Assessment of biocompatibility of selected ferroelectric ceramics

Desde há muitas décadas que é sabido que os organismos vivos, em especial os tecidos, reagem fisicamente a estímulos eléctricos, podendo esses efeitos reproduzirem-se numa libertação de químicos endógenos, ou deformar a sua estrutura física. O tecido ósseo por si só é considerado um material/tecido piezoeléctrico, deformando-se mecanicamente quando lhe é induzido um estímulo eléctrico e vice-versa, ou seja, produz um potencial eléctrico quando sofre uma tracção ou compressão mecânica. A hipótese de que um material ferroeléctrico possa vir a produzir efeitos no desempenho deste tipo de tecidos é então proposta, como por exemplo, para uma melhor, mais rápida e eficaz regeneração óssea. Estes mesmos materiais ferroeléctricos podem porventura alterar as cargas de superfície dos tecidos vivos de modo a atrair, atrasar ou até impedir o fluxo iónico de elementos químicos específicos responsáveis pelo processo de regeneração. São escolhidos então o niobato de lítio e o tantalato de lítio como cerâmicos ferroeléctricos e foi estudada pela primeira vez a sua bioactividade in vitro, esperando-se encontrar pistas relativas à sua bioactividade in vivo. Estes cerâmicos ferroeléctricos foram seleccionados devido às suas importantes propriedades piezoeléctricas e ferroeléctricas. Estas propriedades podem abrir um novo e importante leque de aplicações biomédicas caso estes cerâmicos sejam bioactivos. Este trabalho foi dividido em 3 fases: (i) sintetização dos pós de niobato de lítio e tantalato de lítio, (ii) caracterização dos pós e (iii) preparação das amostras e (iv) estudo da bioactividade destes cerâmicos ferroeléctricos. Os pós foram produzidos através de um processo simples de mistura/moagem seguido de calcinação. Foram estudadas as fases cristalinas presentes através de Difracção de raios-X (DRX) e avaliadas as características morfológicas destes pós, nomeadamente o diâmetro de partículas e área superficial específica. De modo a simular o ambiente do plasma humano, foi produzido sinteticamente um “Simulated Body Fluid” (SBF). Seguidamente as amostras foram imersas nesse ambiente líquido por 1, 3, 7, 15 e 21 dias. Após remoção dos pós foram realizadas uma série de análises de modo a estudar a sua bioactividade. De entre estes testes destacam-se a microscopia electrónica de varrimento (SEM/EDS), DRX e espectroscopia de Infravermelho por transformada de Fourier com reflectância total atenuada (FTIR-ATR). Embora não tenham sido detectadas alterações no DRX realizado aos pós, verificou-se a formação de aglomerados de fosfato de cálcio na superfície dos pós através do SEM, resultados estes, reforçados pelo EDS e FTIR-ATR. Estes precipitados de fosfato de cálcio indiciam a capacidade destes pós cerâmicos ferroeléctricos se comportarem como bioactivos em contacto com tecidos ósseos in vivo.

Evaluation of the matabolic response of osteosarcoma cells to conventional and new anticancer drugs by NMR metabolomics

The main scope of this work was to evaluate the metabolic effects of anticancer agents (three conventional and one new) in osteosarcoma (OS) cells and osteoblasts, by measuring alterations in the metabolic profile of cells by nuclear magnetic resonance (NMR) spectroscopy metabolomics. Chapter 1 gives a theoretical framework of this work, beginning with the main metabolic characteristics that globally describe cancer as well as the families and mechanisms of action of drugs used in chemotherapy. The drugs used nowadays to treat OS are also presented, together with the Palladium(II) complex with spermine, Pd2Spm, potentially active against cancer. Then, the global strategy for cell metabolomics is explained and the state of the art of metabolomic studies that analyze the effect of anticancer agents in cells is presented. In Chapter 2, the fundamentals of the analytical techniques used in this work, namely for biological assays, NMR spectroscopy and multivariate and statistical analysis of the results are described. A detailed description of the experimental procedures adopted throughout this work is given in Chapter 3. The biological and analytical reproducibility of the metabolic profile of MG-63 cells by high resolution magic angle spinning (HRMAS) NMR is evaluated in Chapter 4. The metabolic impact of several factors (cellular integrity, spinning rate, temperature, time and acquisition parameters) on the 1H HRMAS NMR spectral profile and quality is analysed, enabling the definition of the best acquisition parameters for further experiments. The metabolic consequences of increasing number of passages in MG-63 cells as well as the duration of storage are also investigated. Chapter 5 describes the metabolic impact of drugs conventionally used in OS chemotherapy, through NMR metabolomics studies of lysed cells and aqueous extracts analysis. The results show that MG-63 cells treated with cisplatin (cDDP) undergo a strong up-regulation of lipid contents, alterations in phospholipid constituents (choline compounds) and biomarkers of DNA degradation, all associated with cell death by apoptosis. Cells exposed to doxorubicin (DOX) or methotrexate (MTX) showed much slighter metabolic changes, without any relevant alteration in lipid contents. However, metabolic changes associated with altered Krebs cycle, oxidative stress and nucleotides metabolism were detected and were tentatively interpreted at the light of the known mechanisms of action of these drugs. The metabolic impact of the exposure of MG-63 cells and osteoblasts to cDDP and the Pd2Spm complex is described in Chapter 6. Results show that, despite the ability of the two agents to bind DNA, the metabolic consequences that arise from exposure to them are distinct, namely in what concerns to variation in lipid contents (absent for Pd2Spm). Apoptosis detection assays showed that, differently from what was seen for MG-63 cells treated with cDDP, the decreased number of living cells upon exposure to Pd2Spm was not due to cell death by apoptosis or necrosis. Moreover, the latter agent induces more marked alterations in osteoblasts than in cancer cells, while the opposite seemed to occur upon cDDP exposure. Nevertheless, the results from MG-63 cells exposure to combination regimens with cDDP- or Pd2Spm-based cocktails, described in Chapter 7, revealed that, in combination, the two agents induce similar metabolic responses, arising from synergy mechanisms between the tested drugs. Finally, the main conclusions of this thesis are summarized in Chapter 8, and future perspectives in the light of this work are presented.

Loss of Aggressiveness of Phytophthora cinnamomi (Beta-Cinnamomin Silenced Strain) in the Infection of Castanea sativa

Several forest species are severely affected by Phytophthora cinnamomi. The contribution of this oomycete to forest decline and dieback has been broadly reported. In particular, it is consensual that it is the causal agent of ink disease in Castanea sativa. It has been associated with the severe decline of Quercus species, namely the Q. suber and Q. ilex dieback in Portugal and Spain, and has been responsible for the infection of numerous native species and crops. This pathogen persists in the soil or on plant material in the form of chlamydospores allowing the infection of living root tissues when environmental conditions are favorable. © Microscopy Society of America 2012.

Ecologic zonation model of the benthic foraminifera and thecamoebians of Guadiana river estuary and application in paleoenvironmental reconstruction

Tese de doutoramento, Ciências do Mar ( Processos de Ecossistemas Marinhos), Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2012

Topical bacteriophage therapy of the infected diabetic foot

Tese de doutoramento, Medicina (Medicina Interna), Universidade de Lisboa, Faculdade de Medicina, 2014

Telaprevir or boceprevir based therapy for chronic hepatitis C infection: Development of resistance-associated variants in treatment failure

The use of triple-therapy, pegylated-interferon, ribavirin and either of the first generation hepatitis C virus (HCV) protease inhibitors telaprevir or boceprevir, is the new standard of care for treating genotype 1 chronic HCV. Clinical trials have shown response rates of around 70–80%, but there is limited data from the use of this combination outside this setting. Through an expanded access programme, we treated 59 patients, treatment naïve and experienced, with triple therapy. Baseline factors predicting treatment response or failure during triple therapy phase were identified in 58 patients. Thirty seven (63.8%) of 58 patients had undetectable HCV RNA 12 weeks after the end of treatment. Genotype 1a (p = 0.053), null-response to previous treatment (p = 0.034), the rate of viral load decline after 12 weeks of previous interferon-based treatment (p = 0.033) were all associated with triple-therapy failure. The most common cause of on-treatment failure for telaprevir-based regimens was the development of resistance-associated variants (RAVs) at amino acids 36 and/or 155 of HCV protease (p = 0.027) whereas in boceprevir-based regimens mutations at amino acid 54 were significant (p = 0.015). SVR12 rates approaching 64% were achieved using triple therapy outside the clinical trial setting, in a patient cohort that included cirrhotics.

Introducing EbolaCheck: potential for point-of-need infectious disease diagnosis

The 2013–2015 Ebolavirus disease humanitarian crisis has spurred the development of laboratory-free, point-of-care nucleic acid testing solutions. EbolaCheck is an international consortium of public health, academic and biotechnology industry stakeholders aiming to deliver clinical molecular diagnostic standard-of-care testing suitable for the West African milieu within 12 months. In this article, the current status of the EbolaCheck platform is discussed in the context of the current regulatory framework. Presented here are future goals to achieve differential diagnosis of hemorrhagic fever disease from <5-μl of whole blood samples or mucosal biofluids, in a single tube process, under 40 min and with minimal operator training requirements.

Biogeographic and palaeoclimatic relationships of the Middle Pliocene ichthyofauna of the Samoggia Torrent (Bologna, Italy)

New Middle Pliocene ichthyofauna (2.4-2.2 Ma) from central-eastern Italy (Samoggia Torrent, Bologna) are described. These ichthyolites were found in a rather thin laminated layer that was deposited after the 2.4 Ma climatic crisis. The origin of this deposit, in which 31 taxa have been classified, is to be related to anoxic events on a regional and, probably, supraregional scale. This ichthyofaunistic association, which consists of living genera, is characterized by a clearcut predominance of mesopelagic species. The palaeoclimatic characters of these ichthyofauna indicate subtropical-type waters, while from a palaeobiogeographic point of view there is a close relationship with the present-day Atlantic-Mediterranean bioprovince. The Samoggia deposit has yielded six taxa that are absent or only occasionally present in the Mediterranean: one of these, Spratelloides gracilis, is exclusive of the Indo-Pacific bioprovince.

Development of 2-oxazoline-based hydrogels and porous polyester microparticles in scCO2

Thesis submitted to Faculdade de Ciências e Tecnologia from Universidade Nova de Lisboa in partial fulfillment of the requirements for the obtention of the degree of Master of Science in Biotechnology

Initial dilemmas in IST research: design science and design research, a clarification of concepts

The inter-disciplinarity of information systems, applied discipline and activity of design, and the study from different paradigms perspectives explains the diversity of problems addressed. The context is broad and includes important issues beyond technology, as the application, use, effectiveness, efficiency and their organizational and social impacts. In design science, the research interest is in contributing to the improvement of the processes of the design activity itself. The relevance of research in design science is associated with the result obtained for the improvement of living conditions in organizational, inter-organizational and Society contexts. In the research whose results are artifacts, the adoption of design research as a process of research is crucial to ensure discipline, rigor and transparency. Based on a literature review, this paper clarifies the terms of design science and design research. This is the main motivation for presenting this paper, determinant for the phase in research in technologies and information systems which are the three research projects presented. As a result the three projects are discussed in relation to the concepts of design science and design research.

On and off air: an ethnographic exploration of minority radio in Portugal

Taking a Media Anthropology’s approach to dynamics of mediated selfrepresentation in migratory contexts, this thesis starts by mapping radio initiatives produced by, for and/or with migrants in Portugal. To further explore dynamics of support of initial settlement in the country, community-making, cultural reproduction, and transnational connectivity - found both in the mapping stage and the minority media literature (e.g. Kosnick, 2007; Rigoni & Saitta, 2012; Silverstone & Georgiou, 2005) - a case study was selected: the station awarded with the first bilingual license in Portugal. The station in question caters largely to the British population presenting themselves as “expats” and residing in the Algarve. The ethnographic strategy to research it consisted of “following the radio” (Marcus, 1995) beyond the station and into the events and establishments it announces on air, so as to relate production and consumption realms. The leading research question asks how does locally produced radio play into “expats” processes of management of cultural identity – and what are the specificities of its role? Drawing on conceptualizations of lifestyle migration (Benson & O’Reilly, 2009), production of locality (Appadurai 1996) and the public sphere (Butsch, 2007; Calhoun & et al, 1992; Dahlgren, 2006), this thesis contributes to valuing radio as a productive gateway to research migrants’ construction of belonging, to inscribe a counterpoint in the field of minority media, and to debate conceptualizations of migratory categories and flows. Specifically, this thesis argues that the station fulfills similar roles to other minority radio initiatives but in ways that are specific to the population being catered to. Namely, unlike other minority stations, radio facilitates the process of transitioning between categories along on a continuum linking tourists and migrants. It also reflects and participates in strategies of reterritorialization that rest on functional and partial modes of incorporation. While contributing to sustain a translocality (Appadurai, 1996) it indexes and fosters a stance of connection that is symbolically and materially connected to the UK and other “neighborhoods” but is, simultaneously, oriented to engaging with the Algarve as “home”. Yet, besides reifying a British cultural identity, radio’s oral, repetitive and ephemeral discourse particularly trivializes the reproduction of an ambivalent stance of connection with place that is shared by other “expats”. This dynamic is related to migratory projects driven by social imaginaries fostered by international media that stimulate the search for idealized ways of living, which the radio associates with the Algarve. While recurrently localizing and validating the narrative projecting an idealized “good life”, radio amplifies dynamics among migrants that seem to reaffirm the migratory move as a good choice.

Contributions towards smart cities : exploring block level census data for the characterization of change in Lisbon

The interest in using information to improve the quality of living in large urban areas and its governance efficiency has been around for decades. Nevertheless, the improvements in Information and Communications Technology has sparked a new dynamic in academic research, usually under the umbrella term of Smart Cities. This concept of Smart City can probably be translated, in a simplified version, into cities that are lived, managed and developed in an information-saturated environment. While it makes perfect sense and we can easily foresee the benefits of such a concept, presently there are still several significant challenges that need to be tackled before we can materialize this vision. In this work we aim at providing a small contribution in this direction, which maximizes the relevancy of the available information resources. One of the most detailed and geographically relevant information resource available, for the study of cities, is the census, more specifically the data available at block level (Subsecção Estatística). In this work, we use Self-Organizing Maps (SOM) and the variant Geo-SOM to explore the block level data from the Portuguese census of Lisbon city, for the years of 2001 and 2011. We focus on gauging change, proposing ways that allow the comparison of the two time periods, which have two different underlying geographical bases. We proceed with the analysis of the data using different SOM variants, aiming at producing a two-fold portrait: one, of the evolution of Lisbon during the first decade of the XXI century, another, of how the census dataset and SOM’s can be used to produce an informational framework for the study of cities.

Patterns of care in recurrent glioblastoma in Switzerland: a multicentre national approach based on diagnostic nodes.

Despite moderate improvements in outcome of glioblastoma after first-line treatment with chemoradiation recent clinical trials failed to improve the prognosis of recurrent glioblastoma. In the absence of a standard of care we aimed to investigate institutional treatment strategies to identify similarities and differences in the pattern of care for recurrent glioblastoma. We investigated re-treatment criteria and therapeutic pathways for recurrent glioblastoma of eight neuro-oncology centres in Switzerland having an established multidisciplinary tumour-board conference. Decision algorithms, differences and consensus were analysed using the objective consensus methodology. A total of 16 different treatment recommendations were identified based on combinations of eight different decision criteria. The set of criteria implemented as well as the set of treatments offered was different in each centre. For specific situations, up to 6 different treatment recommendations were provided by the eight centres. The only wide-range consensus identified was to offer best supportive care to unfit patients. A majority recommendation was identified for non-operable large early recurrence with unmethylated MGMT promoter status in the fit patients: here bevacizumab was offered. In fit patients with late recurrent non-operable MGMT promoter methylated glioblastoma temozolomide was recommended by most. No other majority recommendations were present. In the absence of strong evidence we identified few consensus recommendations in the treatment of recurrent glioblastoma. This contrasts the limited availability of single drugs and treatment modalities. Clinical situations of greatest heterogeneity may be suitable to be addressed in clinical trials and second opinion referrals are likely to yield diverging recommendations.

The role of molecular, clinical and socioeconomic factors in the long-term survival of axillary node negative breast cancer patients

The overall objective of this study was to investigate factors associated with long-term survival in axillary node negative (ANN) breast cancer patients. Clinical and biological factors included stage, histopathologic grade, p53 mutation, Her-2/neu amplification, estrogen receptor status (ER), progesterone receptor status (PR) and vascular invasion. Census derived socioeconomic (SES) indicators included median individual and household income, proportions of university educated individuals, housing type, "incidence" of low income and an indicator of living in an affluent neighbourhood. The effects of these measures on breast cancer-specific survival and competing cause survival were investigated. A cohort study examining survival among axillary node negative (ANN) breast cancer patients in the greater Toronto area commenced in 1 989. Patients were followed up until death, lost-to-follow up or study termination in 2004. Data were collected from several sources measuring patient demographics, clinical factors, treatment, recurrence of disease and survival. Census level SES data were collected using census geo-coding of patient addresses' at the time of diagnosis. Additional survival data were acquired from the Ontario Cancer Registry to enhance and extend the observation period of the study. Survival patterns were examined using KaplanMeier and life table procedures. Associations were examined using log-rank and Wilcoxon tests of univariate significance. Multivariate survival analyses were perfonned using Cox proportional hazards models. Analyses were stratified into less than and greater than 5 year survival periods to observe whether known markers of short-tenn survival were also associated with reductions in long-tenn survival among breast cancer patients. The 15 year survival probabilities in this cohort were: for breast cancerspecific survival 0.88, competing causes survival 0.89 and for overall survival 0.78. Estrogen receptor (ER) and progesterone receptor (PR) status (Hazard Ratio (HR) ERIPR- versus ER+/PR+, 8.15,95% CI, 4.74, 14.00), p53 mutation (HR, 3.88, 95% CI, 2.00, 7.53) and Her-2 amplification (HR, 2.66, 95% CI, 1.36, 5.19) were associated with significant reductions in short-tenn breast cancer-specific survival «5 years following diagnosis), however, not with long-term survival in univariate analyses. Stage, histopathologic grade and ERiPR status were the clinicallbiologieal factors that were associated with short-term breast cancer specific survival in multivariate results. Living in an affluent neighbourhood (top quintile of median household income compared to the rest of the population) was associated with the largest significant increase in long-tenn breast cancer-specific survival after adjustment for stage, histopathologic grade and treatment (HR, 0.36, 95% CI, 0.12, 0.89).

Youth Living in Residential Care: Implications for Leisure and Identity

The purpose of this study was to explore the intersection of living in residential care, leisure engagement, and adolescent identity development. The investigation included the voices of six youth living in a residential care facility in southern Ontario. The data was collected through participant observations, semi-structured interviews, and document analysis. Moustakas’ (1994) modification of the Stevick-Colaizzi-Keen method was used to analyze the data. The findings determined that living in residential care is rife with dialectical tensions that impact leisure and identity. The youth shared poignant narratives of how living in residential care was a stigmatizing experience that left them feeling restricted and isolated. They also shared their struggles with finding autonomy in a secured facility and managing the violent discourses of their peers. This research contributes to a burgeoning body of literature that explores the experiences of youth living in out-of-home care. Implications for practice and future research are discussed.