Caracterização dos padrões de comportamentos de saúde da população do concelho de Oeiras

Apresentação do projeto encomendado pela Câmara Municipal de Oeiras, no auditório César Batalha, a 13 de Novembro de 2008. Equipa de projeto:Helena Rato (Coord.); Belén Rando, César Madureira, Cláudia Anjos, Conceição Baptista, Luís Lapão, Madalena Antunes, Margarida Martins, Miguel Rodrigues. Consultor: Dr. Mário Pádua. Equipa CMO: Lurdes Poeira, Cristina André, Júlia Cardoso, Marta Camilo, Isabel Matias

Molecular fingerprint of human gastric cell line infected by Helicobacter pylori

Helicobacter pylori infection represents a serious health problem, given its association with serious gastric diseases as gastric ulcers, cancer and MALT lymphoma. Currently no vaccine exists and antibiotic-based eradication therapy is already failing in more than 20% of cases. To increase the knowledge on the infection process diverse gastric cell lines, e.g. the adenocarcinona gastric (AGS) cell line, are routinely used has in vitro models of gastric epithelia. In the present work the molecular fingerprint of infected and non-infected AGS cell lines, by diverse H. pylori strains, was acquired using vibrational infrared spectroscopy. These molecular fingerprints enabled to discriminate infected from non-infected AGS cells, and infection due to different strains, by performing Principal Component Analysis. It was also possible to estimate, from the AGS cells molecular fingerprint, the effect of the infection on diverse biochemical and metabolic cellular status. In resume infra-red spectroscopy enabled the acquisition of infected AGS cells molecular fingerprint with minimal sample preparation, in a rapid, high-throughput, economic process yielding highly sensitive and informative data, most useful for promoting critical knowledge on the H. pylori infection process. © 2015 IEEE.

In Situ Near-Infrared (NIR) Versus High-Throughput Mid- Infrared (MIR) Spectroscopy to Monitor Biopharmaceutical Production

The development of biopharmaceutical manufacturing processes presents critical constraints, with the major constraint being that living cells synthesize these molecules, presenting inherent behavior variability due to their high sensitivity to small fluctuations in the cultivation environment. To speed up the development process and to control this critical manufacturing step, it is relevant to develop high-throughput and in situ monitoring techniques, respectively. Here, high-throughput mid-infrared (MIR) spectral analysis of dehydrated cell pellets and in situ near-infrared (NIR) spectral analysis of the whole culture broth were compared to monitor plasmid production in recombinant Escherichia coil cultures. Good partial least squares (PLS) regression models were built, either based on MIR or NIR spectral data, yielding high coefficients of determination (R-2) and low predictive errors (root mean square error, or RMSE) to estimate host cell growth, plasmid production, carbon source consumption (glucose and glycerol), and by-product acetate production and consumption. The predictive errors for biomass, plasmid, glucose, glycerol, and acetate based on MIR data were 0.7 g/L, 9 mg/L, 0.3 g/L, 0.4 g/L, and 0.4 g/L, respectively, whereas for NIR data the predictive errors obtained were 0.4 g/L, 8 mg/L, 0.3 g/L, 0.2 g/L, and 0.4 g/L, respectively. The models obtained are robust as they are valid for cultivations conducted with different media compositions and with different cultivation strategies (batch and fed-batch). Besides being conducted in situ with a sterilized fiber optic probe, NIR spectroscopy allows building PLS models for estimating plasmid, glucose, and acetate that are as accurate as those obtained from the high-throughput MIR setup, and better models for estimating biomass and glycerol, yielding a decrease in 57 and 50% of the RMSE, respectively, compared to the MIR setup. However, MIR spectroscopy could be a valid alternative in the case of optimization protocols, due to possible space constraints or high costs associated with the use of multi-fiber optic probes for multi-bioreactors. In this case, MIR could be conducted in a high-throughput manner, analyzing hundreds of culture samples in a rapid and automatic mode.

In situ NIR spectroscopy monitoring of plasmid production processes effect of producing strain, medium composition and the cultivation strategy

BACKGROUNDWhile the pharmaceutical industry keeps an eye on plasmid DNA production for new generation gene therapies, real-time monitoring techniques for plasmid bioproduction are as yet unavailable. This work shows the possibility of in situ monitoring of plasmid production in Escherichia coli cultures using a near infrared (NIR) fiber optic probe. RESULTSPartial least squares (PLS) regression models based on the NIR spectra were developed for predicting bioprocess critical variables such as the concentrations of biomass, plasmid, carbon sources (glucose and glycerol) and acetate. In order to achieve robust models able to predict the performance of plasmid production processes, independently of the composition of the cultivation medium, cultivation strategy (batch versus fed-batch) and E. coli strain used, three strategies were adopted, using: (i) E. coliDH5 cultures conducted under different media compositions and culture strategies (batch and fed-batch); (ii) engineered E. coli strains, MG1655endArecApgi and MG1655endArecA, grown on the same medium and culture strategy; (iii) diverse E. coli strains, over batch and fed-batch cultivations and using different media compositions. PLS models showed high accuracy for predicting all variables in the three groups of cultures. CONCLUSIONNIR spectroscopy combined with PLS modeling provides a fast, inexpensive and contamination-free technique to accurately monitoring plasmid bioprocesses in real time, independently of the medium composition, cultivation strategy and the E. coli strain used.

Monitoring the ex-vivo expansion of human mesenchymal stem/stromal cells in xeno-free microcarrier-based reactor systems by MIR spectroscopy

Human mesenchymal stem/stromal cells (MSCs) have received considerable attention in the field of cell-based therapies due to their high differentiation potential and ability to modulate immune responses. However, since these cells can only be isolated in very low quantities, successful realization of these therapies requires MSCs ex-vivo expansion to achieve relevant cell doses. The metabolic activity is one of the parameters often monitored during MSCs cultivation by using expensive multi-analytical methods, some of them time-consuming. The present work evaluates the use of mid-infrared (MIR) spectroscopy, through rapid and economic high-throughput analyses associated to multivariate data analysis, to monitor three different MSCs cultivation runs conducted in spinner flasks, under xeno-free culture conditions, which differ in the type of microcarriers used and the culture feeding strategy applied. After evaluating diverse spectral preprocessing techniques, the optimized partial least square (PLS) regression models based on the MIR spectra to estimate the glucose, lactate and ammonia concentrations yielded high coefficients of determination (R2 ≥ 0.98, ≥0.98, and ≥0.94, respectively) and low prediction errors (RMSECV ≤ 4.7%, ≤4.4% and ≤5.7%, respectively). Besides PLS models valid for specific expansion protocols, a robust model simultaneously valid for the three processes was also built for predicting glucose, lactate and ammonia, yielding a R2 of 0.95, 0.97 and 0.86, and a RMSECV of 0.33, 0.57, and 0.09 mM, respectively. Therefore, MIR spectroscopy combined with multivariate data analysis represents a promising tool for both optimization and control of MSCs expansion processes.

The study of lead white oil paints: a molecular approach to the whites of Amadeo de Souza-Cardoso

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Conservação e Restauro

Determinantes do sucesso das iniciativas empresariais apoiadas pelo microcrédito

Dissertação de Mestrado apresentada ao Instituto de Contabilidade e Administração do Porto para a obtenção do grau de Mestre em Contabilidade e Finanças sob orientação de: Amélia Ferreira da Silva “Esta versão contém as críticas e sugestões dos elementos do júri”

A importância das estratégias argumentativas para o ensino da filiosofia : uma reflexão sobre a relação estabelecida entre estagiário, alunos e professora orientadora

Relatório apresentado para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Ensino da Filosofia no Ensino Secundário.

A comunicação on-line das ONGs e a angariação de fundos

Dissertação de Mestrado apresentado ao Instituto de Contabilidade e Administração do Porto para a obtenção do grau de Mestre em Marketing Digital, sob orientação da Mestre Inês Veiga Pereira “Esta versão contém as críticas e sugestões dos elementos do júri”

Genetic structure of Neisseria meningitidis serogroup C epidemic strains in South Brazil

In the present study we report the results of an analysis, based on serotyping, multilocus enzyme electrophoresis (MEE), and ribotyping of N. meningitidis serogroup C strains isolated from patients with meningococcal disease (MD) in Rio Grande do Sul (RS) and Santa Catarina (SC) States, Brazil, as the Center of Epidemiology Control of Ministry of Health detected an increasing of MD cases due to this serogroup in the last two years (1992-1993). We have demonstrated that the MD due to N.meningitidis serogroup C strains in RS and SC States occurring in the last 4 years were caused mainly by one clone of strains (ET 40), with isolates indistinguishable by serogroup, serotype, subtype and even by ribotyping. One small number of cases that were not due to an ET 40 strains, represent closely related clones that probably are new lineages generated from the ET 40 clone referred as ET 11A complex. We have also analyzed N.meningitidis serogroup C strains isolated in the greater São Paulo in 1976 as representative of the first post epidemic year in that region. The ribotyping method, as well as MEE, could provide useful information about the clonal characteristics of those isolates and also of strains isolated in south Brazil. The strains from 1976 have more similarity with the actual endemic than epidemic strains, by the ribotyping, sulfonamide sensitivity, and MEE results. In conclusion, serotyping with monoclonal antibodies (C:2b:P1.3), MEE (ET 11 and ET 11A complex), and ribotyping by using ClaI restriction enzyme (Rb2), were useful to characterize these epidemic strains of N.meningitidis related to the increased incidence of MD in different States of south Brazil. It is mostly probable that these N.meningitidis serogroup C strains have poor or no genetic corelation with 1971-1975 epidemic serogroup C strains. The genetic similarity of members of the ET 11 and ET 11A complex were confirmed by the ribotyping method by using three restriction endonucleases.

Gestão de produto PREMFIRE – sistema de informação geográfica 3D para apoio ao combate a incêndios.

Dissertação apresentada como requisito parcial para obtenção do grau de Mestre em Ciência e Sistemas de Informação Geográfica.

Hepatic fascioliasis: case report and review

A well documented case of hepatic fascioliasis (HF), successfully treated with triclabendazole, is reported. Predominant clinical manifestations were fever, marked eosinophilia and abdominal pain. Triclabendazole was given as two single oral doses of 10 mg/kg each. Neither side effects nor clinical or parasitological relapses were seen after three months of follow up Based on this experience and few other similar reports in the literature, triclabendazole might be a valid therapeutical alternative in the treatment of human fascioliasis.

Seropositivity for anti-Trypanosoma cruzi antibodies among blood donors of the "Hospital Universitário Regional do Norte do Paraná", Londrina, Brazil

The most frequent form of acquisition of Chagas' disease in endemic areas was the transmission through the feces of contaminated triatominae. However, special attention should be paid in urban areas to transmission by blood transfusion, justifying the compulsory screening of blood donors. Early investigations at blood banks in the town of Londrina, Brazil, demonstrated that the seroprevalence of anti-Trypanosoma cruzi antibodies among blood donors was approximately 7.0% in the fifties9,34. Further studies demonstrated pratically the same seroprevalence until the eighties4,32,41. In an attempt to obtain data about the real dimension of the seropositivity for anti-Trypasonoma cruzi antibodies in the region, the authors carried out a large-scale study on 45,774 serum samples from blood donors of the Hemocentro of Hospital Universitário Regional do Norte do Paraná (HURNP), Universidade Estadual de Londrina. The immunological tests were done at the Division of Clinical Immunology of HURNP from May 1990 to December 1994. The serum samples were studied by the indirect hemagglutination assay (IHA, using kits commercially obtained from EBRAM) and by indirect immunofluorescence (IFI, using kits from LIO SERUM) with anti-human IgG conjugate (LABORCLIN). The results demonstrated that 643 serum samples were positive in both assay corresponding to a seroprevalence of 1.4%, i.e., a significant decrease in anti-Trypanosoma cruzi antibodies in the region in comparison with the previously mentioned rates. Data correlating sex and age of seropositive blood donors are presented, as well as the possible factors that may have contributed to the results observed.

A arte fora da história. Exposições da colecção do museu do Chiado 1994-2009

Dissertação apresentada para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Museologia

Desenvolvimento de apoio à decisão operacional – previsão de curto prazo de geração distribuída de tecnologia eólica ligada à RND

No contexto da penetração de energias renováveis no sistema elétrico, Portugal ocupa uma posição de destaque a nível mundial, muito devido à produção de eólica. Com um sistema elétrico com forte presença de fontes de energia renováveis, novos desafios surgem, nomeadamente no caso da energia eólica pela sua imprevisibilidade e volatilidade. O recurso eólico embora seja ilimitado não é armazenável, surgindo assim a necessidade da procura de modelos de previsão de produção de energia elétrica dos parques eólicos de modo a permitir uma boa gestão do sistema. Nesta dissertação apresentam-se as contribuições resultantes de um trabalho de pesquisa e investigação sobre modelos de previsão da potência elétrica com base em valores de previsões meteorológicas, nomeadamente, valores previstos da intensidade e direção do vento. Consideraram-se dois tipos de modelos: paramétricos e não paramétricos. Os primeiros são funções polinomiais de vários graus e a função sigmoide, os segundos são redes neuronais artificiais. Para a estimação dos modelos e respetiva validação, são usados dados recolhidos ao longo de dois anos e três meses no parque eólico do Pico Alto de potência instalada de 6 MW. De forma a otimizar os resultados da previsão, consideram-se diferentes classes de perfis de produção, definidas com base em quatro e oito direções do vento, e ajustam-se os modelos propostos em cada uma das classes. São apresentados e discutidos resultados de uma análise comparativa do desempenho dos diferentes modelos propostos para a previsão da potência.