Biodiesel production from chicken feather meal, combining biocatalysis and supercritical technology

Dissertação para obtenção do Grau de Mestre em Biotecnologia

Biogas production from potato peel waste

Dissertação para obtenção do Grau de Mestre em Engenharia do Ambiente, perfil de Gestão e Sistemas Ambientais

Some Notes on the Reception of Josquin and of Northern Idioms in Portuguese Music and Culture

Although Josquin is by far the best-represented foreign composer in Gonçalo de Baena's Arte novamente inventada pera aprender a tanger (Lisbon, 1540), his music is undeniably under-represented both in the extant sixteenth-century Portuguese manuscripts containing Franco-Flemish polyphony and in volumes imported from the Netherlands such as Coimbra MM 2 and VienNB 1783. Josquin’s reputation made him, along with Ockeghem, a symbol in Portuguese humanistic culture, but up to at least the late 1530s his name seems to have been much better known than his music. Nevertheless, possible allusions to specific works by Josquin can be found in early- and mid-sixteenth-century Portuguese polyphony. By the 1520s, the general technical and stylistic characteristics of his and the following generation of northerners had begun to permeate locally produced polyphony. This eventually replaced the late-fifteenth- and early-sixteenth-century pan-consonant and homorythmic style associated with the Aragonese and the so-called Spanish court repertory.

Constraint-based modelling of mixed microbial populations: Application to polyhydroxyalkanoates production

Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica

Microbial contribution to biofuels production

Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica

Assessment of Notch1 ligands production - key protein targets in breast cancer

Cell-to-cell communication is required for many biological processes in development and adult life. One of the most common systems utilized by a wide range of eukaryotes is the Notch signalling pathway. Four Notch receptors and five ligands have been identified in mammals that interact via their extracellular domains leading to transcription activation. Studies have shown that the Notch ligands expression is undetectable in normal breast tissues, but moderate to high expression has been detected in breast cancer. Thus, any of the Notch1 ligands can be studied as possible therapeutic targets for breast cancer. To study Notch pathway proteins there is the need to obtain stable protein solutions. E. coli is the host of excellence for recombinant proteins for the ease of use, fast growth and high cell densities. However, the expression of mammalian proteins in such systems may overwhelm the bacterial cellular machinery, which does not possess the ability for post-translational modifications, or dedicated compartments for protein synthesis. Mammalian cells are therefore preferred, despite their technical and financial increased demands. We aim to determine the best expression and purification conditions for the different ligand protein constructs, to develop specific function-blocking antibodies using the Phage Display technology. Moreover, we propose to crystallize the Notch1 ligands alone and in complex with the phage display selected antibodies, unveiling molecular details. hJag2DE3 and hDll1DE6 proteins were purified from refolded inclusion bodies or mammalian cell culture supernatants, respectively, and purity was confirmed by SDS-PAGE (>95%). Protein produced in mammalian cells showed to be more stable, apparently with the physiological disulfide pattern, contrary to what was observed in the refolded protein. Several nano-scale crystallization experiments were set up in 96-well plates, but no positive result was obtained. We will continue to pursue for the best expression for the Notch ligand constructs in both expression systems.

Popular music exhibitions in Portugal: practices of representation from 2007 to 2013

This thesis focuses on the representation of Popular Music in museums by mapping, analyzing, and characterizing its practices in Portugal at the beginning of the 21st century. Now that museums' ability to shape public discourse is acknowledged, the examination of popular music's discourses in museums is of the utmost importance for Ethnomusicology and Popular Music Studies as well as for Museum Studies. The concept of 'heritage' is at the heart of this processes. The study was designed with the aim of moving the exhibiting of popular music in museums forward through a qualitative inquiry of case studies. Data collection involved surveying pop-rock music exhibitions as a qualitative sampling of popular music exhibitions in Portugal from 2007 to 2013. Two of these exhibitions were selected as case studies: No Tempo do Gira-Discos: Um Percurso pela Produção Fonográfica Portuguesa at the Museu da Música in Lisbon in 2007 (also Faculdade de Letras, 2009), and A Magia do Vinil, a Música que Mudou a Sociedade at the Oficina da Cultura in Almada in 2008 (and several other venues, from 2009 to 2013). Two specific domains were observed: popular music exhibitions as instances of museum practice and museum professionals. The first domain encompasses analyzing the types of objects selected for exhibition; the interactive museum practices fostered by the exhibitions; the concepts and narratives used to address popular music discursively, as well as the interpretative practices they allow. The second domain, focuses museum professionals and curators of popular music exhibitions as members of a group, namely their goals, motivations and perspectives. The theoretical frameworks adopted were drawn from the fields of ethnomusicology, popular music studies, and museum studies. The written materials of the exhibitions were subjected of methods of discourse analysis methods. Semi-structured interviews with curators and museum professional were also conducted and analysed. From the museum studies perspective, the study research suggests that the practice adopted by popular music museums largely matches that of conventional museums. From the ethnomusicological and popular music studies stand point, the two case studies reveal two distinct conceptual worlds: the first exhibition, curated by an academic and an independent researcher, points to a mental configuration where popular music is explained through a framework of genres supported by different musical practices. Moreover, it is industry actors such as decision makers and gatekeepers that govern popular music, which implies that the visitors' romantic conception of the musician is to some extent dismantled; the second exhibition, curated by a record collector and specialist, is based on a more conventional process of the everyday historical speech that encodes a mismatch between “good” and “bad music”. Data generated by a survey shows that only one curator, in fact that of my first case study, has an academic background. The backgrounds of all the others are in some way similar to the curator of the second case study. Therefore, I conclude that the second case study best conveys the current practice of exhibiting Popular Music in Portugal.

Establishment and evaluation of flexible insect cell lines for rapid production of recombinant proteins

Fundação para a Ciência e a Tecnologia (FCT) - (PTDC/EBB-EBI/102266/2008 and SFRH/BD/43830/2008, respectively) and by European Community’s FP7/2007-2013 (grant agreement nº 270089)

Impact of carbon/nitrogen feeding strategy on polyhydroxyalkanoates production using mixed microbial cultures

Polyhydroxyalkanoates (PHA) production using mixed microbial cultures (MMC) requires a multi-stage process involving the microbial selection of PHA-storing microorganisms, typically operated in sequencing batch reactors (SBR), and an accumulation reactor. Since low-cost renewable feedstocks used as process feedstock are often nitrogen-deficient, nutrient supply in the selection stage is required to allow for microbial growth. In this context, the possibility to uncouple nitrogen supply from carbon feeding within the SBR cycle has been investigated in this study. Moreover, three different COD:N ratios (100:3.79, 100:3.03 and 100:2.43) were tested in three different runs which also allowed the study of COD:N ratio on the SBR performance. For each run, a synthetic mixture of acetic and propionic acids at an overall organic load rate of 8.5 gCOD L-1 d-1 was used as carbon feedstock, whereas ammonium sulfate was the nitrogen source in a lab-scale sequence batch reactor (SBR) with 1 L of working volume. Besides, a sludge retention time (SRT) of 1 d was used as well as a 6 h cycle length. The uncoupled feeding strategy significantly enhanced the selective pressure towards PHA-storing microorganisms, resulting in a two-fold increase in the PHA production (up to about 1.3 gCOD L-1). A high storage response was observed for the two runs with the COD:N ratios (gCOD:gN) of 100:3.79 and 100:3.03, whereas the lowest investigated nitrogen load resulted in very poor performance in terms of polymer production. In fact, strong nitrogen limitation caused fungi to grow and a very poor storage ability by microorganisms that thrived in those conditions. The COD:N ratio also affected the polymer composition, indeed the produced poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) showed a variable HV content (1-20 %, w/w) among the three runs, lessening as the COD:N increased. This clearly suggests the possibility to use the COD:N ratio as a tool for tuning polymer properties regardless the composition of the feedstock.

Exploring the relationship between toxin and spore production in the human enteric pathogen Clostridium difficile

RESUMO: Clostridium difficile é presentemente a principal causa de doença gastrointestinal associada à utilização de antibióticos em adultos. C. difficile é uma bactéria Gram-positiva, obrigatoriamente anaeróbica, capaz de formar endósporos. Tem-se verificado um aumento dos casos de doença associada a C. difficile com sintomas mais severos, elevadas taxas de morbilidade, mortalidade e recorrência, em parte, devido à emergência de estirpes mais virulentas, mas também devido à má gestão do uso de antibióticos. C. difficile produz duas toxinas, TcdA e TcdB, que são os principais fatores de virulência e responsáveis pelos sintomas da doença. Estas são codificadas a partir do Locus de Patogenicidade (PaLoc) que codifica ainda para um regulador positivo, TcdR, uma holina, TcdE, e um regulador negativo, TcdC. Os esporos resistentes ao oxigénio são essenciais para a transmissão do organismo e recorrência da doença. A expressão dos genes do PaLoc ocorre em células vegetativas, no final da fase de crescimento exponencial, e em células em esporulação. Neste trabalho construímos dois mutantes de eliminação em fase dos genes tcdR e tcdE. Mostrámos que a auto-regulação do gene tcdR não é significativa. No entanto, tcdR é sempre necessário para a expressão dos genes presentes no PaLoc. Trabalho anterior mostrou que, com a exceção de tcdC, os demais genes do PaLoc são expressos no pré-esporo. Mostrámos aqui que TcdA é detectada à superfície do esporo maduro e que a eliminação do tcdE não influencia a acumulação de TcdA no meio de cultura ou em associação às células ou ao esporo. Estas observações têm consequências para o nosso entendimento do processo infecioso: sugeremque o esporo possa ser também um veículo para a entrega da toxina nos estágios iniciais da infecção, que TcdA possa ser libertada durante a germinação do esporo, e que o esporo possa utilizar o mesmo receptor reconhecido por TcdA para a ligação à mucosa do cólon.---------------------------ABSTRACT: Clostridium difficile is currently the major cause of antibiotic-associated gastrointestinal diseases in adults. This is a Gram-positive bacterium, endospore-forming and an obligate anaerobe that colonizes the gastrointestinal tract. Recent years have seen a rise in C. difficile associated disease (CDAD) cases, associated with more severe disease symptoms, higher rates of morbidity, mortality and recurrence, which were mostly caused due to the emergence of “hypervirulent” strains but also due to changing patterns of antibiotics use. C. difficile produces two potent toxins, TcdA and TcdB, which are the main virulence factors and the responsible for the disease symptoms. These are codified from a Pathogenicity Locus (PaLoc), composed also by the positive regulator, TcdR, the holin-like protein, TcdE, and a negative regulator, TcdC. Besides the toxins, the oxygen-resistant spores are also essential for transmission of the organism through diarrhea; moreover, spores can accumulate in the environment or in the host, which will cause disease recurrence. The expression of the PaLoc genes occurs in vegetative cells, at the end of the exponential growth phase, and in sporulating cells. In this work, we constructed two in-frame deletion mutants of tcdR and tcdE. We showed that the positive auto regulation of tcdR is not significant. However, tcdR is always necessary for the expression of the PaLoc genes. A previous work showed that, except tcdC, all the PaLoc genes are expressed in the forespore. Here, we detected TcdA at the spore surface. Furthermore, we showed that the in-frame deletion of tcdE does not affect the accumulation of TcdA in the culture medium or in association with cells or spores. This data was important for us to conclude about the infeccious process: it suggests that the spore may be the vehicle for the delivery of TcdA in early stages of infection, that TcdA may be released during spores germination and that this spore may use the same receptor recognized by TcdA to bind to the colonic mucosa.

Satisfação dos utilizadores de sistemas de informação e documentação de enfermagem em suporte eletrónico no Centro Hospitalar Lisboa Norte, E.P.E.

RESUMO - Os registos de enfermagem no Centro Hospitalar Lisboa Norte, E.P.E. (CHLN) são feitos em suporte de papel ou através de sistemas de informação (SI) próprios de cada serviço, com a utilização de várias aplicações como o Alert, Picis, etc. Esta diversidade gera alguns constrangimentos em termos de fluxo de informação, em virtude da falta de interoperabilidade dos respetivos sistemas. Esta realidade pode ter impactos na área da qualidade e segurança do utente, com a possibilidade de ocorrência de erros e/ou eventos adversos. Podem ainda ser notórios na área da privacidade e confidencialidade dos dados clínicos, na tomada de decisão, na gestão clínica e financeira e na produção de informação útil para a investigação científica. No CHLN está em curso a implementação de um SI capaz de dar resposta aos registos de enfermagem, integrados num registo de saúde eletrónico focado no utente que obedece à metodologia do processo de enfermagem e utiliza a linguagem codificada da Classificação Internacional para a Prática de Enfermagem (CIPE). Com o desenvolvimento desta investigação, devidamente autorizada pelo Conselho de Administração do CHLN, pretendeu-se dar resposta à pergunta de partida:  Estarão os enfermeiros, utilizadores do Desktop de Enfermagem do CHLN, satisfeitos com esse sistema de informação? Com esse propósito, foi elaborada uma abordagem exploratória com recurso a pesquisa bibliográfica sobre os sistemas de informação de enfermagem e a sua avaliação, com base no “Modelo de Sucesso dos Sistemas de Informação de DeLone e McLean”, tendo sido desenvolvido um estudo de caso com uma abordagem quantitativa, mediante a aplicação de um inquérito por questionário aos 262 enfermeiros do CHLN, nos serviços onde já utilizavam o referido SI, entre maio e junho de 2014, com uma taxa de resposta de 84%. Os resultados da aplicação do questionário, objeto de análise estatística univariada e bivariada com recurso a procedimentos descritivos e inferenciais, visando a produção de sínteses dirigidas aos objetivos do estudo, permitiram caracterizar o nível de satisfação dos enfermeiros, enquanto utilizadores do “desktop de enfermagem”, suportados por Tecnologias de Informação e Comunicação. Na escala utilizada (de 1 a 5), o nível médio de satisfação global (2,78) foi ligeiramente inferior ao seu ponto médio (3). No entanto, a maioria dos inquiridos (81,5%) não pretende abandonar o SI que utilizam. Os resultados obtidos permitem demonstrar que a satisfação dos enfermeiros face à implementação e utilização do SIE se trata de uma estratégia bem sucedida do CHLN, ainda que haja áreas onde foram evidenciados menores níveis de satisfação, tais como a “velocidade de processamento”, o “equipamento informático” e o “apoio técnico”, que podem ser alvo de uma maior atenção e reflexão pela gestão de topo, numa estratégia de melhoria contínua da qualidade, com importantes benefícios para a governação da instituição, para os profissionais e para os utentes, no futuro.