Cell-to cell Communication in Pectobacterium wasabiae

Dissertation presented to obtain the PhD degree in Biology

Non-cell autonomous neuroprotective effect of carbon monoxide : microglia-to-neuron communication

RESUMO: A isquémia cerebral é uma das doenças mais predominantes a nivel mundial, sendo uma das principais causas de mortalidade e invalidez. Parte da propagação de dano no cérebro é causado por inflamação descontrolada, causada principalmente por disfunção da microglia. Desta forma, existe a necessidade de tentar desenvolver estratégias para melhor compreender e modular as acções destas células. O monóxido de carbono (CO), é uma molécula endógena com provas dadas como anti-neuroinflamatório em vários modelos. Assim, o principal objectivo do trabalho foi o estudo do CO como um modulador da acção da microglia, com principal foco dado à comunicação entre estas células e neurónios, tentando entender se existe um efeito neuroprotector por inibição da inflamação. Um protocolo de meio condicionado foi estabelecido usando as linhas celulares BV2 e SH-SY5Y, de microglia e neurónio. A molécula CORM-A1, que liberta expontaniamente CO, foi usada como método de entrega da molécula às celulas. Demonstrámos que o pre-tratamento de células BV2 com CORM-A1 gera neuroprotecção já que reduz a morte celular de neurónios SH-SY5Y quando são incubados com meio condicionado de microglia activada em conjunto com o pró-oxidante t-BHP (tert-butil hidroperóxido). Assim, considerámos que o CO promove neuroprotecção ao inibir as acções inflamatórias da microglia. O papel anti-inflamatório da molécula CORM-A1 foi confirmado quando se verificou que pré-tratamento desta molécula em microglia BV2 limita a secreção de TNF-α mas estimula a secreção de IL-10. Por último, a CORM-A1 induziu a expressão do receptor da microglia CD200R1, molécula que participa na comunicação neurónio-microglia e fundamental para a modulação das acções inflamatórias destas últimas. Em suma, o nosso trabalho reforçou as propriedades anti-neuroinflamatórias do CO e uma capacidade de modular viabilidade neuronal através do seu efeito a nível de comunicação célula-célula. ---------------------------- ABSTRACT: Brain ischemia is a widespread disease worldwide, being one of the main causes of mortality and permanent disability. A portion of the damage that ensues following the ischemic event is caused by unrestrained inflammation, which is mainly orchestrated by exacerbated microglial activity. Hence, developing strategies for modulating microglial inflammation is a major concern nowadays. The endogenous molecule carbon monoxide (CO) has been shown to possess anti-neuroinflammatory properties using in vitro and in vivo approaches. Thus, our objective was to study CO as modulator of microglial activity, in particular in what concerns their communication with neurons, by promoting neuronal viability and limiting inflammatory output of activated microglia. A conditioned media strategy was established with BV2 microglia and SH-SY5Y neurons as cell models. CO-releasing molecule A1 (CORM-A1), a compound that releases CO spontaneously, was used as method of CO delivery to cells. We found that CORM-A1 pre-treatment in BV2 cells yields neuroprotective results, as it limits cell death when SH-SY5Y neurons are challenged with conditioned media from LPS-activated microglia and the pro-oxidant t-BHP (tert-butyl-hydroperoxide). Thus, we assumed carbon monoxide promotes neuroprotection via inhibition of microglial inflammation, displaying a non-cell autonomous role. CORM-A1 pre-treatment limited inflammation by inhibiting BV2 secretion of TNF-α and stimulating IL-10 production. These results reinforce that CO’s anti-inflammatory role confers neuroprotection, as the alterations in these cytokines occur concurrently with the increase in SH-SY5Y viability. Finally, we showed for the first time that carbon monoxide promotes the expression of CD200R1, a microglial receptor involved in neuron-glia communication and modulation of microglia inflammation. Further studies are necessary to clarify this role. Altogether, other than just highlighting CO as an anti-inflammatory and neuroprotective molecule, this work set the foundation for disclosing its involvement in cell-to-cell communication.

Geodupa - a geography-driven partial membership algorithm

Trabalho apresentado no âmbito do Mestrado em Engenharia Informática, como requisito parcial para obtenção do grau de Mestre em Engenharia Informática

Bits, Atoms, and Information Sharing: new opportunities for participation

The particular characteristics and affordances of technologies play a significant role in human experience by defining the realm of possibilities available to individuals and societies. Some technological configurations, such as the Internet, facilitate peer-to-peer communication and participatory behaviors. Others, like television broadcasting, tend to encourage centralization of creative processes and unidirectional communication. In other instances still, the affordances of technologies can be further constrained by social practices. That is the case, for example, of radio which, although technically allowing peer-to-peer communication, has effectively been converted into a broadcast medium through the legislation of the airwaves. How technologies acquire particular properties, meanings and uses, and who is involved in those decisions are the broader questions explored here. Although a long line of thought maintains that technologies evolve according to the logic of scientific rationality, recent studies demonstrated that technologies are, in fact, primarily shaped by social forces in specific historical contexts. In this view, adopted here, there is no one best way to design a technological artifact or system; the selection between alternative designs—which determine the affordances of each technology—is made by social actors according to their particular values, assumptions and goals. Thus, the arrangement of technical elements in any technological artifact is configured to conform to the views and interests of those involved in its development. Understanding how technologies assume particular shapes, who is involved in these decisions and how, in turn, they propitiate particular behaviors and modes of organization but not others, requires understanding the contexts in which they are developed. It is argued here that, throughout the last century, two distinct approaches to the development and dissemination of technologies have coexisted. In each of these models, based on fundamentally different ethoi, technologies are developed through different processes and by different participants—and therefore tend to assume different shapes and offer different possibilities. In the first of these approaches, the dominant model in Western societies, technologies are typically developed by firms, manufactured in large factories, and subsequently disseminated to the rest of the population for consumption. In this centralized model, the role of users is limited to selecting from the alternatives presented by professional producers. Thus, according to this approach, the technologies that are now so deeply woven into human experience, are primarily shaped by a relatively small number of producers. In recent years, however, a group of three interconnected interest groups—the makers, hackerspaces, and open source hardware communities—have increasingly challenged this dominant model by enacting an alternative approach in which technologies are both individually transformed and collectively shaped. Through a in-depth analysis of these phenomena, their practices and ethos, it is argued here that the distributed approach practiced by these communities offers a practical path towards a democratization of the technosphere by: 1) demystifying technologies, 2) providing the public with the tools and knowledge necessary to understand and shape technologies, and 3) encouraging citizen participation in the development of technologies.

Which communication strategy should a male luxury (fashion) brand – associated with a luxury car brand – develop, in order to increase its awareness in Europe

This work project explores how a male luxury (fashion) brand (subsidiary) that is associated with a luxury car brand (parent company) should develop its communication strategy in order to increase awareness in Europe. For this purpose a quantitative research was conducted. The aim was to find out whether the company in question had low brand awareness among European luxury consumers. Hereafter, a qualitative research revealed important insights in regard to luxury communication among male luxury consumers. Both the results of the research and the recommendations of luxury experts laid the foundation for the development of a solution-oriented communication strategy. The result of the analysis crystallizes the importance of the shared heritage and the synergistic effects, of which the subsidiary should make vast use when communicating.

Development of web services to help risk management of construction projects

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Engenharia Electrotécnica e de Computadores

Heterogeneous sensor database framework for the sensor observation service: integrating remote and in-situ sensor observations at the database backend

Dissertation submitted in partial fulfillment of the requirements for the Degree of Master of Science in Geospatial Technologies.

A technology market transfer for the Zonadvanced video technology

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

Investigation of the regulation mechanisms for bioplastics production from industrial residues

Dissertação para obtenção do Grau de Mestre em Biotecnologia

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

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Management from the NOVA – School of Business and Economics

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.

How do Facebook users interact and what are the underlying reasons that trigger these interactions

This research was conducted to understand how Facebook users interact and the underlying reasons for doing so with a focus on one-to-mass communication interactions. Different methods and sources were used to generate accurate and valid insights. It was discovered that liking, groups, commenting, events and sharing are essential interactions, whereby liking, commenting and sharing were investigated in more detail. This investigations proves that emotions do trigger these three interactions; The most influencing emotions are Surprise/Wonder, Deep Respect/ Impressiveness and Fun/Joy. Moreover a variety of specific factors that trigger each of the interactions are revealed.

Video metadata extraction in a videoMail system

Currently the world swiftly adapts to visual communication. Online services like YouTube and Vine show that video is no longer the domain of broadcast television only. Video is used for different purposes like entertainment, information, education or communication. The rapid growth of today’s video archives with sparsely available editorial data creates a big problem of its retrieval. The humans see a video like a complex interplay of cognitive concepts. As a result there is a need to build a bridge between numeric values and semantic concepts. This establishes a connection that will facilitate videos’ retrieval by humans. The critical aspect of this bridge is video annotation. The process could be done manually or automatically. Manual annotation is very tedious, subjective and expensive. Therefore automatic annotation is being actively studied. In this thesis we focus on the multimedia content automatic annotation. Namely the use of analysis techniques for information retrieval allowing to automatically extract metadata from video in a videomail system. Furthermore the identification of text, people, actions, spaces, objects, including animals and plants. Hence it will be possible to align multimedia content with the text presented in the email message and the creation of applications for semantic video database indexing and retrieving.

A importância da comunicação estratégica na alteração de comportamentos na sociedade : o caso Sociedade Ponto Verde

A comunicação estratégica poderá ser entendida como a comunicação alinhada com a estratégia global de uma organização para alcançar o seu posicionamento estratégico. Ou seja, para alcançar o sucesso, uma organização precisa de ter uma comunicação consistente com a sua identidade e, ao mesmo tempo, adequada ao seu público-alvo e ao tipo de sector onde se insere. Só desta forma consegue conquistar a confiança e fidelidade dos consumidores, essenciais para o alcance dos objectivos. Em relação à alteração comportamentos, é essencial que as empresas costruam uma estratégia de comunicação coerente e consistente, adaptando as mensagens e os públicos-alvo escolhidos. Neste âmbito da alteração de comportamentos, estudou-se neste trabalho a Sociedade Ponto Verde (SPV), uma associação portuguesa sem fins lucrativos, que actua no âmbito da gestão e reciclagem de resíduos de embalagens. Procurou estudar-se a evolução da sua comunicação ao longo dos anos e a forma como adaptou as suas mensagens, de forma a conseguir incutir na sociedade portuguesa a importância do comportamento de reciclagem de resíduos. Neste estudo, procedeu-se a uma análise individualizada de cada uma das campanhas de comunicação da SPV e, posteriormente, aplicou-se um inquérito por questionário, para verificar a eficácia das mesmas campanhas perante o público-alvo. A metodologia utilizada neste trabalho consistiu na distribuição de um inquérito por questionário online, que permitiu recolher uma amostra probabilística por conveniência. Em seguida, recolhidos e tratados os dados referentes às 482 respostas recebidas, utilizou-se o programa SPSS (Stastitical Package for the Social Sciences) para analisar, tratar e discutir os resultados obtidos. Estes resultados demonstraram que, de facto, não houve uma grande percentagem de inquiridos que se lembram das campanhas da SPV. Os resultados são positivos e não desmentem a qualidade e coerência das mensagens, mas a verdade é que a percentagem de indivíduos que não se lembra de algumas campanhas chega a ser elevada (em especial nas campanhas iniciais da organização, quando existia falta de uma estratégia coerente). Justifica-se este facto, em parte, pelo excesso de informação e estímulos a que as pessoas estão sujeitas, pelo que a informação que retêm não fica para sempre, acabando por dar lugar a outras informações. Os indivíduos são inconscientemente selectivos com a informação que processam e guardam, pois não seria possível guardar toda a informação que lhes é transmitida. No que diz respeito à comunicação estratégica que visa alterar uma atitude e, posteriormente, um comportamento, conclui-se que, no geral, o comportamento das pessoas inquiridas aparenta ter sido influenciado pela comunicação da SPV, caso contrário, as respostas afirmativas quanto à reciclagem e de concordância com algumas das afirmações teriam sido inferiores. Ou seja, mesmo reconhecendo o carácter um pouco enviesado da amostra (não deverá ser considerada representativa da população portuguesa), é possível dizer que a estratégia de comunicação da SPV (em especial a partir de 2005) foi eficiente. Palavras-

A lightweight distributed super peer election algorithm for unstructured dynamic P2P systems

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Engenharia Electrotécnica e de Computadores