M&A : Deutsche Post DHL and Austrian Post

The goal of this thesis is to assess how would a potential deal between Deutsche Post DHL (also referred to as DPDHL or simply DHL) and Austrian Post (also referred to as Österreichische Post or simply OP) result in terms of value creation for both companies. In an era of abrupt economical and technological changes, the Transportation and Logistics sector is one of the more affected, and thus the urge to constantly adapt. The top capabilities of DHL as well as geographic proximity with OP could leverage very important efficiencies of many kinds. Common practices on valuations are used to assess the individual value of both companies, which serve as basis to project future trends based in case a merger would take place. Decreasing traditional Mail volume and opportunities in Express deliveries due to e-commerce are found to be the main factors affecting the value of these companies. Following the cautious estimation of resulting synergies, an all-cash offer of €39.2 per share was proposed, resulting in a premium on average share price of 25.1% to OP’s shareholders

A revamped business strategy for the health business segment of EA: status quo and strategic alignment

EuropAssistance is currently facing a decisive moment to diversify its business model which is very reliant on the auto business line. The purpose of this part of the paper is to analyse the status quo and strategic alignmentof the firm. An extensive literature review, a broad range of interviews with all relevant external and internal stakeholders as well as a global industry benchmarking had been conducted to identify the market needs and potential solutions

Understanding chromatin dynamics during antigenic variation in trypanosoma brucei

Tese de doutoramento, Ciências Biomédicas (Microbiologia e Parasitologia), Universidade de Lisboa, Faculdade de Medicina, 2016African trypanosomiasis is a disease restricted to sub-Saharian Africa and it assumes two forms: Human African trypanosomiasis (HAT or sleeping sickness) or Animal African trypanosomiasis (AAT or n’gana), both being fatal if untreated. While HAT causes an estimated number of 15000 new cases each year and 70 million people are at risk, AAT causes an enormous economic burden due to livestock elimination [1]. HAT is caused by the unicellular Trypanosoma brucei parasite, which is transmitted by the tsetse transmitting vector. Upon a tsetse blood meal, parasites are delivered into a mammalian host where they proliferate in the bloodstream, lymphatic system and inside preferred tissues, such as the adipose tissue [2]. The parasites can progressively migrate to the central nervous system, leading to neurological disorders, coma and death [3]. Each Trypanosoma brucei parasite expresses a single type of Variant Surface Glycoprotein (VSG) at its cell surface. To avoid being eliminated by the host immune system, parasites use antigenic variation, which consists in periodically changing to a new VSG coat, obligating the immune system to reinitiate a new adaptive response and generate new antibodies. VSG genes are transcribed by RNA polymerase I (Pol I) from specialized subtelomeric loci termed Bloodstream Expression Sites (BESs) [4]. Although there are around 15 BESs in the genome, only one is active at any given time, thus ensuring VSG monoallelic expression. Replacement of the VSG coat starts in the nucleus by changing the actively transcribed VSG. This process takes place through: i) switching by recombination, which involves DNA recombination between the active and a silent BES encompassing the VSG gene or by copying a VSG sequence from archival copies present in the genome into the active BES [5, 6]; ii) in situ switching, where the active BES becomes silenced with the concomitant activation of a silent BES [7, 8]. VSG in situ switching is very poorly understood. We do not know the key players or the sequence of events that regulate it. As the actively transcribed BES possesses a very open chromatin conformation relative to transcriptionally silent BESs [9, 10], we proposed to understand how chromatin and transcription interweave during an in situ switching. Another event in which the active BES becomes silenced is during differentiation from bloodstream to procyclic (present in the tsetse) forms [11]. In this situation, we observed that transcriptional silencing precedes chromatin changes. Thus, we postulated that during an in situ switching event transcription is probably also halted prior to closing chromatin structure. We chose to use a tetracycline-inducible BES transcriptional silencing system from the Horn lab that induces VSG silencing [12], resulting in an increase of the switching frequency to 8%. Blockage of transcription was confirmed by the absence of Pol I in the active BES and led to a rapid decrease in transcript levels of this locus within the first 8 hr after BES silencing. Despite such significant changes at the transcriptional level, chromatin of the previously active BES remained in an open conformation. We hypothesized that the cell-cycle was necessary to induce chromatin remodeling as in Pol I-transcribed ribosomal DNA (rDNA) genes in yeast, but this revealed unfruitful. We observed that TDP1 (Trypanosome DNA-binding protein 1), an essential high mobility group box protein [13, 14], remained in the active BES even after BES silencing had been triggered. Depletion of TDP1 in these conditions led to closure of chromatin indicating that TDP1 is a key factor in stabilizing open chromatin conformation under transcriptional switching. We showed that during in situ switching, cells probe more than one silent BES, inclusively expressing other VSGs. We determined that after 24 hr of BES silencing, roughly half of the cells were committed to switch while the remaining could revert to transcribe the initial BES. Interestingly, cells that at 24 hr of BES silencing were not probing a specific BES (and VSG) did not switch to that BES. Overall, these results led us to propose a model in which under an in situ switching, transcription of the active BES is halted but its chromatin is maintained in an open conformation by TDP1. This allows parasites to probe silent BES to make a decision of switching or returning to the initial BES. The importance of TDP1 in Pol I transcription and in the switching process led us to ask if TDP1 overexpression could interfere with VSG monoallelic expression. Using a TDP1-overexpressing cell-line, we observed that genome-wide chromatin becomes more open (at different extent) except in the active BES and rDNA genes, suggesting that these two loci already have the chromatin fully open. Furthermore, the mRNA transcript levelsonly increased in silent BES, MES (Metacyclic Expression Sites) and procyclin loci confirming the role of TDP1 as a Pol I transcription facilitator [14]. Importantly, we observed the expression of a silent VSG upon TDP1 overexpression confirming the disruption of monoallelic expression. Overall, this dissertation enlightens the relevance of chromatin during an in situ switching and elucidates the roles of TDP1 as key factor for antigenic variation

A cidade e a música: a escola de música do conservatório nacional

É reconhecida na cidade contemporânea uma dispersão urbana e uma falta de sentido “humano”, esta aparece dividida entre o que são as áreas formais e as áreas informais resultando num limite que estimula a exclusão social. Neste limite, surge espaço público, o espaço entre e que congrega os opostos. Na procura de uma humanização da cidade e diluição dos limites delineados considera-se necessária, tal como Jorge Jáuregui aponta, a existência de estratégias de intervenção que conciliem as necessidades e interesses locais e da cidade a fim de uma cidade contemporânea conectada1, como Aldo Van Eyck defende “(…) a cidade como uma grande casa (…)”2. A música é considerada como uma possível ferramenta de intervenção na cidade com qualidades sociais e urbanas. As suas qualidades sociais são reconhecidas já desde Platão e Aristóteles pelo seu poder de formação e transformação do indivíduo e utilizadas em projetos inseridos em comunidades desfavorecidas que beneficiam do poder da música e da educação musical a fim de se conseguirem resultados a nível social, podendo-se considerar um novo paradigma da educação musical, as “orquestras de rua”. Quanto às suas qualidades urbanas, é defendido que a música é uma importante ferramenta para a criação de “lugar”, promovendo a interação social e diferentes perceções do espaço, qualidades reconhecidas em projetos de arquitetura de intervenção que utilizam a música como ferramenta de revitalização e regeneração de espaços da cidade. Procura-se uma possível articulação destas qualidades a fim de estas se beneficiarem mutuamente e de “a cidade ser uma grande casa”3.It is recognized in the contemporary city a urban dispersion and a lack of “human” meaning, the city is divided between the formal and the informal areas resulting in a borderline that stimulates the social exclusion. In this borderline line, comes public space, the space between and that brings together the opposites. In order to the humanization and the thinning of the limits is considered necessary, as Jorge Jáuregui points, a existence of intervention strategies that balance the local and city needs and interests for a connected contemporary city4, as Aldo Van Eyck says “(…) a city like a large house (…)5. Music is considered a possible tool of intervention in the city, with social and urban qualities. Its social qualities are recognized since Plato and Aristotle for its power of formation and transformation of the individual and used in projects placed in deprived communities that benefit from the power of music and musical education in order to achieve social outcomes, turning possible to be considered a new paradigm of music education, the “street orchestras”. Referring to its urban qualities, it is argued that music is an important tool to the creation of “place”, promoting social interaction and different perceptions of space, qualities recognized in intervention architecture projects that use music as a tool for the revitalization and regeneration of city spaces. A possibility of articulation of these qualities could benefit each other and turn the “(…) city like a large house (…)”6

Expansão do sector oleícola e os factores críticos de sucesso segmento gourmet nas estratégias de internacionalização das empresas Portuguesas

Mestrado em MarketingApós o acentuado decréscimo na produção de azeite verificado nas décadas de 60 a 80, é notório que os últimos anos têm sido de recuperação em Portugal: nos últimos 5 anos a produção nacional anual tem demonstrado um crescimento de 50%. Num país como Portugal, em que o mercado interno é de apenas 10 milhões de consumidores, a exportação surge como uma via de sentido obrigatório indispensável para o crescimento económico. Neste sentido, apesar de os países do mediterrâneo serem os principais consumidores de azeite, esta tendência tem vindo a expandir-se e existem oportunidades significativas em novas partes do mundo como sudeste da Ásia, China e países da América Latina. Para tal, uma das estratégias que tem vindo a ser adoptada pelas empresas Portuguesas é a introdução do segmento gourmet, que sendo um posicionamento diferente tem inúmeras características de marketing que vão para além da qualidade do produto e que precisam de ser analisadas e trabalhadas. Ter o produto certo não é suficiente, é necessário dá-lo a conhecer. Assim, o presente estudo vem analisar as estratégias que as empresas portuguesas de azeite utilizam na exportação deste novo segmento.After the significant decrease in the production of Olive Oil that was observed in the 60's and the 80's, it's visible that the last years have been of recovery for Portugal: in the last 5 years the annual national production report shows that there's a growth of 50%. In a country like Portugal, in which the internal market is just about 10 million consumers, exportation urges as a way that must be followed for economical growth. In this sense, although Mediterranic countries represent the main consumers of Olive Oil, this tendency is expanding to significant opportunities in new regions of the world such as Southeast Asia, China and South America countries. To do that, one of the strategies that is being used by Portuguese companies, is the introduction of the Gourmet segment. Having a different positioning, this segment has several specific marketing characteristics that needs to be addressed, analyzed and worked out. Have the right product is not enough, it?s necessary to give it to know. This way, these studies analyze strategies of internationalization of Olive oil Portuguese companies, for this segment

Construir o vazio: centro desportivo náutico

A presente dissertação tem como objeto de estudo quatro obras do arquiteto/autor Peter Zumthor. Duas escritas, Atmosferas e Pensar a Arquitectura, e duas construídas, Therme Vals e Bruder Klaus Field Chapel. A análise destas obras pretende demonstrar como o seu processo escrito (linguagem) está presente na sua obra física (objeto construído) e estabelecer as pontes entre o processo escrito, gráfico e expressivo tendo como objeto final a obra concluída. Com a sua obra escrita, Peter Zumthor resume as suas introspeções que expressam uma vontade de melhor entender aquilo que põe em prática no exercício da arquitetura. Ao analisar as obras construídas em paralelo com as escritas pretende-se uma leitura transversal desde a conceção à obra construída. O estudo desenvolve-se em duas partes. Uma primeira dedicada à obra Pensar a Arquitetura que se debruça sobre os processos vários dentro do pensamento do projeto pretendendo clarificar os percursos desde o imaginado ao realizado. Numa segunda parte, a interpretação das obras Therme Vals e Bruder Klaus Field Chapel através da análise da obra Atmosferas, fazendo paralelos entre as descrições escritas e as respetivas formalizações físicas no objeto construído.This dissertation studies four works by the architect/author Peter Zumthor. Two books, Atmospheres and Thinking Architecture, and two buildings, Therme Vals and Bruder Klaus Field Chapel. The analysis of these works intends to demonstrate how their written process (language) is present in their physical work (constructed object) and establishes the bridges between the written, graphic and expressive process having as final object the finished work. With his written work, Peter Zumthor summarizes his introspections that expresses a desire to better understand what he puts into practice in the exercise of architecture. Analyzing the works constructed in parallel with the written ones intends to reach a transversal reading from the conception to the constructed work. The study has two parts. The first one, dedicated to Thinking Architecture, a book focused on the various processes within the project thinking aiming to clarify the paths from the imagined to the realized. In a second part, the interpretation of the works Therme Vals and Bruder Klaus Field Chapel through the analysis of the book Atmospheres, making parallels between the written descriptions and the respective physical formalizations in the constructed object

Caracterização biofísica da membrana plasmática da levedura

Tese de mestrado, Bioquímica (Bioquímica Médica), Universidade de Lisboa, Faculdade de Ciências, 2009A membrana plasmática de Saccharomyces cerevisiae foi estudada por espectroscopia de fluorescência, utilizando as sondas de membrana ácido transparinárico e difenil-hexatrieno, de modo a compreender os princípios biofisicos subjacentes à formação e função de compartimentos membranares, cuja importância foi reconhecida recentemente. O estudo foi realizado em (i) células wt; (ii) esferoblastos (celulas wt com remoção completa da parede celular); (iii) lipossomas preparados a partir de extractos lipídicos totais de células wt; (iv) células erg6, que acumulam zimosterol em vez de ergosterol; (v) células scs7, que não sintetizam esfingolípidos com ácidos gordos C26:0 -hidroxilados. As principais observações foram: (a) a membrana plasmática da levedura possui domínios de gel ricos em esfingolípidos com conteúdo baixo ou nulo de esteróis; (b) os lípidos de levedura possuem a capacidade de formar domínios mais rígidos na ausência de proteínas do que os detectados na membrana plasmática; (c) os sistemas erg6e esferoblastos possuem uma maior ordem global de membrana; (d) modificações na biossíntese do ergosterol e a remoção da parede celular não alteram significativamente a rigidez dos domínios ricos em esfingolípidos, mas em ambos os casos há uma diminuição da sua abundância relativa. Os resultados obtidos sugerem que a parede celular desempenha um papel na estabilização dos domínios ricos em esfingolípidos que poderá ocorrer por interacção da mesma com proteínas ancoradas a glucosilfosfatidilinositol. A partir dos resultados também foi proposto um modelo em que nas células onde os domínios ricos em esfingolípidos sao menos abundantes ocorre uma distribuição mais homogénea de esfingolípidos por toda a membrana, responsável pelo aumento de ordem global da mesma. Concluindo, este trabalho comprova a existência de domínios ordenados tipo gel na membrana plasmática de organismos vivos, reforçando a ideia de que os esfingolípidos são componentes essenciais na constituição da membrana e na resposta a alterações fisiológicas que ponham em causa a integridade da célula.The plasma membrane of Saccharomyces cerevisiae was studied by fluorescence spectroscopy with the membrane probes trans-parinaric acid and diphenylhexatriene to understand biophysical principles underlying the formation and role of membrane compartments, with relevance recently recognized. The study included: (i) wt cells; (ii) spheroplasts (wt cells with complete cell wall removal); (iii) liposomes from total lipid extracts of wt cells; (iv) erg6Δ cells, which accumulate zymosterol instead of ergosterol; (v) scs7Δ cells, which lack synthesis of α- hydroxylated sphingolipid-associated C26:0 fatty acids. The main observations were: (a) the yeast plasma membrane contains sphingolipid-enriched gel domains with low or none sterol content; (b) yeast lipids have the ability to form, in the absence of proteins, domains more rigid than those detected in the plasma membrane; (c) erg6 cells and spheropasts have a higher membrane order; (d) alterations in sterol biosynthesis and cell wall removal do not significantly affect sphingolipid-enriched domains rigidity, although significantly reducing their relative abundance. The results obtained suggest that the cell wall has an important role in the stabilization of sphingolipid-enriched domains which might occur through its interaction with glucosylphosphatidylinositol anchored proteins. Furthermore, a model is proposed that explains the lower abundance of sphingolipid-enriched domains by a more homogeneous distribution of those lipids throughout the whole plasma membrane, thereby increasing the membrane global order. In conclusion, this work shows the existence of gel-like ordered domains in the plasma membrane of living cells, supporting the idea that sphingolipids are essential components for the constitution of biomembranes and for the response to physiological changes that are dangerous for the cell integrity

Spatial mapping of ectonucleotidase gene expression in the murine urinary bladder

Purinergic signaling is important for normal bladder function, as it is thought to initiate the voiding reflex and modulate smooth muscle tone. The availability of adenine nucleotides and nucleosides (aka purines) at receptor sites of various cell types in the bladder wall is regulated by ectonucleotidases (ENTDs). ENTDs hydrolyze purines such as adenosine 5′-triphosphate (ATP) and adenosine 5′-diphosphate (ADP) with varying preference for the individual substrate. Therefore, the end effect of extracellular purines may depend significantly on the type of ENTD that is expressed in close proximity to the target cells. ENTDs likely have distinct cellular associations, but the specific locations of individual enzymes in the bladder wall are poorly understood. We used RNAscope™, an RNA in situ hybridization (ISH) technology, to visualize the distribution and measure the levels of gene expression of the main recognized ectonucleotidases in large high-resolution images of murine bladder sections. The relative gene expression of ENTDs was Entpd3 > Alpl >> Enpp1 = Entpd2 >> Enpp3 > Entpd1 (very low to no signal) in the urothelium, Entpd1 ≥ Entpd2 >> Enpp3 > Enpp1 = Alpl ≥ Nt5e (very low to no signal) in the lamina propria, and Entpd1 >> Nt5e = Entpd2 >> Enpp1 > Alpl = Enpp3 in the detrusor. These layer-specific differences might be important in compartmentalized regulation of purine availability and subsequent functions in the bladder wall and may explain reported asymmetries in purine availability in the bladder lumen and suburothelium/lamina propria spaces

Trypanosoma brucei parasites occupy and functionally adapt to the adipose tissue in mice

This work was supported by 55007419 (HHMI) and 2151 (EMBO) to L.M.F., D.P.-N., F.B., and F.G.; FCT fellowships to S.T., F.R.-F., and F.A.-B. (SFRH/BPD/89833/2012, SFRH/BD/51286/2010, and SFRH/BD/80718/2011, respectively); Wellcome Trust grant (093228), MRC MR/M020118/1, and European Community Seventh Framework Programme under grant agreement No. 602773 (Project KINDRED) to S.A.Y. and T.K.S.; and PAI 7/41 (Belspo) and ERC-NANOSYM to J.V.D.A.Trypanosoma brucei is an extracellular parasite that causes sleeping sickness. In mammalian hosts, trypanosomes are thought to exist in two major niches: early in infection, they populate the blood; later, they breach the blood-brain barrier. Working with a well-established mouse model, we discovered that adipose tissue constitutes a third major reservoir for T. brucei. Parasites from adipose tissue, here termed adipose tissue forms (ATFs), can replicate and were capable of infecting a naive animal. ATFs were transcriptionally distinct from bloodstream forms, and the genes upregulated included putative fatty acid β-oxidation enzymes. Consistent with this, ATFs were able to utilize exogenous myristate and form β-oxidation intermediates, suggesting that ATF parasites can use fatty acids as an external carbon source. These findings identify the adipose tissue as a niche for T. brucei during its mammalian life cycle and could potentially explain the weight loss associated with sleeping sickness.Publisher PDFPeer reviewe