Biochemical behavior of Trypanosoma cruzi strains isolated from mice submitted to specific chemotherapy

To investigate the influence of chemotherapy on the biochemical beha vior of Trypanosoma cruzi strains, three groups of mice were infected with one of three strains of T. cruzi of different biological and isoenzymic patterns (Peruvian, 21 SF and Colombian strains). Each group was subdivided into subgroups: 1 - treated with nifurtimox; 2 - treated with benznidazole and 3 - untreated infected controls. At the end of treatment, that lasted for 90 days, xenodiagnosis, sub inoculation of blood into new born mice and haemoculture were performed as tests of cure. From the positive tests, 22 samples of T. cruzi were isolated from all subgroups. Electrophoretic analysis of the isoenzymes PGM, GP1, ALAT and AS AT failed to show any difference between parasite strains isolated from treated and untreated mice, which indicates that no detectable clonal selection or parasite genetic markers alterations concerning the isoenzymes analysed have been determined by treatment with drugs of recognized antiparasitic effect, suggesting stability of the phenotypic characteristics of the three biological types of T. cruzi strains.

Evaluation of a direct immunofluorescent antibody (difma) test using Leishmania genus - specific monoclonal antibody in the routine diagnosis of cutaneous leishmaniasis

A direct immunofluorescent antibody (DIFMA) test using a Leishmania genus- specific monoclonal antibody was evaluated in the routine diagnosis of cutaneous leishmaniasis (CL) in Ecuador. This test was compared with the standard diagnostic techniques of scrapings, culture and histology. Diagnostic samples were taken from a total of 90 active dermal ulcers from patients from areas of Ecuador known to be endemic for cutaneous leishmaniasis. DIFMA was positive in all lesions. It was shown to be significantly superior to standard diagnostic methods either alone or in combination. The sensitivity of DIFMA did not diminish with chronicity of lesions. This test proved to be extremely useful in the routine diagnosis of CL because it is highly sensitive, is easy to use and produces rapid results.

Cancer theranostics: multifunctional gold nanoparticles for diagnostics and therapy

Doctorate in Biology, Specialty in Biotechnology

Target-specific multiphysics modeling for thermal medicine applications

Dissertation to obtain the degree of Doctor of Philosophy in Biomedical Engineering

Diagnosis of rubella infection by detecting specific immunoglobulin M antibodies in saliva samples: a clinic-based study in Niterói, RJ, Brazil

This study was designed to investigate whether saliva could be a feasible alternative to serum for the diagnosis of recent rubella infection in a clinic setting. Forty-five paired blood and saliva samples collected 1 to 29 days after onset of illness were tested for specific immunoglobulin (Ig) M by antibody-capture radioimmunoassay (MACRIA). Rubella IgM was detected in all serum samples and in 38 (84.4%) saliva specimens. Forty-six serum and saliva samples from other patients with rash diseases were tested by MACRIA for control purposes and two saliva specimens were reactive. The saliva test had specificity of 96%. These results indicate that salivary IgM detection may be a convenient non-invasive alternative to serum for investigation of recent rubella cases, especially for disease surveillance and control programmes.

Optimization of investment promotion tools for Portugal: Specific recommendations to attract investments from Germany

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

Attracting investments from Germany to Portugal: A specific value-proposition for automotive suppliers

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

Child programming: an adequate domain specific language for programming specific robots

Dissertação para obtenção do Grau de Mestre em Engenharia Informática

Development of population-specific function assessment intrument in Lebanon

RESUMO: Contexto: O funcionamento tem sido reconhecido como um dos principais indicadores de resultados para avaliar se as pessoas beneficiam das intervenções destinadas a melhorar a sua saúde mental. O funcionamento refere-se à forma como um indivíduo consegue responder às suas tarefas e solicitações, dos seus familiares e da sua comunidade, de acordo com os requisitos do local e a cultura em que vive (eg, tarefa de cozinhar e limpar para as mulheres em algumas culturas ). O funcionamento é altamente dependente da cultura - por isso, tem sido recomendado o desenvolvimento de medidas de funcionamento específicas de cada cultura. Desenvolver localmente os instrumentos de medida evita problemas de adequação, associados com a adaptação de instrumentos ocidentais. Embora os instrumentos criados desta forma sejam específicos de um meio cultural, eles são simultaneamente "transculturais", no sentido em que cada um se refere às tarefas mais importantes para a população local . Esta abordagem mostrou-se útil para investigadores e agências de ajuda (eg, ONGs) que trabalham em países não-ocidentais . Este estudo descreve o trabalho da agência International Medical Corps (IMC) na criação e validação de um questionário de funcionamento específico nas dimensões cultura e gênero, no Líbano, destinado a avaliar eventuais melhorias em pessoas que receberam intervenções de para problemas de saúde mental, a nível dos cuidados primários de saúde. Método: O instrumento foi desenvolvido usando um método que é uma alternativa à abordagem existente de adaptação de instrumentos ocidentais a outras culturas e situações; esta abordagem é rápida e exequível, tendo já demonstrado ser útil no desenvolvimento de instrumentos válidos e fidedignos. Inicialmente, foi solicitado que as pessoas identificassem, de uma lista livre, as tarefas mais importantes para cuidar de si próprias, da sua família e da sua comunidade; as tarefas identificadas foram posteriormente usadas como base para um instrumento de avaliação de funcionamento culturalmente válido. A partir daqui, foram desenvolvidos questionários específicos da comunidade em questão, posteriormente testados no terreno nas vertentes da validade (de conteúdo, facial e de constructo) e da fiabilidade (teste-reste e inter-entrevistadores). Resultados. O estudo resultou na criação e validação de um questionário de funcionamento específico de cultura e gênero capaz de medir efectivamente a capacidade de execução de tarefas importantes do quotidiano,como parte da avaliação de resultados levada a cabo por profissionais da CSP previamente treinados na identificação, suporte e encaminhamento de pessoas com problemas de saúde mental no Líbano. Conclusão. Neste trabalho descreve-se o desenvolvimento de um questionário de funcionamento específico de cultura e gênero, orientado para a avaliação de resultados, num contexto mais lato de um sistema abrangente de avaliação e monitorização de um serviço comunitário. --------------ABSTRACT: Background. Functioning has been recognized as one of the most important key outcomes to assess whether people benefit from interventions aimed to improve their mental health. Functioning refers to how well na individual can complete the tasks and demands for themselves, their family, and their community which are required by them depending on the setting and the culture they live in (e.g. task of cooking and cleaning for women in some cultures). Functioning is highly dependent on culture. Therefore, it has been recommended to develop culture-specific measures of function. Developing instruments locally avoids the problems of limited local relevance and appropriateness associate with adapting western instruments. Although each instrument created in this way is culturally bound, they are “cross cultural” in the sense that each refers to the tasks most important to local people. This approach proves useful for both researchers and aid agencies working in non-western countries. This study describes International Medical Corps’ (IMC) work in Lebanon to create and validate a culture and gender specific functioning questionnaire to assess improvements in people who received treatment interventions for mental health problems at the primary health care (PHC) level. Method. The measure was developed using a method that is an alternative to the existing approach of adapting western function instruments to other cultures and situations; an approach which has been demonstrated as rapid, feasible and which can yield valid and reliable instruments. Function was assessed by first asking local people what tasks are important to care for themselves, their family and their community using free listing, then using these tasks as the basis for a culturally valid function assessment instrument. Community specific function questionnaires based on these tasks were then created, and field-tested for validity using content, face and construct validity methods, and also field tested for reliability using inter-rater and test retest reliability methods. Results. The study resulted in the creation and validation of a culture and gender specific functioning questionnaire that would effectively measure the ability to do tasks important to daily existence, as part of assessing client level outcomes where PHC providers were trained in the identification, management and referral of people with mental health problems in Lebanon. Conclusion. The paper describes a successful pilot for developing culture and gender specific functioning questionnaires that evaluate client level outcomes as part of a more comprehensive system for monitoring and evaluation of community based case management supports and services.

Making connections: engineering and language issues in teaching english for specific purposes

Tese apresentada para cumprimento dos requisitos necessários à obtenção do grau de Doutor em Línguas, Literaturas e Culturas

Evaluation of intrathecal synthesis of specific IgG antibodies against Toxoplasma gondii in the diagnosis assessment of presumptive toxoplasma encephalitis in aids patients

The diagnosis of neurotoxoplasmosis in patients with acquired immunodeficiency syndrome is mainly based on tomographic or magnetic resonance findings and on the response to specific treatment. We studied 55 patients with AIDS and neurotoxoplasmosis according to these diagnostic criteria (group 1), 37 patients with AIDS and neurological involvement of other etiology (group 2), and 16 anti-HIV-negative individuals with neurological manifestations (group 3). Serum and cerebrospinal fluid were examined for the presence of anti-T. gondii IgG, by indirect immunofluorescence. In 72 of them, the total amounts of these antibodies were determined in order to assess local production of anti-T. gondii antibodies in the central nervous system and to correlate their titers with infection activity in patients with AIDS and neurotoxoplasmosis. IgG titers > 1/64 in cerebrospinal fluid reached 100% specificity for the diagnosis of neurotoxoplasmosis in AIDS. Evidence of local synthesis of these antibodies was detected in 42.8% of patients of group 1, in 29.1% of patients of group 2 and in no patient of group 3. The test showed 70.8% specificity and therefore was not useful in our study for the differential diagnosis of neurotoxoplasmosis in patients with AIDS.

Development of novel active pharmaceutical ionic liquids and salts based on antibiotics and anti-fungal drugs

Ionic Liquids (ILs) are class of compounds, which have become popular since the mid-1990s. Despite the fact that ILs are defined by one physical property (melting point), many of the potential applications are now related to their biological properties. The use of a drug as a liquid can avoid some problems related to polymorphism which can influence a drug´s solubility and thus its dosages. Also, the arrangement of the anion or cation with a specific drug might be relevant in order to: a) change the correspondent biopharmaceutical drug classification system; b) for the drug formulation process and c) the change the Active Pharmaceutical Ingredients’ (APIs). The main goal of this Thesis is the synthesis and study of physicochemical and biological properties of ILs as APIs from beta-lactam antibiotics (ampicillin, penicillin G and amoxicillin) and from the anti-fungal Amphotericin B. All the APIs used here were neutralized in a buffer appropriate hydroxide cations. The cation hydroxide was obtained on Amberlite resin (in the OH form) in order to exchange halides. The biological studies of these new compounds were made using techniques like the micro dilution and colorimetric methods. Overall a total of 19 new ILs were synthesised (6 ILs based on ampicillin, 4 ILs, based on amoxicillin, 6 ILs based on penicillin G and 4 ILs based on amphotericin B) and characterized by spectroscopic and analytical methods in order to confirm their structure and purity. The study of the biological properties of the synthesised ILs showed that some have antimicrobial activity against bacteria and yeast cells, even in resistant bacteria. Also this work allowed to show that ILs based on ampicillin could be used as anti-tumour agents. This proves that with a careful selection of the organic cation, it is possible to provoke important physico-chemical and biological alteration in the properties of ILs-APIs with great impact, having in mind their applications.

Thomsen-Friedenreich antigens in bladder cancer: evaluation of their prognostic value

RESUMO: Introdução. O cancro de bexiga é uma patologia comum que representa o 6° e o 5° cancro mais incidente em Portugal e na Itália, respetivamente. Em mais de metade dos casos ocorre reincidência durante o primeiro ano, requerendo acompanhamento clínico ao longo da vida. A instilação intravesical de Bacillus Calmette-Guérin (BCG) (uma estirpe atenuada do Mycobacterium bovis) representa uma imunoterapia eficaz no combate ao cancro de bexiga, no entanto, muitos aspetos da interação de BCG com as células tumorais bem como com as células do sistema imunitário permanecem por desvendar. As células tumorais de bexiga expressam frequentemente as formas sialiladas dos antigénios de Thomsen-Friedenreich (TF), i.e., sialil-T (sT) e sialil-Tn (sTn). Contudo ainda se desconhece o significado da sua expressão na malignidade tumoral e se afeta a eficácia da terapêutica BCG. Objetivo do estudo. Investigar o papel dos antigénios sT e sTn no fenótipo maligno de células de cancro de bexiga bem como na resposta mediada pelo sistema imunitário à terapia com BCG. Metodologia. Para tal, foram utilizadas as linhas celulares de cancro da bexiga HT1376 e MCR, geneticamente modificadas por transdução com vetores codificantes para as sialiltransferases ST3GAL1 ou ST6GALNAC1, de forma a expressar homogeneamente os antigénios sT ou sTn respetivamente. Estes modelos celulares foram estudados após confronto com BCG. O nível de BCG internalizado foi avaliado por citometria de fluxo. O perfil global de expressão genética dos modelos celulares antes e após incubação com BCG foi analisado pela tecnologia de microarray. O perfil de citocinas secretadas pelos modelos celulares após incubação com BCG, bem como de macrófagos estimulados pelo secretoma de células de cancro de bexiga que por sua vez foram estimuladas previamente por BCG, foi estudado pelo sistema multiplex de “imuno-esferas”. Resultados. A análise do transcritoma dos modelos celulares revelou que grupos de genes envolvidos em funções específicas foram modulados em paralelo nos dois modelos celulares, após transdução, independentemente da sialiltransferase expressa. Ou seja, em células que expressavam a sialiltransferase ST3GAL1 ou ST6GALNAC1, os genes envolvidos na regulação da segregação cromossómica e na reparação do DNA foram consistentemente regulados negativamente. Genes descritos na literatura como marcadores para o cancro de bexiga foram também modulados. A incubação com BCG resultou numa tendência ao aumento da expressão de genes relevantes na preservação e estabilidade genómica e menor malignidade, no entanto, apenas em células que expressavam sT ou sTn. Entre as dez citocinas testadas, apenas a IL-6 e IL-8 foram expressas pelas linhas celulares de cancro da bexiga, com indução destas após estimulação com BCG, e principalmente em células que expressavam ST3GAL1 ou ST6GALNAC1. Em macrófagos, citocinas inflamatórias, tais como IL-1β, IL-6 e TNFα, e a citocina anti-inflamatória IL-10, foram induzidas apenas pelo secretoma de células de cancro da bexiga confrontadas com BCG, com maior relevância quando estas expressavam ST3GAL1 ou ST6GALNAC1, prevendo a estimulação de macrófagos semelhantes aos de tipo M1 e uma melhor resposta à terapia com BCG. Conclusões. O efeito geral da expressão destas sialiltransferases e dos produtos enzimáticos sT ou sTn nas células de cancro de bexiga conduz a um fenótipo de maior malignidade. Contudo, a maior avidez de estas na produção de citocinas inflamatórias após confronto com BCG, bem como a maior capacidade de estimulação de macrófagos, predirá uma resposta à terapia com BCG mais eficaz em tumores que expressem os antigénios de TF sialilados. Tais conclusões são totalmente concordantes com os nossos mais recentes dados clínicos obtidos em colaboração, que mostram que em doentes com cancro de bexiga que expressam sTn respondem melhor a terapia BCG. ----------ABSTRACT: Background. Bladder cancer is a common malignancy representing the 6th and the 5th most incident cancer in Portugal and in Italy, respectively. More than half of the cases relapse within one year, requiring though a lifelong follow-up. Intravesical instillation of Bacillus Calmette-Guérin (BCG) (an attenuated strain of Mycobacterium bovis) represents an effective immunotherapy of bladder cancer, although many aspects of the interaction of BCG with cancer cells and host immune cells remain obscure. Bladder cancer cells often express the sialylated forms of the Thomsen-Friedenreich (TF), i.e., sialil-T (sT) e sialil-Tn (sTn). However, it’s still unknown the sense of such expression in tumour malignancy and in the BCG therapy efficacy. Aim of the study. To investigate the role of the sT and sTn antigens on the malignant phenotype of bladder cancer cells and the immune mediated response to BCG therapy. Experimental. We have utilized populations of the bladder cancer cell lines HT1376 and MCR, genetically modified by transduction with the sialyltransferases ST3GAL1 or ST6GALNAC1 to express homogeneously sT or sTn antigens. The level of BCG internalized was assessed by flow cytometry. The whole gene expression profile of BCG-challenged or unchallenged bladder cancer cell lines was studied by microarray technology. The profile of cytokines secreted by BCG-challenged bladder cancer cells and that of macrophages challenged by the secretome of BCG-challenged bladder cancer cells was studied by multiplex immune-beads assay. Results. Transcriptome analysis of the sialyltransferase-transduced cells revealed that groups of genes involved in specific functions were regulated in parallel in the two cell lines, regardless the sialyltransferase expressed. Namely, in sialyltransferase-expressing cells, genes involved in the proper chromosomal segregation and in the DNA repair were consistently down-regulated, while genes reported in literature as markers for bladder cancer were modulated. BCG-challenging induced a tendency to up-regulation of the genes preserving genomic stability and reducing malignancy, but only in cells expressing either sT or sTn. Among the ten cytokines tested, only IL-6 and IL-8 were expressed by bladder cancer cell lines and up-regulated by BCG-challenging, mainly in sialyltransferases-expressing cells. In macrophages, inflammatory cytokines, such as IL-1β, IL-6 and TNFα, and the antinflammatory IL-10 were induced only by the secretome of BCG-challenged bladder cancer cells, particularly when expressing either sialyltransferase, predicting the stimulation of M1-like macrophages and a better response to BCG therapy. Conclusions. The general effect of the expression of the two sialyltransferases and their products in the bladder cancer cells is toward a more malignant phenotype. However, the stronger ability of sialyltransferase expressing cells to produce inflammatory cytokines upon BCG-challenging and to stimulate macrophages predicts a more effective response to BCG in tumours expressing the sialylated TF antigens. This is fully consistent with our recent clinical data obtained in collaboration, showing that patients with bladder cancer expressing sTn respond better to BCG therapy.

Specific features of endothelial cells in primary tumors

RESUMO: As células endoteliais definem e delineiam todo o sistema vascular...Nesta tese procurámos explorar o papel que o ambiente tumoral exerce sobre as células endoteliais. ... Avaliamos também a capacidade anti-angiogénica de alguns derivados do estrogénio... Em suma os nossos resultados mostram a importância de um controlo rigoroso da regulação transcricional...

A domain specific language for domotic systems

To cope with modernity, the interesting of having a fully automated house has been increasing over the years, as technology evolves and as our lives become more stressful and overloaded. An automation system provides a way to simplify some daily tasks, allowing us to have more spare time to perform activities where we are really needed. There are some systems in this domain that try to implement these characteristics, but this kind of technology is at its early stages of evolution being that it is still far away of empowering the user with the desired control over a habitation. The reason is that the mentioned systems miss some important features such as adaptability, extension and evolution. These systems, developed from a bottom-up approach, are often tailored for programmers and domain experts, discarding most of the times the end users that remain with unfinished interfaces or products that they have difficulty to control. Moreover, complex behaviors are avoided, since they are extremely difficult to implement mostly due to the necessity of handling priorities, conflicts and device calibration. Besides, these solutions are only reachable at very high costs, yet they still have the limitation of being difficult to configure by non-technical people once in runtime operation. As a result, it is necessary to create a tool that allows the execution of several automated actions, with an interface that is easy to use but at the same time supports all the main features of this domain. It is also desirable that this tool is independent of the hardware so it can be reused, thus a Model Driven Development approach (MDD) is the ideal option, as it is a method that follows those principles. Since the automation domain has some very specific concepts, the use of models should be combined with a Domain Specific Language (DSL). With these two methods, it is possible to create a solution that is adapted to the end users, but also to domain experts and programmers due to the several levels of abstraction that can be added to diminish the complexity of use. The aim of this thesis is to design a Domain Specific Language (DSL) that uses the Model Driven Development approach (MDD), with the purpose of supporting Home Automation (HA) concepts. In this implementation, the development of simple and complex scenarios should be supported and will be one of the most important concerns. This DSL should also support other significant features in this domain, such as the ability to schedule tasks, which is something that is limited in the current existing solutions.