Telecommunication Company. In that sense the credit exposition analysis must took into consideration all financial fluxes, both inbound and outbound. The current strain on the Financial Sector in Portugal, and other peripheral European economies, combined with the high leverage situation of most companies, generates an environment prone to credit default risk. Due to these circumstances managing credit risk exposure is becoming increasingly a critical function for every company Financial Department. The approach designed in the current study combined two traditional risk monitoring tools: credit risk scoring and credit limitation policies. The objective was to design a new credit monitoring framework that is more flexible, uses both external and internal relationship history to assess risk and takes into consideration commercial objectives inside the agents��� network. Although not explored at length, the blueprint of a Credit Governance model was created for implementing the new credit monitoring framework inside the telecom firm. The Telecom Company that served as a model for the present work decided to implement the new Credit Monitoring framework after this was presented to its Executive Commission.