Molecular genomics of a genetic code alteration

The genetic code is not universal. Alterations to its standard form have been discovered in both prokaryotes and eukaryotes and demolished the dogma of an immutable code. For instance, several Candida species translate the standard leucine CUG codon as serine. In the case of the human pathogen Candida albicans, a serine tRNA (tRNACAGSer) incorporates in vivo 97% of serine and 3% of leucine in proteins at CUG sites. Such ambiguity is flexible and the level of leucine incorporation increases significantly in response to environmental stress. To elucidate the function of such ambiguity and clarify whether the identity of the CUG codon could be reverted from serine back to leucine, we have developed a forced evolution strategy to increase leucine incorporation at CUGs and a fluorescent reporter system to monitor such incorporation in vivo. Leucine misincorporation increased from 3% up to nearly 100%, reverting CUG identity from serine back to leucine. Growth assays showed that increasing leucine incorporation produced impressive arrays of phenotypes of high adaptive potential. In particular, strains with high levels of leucine misincorporation exhibited novel phenotypes and high level of tolerance to antifungals. Whole genome re-sequencing revealed that increasing levels of leucine incorporation were associated with accumulation of single nucleotide polymorphisms (SNPs) and loss of heterozygozity (LOH) in the higher misincorporating strains. SNPs accumulated preferentially in genes involved in cell adhesion, filamentous growth and biofilm formation, indicating that C. albicans uses its natural CUG ambiguity to increase genetic diversity in pathogenesis and drug resistance related processes. The overall data provided evidence for unantecipated flexibility of the C. albicans genetic code and highlighted new roles of codon ambiguity on the evolution of genetic and phenotypic diversity.

Educação para o desenvolvimento sustentável no ensino básico : contributos da integração das tecnologias da informação e comunicação e do envolvimento familiar nos primeiros anos de escolaridade

A necessidade de se educar para o desenvolvimento sustentável é um ponto assente nas atuais políticas educativas internacionais. Considera-se fulcral desenvolver nos alunos as competências necessárias para analisar criticamente e tomar de decisão frente às atuais problemáticas relacionadas com as dimensões ambiental, social e económica, sob uma perspetiva sistémica. No entanto, a literatura de referência aponta a escassez de recursos intencionalmente concebidos para apoiar práticas didático-pedagógicas dos professores mais coerentes com estas diretrizes. Assim, o presente estudo tem por finalidade desenvolver estratégias, conceber e explorar recursos didáticos multimédia para promover a Educação para o Desenvolvimento Sustentável (EDS), no contexto da Educação em Ciência com orientação Ciência-Tecnologia-Sociedade (CTS), através da integração das Tecnologias da Informação e Comunicação (TIC) e do envolvimento familiar (EF) nos primeiros anos de escolaridade, assim como compreender os impactes dos mesmos na alteração das práticas dos professores tornando-as mais coerentes com as orientações EDS/CTS. Face a esta finalidade, adotámos uma metodologia de natureza qualitativa, nomeadamente o Estudo de Caso na vertente multicasos, ao longo da investigação. O estudo envolveu o 1º Ciclo do Ensino Básico de um Agrupamento de Escolas de Aveiro (Portugal), nomeadamente professores, alunos e respetivos familiares. Por entender serem os professores um elemento-chave de mudança do processo educativo, o estudo envolveu a conceção e implementação de uma Oficina de Formação de professores e uma comunidade online, a servir de plataforma de comunicação entre as escolas e as famílias. Assim, para a recolha de dados recorremos às técnicas de inquérito, observação e análise documental. Os respetivos instrumentos utilizados para cada técnica foram questionários, diário da investigadora, e-portefólios reflexivos e registos de interação na comunidade online. Os dados qualitativos e quantitativos foram analisados através da técnica de análise de conteúdo e análise estatística descritiva, respetivamente. Os resultados apontam que a integração das TIC e o incentivo ao envolvimento familiar contribuem positivamente para a promoção da EDS nos primeiros anos de escolaridade. Contudo, considerando o contexto em estudo onde as escolas atendem muitas famílias provenientes de meios socioeconómicos menos favorecidos, é preciso repensar a integração das TIC, para que estas não se configurem como mais uma barreira para o envolvimento familiar. Assim, trabalhar a EDS/CTS para além dos muros da escola, envolvendo as famílias dos alunos e colocando as TIC como uma ponte de comunicação é apresentado nesta investigação como o principal contributo para a consecução da EDS.

O quadro interativo na aula de inglês : desenvolvimento de práticas comunicativas

Na sociedade da informação, o papel da escola como centro transmissor de conhecimentos está a perder relevância face ao crescente número de fontes de informação alternativas, ricas em conteúdos e de fácil acesso. A alteração de paradigmas cria a necessidade de se introduzirem nos contextos educacionais novas ferramentas e que se adequem estratégias que permitam motivar e ensinar os aprendentes a procurarem e a selecionarem essa informação, participando, desta forma, ativamente na construção do conhecimento. Neste contexto, consideramos que o quadro interativo, pela sua capacidade de funcionar como hub digital, permite integrar recursos dinâmicos e interativos facilitadores do acesso ao conhecimento e à informação. A dissertação que apresentamos, “O Quadro Interativo na aula de inglês: desenvolvimento de práticas comunicativas”, situa-se na área do Multimédia em Educação e, mais especificamente, no domínio da tecnologia e pedagogia em sala de aula. A investigação foi desenhada em torno de um projeto de implementação de quadros interativos (Inov@r com QI) em agrupamentos/escolas do interior do país, associado a um Centro de Formação. A partir de uma investigação qualitativa mista, baseada no tracer study (análise documental, entrevistas e questionários) e no estudo etnográfico (aulas observadas, entrevistas e questionários), procurámos evidenciar a forma como essa tecnologia levou à integração de recursos e à adoção de estratégias promotoras de um ensino e aprendizagem ativo, centrado no aprendente e criando dinâmicas de interação e comunicação facilitadoras do desenvolvimento da competência comunicativa. Os resultados obtidos revelam que a introdução do quadro interativo produz efeitos positivos no ensino e aprendizagem do inglês. A sua integração em contexto de sala de aula abre espaço a um conjunto de potencialidades pedagógicas, promove o uso de recursos digitais variados; leva a uma diversificação de abordagens metodológicas que promovem a interação, criando espaços que levam ao desenvolvimento da competência comunicativa. No entanto, para tornar a introdução do quadro interativo mais eficaz, como ferramenta educativa no processo de ensino e aprendizagem, devemos ter em conta que esse processo de introdução do quadro deve ser planeada e refletida, devendo ter em conta uma nova realidade de conceitos, a tecnologia e as literacias necessárias para uma implementação eficaz. Os docentes devem ser acompanhados, ter acesso a formação técnica e pedagógica; e a possibilidade de integrarem um espaço de colaboração e partilha, como forma mais eficaz de se libertarem de práticas centradas no docente e orientadas apenas para a aquisição de conteúdos. Assim será possível dar lugar a um discurso pedagógico que inclua os vários intervenientes no processo de aprendizagem e a construção de conhecimento, num espaço de interação suportado pela tecnologia

Contributos para a eficácia do clustering usando o tagging social

Nos últimos anos temos vindo a assistir a uma mudança na forma como a informação é disponibilizada online. O surgimento da web para todos possibilitou a fácil edição, disponibilização e partilha da informação gerando um considerável aumento da mesma. Rapidamente surgiram sistemas que permitem a coleção e partilha dessa informação, que para além de possibilitarem a coleção dos recursos também permitem que os utilizadores a descrevam utilizando tags ou comentários. A organização automática dessa informação é um dos maiores desafios no contexto da web atual. Apesar de existirem vários algoritmos de clustering, o compromisso entre a eficácia (formação de grupos que fazem sentido) e a eficiência (execução em tempo aceitável) é difícil de encontrar. Neste sentido, esta investigação tem por problemática aferir se um sistema de agrupamento automático de documentos, melhora a sua eficácia quando se integra um sistema de classificação social. Analisámos e discutimos dois métodos baseados no algoritmo k-means para o clustering de documentos e que possibilitam a integração do tagging social nesse processo. O primeiro permite a integração das tags diretamente no Vector Space Model e o segundo propõe a integração das tags para a seleção das sementes iniciais. O primeiro método permite que as tags sejam pesadas em função da sua ocorrência no documento através do parâmetro Social Slider. Este método foi criado tendo por base um modelo de predição que sugere que, quando se utiliza a similaridade dos cossenos, documentos que partilham tags ficam mais próximos enquanto que, no caso de não partilharem, ficam mais distantes. O segundo método deu origem a um algoritmo que denominamos k-C. Este para além de permitir a seleção inicial das sementes através de uma rede de tags também altera a forma como os novos centróides em cada iteração são calculados. A alteração ao cálculo dos centróides teve em consideração uma reflexão sobre a utilização da distância euclidiana e similaridade dos cossenos no algoritmo de clustering k-means. No contexto da avaliação dos algoritmos foram propostos dois algoritmos, o algoritmo da “Ground truth automática” e o algoritmo MCI. O primeiro permite a deteção da estrutura dos dados, caso seja desconhecida, e o segundo é uma medida de avaliação interna baseada na similaridade dos cossenos entre o documento mais próximo de cada documento. A análise de resultados preliminares sugere que a utilização do primeiro método de integração das tags no VSM tem mais impacto no algoritmo k-means do que no algoritmo k-C. Além disso, os resultados obtidos evidenciam que não existe correlação entre a escolha do parâmetro SS e a qualidade dos clusters. Neste sentido, os restantes testes foram conduzidos utilizando apenas o algoritmo k-C (sem integração de tags no VSM), sendo que os resultados obtidos indicam que a utilização deste algoritmo tende a gerar clusters mais eficazes.

Dependable wireless real-time communications for open environments

Wireless communication technologies have become widely adopted, appearing in heterogeneous applications ranging from tracking victims, responders and equipments in disaster scenarios to machine health monitoring in networked manufacturing systems. Very often, applications demand a strictly bounded timing response, which, in distributed systems, is generally highly dependent on the performance of the underlying communication technology. These systems are said to have real-time timeliness requirements since data communication must be conducted within predefined temporal bounds, whose unfulfillment may compromise the correct behavior of the system and cause economic losses or endanger human lives. The potential adoption of wireless technologies for an increasingly broad range of application scenarios has made the operational requirements more complex and heterogeneous than before for wired technologies. On par with this trend, there is an increasing demand for the provision of cost-effective distributed systems with improved deployment, maintenance and adaptation features. These systems tend to require operational flexibility, which can only be ensured if the underlying communication technology provides both time and event triggered data transmission services while supporting on-line, on-the-fly parameter modification. Generally, wireless enabled applications have deployment requirements that can only be addressed through the use of batteries and/or energy harvesting mechanisms for power supply. These applications usually have stringent autonomy requirements and demand a small form factor, which hinders the use of large batteries. As the communication support may represent a significant part of the energy requirements of a station, the use of power-hungry technologies is not adequate. Hence, in such applications, low-range technologies have been widely adopted. In fact, although low range technologies provide smaller data rates, they spend just a fraction of the energy of their higher-power counterparts. The timeliness requirements of data communications, in general, can be met by ensuring the availability of the medium for any station initiating a transmission. In controlled (close) environments this can be guaranteed, as there is a strict regulation of which stations are installed in the area and for which purpose. Nevertheless, in open environments, this is hard to control because no a priori abstract knowledge is available of which stations and technologies may contend for the medium at any given instant. Hence, the support of wireless real-time communications in unmanaged scenarios is a highly challenging task. Wireless low-power technologies have been the focus of a large research effort, for example, in the Wireless Sensor Network domain. Although bringing extended autonomy to battery powered stations, such technologies are known to be negatively influenced by similar technologies contending for the medium and, especially, by technologies using higher power transmissions over the same frequency bands. A frequency band that is becoming increasingly crowded with competing technologies is the 2.4 GHz Industrial, Scientific and Medical band, encompassing, for example, Bluetooth and ZigBee, two lowpower communication standards which are the base of several real-time protocols. Although these technologies employ mechanisms to improve their coexistence, they are still vulnerable to transmissions from uncoordinated stations with similar technologies or to higher power technologies such as Wi- Fi, which hinders the support of wireless dependable real-time communications in open environments. The Wireless Flexible Time-Triggered Protocol (WFTT) is a master/multi-slave protocol that builds on the flexibility and timeliness provided by the FTT paradigm and on the deterministic medium capture and maintenance provided by the bandjacking technique. This dissertation presents the WFTT protocol and argues that it allows supporting wireless real-time communication services with high dependability requirements in open environments where multiple contention-based technologies may dispute the medium access. Besides, it claims that it is feasible to provide flexible and timely wireless communications at the same time in open environments. The WFTT protocol was inspired on the FTT paradigm, from which higher layer services such as, for example, admission control has been ported. After realizing that bandjacking was an effective technique to ensure the medium access and maintenance in open environments crowded with contention-based communication technologies, it was recognized that the mechanism could be used to devise a wireless medium access protocol that could bring the features offered by the FTT paradigm to the wireless domain. The performance of the WFTT protocol is reported in this dissertation with a description of the implemented devices, the test-bed and a discussion of the obtained results.

Prevenção e detecção de incêndios

Os fogos florestais, embora possam ser encarados como um fenómeno natural, constituem igualmente uma ameaça com impactos negativos a vários níveis. A alteração dos regimes naturais do fogo por acção antrópica tem vindo a maximizar o potencial catastrófico do fenómeno em diferentes regiões do planeta, destacando-se, em particular, a Região Mediterrânica, onde o fogo constitui uma perturbação recorrente associada, por um lado, às características climáticas desta região e, por outro, às práticas tradicionais decorrentes da presença humana nessa área, numa extensão temporal de milhares de anos. Em Portugal, o problema dos fogos florestais assume uma expressividade de particular relevo, colocando-o entre os países do sul da Europa mais atingidos pelo fogo a cada Verão. Reconhecendo o papel das condições meteorológicas nas interacções envolvidas no processo de deflagração e propagação de incêndios, a identificação de condições meteorológicas propícias à ocorrência de incêndios, expressa sob a forma de índices de risco reveste-se de crucial importância para as operações de prevenção e combate aos incêndios, para além da mais-valia que constituem para os restantes sectores da sociedade. Adicionalmente, a detecção e monitorização de ocorrências de incêndios a partir de satélites tem vindo a assumir um papel preponderante, revelando-se fundamental enquanto fonte de alerta e pelas potencialidades que as bases de dados resultantes da observação contínua da Terra pelos satélites oferecem, no que às mais diversas áreas de investigação diz respeito. O presente trabalho explora, nas vertentes da prevenção de incêndios, a utilização de previsões de tempo de alta resolução espacial obtidas com o modelo WRF para a determinação das componentes do sistema FWI canadiano e sua disponibilização operacional. Os resultados apontam para um impacto positivo, em virtude do incremento da resolução espacial, da representação espacial do risco meteorológico de incêndio no território português. Paralelamente, apresentam-se os resultados do desenvolvimento de produtos de detecção de incêndios activos a partir da infra-estrutura de recepção de dados de satélite implementadas na Universidade de Aveiro, destacando a sua adequabilidade para a monitorização e identificação, em tempo quase-real, da ocorrência de fogos florestais em Portugal Continental.

Asymmetric rolling of 5182 aluminium alloy and interstitial free steel sheets

This Ph.D. research focuses on asymmetric rolling (ASR), as an alternative method for improving mechanical responses of aluminium-magnesium alloy and interstitial free (IF) steel regarding industrial requirements. Aluminium alloys are attractive materials in various industries due to their appropriate properties such as low density and corrosion resistance; however, their low formability has limited their applications. As formability of aluminium alloys can be improved through texture development, part of this dissertation is dedicated to producing the desired crystallographic texture with the ASR process. Two types of ASR (i.e. reverse and continuous asymmetric rolling) were investigated. The impact of shear deformation imposed by ASR processes on developing the desirable texture and consequently on mechanical behaviours was observed. The developed shear texture increased the normal and also planar anisotropy. Texture evolution during plastic deformation as well as induced mechanical behaviour were simulated using the “self-consistent” and Taylor models. Interstitial free (IF) steel was the second material selected in this dissertation. Since IF steel is one of the most often used materials in automotive industries it was chosen to investigate the effect of shear deformation through ASR on its properties. Two types of reverse and continuous asymmetric rolling were carried out to deform IF steel sheets. The results of optical microscopy and atomic force microscopy observations showed no significant difference between the grains’ morphology of asymmetric and conventionally rolled samples, whereas the obtained results of transmission electron microscopy indicated that fine and equiaxed dislocation cells were formed through the asymmetric rolling process. This structure is due to imposed shear deformation during the ASR process. Furthermore, the mechanical behaviour of deformed and annealed sheets was evaluated through uniaxial tensile tests. Results showed that at low thickness reductions (18%) the asymmetric rolled sample presented higher stress than that of the conventionally rolled sheet; while for higher thickness reductions (60%) the trend was reversed. The texture analyses indicated that intense rolling texture components which developed through 60% thickness reduction of conventional rolling cause a relatively higher stress; on the contrary the fine structure resulting from ASR appears to be the source of higher stress observed after pre-deformation of 18%.

Novel hardware and software combination for automotive motorsport vehicles based on direct processing of graphical functions

The main motivation for the work presented here began with previously conducted experiments with a programming concept at the time named "Macro". These experiments led to the conviction that it would be possible to build a system of engine control from scratch, which could eliminate many of the current problems of engine management systems in a direct and intrinsic way. It was also hoped that it would minimize the full range of software and hardware needed to make a final and fully functional system. Initially, this paper proposes to make a comprehensive survey of the state of the art in the specific area of software and corresponding hardware of automotive tools and automotive ECUs. Problems arising from such software will be identified, and it will be clear that practically all of these problems stem directly or indirectly from the fact that we continue to make comprehensive use of extremely long and complex "tool chains". Similarly, in the hardware, it will be argued that the problems stem from the extreme complexity and inter-dependency inside processor architectures. The conclusions are presented through an extensive list of "pitfalls" which will be thoroughly enumerated, identified and characterized. Solutions will also be proposed for the various current issues and for the implementation of these same solutions. All this final work will be part of a "proof-of-concept" system called "ECU2010". The central element of this system is the before mentioned "Macro" concept, which is an graphical block representing one of many operations required in a automotive system having arithmetic, logic, filtering, integration, multiplexing functions among others. The end result of the proposed work is a single tool, fully integrated, enabling the development and management of the entire system in one simple visual interface. Part of the presented result relies on a hardware platform fully adapted to the software, as well as enabling high flexibility and scalability in addition to using exactly the same technology for ECU, data logger and peripherals alike. Current systems rely on a mostly evolutionary path, only allowing online calibration of parameters, but never the online alteration of their own automotive functionality algorithms. By contrast, the system developed and described in this thesis had the advantage of following a "clean-slate" approach, whereby everything could be rethought globally. In the end, out of all the system characteristics, "LIVE-Prototyping" is the most relevant feature, allowing the adjustment of automotive algorithms (eg. Injection, ignition, lambda control, etc.) 100% online, keeping the engine constantly working, without ever having to stop or reboot to make such changes. This consequently eliminates any "turnaround delay" typically present in current automotive systems, thereby enhancing the efficiency and handling of such systems.

Regional and urban air quality modelling under a climate change scenario

The better understanding of the interactions between climate change and air quality is an emerging priority for research and policy. Climate change will bring changes in the climate system, which will affect the concentration and dispersion of air pollutants. The main objective of the current study is to assess the impacts of climate change on air quality in 2050 over Portugal and Porto urban area. First, an evaluation and characterization of the air quality over mainland Portugal was performed for the period between 2002 and 2012. The results show that NO2, PM10 and O3 are the critical pollutants in Portugal. Also, the influence of meteorology on O3, NO2 and PM10 levels was investigate in the national main urban areas (Porto and Lisboa) and was verified that O3 has a statistically significant relationship with temperature in most of the components. The results also indicate that emission control strategies are primary regulators for NO2 and PM10 levels. After, understanding the national air quality problems and the influence that meteorology had in the historical air quality levels, the air quality modelling system WRF-CAMx was tested and the required inputs for the simulations were prepared to fulfil the main goal of this work. For the required air quality modelling inputs, an Emission Projections under RCP scenarios (EmiPro-RCP) model was developed to assist the estimation of future emission inventories for GHG and common air pollutants. Also, the current emissions were estimated for Portugal with a higher detailed disaggregation to improve the performance of the air quality simulations. The air quality modelling system WRF/CAMx was tested and evaluated over Portugal and Porto urban area and the results point out that is an adequate tool for the analysis of air quality under climate change. For this purpose, regional simulations of air quality during historical period and future (2045-2050) were conducted with CAMx version 6.0 to evaluate the impacts of simulated future climate and anthropogenic emission projections on air quality over the study area. The climate and the emission projections were produced under the RCP8.5 scenario. The results from the simulations point out, that if the anthropogenic emissions keep the same in 2050, the concentrations of NO2, PM10 and O3 will increase in Portugal. When, besides the climate change effects, is consider the projected anthropogenic emissions the annual mean concentrations of NO2 decrease significantly in Portugal and Porto urban area, and on the contrary the annual mean PM10 concentrations increases in Portugal and decrease in Porto urban area. The O3 results are mainly caused by the reduction of ozone precursors, getting the higher reductions in urban areas and increases in the surrounding areas. All the analysis performed for both simulations for Porto urban area support that, for PM10 and O3, there will be an increase in the occurrence of extreme values, surpassing the annual legislated parameters and having more daily exceedances. This study constitutes an innovative scientific tool to help in future air quality management in order to mitigate future climate change impacts on air quality.

Oxidation and glycoxidation of phosphatidylserine and their effect in immune response : a lipidomic approach

Phosphatidylserine (PS) is a member of the class of phospholipids, and is distributed among all cells of mammalians, playing important roles in diverse biological processes, including blood clotting and apoptosis. When externalized, PS is a ligand that is recognized on apoptotic cells. It has been considered that before externalization PS is oxidized and oxPS enhance the recognition by macrophages receptors, however the knowledge about oxidation of PS is still limited. PS, like others phospholipids, has two fatty acyl chains and one polar head group, in this case is the amino acid serine. The modifications in PS structure can occur by oxidation of the unsaturated fatty acyl chains and by glycation of the polar head group, due to free amine group, thus increasing the susceptibility to oxidative events. The main goal of this work was to characterize and identify oxidized and glycoxidized PS, contributing to the knowledge of the biological role of oxidation products of PS, as well as of glycated PS, in immune and inflammatory processes. To achieve this goal, PS standards (1-palmitoyl-2-oleoyl-sn-glycero-3-phospho- L-serine (POPS), 1,2-dipalmitoyl-sn-glycero-3-phospho-L-serine (DPPS), 1- palmitoyl-2-linoleoyl-sn-glycero-3-phospho-L-serine (PLPS) and 1-palmitoyl-2- arachidonoyl-sn-glycero-3-phospho-L-serine (PAPS)) and glycated PS (PAPS and POPS) were induced to oxidize in model systems, using different oxidant reagents: HO• and 2,2'-azobis-2-methyl-propanimidamide dihydrochloride (AAPH) . The detailed structural characterization of the oxidative products was performed by ESI-MS and MS/MS coupled to separation techniques such as off line TLC-MS and on line LC-MS, in order to obtained better characterization of the larger number of PS and glycated PS oxidation products. The results obtained in this work allowed to identify several oxidation products of PS and glycated PS with modifications in unsaturated fatty acyl chain. Also, oxidation products formed due to structural changes in the serine polar head with formation of terminal acetamide, terminal hydroperoxyacetaldehyde.and terminal acetic acid (glycerophosphacetic acid, GPAA) were identified. The mass spectrometric specific fragmentation pathway of each type of oxidation product was determined using different mass spectrometry approaches. Based on the identified fragmentation pathways, targeted lipidomic analysis was performed to detect oxidation products modified in serine polar head in HaCaT cell line treated with AAPH. The GPAA was detected in HaCaT cells treated with AAPH to induce oxidative stress, thus confirming that modifications in PS polar head is possible to occur in biological systems. Furthermore, it was found that glycated PS species are more prone to oxidative modifications when compared with non glycated PS. During oxidation of glycated PS, besides the oxidation in acyl chains, new oxidation products due to oxidation of the glucose moiety were identified, including PS advanced glycation end products (PSAGES). To investigate if UVA oxidative stress exerted changes in the lipidome of melanoma cell lines, particularly in PS profile, a lipidomic analysis was performed. The lipid profile was obtained using HILIC-LC-MS and GC-MS analysis of the total lipid extracts obtained from human melanoma cell line (SKMEL- 28) after UVA irradiation at 0, 2 and 24 hours. The results did not showed significant differences in PS content. At molecular level, only PS (18:0:18:1) decreased at the moment of irradiation. The most significant changes in phospholipids content occurred in phosphatidylcholines (PC) and phosphatidylinositol (PI) classes, with an increase of mono-unsaturated fatty acid (MUFA), similarly as observed for the fatty acid analysis. Overall, these data indicate that the observed membrane lipid changes associated with lipogenesis after UVA exposure may be correlated with malignant transformations associated with cancer development and progression. Despite of UVA radiation is associated with oxidative damage, in this work was not possible observe oxidation phospholipids. The anti/pro-inflammatory properties of the oxidized PLPS (oxPLPS) versus non-oxidized PLPS were tested on LPS stimulated RAW 264.7 macrophages. The modulation of intracellular signaling pathways such as NF-kB and MAPK cascades by oxPLPS and PS was also examined in this study. The results obtained from evaluation of anti/pro-inflammatory properties showed that neither PLPS or oxPLPS species activated the macrophages. Moreover only oxidized PLS were found to significantly inhibit NO production and iNOS and il1β gene transcription induced by LPS. The analysis at molecular level showed that this was the result of the attenuation of LPS-induced c-Jun-N-terminal kinase (JNK) and p65 NF-kB nuclear translocation. Overall these data suggest that oxPLPS, but not native PLPS, mitigates pro-inflammatory signaling in macrophages, contributing to containment of inflammation during apoptotic cell engulfment. The results obtained in this work provides new information on the modifications of PS, facilitating the identification of oxidized species in complex samples, namely under physiopathologic conditions and also contributes to a better understanding of the role of oxPS and PS in the inflammatory response, in the apoptotic process and other biological functions.

Development of polymeric materials to inhibit bacterial quorum sensing

Bacterial infections are an increasing problem for human health. In fact, an increasing number of infections are caused by bacteria that are resistant to most antibiotics and their combinations. Therefore, the scientific community is currently searching for new solutions to fight bacteria and infectious diseases, without promoting antimicrobial resistance. One of the most promising strategies is the disruption or attenuation of bacterial Quorum Sensing (QS), a refined system that bacteria use to communicate. In a QS event, bacteria produce and release specific small chemicals, signal molecules - autoinducers (AIs) - into the environment. At the same time that bacterial population grows, the concentration of AIs in the bacterial environment increases. When a threshold concentration of AIs is reached, bacterial cells respond to it by altering their gene expression profile. AIs regulate gene expression as a function of cell population density. Phenotypes mediated by QS (QSphenotypes) include virulence factors, toxin production, antibiotic resistance and biofilm formation. In this work, two polymeric materials (linear polymers and molecularly imprinted nanoparticles) were developed and their ability to attenuate QS was evaluated. Both types of polymers should to be able to adsorb bacterial signal molecules, limiting their availability in the extracellular environment, with expected disruption of QS. Linear polymers were composed by one of two monomers (itaconic acid and methacrylic acid), which are known to possess strong interactions with the bacterial signal molecules. Molecularly imprinted polymer nanoparticles (MIP NPs) are particles with recognition capabilities for the analyte of interest. This ability is attained by including the target analyte at the synthesis stage. Vibrio fischeri and Aeromonas hydrophila were used as model species for the study. Both the linear polymers and MIP NPs, tested free in solutions and coated to surfaces, showed ability to disrupt QS by decreasing bioluminescence of V. fischeri and biofilm formation of A. hydrophila. No significant effect on bacterial growth was detected. The cytotoxicity of the two types of polymers to a fibroblast-like cell line (Vero cells) was also tested in order to evaluate their safety. The results showed that both the linear polymers and MIP NPs were not cytotoxic in the testing conditions. In conclusion, the results reported in this thesis, show that the polymers developed are a promising strategy to disrupt QS and reduce bacterial infection and resistance. In addition, due to their low toxicity, solubility and easy integration by surface coating, the polymers have potential for applications in scenarios where bacterial infection is a problem: medicine, pharmaceutical, food industry and in agriculture or aquaculture.

A importância dos processos físicos no controlo da eutrofização em estuários

Muitos dos problemas tróficos existentes nos estuários e zonas costeiras têm origem no excesso de nutrientes, em particular Azoto (N) e Fósforo (P), resultante do sucessivo enriquecimento dos rios durante o seu percurso até ao mar e também de descargas directas nestes sistemas. Nixon (1995) definiu eutrofização como o aumento da quantidade de matéria orgânica disponível na massa de água. Esta é uma definição assumida neste estudo, considerando que a eutrofização é o primeiro, de uma sequência de fenómenos resultantes do sobre-enriquecimento de nutrientes, que inclui a alteração de espécies dominantes e o aumento dos níveis de turbidez da água.

Caracterização química e bioquímica dos frutos de ARBUTUS UNEDO L. e de RUBUS FRUTICOSUS AGG.: contribuição para a sua valorização

Dissertação mest., Agricultura sustentável, Universidade do Algarve, 2007

A coroa, os mestres e os comendadores: as Ordens Militares de Avis de de Santiago (1330 - 1449)

Tese de Doutoramento , História, especialidade de História Medieval, Faculdade de Ciências Humanas e Sociais, Universidade do Algarve, 2006

Arqueologia de emergência em Portugal

Dissertação mest., Teoria e Métodos da Arqueologia, Universidade do Algarve, 2007