As comunidades de Porífera do litoral Norte

Mestrado em Biologia MarinhaOs indivíduos do filo Porifera possuem um papel essencial nas comunidades bentónicas, não só por serem animais filtradores e detritívoros, como pelo facto de estabelecerem relações simbióticas com vários organismos, nomeadamente algas, e servirem de abrigo e alimento a muitos animais. A estrutura das comunidades Porifera é influenciada por vários factores ambientais, entre os quais os padrões de sedimentação, a velocidade da corrente, e o tamanho e inclinação do substrato. Dado que as espécies pertencentes a este filo são animais sésseis, com taxas de crescimento reduzidas e esperança média de vida relativamente longa, a estrutura das suas comunidades poderá ajudar não só a estudar os padrões de circulação local, mas também a monitorizar possíveis impactos sobre estes. Assim, ao longo deste trabalho, foram estudadas quer a riqueza específica quer a diversidade morfológica das esponjas e a sua distribuição na área entre Esposende e a Apúlia e até 3Km da costa. Tal permitiu não só verificar que esta zona têm um número de espécies muito elevado, quando comparada com outros estudos com esforços de amostragem semelhantes, mas também inferir características ambientais dos locais e quais os parâmetros que mais fortemente influenciam a colonização e crescimento das esponjas no local. Uma continuação deste trabalho e, quem sabe, a sua aplicação em maior escala, permitiria um melhor conhecimento dos factores que influenciam a distribuição das espécies de Porifera e determinam o seu desenvolvimento tridimensional.Phylum Porifera have an essential role in benthic communities, not only due to their filtrating and detritic nature, but also for the establishment of symbiotic relationships with several organisms, namely algae. Sponges are also the source of nutrients and shelter for many animals. Its community’s structure is influenced by several environmental factors such as sedimentation patterns, current’s speed and substrate’s dimension and slope. The members of this phylum are sessile animals, with low growth rates and relatively long life expectancy. Because of that, the structure of communities dominated by sponges might help not only to study local circulation patterns, but also to assess possible impacts over them. In this work, both specific and morphological richness of sponges and its distribution were studied in the area between Esposende and Apulia, in the infralittoral zone till a maximum depth of 30 meters. This allowed not only to verify the great number of species in this area, when compared to other studies with similar sampling efforts, but also to deduce local environmental characteristics that may influence sponge’s colonization and growth. This work’s extension and, maybe, its application in greater scale, would allow the gathering of information in environmental parameters’ influence over Porifera’s distribution and three-dimensional development

Studies on resistance and response to vancomycin in Enterococcus faecalis: a last resort antibiotic

Dissertation presented to obtain a Ph.D. degree in Biochemistry by Instituto de Tecnologia Química e Biológica Universidade Nova de Lisboa.Enterococci are part of the normal human and animal gut microbiota and hardly cause infections in healthy individuals. In the last 20 years enterococci have emerged as common causes of hospital-acquired infections. One of the major reasons why these microorganisms easily survive in the hospital environment is their intrinsic resistance to several commonly used antibiotics, and more importantly, their ability to acquire resistance to many currently used antibiotics, including glycopeptides. Development of resistance to the glycopeptide vancomycin in the Enterococcus genus presents a worldwide major problem. Infections with vancomycin resistant enterococci are not only difficult to treat but the organisms show a strong propensity to disseminate and spread from patient to patient in the hospital setting. Accurate knowledge of the real scenario of vancomycin resistance is essential to design national and global strategies and prevent community and nosocomial transmission of vancomycin resistant organisms.(...)Financial Support from Fundação para a Ciência e Tecnologia (FCT) – Ph.D: grant - SFRH/BD/21535/2005

Análise de projetos pessoais de alunos do 9º ano de escolaridade do ensino básico

Dissertação de mestrado em Psicologia da Educação, Desenvolvimento e Aconselhamento, apresentada à Faculdade de Psicologia e de Ciências da Educação da Universidade de CoimbraEste estudo incide sobre os projetos pessoais numa amostra de alunos do 9º ano de escolaridade. A literatura revista incide na Psicologia da Personalidade, nos projetos pessoais e na adolescência, e na relação entre estes assuntos. Na segunda parte deste estudo são apresentados os procedimentos e resultados da aplicação do instrumento “Análise dos Projetos Pessoais” (Personal Project Analysis – PPA; Little, 1983) a 66 sujeitos. As conclusões indicam que os resultados obtidos neste estudo são, em geral, semelhantes aos revistos na literatura, sendo de destaque a importância superior que os projetos pessoais interpessoais assumem para os adolescentes.This study addresses the personal projects in a sample of students from 9th grade. The reviewed literature focuses in the Personality Psychology, in the personal projects and in the adolescence, and in the relationship between these subjects. In the second part of this study the procedures and results from the application of the Personal Project Analysis – PPA (Little, 1983) to 66 individuals are presented. The conclusions indicate that the the results obtained in this study are generally similar to those reviewed in the literature, and the importance of personal projects about interpersonal relationships to adolescents is highlighted

Intracellular pH as a new mechanism of signal transduction

El pH juega un papel clave en el control de la patogénesis fúngica. En el patógeno del suelo Fusarium oxysporum, causante de la marchitez vascular, el pH actúa como regulador maestro de Fmk1, una MAPK altamente conservada, esencial para el crecimiento invasivo y patogénesis en raíces de tomate. Estudios previos demostraron que el pH también regula las MAPKs Mpk1 y Hog1, de manera opuesta a Fmk1. Sin embargo, se desconocen los mecanismos moleculares de la respuesta al pH. En el presente trabajo se ha investigado, si el pH del medio afecta las rutas MAPK a través de un cambio en el pH intracelular (pHi). Se ha determinado que el pHi funciona como una señal para la regulación de las rutas MAPK en F. oxysporum. El tratamiento de las células con diethylstilbestrol (DES), un inhibidor especifico de Pma1, la principal bomba de protones de la membrana plasmática, conlleva una rápida y sostenida bajada del pHi, acompañada de una rápida y transitoria activación de las MAPKs Mpk1 y Hog1, y la inactivación de Fmk1. Esta respuesta esta conservada en la levadura panadera Saccharomyces cerevisiae, lo que indica que está conservado en hongos. Por ello se ha utilizado la levadura como modelo para investigar el mecanismo de activación de Mpk1 por acidificación del pHi. Se identificaron: una ruta alternativa de la respuesta a stress de pared, mediada por la quinasa Ypk1/2; la rama Sln1 de la ruta Hog1; y la MAPK fosfatasa Msg5 cuya abundancia es afectada por el pHi. En segundo lugar, un escrutinio realizado con mutantes knockout sensibles a ácido identificó un gran número de proteínas candidatas implicadas en la activación de Mpk1 inducida por DES. Muchos de ellos poseen ortólogos en F. oxysporum. Las funciones celulares más relevantes en este proceso son tráfico celular, homeostasis de iones (principalmente V-ATPase) y metabolismo de lípidos. Los resultados revelan que el pHi es un nuevo mecanismo de regulación de MAPKs en hongos, y sugieren que existe una compleja interacción entre rutas de señalización y funciones celulares en la regulación de este proceso.pH is as a key player in the control of fungal pathogenicity. In the soilborne vascular wilt fungus Fusarium oxysporum, ambient pH acts as a master switch for regulation of the conserved Fmk1 MAPK cascade, essential for invasive growth and pathogenicity toward tomato roots. Previous studies indicated that pH also modulates the activity of the two other MAPKs, Mpk1 and Hog1, in an opposite way to Fmk1. The molecular events underlying the pH response are currently unknown. Here we investigated whether ambient pH modulates MAPK signaling by affecting the intracellular pH (pHi). We found that pHi acts as a signal regulating MAPK activity in F. oxysporum. Diethylstilbestrol (DES), a specific inhibitor of the plasma membrane H+-ATPase Pma1, induced a rapid and sustained decrease of pHi accompanied by rapid and transitory phosphorylation of the MAPKs Mpk1 and Hog1, and inactivation of Fmk1. Analogously, pHi also regulates MAPK signaling in the budding yeast Saccharomyces cerevisiae, suggesting that this mechanism is conserved in fungi. We initially investigated the mechanism of pHi-induced Mpk1 activation in the yeast model, using a candidate gene approach. Our studies revealed three potential mechanisms: an alternative branch of the canonical cell wall integrity pathway mediated by the Ypk1/2 kinase; the Sln1 branch of the Hog1 cascade; and the dual specificity MAPK phosphatase Msg5 whose abundance is affected by pHi. Moreover, a screen with a subset of acid-sensitive yeast mutants identified a number of candidates affected in DES-triggered Mpk1 activation, many of which have predicted orthologues in F. oxysporum. These candidate proteins have conserved cellular functions including cellular trafficking, ion homeostasis (mainly V-ATPase function) and lipid metabolism. Our results discover pHi as a new mechanism for MAPK signaling in fungi and reveal a complex interplay between signaling pathways and cellular functions in the regulation of this process

Detection of Pasteurellaceae in Laboratory Mice by Fecal PCR

A utilização de animais de laboratório microbiologicamente definidos na investigação biomédica tem-se tornado prática comum nas últimas décadas. A uniformização microbiológica, com base na realização de testes de rotina aos animais em intervalos regulares, tem contribuído para a política dos 3Rs (Refinement, Reduction e Replacement). Permite a redução do número de animais utilizados na medida em que diminui a variação dentro e entre grupos experimentais, e desempenha um papel importante no refinamento do estado de saúde dos animais melhorando assim o seu bem-estar. Adicionalmente, tem reduzido os riscos para a saúde humana devido a zoonoses. O estatuto sanitário SPF (Specific Pathogen Free) foi desenvolvido para garantir a ausência de patogénicos específicos em animais de laboratório enquanto o estatuto sanitário SOPF (Specific Opportunistic and Pathogen Free) garante a ausência dos principais agentes oportunistas para além dos patogénicos específicos. Estas definições têm base numa detalhada lista de exclusão de agentes susceptíveis de infectar roedores e confundir resultados de experiências com animais, tal como os membros da família Pasteurellaceae. Entre estes, a Pasteurella pneumotropica é considerada o patogénico oportunista mais frequente e é geralmente isolada do tracto respiratório superior, pulmões, tracto genital e gastrointestinal. A patogenicidade deste organismo em ratos e murganhos imunocompetentes é considerada baixa, mas em animais imunodeprimidos pode levar ao desenvolvimento de pneumonia, conjuntivite e infecção dos tractos respiratório e genital. Contudo, graças à estrutura taxonómica irresoluta da família Pasteurellaceae e à ocorrência de outros taxa para além da P. pneumotropica, a FELASA recomenda a monitorização de roedores SPF para todos os membros desta família. As técnicas de diagnóstico disponíveis para a detecção de Pasteurellaceae incluem métodos bacteriológicos e caracterização imunológica e bioquímica. Estes procedimentos são morosos e por vezes produzem resultados indeterminados dada a diversidade fenotípica desta família bacteriana. Ensaios de PCR com base na sequência do gene 16S rRNA foram recentemente descritos como alternativas para a detecção de Pasteurellaceae. No entanto, os protocolos utilizados baseiam-se em métodos de amostragem invasivos que requerem o sacrifício dos animais. Neste estudo desenvolvemos um ensaio de PCR simples, não invasivo e específico para a detecção de Pasteurellaceae usando DNA isolado de fezes de murganhos. Discutimos ainda o impacto desta técnica não invasiva na avaliação da prevalência de Pasteurellaceae em roedores de laboratório.The use of microbiologically defined laboratory animals in biomedical research has become standard practice in the last few decades. Microbiological standardization, based upon routine testing of the animals at regular intervals, has contributed to the 3Rs policy (Refinement, Reduction and Replacement). It allows the reduction of the number of animals used as it decreases the variation within and between test groups, and it plays an important role in the refinement of the overall health of laboratory animals thus improving their welfare. Additionally, it has reduced human health risks due to zoonotic diseases. Specific pathogen free (SPF) health status was developed to guarantee the absence of specific pathogens in laboratory animals whereas Specific Opportunistic and Pathogen-Free (SOPF) health status guarantees the absence of the major interfering opportunistic agents in addition to the specific pathogens. These negative definitions are based on a detailed exclusion list of agents which are likely to infect laboratory rodents and confound results from animal experiments, such as members of the Pasteurellaceae family. Among these, Pasteurella pneumotropica is considered to be the most frequently occurring opportunistic pathogen in laboratory rodents and it is usually isolated from the upper respiratory tract, lungs, genital and gastrointestinal tracts. The pathogenicity of this organism in immunocompetent laboratory mice and rats is regarded as low, but in immunodeficient animals it may lead to pneumonia, conjunctivitis, and respiratory and genital tract infections. However, due to the unsettled taxonomic structure of the Pasteurellaceae family and the occurrence of taxa other than Pasteurella pneumotropica, FELASA recommends the monitoring of SPF rodents for all Pasteurellaceae. The available diagnostic techniques for Pasteurellaceae screening traditionally include bacteriological methods, immunological and biochemical characterization. These procedures are time-consuming and sometimes yield indeterminate results due to the phenotypical diversity of this bacterial family. PCR assays based on the 16S rRNA gene sequence have recently been reported as alternatives to biochemical and culture methods for Pasteurellaceae detection. However, the protocols used are based on invasive sampling methods that require the sacrifice of animals. In this study we developed a simple, non invasive and specific PCR assay to detect Pasteurellaceae by using DNA isolated from mice feces. Furthermore we discuss the impact of this non-invasive technique in assessing the prevalence of Pasteurellaceae on laboratory rodents

Moisture buffering capacity of earth mortar plasters and hemp concrete. Effect of temperature and thickness

Earth mortar plasters and hemp concrete are hygroscopic materials since they have the ability to uptake and release water vapour. These materials can be used for a passive control of relative humidity (RH) inside buildings, with the benefits of improving the health and comfort of the occupants, as well as reducing the energy consumption of buildings. The quantification of this passive control potential for buildings material has been extensively studied by the scientific community since the last decade. These studies notably lead to the Nordtest project (Rode et al. 2005) where the concept of Moisture Buffering Value (MBV) was developed, as well as a test protocol in order to quantify this characteristic of the material. The main objective of this dissertation is to study the impact of temperature and thickness of a coating material on its passive regulation potential of indoor air RH. For that purpose, two types of hygroscopic coatings materials are studied: earth mortar plasters and hemp concretes. The influence of the thickness is analysed through dynamic MBV test (“Moisture Buffering Value“ test) at 23°C on three formulations of each material (earth and hemp concrete) at three different thicknesses. The influence of temperature is performed only for hemp concrete formulations on static (sorption curves) and dynamic (MBV tests) tests at 23°C and 40°C. The analysis of the results, made using the support of theoretical models, allows underlining that the thickness involved in the daily passive control of RH remains most of the time limited (only the first centimetre of the earth plasters and 5cm for hemp concretes). However, some surface irregularities may have a strong impact on this active thickness. The other main result of this dissertation is the strong impact of temperature on both static and dynamic hygroscopic characteristics of the material. Indeed, the increase of temperature at a constant RH tends to significantly reduce the water content of the material. But, on the other side, due to higher vapour pressure at saturation and water vapour diffusion coefficient, the dynamic sorption-desorption properties increased with temperature by approximately 65%.Developed within the IBIS project and BIOTERRA project with the collaboration of the "Direction Territoriale Centre-Est" of the CEREM

Development and characterization of functional ingredients from olive pomace : bioactivity and potential application

Atualmente, grande volume dos resíduos gerados ao longo da cadeia alimentar Europeia é produzido durante o processamento alimentar (19-39%). A indústria de extração de azeite não é exceção, gerando anualmente grandes quantidades de subprodutos e resíduos com elevado impacto fitotóxico, mas que são também ricos em diversos compostos bioativos e nutrientes com potencial aplicação como ingredientes alimentares. Assim sendo, é imprescindível a adoção de estratégias de redução do desperdício e de valorização dos subprodutos, no sentido de melhorar a sustentabilidade económica e ambiental da indústria alimentar. Atualmente, o bagaço de azeitona é o subproduto de maior relevo da indústria de azeite e a sua valorização adequada é também o maior desafio do sector para alcançar uma cadeia produtiva sustentável alinhada com os princípios da Bioeconomia Circular. Esta tese visou alcançar uma valorização “zero desperdício” para o bagaço de azeitona, priorizando os produtos de alto valor acrescentado sobre os de baixo valor através da produção e caracterização de ingredientes alimentares, nomeadamente ingredientes em pó ricos em vários compostos bioativos, seguida da validação in vitro das suas atividades biológicas ligadas a benefícios para a saúde, como a prevenção e redução de doenças cardiovasculares, diabetes, problemas intestinais e posterior aplicação no desenvolvimento de alimentos funcionais. Uma estratégia de “desperdício zero” foi alcançada para o bagaço de azeitona. Esta estratégia consiste num procedimento simples de fracionamento, em que se obteve uma fração líquida que revelou um alto teor de hidroxitirosol (513,61-625,76 mg/100 g de peso seco) e uma fração de polpa caracterizada por ser uma boa fonte de fibra alimentar antioxidante. Adicionalmente, obteve-se ainda uma fração de caroços que exibiu valores caloríficos significativamente elevados (18,65-18,94 MJ/kg). Estes resultados validaram o potencial valor acrescentado das frações líquidas e de polpa como ingredientes alimentares e como biocombustível no caso da fração de caroços. Após validação do potencial da fração líquida e de polpa do bagaço de azeitona como ingredientes alimentares, dois novos ingredientes em pó, estáveis e seguros, foram desenvolvidos sem qualquer etapa de extração, nomeadamente um pó enriquecido na fração líquida (LOPP) e um pó enriquecido na fração de polpa (POPP). O LOPP exibiu uma quantidade significativa de manitol (141 g/kg), potássio (54 g/kg) e hidroxitirosol/derivados (5 mg/g). O POPP exibiu uma grande quantidade de fibra alimentar (620 g/kg) associada a um teor significativo de fenólicos (7,41 mg equivalentes de ácido gálico/g fibra) com atividade antioxidante substancial. O POPP também revelou uma composição de ácidos gordos insaturados semelhante à do azeite (76% do total de ácidos gordos), sendo ainda uma fonte razoável de proteína (12%). As suas propriedades funcionais (solubilidade, capacidade de retenção de água e de óleo), capacidade antioxidante e atividade antimicrobiana foram também avaliadas, assim como a sua segurança biológica. Em síntese, a multifuncionalidade e aplicabilidade dos pós de bagaço de azeitona como fortificantes nutricionais, promotores de benefícios para a saúde e conservantes foram atestadas. A digestão simulada in vitro permitiu concluir que os fenólicos e minerais do LOPP foram afetados negativamente pela digestão, no entanto, uma bioacessibilidade significativa de potássio e hidroxitirosol foi verificada ( 57%) e as suas bioatividades foram apenas moderadamente afetadas. Em relação às suas bioatividades, pelo menos metade da atividade antioxidante (capacidade de Resumo x absorção do radical de oxigénio), de inibição da -glucosidase e de inibição da enzima conversora de angiotensina (91,98%) foram retidas e bioacessíveis após a digestão. Por sua vez, a digestão do POPP revelou uma perda significativa de fenólicos na digestão oral (62,48%), mas o papel da fibra como veículo de fenólicos permitiu recuperar uma quantidade significativa de fenólicos no estômago (77,11%) e um índice de bioacessibilidade de pelo menos 50% (tirosol e o seu glucosídeo). A fibra do POPP também interagiu positivamente com os ácidos gordos, diminuindo e facilitando bioacessibilidade dos saturados (5-6%) e dos insaturados (4-11%), respetivamente. Após realização da digestão in vitro dos pós de bagaço de azeitona, as frações disponíveis no cólon de ambos os pós exibiram uma composição abundante em compostos associados a potenciais benefícios para a saúde intestinal, nomeadamente fibra e compostos fenólicos e fibra. Assim sendo, procedeu-se a fermentação fecal in vitro das frações não digeridas retidas no cólon após a digestão gastrointestinal simulada in vitro do LOPP e POPP. Os resultados da análise do gene 16S rRNA e dos ácidos gordos de cadeia curta mostraram que estes pós não tiveram um impacto negativo na diversidade da microbiota intestinal e que promoveram uma maior produção de ácidos de cadeia curta (acetato>butirato>propionato) do que os frutooligossacarídeos. A identificação dos metabólitos fenólicos dos pós de bagaço de azeitona por LC-ESI-UHR-QqTOF-MS seguida das análises de clustering supervisionada e clustering hierárquico permitiram avaliar os padrões destes metabólitos ao longo da fermentação fecal in vitro, bem como explicar as potenciais atividades biológicas destes pós no intestino. Ambos os pós apresentaram um conteúdo fenólico e uma capacidade de absorção de radical de oxigénio significativas e capacidade de inibição da adesão de patógenos como o Bacillus cereus (22,03%) e Listeria monocytogenes (20,01%), sobretudo pelo POPP. Por fim, para validar o desempenho dos pós de bagaço de azeitona como ingredientes funcionais, LOPP e POPP foram incorporados em iogurte para aumentar o seu conteúdo em fibra alimentar, hidroxitirosol e ácidos gordos insaturados. A adição de POPP (2%) ao iogurte permitiu cumprir a condição de “fonte de fibra”, por sua vez LOPP (1%) forneceu 5 mg de hidroxitirosol/ derivados num iogurte (120 g). A adição de POPP e azeite juntamente com LOPP ou POPP permitiu uma melhoria do perfil de ácidos gordos insaturados dos iogurtes. Além disso, todos os iogurtes fortificados apresentaram um maior teor de fenólicos e atividade antioxidante. A digestão in vitro reduziu a atividade antioxidante de todas as formulações e mostrou que a incorporação de azeite juntamente com o LOPP e POPP aumentou a bioacessibilidade de hidroxitirosol e do hidroxitirosol glucosídeo, respetivamente. Contudo, todas as formulações com os pós de bagaço de azeitona podem ser consideradas boas fontes de hidroxitirosol, lípidos insaturados e fibras para os consumidores. Em suma, os resultados deste trabalho irão contribuir para a sustentabilidade da indústria do azeite no contexto da bioeconomia circular, através do desenvolvimento de ingredientes de valor acrescentado destinados à indústria alimentar com potencial impacto positivo na saúde, proporcionando ainda um biocombustível sólido.Currently, a large amount of the total European food supply chain waste (between 19 and 39%) is produced during food processing. The olive oil industry is no exception, generating large quantities of by-products and wastes annually with high phytotoxic impact, but also rich in several bioactive compounds and nutrients with potential application as food ingredients. Thus, it is imperative to implement strategies to reduce waste and valorise by-products to improve the food industry's economic and environmental sustainability. Olive pomace is the most prominent by-product of the modern olive oil industry, and its proper valorisation is one of the major obstacles to olive oil industry achieve a sustainable production chain aligned with the principles of the Circular Bioeconomy concept. This project aimed at achieving a “zero waste” valorising approach to olive pomace that prioritised the high over low added-value products, through the production and characterisation of food ingredients, namely high-value powdered ingredients rich in several bioactive compounds, followed by the validation of their biological activities related with health benefits as prevention and reduction of the prevalence of cardiovascular diseases, diabetes and gut disorders and, their application in the development of functional foods. A “zero waste” strategy was achieved for olive pomace. This strategy is based on a feasible fraction approach, and it was obtained a liquid fraction that revealed a high amount of hydroxytyrosol (513.61- 625.76 mg/100 g dry weight) and pulp fraction that was characterised to be a good source of antioxidant dietary fibre. Additionally, it was achieved a stones fraction that exhibited substantial higher heating values (18.65-18.94 MJ/kg). These results supported the potential value-added of the liquid and pulp fractions from olive pomace as functional food ingredients and as biofuel to stones. After validating the liquid and pulp fraction potential as food ingredients, new two stable and safe, powdered ingredients without any extraction step were developed, namely a liquid-enriched powder (LOPP) and a pulp-enriched powder (POPP). LOPP exhibited a significant amount of mannitol (141 g/kg), potassium (54 g/kg) and hydroxytyrosol/derivatives (5 mg/g). POPP exhibited a high amount of dietary fibre (620 g/kg) associated with a significant amount of bound phenolics (7.41 mg gallic acid equivalents/g fibre dry weight) with substantial antioxidant activity. POPP also revealed an unsaturated fatty acid composition similar to that of olive oil (76% of total fatty acids) and a reasonable source of protein (12%). Their functional properties (solubility, water‐holding and oil‐ holding capacity), antioxidant capacity and antimicrobial activity were also assessed, and their biological safety was validated. The multifunctionality and applicability as nutritional enhancers, health-benefits promoters and preservatives of the olive pomace powders were recognised. The in vitro simulated digestion allowed concluding that phenolics and minerals from LOPP were negatively affected by digestion. However, a significant bioaccessibility of potassium and hydroxytyrosol was verified ( 57%) and its bioactivities were only moderately affected. Regarding its bioactivities, at least half of the antioxidant activity (measured by Oxygen Radical Absorbance Capacity assay), -glucosidase inhibition activity and angiotensin-converting enzyme inhibitory activity (91.98%) were retained and bioaccessible after in vitro digestion. In its turn, POPP digestion Abstract xii revealed a significant loss of phenolics in the oral step (62.48%), but the dietary fibre role as phenolics’ carrier allowed recovering a significant amount of phenolics in the stomach (77.11%) and a bioaccessibility index of at least 50% (mainly for tyrosol and its glucoside). POPP dietary fibre also positively interacts with lipids decreasing the saturated fatty acids bioaccessibility (5-6%) and facilitating the unsaturated fatty acid bioaccessibility (4-11%). After the in vitro digestion system of both olive pomace powders, the colon-available fractions LOPP and POPP exhibited an abundant composition in phenolics, but also dietary fibre with potential gut health benefits as antioxidants, antimicrobial and prebiotic agents. So, the gut's potential beneficial effects were analysed by in vitro faecal fermentation of the undigested fractions retained in the colon after in vitro simulated gastrointestinal digestion. The 16S rRNA gene analysis results showed that LOPP and POPP did not negatively impact gut microbiota diversity. The short-chain fatty acids analysis showed a higher production of these fatty acids (acetate > butyrate > propionate) by LOPP and POPP than by fructooligosaccharides. The identification of phenolic metabolites by LC-ESIUHR-QqTOF-MS followed by supervised cluster and hierarchical clustering analysis allowed to evaluate the metabolite patterns of olive pomace powders phenolics throughout faecal fermentation, explaining the potential biological activities exhibited in the gut by these powders. Both powders showed significant total phenolic content and oxygen radical absorbance capacity during faecal fermentation. But also, mucin‐adhesion inhibition ability against pathogens as Bacillus cereus (22.03%) and Listeria monocytogenes (20.01%), specially POPP. Finally, to validate olive pomace powders' performance as functional ingredients, LOPP and POPP were incorporated into yoghurt to increase its content in dietary fibre, hydroxytyrosol, and unsaturated fatty acids. POPP (2%) and LOPP (1%) addition to yoghurt allowed fulfilling the condition on being a “source of fibre” and provides 5 mg of hydroxytyrosol and derivatives in a standard yoghurt (120 g), respectively. The addition of olive oil, together with olive pomace powders, was also investigated in order to understand the lipids-phenolics interaction, and not only POPP addition improved the yoghurts’ unsaturated fatty acids profile, but the olive oil addition together both olive pomace powders also improved too. Besides that, all yoghurts fortified with olive pomace powders exhibited higher total phenolic content and antioxidant activity. The in vitro digestion of yoghurts showed that olive oil incorporation together with olive pomace powders increased the bioaccessibility of hydroxytyrosol (LOPP + olive oil) and hydroxytyrosol glucoside (POPP + olive oil). Despite the reduction of antioxidant activity after digestion, the yoghurts formulations with olive pomace powders could be considered suitable carriers to deliver hydroxytyrosol, unsaturated fatty acids and fibre to the consumers. In short, the results from this work will contribute to the sustainability of the olive oil industry in circular bioeconomy context, through the development of value-added ingredients to the food industry with a positive impact in health, offering also a solid biofuel

UEFA Euro 2004 Visitors Analysis

The purpose of this paper is to establish the profile of the foreign visitor that attended the UEFA Euro 2004 Championship in Portugal, namely in the Minho region. Data were collected through a one-to-one inquiry carried out before the matches that took place in Braga e Guimarães. The survey instrument included aspects like the visitors´past comsumption behaviour, media audience and live attendance sport habits. The results we got show that sport event tourists living in countries with higher per capita income spent more in the UEFA Euro 2004. They also stayed in country longer than other visitors. Other valuable information for both tourism operators and tournament managers we got is that most sport event tourists decided to overnight on sites with efficient and direct accesses to the matches. Otherwise, they may be characterized as usual sport consumers in terms of active and passive sport comsumption behaviour. The return of visitors to the sites where the UEFA Euro 2004 took place remains unsolved. Future studies should concentrate on community reimbursement and mid/long term benefits.UEFA Euro 2004, economic impact of major sport events, sports tourism.

UEFA Euro 2004 Tourism Impact Analysis

The purpose of this paper is exactly to approximate the tourism economic impact of the UEFA Euro 2004 in Portugal. The survey concentrates on the immediate, direct and short-term additional revenue brought into one region by the foreign sport event spectators. One innovating aspect of this paper is the direct data collection on the foreign visitors spending, contouring simulation and forecast problems. Results show an immediate short-term return investment costs generated an immediate and short-term revenue of less than one tenth of the investment costs and overall revenue leakage (or income export) of nearly 325% (50,142 million Euro) of the revenue within the defined regional spatial circuit.UEFA Euro 2004, tourism impact of major sport events, sport event tourism.

Improving the early diagnostic of prostate cancer by multiple biomarker detection with new biosensing devices

Prostate cancer (PCa) is the most common form of cancer in men, in Europe (World Health Organization data). The most recent statistics, in Portuguese territory, confirm this scenario, which states that about 50% of Portuguese men may suffer from prostate cancer and 15% of these will die from this condition. Its early detection is therefore fundamental. This is currently being done by Prostate Specific Antigen (PSA) screening in urine but false positive and negative results are quite often obtained and many patients are sent to unnecessary biopsy procedures. This early detection protocol may be improved, by the development of point-of-care cancer detection devices, not only to PSA but also to other biomarkers recently identified. Thus, the present work aims to screen several biomarkers in cultured human prostate cell lines, serum and urine samples, developing low cost sensors based on new synthetic biomaterials. Biomarkers considered in this study are the following: prostate specific antigen (PSA), annexin A3 (ANXA3), microseminoprotein-beta (MSMB) and sarcosine (SAR). The biomarker recognition may occurs by means of molecularly imprinted polymers (MIP), which are a kind of plastic antibodies, and enzymatic approaches. The growth of a rigid polymer, chemically stable, using the biomarker as a template allows the synthesis of the plastic antibody. MIPs show high sensitivity/selectivity and present much longer stability and much lower price than natural antibodies. This nanostructured material was prepared on a carbon solid. The interaction between the biomarker and the sensing-material produces electrical signals generating quantitative or semi-quantitative data. These devices allow inexpensive and portable detection in point-of-care testing