An intertemporal pricing model for CO2 allowances: The impact of the clean development mechanism

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

Avatars and the «Imitation Game» – can machines smile?

Considering Alan Turing’s challenge in «Computing Machinery and Intelligence» (1950) – can machines play the «imitation game»? – it is proposed that the requirements of the Turing test are already implicitly being used for checking the credibility of virtual characters and avatars. Like characters, Avatars aim to visually express emotions (the exterior signs of the existence of feeling) and its creators have to resort to emotion codes. Traditional arts have profusely contributed for this field and, together with the science of anatomy, shaped the grounds for current Facial Action Coding System (FACS) and their databases. However, FACS researchers have to improve their «instruction tables» so that the machines will be able, in a near future, to be programmed to carry out the operation of recognizing human expressions (face and body) and classify them adequately. For the moment, the reproductions have to resort to the copy of real life expressions, and the presente smile of avatars comes from mirroring their human users.

Exploratory analysis of OpenStreetMap for land use classification

In the last years, volunteers have been contributing massively to what we know nowadays as Volunteered Geographic Information. This huge amount of data might be hiding a vast geographical richness and therefore research needs to be conducted to explore their potential and use it in the solution of real world problems. In this study we conduct an exploratory analysis of data from the OpenStreetMap initiative. Using the Corine Land Cover database as reference and continental Portugal as the study area, we establish a possible correspondence between both classification nomenclatures, evaluate the quality of OpenStreetMap polygon features classification against Corine Land Cover classes from level 1 nomenclature, and analyze the spatial distribution of OpenStreetMap classes over continental Portugal. A global classification accuracy around 76% and interesting coverage areas’ values are remarkable and promising results that encourages us for future research on this topic.

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.

Melanosome transfer between melanocytes and keratinocytes

RESUMO: A pele é o maior órgão do corpo humano e a sua pigmentação é essencial para a sua coloração e proteção contra os efeitos nocivos da radiação ultravioleta (UV). A pigmentação da pele resulta essencialmente de três processos: a síntese e o armazenamento de melanina pelos melanócitos, em organelos especializados denominados melanossomas; o transporte dos melanossomas dentro dos melanócitos; e finalmente, a transferência dos melanossomas para os queratinócitos adjacentes. Nos queratinócitos, a melanina migra para a região perinuclear apical da célula para formar um escudo protetor,responsável pela proteção do DNA dos danos causados pela radiação UV. Os melanócitos estão localizados na camada basal da epiderme e contactam com 30-40 queratinócitos. Em conjunto, estas células formam a “unidade melano-epidérmica”. Apesar dos processos de síntese e transporte de melanina nos melanócitos estarem bastante bem caracterizados, os mecanismos moleculares subjacentes à transferência inter-celular de melanina são menos conhecidos e ainda controversos. Dados preliminares obtidos pelo nosso grupo, que se basearam na observação de amostras de pele humana por microscopia electrónica, indicam que a forma predominante de transferência de melanina na epiderme consiste na exocitose dos melanossomas pelos melanócitos e subsequente endocitose da melanina por queratinócitos. Para além disso sabe-se que as proteínas Rab, que controlam o tráfego membranar, estão envolvidas em várias etapas de pigmentação da pele, nomeadamente na biogénese e no transporte de melanina. Assim, dado o seu papel fundamental nestes processos, questionámo-nos sobre o seu envolvimento na transferência de melanina. Com este trabalho, propomo-nos a expandir o conhecimento atual sobre a transferência de melanina na pele, através do estudo detalhado dos seus mecanismos moleculares, identificando as proteínas Rab que regulam o processo. Pretendemos também confirmar o modelo de exo/endocitose como sendo o mecanismo principal de transferência de melanina. Primeiro, explorámos a regulação da secreção de melanina pelos melanócitos e analisámos o papel de proteínas Rab neste processo. Os resultados foram obtidos recorrendo a um método in vitro, desenvolvido previamente no laboratório, que avalia a quantidade de melanina segregada para o meio de cultura por espectrofotometria, e ainda por microscopia, contando o número de melanossomas transferidos para os queratinócitos. Através de co-culturas de melanócitos e queratinócitos, verificou-se que os queratinócitos estimulam a libertação de melanina dos melanócitos para o meio extra-celular, bem como a sua transferência para os queratinócitos. Além disso, a proteína Rab11b foi identificada como um regulador da exocitose de melanina e da sua transferência para os queratinócitos. De facto, a diminuição da expressão de Rab11b em melanócitos provocou a redução da secreção de melanina estimulada por queratinócitos, bem como da transferência desta. Em segundo lugar, para complementar o nosso estudo, centrámos a nossa investigação na internalização de melanina por queratinócitos. Especificamente, usando uma biblioteca de siRNA, explorámos o envolvimento de proteínas Rab na captação de melanina por queratinócitos. Como primeira abordagem, usámos esferas fluorescentes como substituto de melanina, avaliando os resultados por citometria de fluxo. No entanto, este método revelou-se ineficaz uma vez que a internalização destas esferas é independente do recetor PAR-2 (recetor 2 ativado por protease), que foi previamente descrito como essencial na captação de melanina por queratinócitos Posteriormente, foi desenvolvido um novo protocolo de endocitose baseado em microscopia, usando melanossomas sem a membrana envolvente (melanocores) purificados do meio de cultura de melanócitos, incluindo um programa informático especialmente desenhado para realizar uma análise semi-automatizada. Após internalização, os melanocores acumulam-se na região perinuclear dos queratinócitos, em estruturas que se assemelham ao escudo supranuclear observado na pele humana. Seguidamente, o envolvimento do recetor PAR-2 na captação de melanocores por queratinócitos foi confirmado, utilizando o novo protocolo de endocitose desenvolvido. Para além disso, a necessidade de quatro proteínas Rab foi identificada na internalização de melanocores por queratinócitos. A redução da expressão de Rab1a ou Rab5b em queratinócitos diminuiu significativamente o nível de internalização de melanocores, enquanto o silenciamento da expressão de Rab2a ou Rab14 aumentou a quantidade de melanocores internalizados por estas células. Em conclusão, os resultados apresentados corroboram as observações anteriores, obtidas em amostras de pele humana, e sugerem que o mecanismo de transferência predominante é a exocitose de melanina pelos melanócitos, induzida por queratinócitos, seguida por endocitose pelos queratinócitos. A pigmentação da pele tem implicações tanto ao nível da cosmética, como ao nível médico, relacionadas com foto-envelhecimento e com doenças pigmentares. Assim sendo, ao esclarecer quais os mecanismos moleculares que regulam a transferência de melanina na pele, este trabalho pode conduzir ao desenvolvimento de novas estratégias para modular a pigmentação da pele.----------------ABSTRACT: Skin pigmentation is achieved through the highly regulated production of the pigment melanin in specialized organelles, termed melanosomes within melanocytes. These are transported from their site of synthesis to the melanocyte periphery before being transferred to keratinocytes where melanin forms a supra-nuclear cap to protect the DNA from UVinduced damage. Together, melanocytes and keratinocytes form a functional complex, termed “epidermal-melanin unit”, that confers color and photoprotective properties to the skin. Skin pigmentation requires three processes: the biogenesis of melanin; its intracelular transport within the melanocyte to the cell periphery; and the melanin transfer to keratinocytes. The first two processes have been extensively characterized. However, despite significant advances that have been made over the past few years, the mechanisms underlying inter-cellular transfer of pigment from melanocytes to keratinocytes remain controversial.Preliminary studies from our group using electron microscopy and human skin samples found evidence for a mechanism of coupled exocytosis-endocytosis. Rab GTPases are master regulators of intracellular trafficking and have already been implicated in several steps of skin pigmentation. Thus, we proposed to explore and characterize the molecular mechanisms of melanin transfer and the role of Rab GTPases in this process. Moreover, we investigated whether the exo/endocytosis model is the main mechanism of melanin transfer. We first focused on melanin exocytosis by melanocytes. Then, we started to investigate the key regulatory Rab proteins involved in this step by establishing an in vitro tissue culture model of melanin secretion. Using co-cultures of melanocytes and keratinocytes, we found that keratinocytes stimulate melanin release and transfer. Moreover, depletion of Rab11b decreases keratinocyte-induced melanin exocytosis by melanocytes. In order to determine whether melanin exocytosis is a predominant mechanism of melanin transfer, the amount of melanin transferred to keratinocytes was then assayed in conditions where melanin exocytosis was inhibited. Indeed, Rab11b depletion resulted in a significant decrease in melanin uptake by keratinocytes. Taken together, these observations suggest that Rab11b mediates melanosome exocytosis from melanocytes and transfer to keratinocytes. To complement and extend our study, we of melanin by keratinocytes. Thus, we aimed to explore the effect of depleting Rab GTPases on melanin uptake and trafficking within keratinocytes. As a first approach, we used fluorescent microspheres as a melanin surrogate. However, the uptake of microspheres was observed to be independent of PAR-2, a receptor that is required for melanin uptakecentred our attention in the internalization of melanin by keratinocytes. Thus, we aimed to explore the effect of depleting Rab GTPases on melanin uptake and trafficking within keratinocytes. As a first approach, we used fluorescent microspheres as a melanin surrogate. However, the uptake of microspheres was observed to be independent of PAR-2, a receptor that is required for melanin uptake.Therefore, we concluded that microspheres were uptaken by keratinocytes through a different pathway than melanin. Subsequently, we developed a microscopy-based endocytosis assay using purified melanocores (melanosomes lacking the limiting membrane) from melanocytes, including a program to perform a semi-automated analysis. Melanocores are taken up by keratinocytes and accumulate in structures in the perinuclear area that resemble the physiological supranuclear cap observed in human skin. We then confirmed the involvement of PAR-2 receptor in the uptake of melanocores by keratinocytes, using the newly developed assay. Furthermore, we identified the role of four Rab GTPases on the uptake of melanocores by keratinocytes. Depletion of Rab1a and Rab5b from keratinocytes significantly reduced the uptake of melanocores, whereas Rab2a, and Rab14 silencing increased the amount the melanocores internalized by XB2 keratinocytes. In conclusion, we present evidence supporting keratinocyte-inducedmelanosome exocytosis from melanocytes, followed by endocytosis of the melanin core by keratinocytes as the predominant mechanism of melanin transfer in skin. Although advances have been made, there is a need for more effective and safer therapies directed at pigmentation disorders and also treatments for cosmetic applications. Hence, the understanding of the above mechanisms of skin pigmentation will lead to a greater appreciation of the molecular machinery underlying human skin pigmentation and could interest the pharmaceutical and cosmetic industries.

As vias de sucessão no estado português da Índia (1524-1581)

A presente dissertação procura esclarecer as origens das “vias de sucessão”, o respectivo modelo de funcionamento e qual a sua importância no sistema político do Estado Português da Índia, desde a data da sua implementação por D. João III, em 1524, até ao início da dinastia Filipina. Na base deste sistema estava o pressuposto de que um governador do Estado Português da Índia podia falecer durante o seu mandato, pelo que era necessário o rei assegurar, a priori, a sua hipotética sucessão, sem deixar a mesma nas mãos dos nobres presentes na Índia, que, provavelmente, nunca conseguiriam chegar a um consenso sobre quem deveria assumir o poder, por aspirarem ao mesmo. Procuramos, assim, compreender de que modo este sistema funcionava, não sem antes recuarmos ao reinado de D. Manuel I, para tentarmos indagar de que forma o Venturoso asseguraria uma hipotética sucessão de um governador do Estado Português da Índia, caso esta tivesse sido necessária. Deste modo, tentaremos perceber se o sistema criado em 1524 resultou de uma ideia inovadora de D. João III ou se, pelo contrário, este sistema já existia anteriormente e em outros espaços, limitando-se este monarca a transplantar o mesmo para a Índia. Com a questão do surgimento deste sistema de sucessão esclarecida, procedermos, então, à análise do funcionamento do mesmo e tentar-se-á concluir se este sistema se revelou, a longo prazo, eficaz para assegurar a sucessão no governo do Estado Português da Índia. Partindo de uma análise comparativa dos sistemas de sucessão existentes nas outras zonas do Império Português, no momento da entronização de D. João III, em 1521, pretendemos explicar de que forma o sistema das vias de sucessão surgiu na Índia e se desenvolveu e contribuir, assim, para um maior conhecimento do funcionamento da administração do Estado Português da Índia.

Analyzing Geospatial patterns of syrian refugee flows in southeastern Turkey by use of remote sensing and complementary data

Crisis-affected communities and global organizations for international aid are becoming increasingly digital as consequence geotechnology popularity. Humanitarian sector changed in profound ways by adopting new technical approach to obtain information from area with difficult geographical or political access. Since 2011, turkey is hosting a growing number of Syrian refugees along southeastern region. Turkish policy of hosting them in camps and the difficulty created by governors to international aid group expeditions to get information, made such international organizations to investigate and adopt other approach in order to obtain information needed. They intensified its remote sensing approach. However, the majority of studies used very high-resolution satellite imagery (VHRSI). The study area is extensive and the temporal resolution of VHRSI is low, besides it is infeasible only using these sensors as unique approach for the whole area. The focus of this research, aims to investigate the potentialities of mid-resolution imagery (here only Landsat) to obtain information from region in crisis (here, southeastern Turkey) through a new web-based platform called Google Earth Engine (GEE). Hereby it is also intended to verify GEE currently reliability once the Application Programming Interface (API) is still in beta version. The finds here shows that the basic functions are trustworthy. Results pointed out that Landsat can recognize change in the spectral resolution clearly only for the first settlement. The ongoing modifications vary for each case. Overall, Landsat demonstrated high limitations, but need more investigations and may be used, with restriction, as a support of VHRSI.

Characterization of molecular damage induced by UV photons and carbon ions on biomimetic heterostructures

The study of the effect of radiation on living tissues is a rather complex task to address mainly because they are made of a set of complex functional biological structures and interfaces. Particularly if one is looking for where damage is taking place in a first stage and what are the underlying reaction mechanisms. In this work a new approach is addressed to study the effect of radiation by making use of well identified molecular hetero-structures samples which mimic the biological environment. These were obtained by assembling onto a solid support deoxyribonucleic acid (DNA) and phospholipids together with a soft water-containing polyelectrolyte precursor in layered structures and by producing lipid layers at liquid/air interface with DNA as subphase. The effects of both ultraviolet (UV) radiation and carbon ions beams were systematically investigated in these heterostructures, namely damage on DNA by means vacuum ultraviolet (VUV), infrared (IR), X-Ray Photoelectron (XPS) and impedance spectroscopy. Experimental results revealed that UV affects furanose, PO2-, thymines, cytosines and adenines groups. The XPS spectrometry carried out on the samples allowed validate the VUV and IR results and to conclude that ionized phosphate groups, surrounded by the sodium counterions, congregate hydration water molecules which play a role of UV protection. The ac electrical conductivity measurements revealed that the DNA electrical conduction is arising from DNA chain electron hopping between base-pairs and phosphate groups, with the hopping distance equal to the distance between DNA base-pairs and is strongly dependent on UV radiation exposure, due loss of phosphate groups. Characterization of DNA samples exposed to a 4 keV C3+ ions beam revealed also carbon-oxygen bonds break, phosphate groups damage and formation of new species. Results from radiation induced damage carried out on biomimetic heterostructures having different compositions revealed that damage is dependent on sample composition, with respect to functional targeted groups and extent of damage. Conversely, LbL films of 1,2-dipalmitoyl-sn-Glycero-3-[Phospho-rac-(1-glycerol)] (Sodium Salt) (DPPG) liposomes, alternated with poly(allylamine hydrochloride) (PAH) revealed to be unaffected, even by prolonged UV irradiation exposure, in the absence of water molecules. However, DPPG molecules were damaged by the UV radiation in presence of water with cleavage of C-O, C=O and –PO2- bonds. Finally, the study of DNA interaction with the ionic lipids at liquid/air interfaces revealed that electrical charge of the lipid influences the interaction of phospholipid with DNA. In the presence of DNA in the subphase, the effects from UV irrladiation were seen to be smaller, which means that ionic products from biomolecules degradation stabilize the intact DPPG molecules. This mechanism may explain why UV irradiation does not cause immediate cell collapse, thus providing time for the cellular machinery to repair elements damaged by UV.

Optimization of export support systems: How to promote exports from Portugal to Germany in the area of information technology and electronics

This work project focuses on developing new approaches which enhance Portuguese exports towards a defined German industry sector within the information technology and electronics fields. Firstly and foremost, information was collected and a set of expert and top managers’ interviews were performed in order to acknowledge the demand of the German market while identifying compatible Portuguese supply capabilities. Among the main findings, Industry 4.0 presents itself as a valuable opportunity in the German market for Portuguese medium sized companies in the embedded systems area of expertise for machinery and equipment companies. In order to achieve the purpose of the work project, an embedded systems platform targeting machinery and equipment companies was suggested as well as it was developed several recommendations on how to implement it. An alternative approach for this platform was also considered within the German market namely the eHealth sector having the purpose of enhancing the current healthcare service provision.

Optimization of export support systems

The present paper is a personal reflection on a work project carried out to promote exports from Portugal to Germany in the IT area, under consideration of the deliverables required by the clients CCILA and Anetie. The project outcome approaches the fact that the majority of the Portuguese market players has disadvantages in size and does rarely coordinate activities among each other, which hinders them to export successfully on a broad scale. To bring together Portuguese delivery potential and German market demand, expert interviews were conducted. Based on the findings, a concept was developed to overcome the domestic collaboration issues in order to strengthen the national exports in the identified sector - embedded systems implementation services for machinery and equipment companies.

Study of in vivo interactions between penicillin-binding proteins of Staphylococcus aureus

Staphylococcus aureus (S. aureus) is a major human pathogen that has acquired resistance to practically all classes of β-lactam antibiotics, being responsible of Multidrug resistant S. aureus (MRSA) associated infections both in healthcare (HA-MRSA) and community settings (CA-MRSA). The emergence of laboratory strains with high-resistance (VRSA) to the last resort antibiotic, vancomycin, is a warning of what is to come in clinical strains. Penicillin binding proteins (PBPs) target β-lactams and are responsible for catalyzing the last steps of synthesis of the main component of cell wall, peptidoglycan. As in Escherichia coli, it is suggested that S. aureus uses a multi-protein complex that carries out cell wall synthesis. In the presence of β-lactams, PBP2A and PBP2 perform a joint action to build the cell wall and allow cell survival. Likewise, PBP2 cooperates with PBP4 in cell wall cross-linking. However, an actual interaction between PBP2 and PBP4 and the location of such interaction has not yet been determined. Therefore, investigation of the existence of a PBP2-PBP4 interaction and its location(s) in vivo is of great interest, as it should provide new insights into the function of the cell wall synthesis machinery in S. aureus. The aim of this work was to develop Split-GFPP7 system to determine interactions between PBP2 and PBP4. GFPP7 was split in a strategic site and fused to proteins of interest. When each GFPP7 fragment, fused to proteins, was expressed alone in staphylococcal cells, no fluorescence was detectable. When GFPP7 fragments fused to different peptidoglycan synthesis (PBP2 and PBP4) or cell division (FtsZ and EzrA) proteins were co-expressed together, fluorescent fusions were localized to the septum. However, further analysis revealed that this positive result is mediated by GFPP7 self-association. We then interpret the results in light of such event and provide insights into ways of improving this system.

Hospital-empresa : crónica de um nascimento retardado : Políticas de saúde : Em foco

RESUMO - A gestão empresarial dos hospitais é uma velha aspiração do sistema e dos profissionais da saúde em Portugal. Já o Estatuto Hospitalar de 1968 previa a organização e a gestão dos hospitais «em termos de gestão empresarial». A Lei de Bases da Saúde, de 1990, relembrava que a administração das unidades de saúde deveria obedecer a «regras de gestão empresarial». O Hospital Fernando da Fonseca, criado desde 1991, foi objecto de concessão de gestão por contrato, precedendo concurso público, a uma entidade privada, em 1995. Em 1997, o relatório do Grupo de Trabalho sobre o Estatuto Jurídico do Hospital recomendava a adopção da figura de instituto público com natureza empresarial, adequada autonomia de gestão e forte responsabilidade, podendo regular-se, em alguns domínios, por normas de direito privado. Em 1998 foi criado o Hospital de São Sebastião, em Santa Maria da Feira, com formas inovadoras de gestão, utilizando meios de gestão maleáveis. Em 1999 foi criada a Unidade Local de Saúde de Matosinhos, englobando não apenas o Hospital de Pedro Hispano, naquela cidade, mas também os quatro centros de saúde da sua área de atracção. Em 2001 foi criado o Hospital do Barlavento Algarvio, em moldes semelhantes aos do Hospital de São Sebastião. Os restantes hospitais públicos mantiveram a estrutura e regras de funcionamento convencionais. Observa-se que o modelo de gestão convencional do hospital público tem hoje consequências desfavoráveis para os cidadãos, para os profissionais que nele trabalham e também para o sistema de saúde no seu conjunto. Em 2002, uma nova lei alterou disposições da Lei de Bases da Saúde de 1990 e aprovou um novo regime jurídico de gestão hospitalar. De acordo com ele, a rede de prestação de cuidados de saúde passou a integrar vários modelos de hospitais: hospitais SPA, hospitais EPE, hospitais SA, clínicas privadas com ou sem nome de hospital, instituições e serviços geridos por entidades públicas ou privadas, mediante contrato de gestão e hospitais PPP. Analisam-se os ganhos introduzidos pelo modelo inovador de hospital SA, no que respeita ao estatuto, dotação de capital, poderes especiais, regras de controlo financeiro, regimes laborais, órgãos sociais, instrumentos de gestão e direcção técnica. Finalmente, antecipa-se um quadro analítico de oportunidades e riscos sobre este modelo. As críticas têm-se concentrado sobre a estratégia de mudança e sobre o mecanismo de escolha dos dirigentes e das respectivas chefias intermédias. Em relação à estratégia, conclui-se ser a questão mais empírica do que conceptual. Em relação à forma de identificação dos dirigentes, recomenda-se o acompanhamento crítico da experiência, salientando-se, a par do que ela pode trazer de positivo, os riscos de partidarização e instabilidade.

Afonso de Albuquerque e a primeira expedição portuguesa ao Mar Vermelho (1513)

Este artigo pretende analisar a primeira expedição portuguesa ao Mar Vermelho, capitaneada pelo governador Afonso de Albuquerque, em 1513. Quinhentos anos depois desta viagem, reavaliamos a política manuelina para o Mar Roxo, observando o ataque à cidade de Adém e a consecutiva entrada naquele mar. Através da correspondência de Albuquerque e da cronística da época, tentamos compreender os motivos da derrota no assalto àquela cidade portuária e a importância desta expedição na acção daquele governador e na estratégia imperial de D. Manuel I.