Recombinant expression and functional characterisation of regiospecific flavonoid glucosyltransferases from Hieracium pilosella L.

Five glucosyltransferases were cloned by RT-PCR amplification using total RNA from Hieracium pilosella L. (Asteraceae) inflorescences as template. Expression was accomplished in Escherichia coli, and three of the HIS-tagged enzymes, UGT90A7, UGT95A1, and UGT72B11 were partially purified and functionally characterised as UDP-glucose:flavonoid O-glucosyltransferases. Both UGT90A7 and UGT95A1 preferred luteolin as substrate, but possessed different regiospecificity profiles. UGT95A1 established a new subgroup within the UGT family showing high regiospecificity towards the C-3' hydroxyl group of luteolin, while UGT90A7 primarily yielded the 4'-O-glucoside, but concomitantly catalysed also the formation of the 7-O-glucoside, which could account for this flavones glucoside in H. pilosella flower heads. Semi quantitative expression profiles revealed that UGT95A1 was expressed at all stages of inflorescence development as well as in leaf and stem tissue, whereas UGT90A7 transcript abundance was nearly limited to flower tissue and started to develop with the pigmentation of closed buds. Other than these enzymes, UGT72B11 showed rather broad substrate acceptance, with highest activity towards flavones and flavonols which have not been reported from H. pilosella. As umbelliferone was also readily accepted, this enzyme could be involved in the glucosylation of coumarins and other metabolite

Intra- and inter-metabolite correlation spectroscopy of tomato metabolomics data obtained by liquid chromatography-mass spectrometry and nuclear magnetic resonance

Nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LCMS) are frequently used as technological platforms for metabolomics applications. In this study, the metabolic profiles of ripe fruits from 50 different tomato cultivars, including beef, cherry and round types, were recorded by both 1H NMR and accurate mass LC-quadrupole time-of-flight (QTOF) MS. Different analytical selectivities were found for these both profiling techniques. In fact, NMR and LCMS provided complementary data, as the metabolites detected belong to essentially different metabolic pathways. Yet, upon unsupervised multivariate analysis, both NMR and LCMS datasets revealed a clear segregation of, on the one hand, the cherry tomatoes and, on the other hand, the beef and round tomatoes. Intra-method (NMR¿NMR, LCMS¿LCMS) and inter-method (NMR¿LCMS) correlation analyses were performed enabling the annotation of metabolites from highly correlating metabolite signals. Signals belonging to the same metabolite or to chemically related metabolites are among the highest correlations found. Inter-method correlation analysis produced highly informative and complementary information for the identification of metabolites, even in de case of low abundant NMR signals. The applied approach appears to be a promising strategy in extending the analytical capacities of these metabolomics techniques with regard to the discovery and identification of biomarkers and yet unknown metabolites

Factors Associated with Marburg Hemorrhagic Fever:

Background. Reliable on-site polymerase chain reaction (PCR) testing for Marburg hemorrhagic fever (MHF) is not always available. Therefore, clinicians triage patients on the basis of presenting symptoms and contact history. Using patient data collected in Uige, Angola, in 2005, we assessed the sensitivity and specificity of these factors to evaluate the validity of World Health Organization (WHO)–recommended case definitions for MHF. Methods. Multivariable logistic regression was used to identify independent predictors of PCR confirmation of MHF. A data-derived algorithm was developed to obtain new MHF case definitions with improved sensitivity and specificity. Results. A MHF case definition comprising (1) an epidemiological link or (2) the combination of myalgia or arthralgia and any hemorrhage could potentially serve as an alternative to current case definitions. Our dataderived case definitions maintained the sensitivity and improved the specificity of current WHO-recommended case definitions. Conclusions. Continued efforts to improve clinical documentation during filovirus outbreaks would aid in the refinement of case definitions and facilitate outbreak control

All Under Control: How Trait Emotional Intelligence May Moderate the Effect of State Arousal on Implicit Attitudes Towards Academic Dishonesty

The current study examined predictors in the changing of implicit attitudes toward academic dishonesty (AD). The primary predictor measured was self-reported stress and arousal, the strength of which being a possible influence on one’s change in implicit attitudes. Trait emotional intelligence (EI) was measured as a covariate and potential moderator of the interaction between stress and arousal, and one’s implicit attitudes toward AD. One hundred nine undergraduate psychology students from a public university were recruited to take assessments for trait EI, stress and arousal, and implicit attitudes toward AD. To manipulate the induction of stress and arousal, participants were randomly assigned between two groups. One of which played Geometry Dash to induce stress and arousal, and the other played Patchworkz as a control condition. Results of the study found that those who played Geometry Dash experienced greater stress and arousal than those who played Patchworkz. Trait EI was negatively correlated with stress and arousal before intervention. There were no significant differences in implicit attitudes toward AD between groups, but significant differences between times, with decreases for both groups after intervention. Stress and arousal after intervention did not significantly predict implicit attitudes toward AD, nor did any interactions between group, stress and arousal, and trait EI. Implications of the study include how gaming may be used as a subtle intervention for inducing stress and arousal, how trait EI may facilitate emotion regulation, and how the interactions between stress and arousal, trait EI, and implicit attitudes toward AD require further investigation

Successful Corporate Social Responsibility Initiative Incorporation Strategies for Small Business Leaders

Some small business leaders lack strategies to incorporate corporate social responsibility (CSR) initiatives into their organizations to manage financial risk. Small business leaders who do not successfully use CSR to balance stakeholder interests may face additional or avoidable financial hardships. Grounded in stakeholder theory, the purpose of this qualitative single case study was to explore successful strategies small business leaders use to incorporate CSR initiatives into their organizations to manage financial risk. The participants were four business leaders from a small business in the mid-Atlantic and Midwest regions of the United States. Data were gathered from organizational documents, archival records, and semistructured interviews. The framework method of thematic coding was used to analyze the data. Key themes were meaningful communication, using voice-of-the-customer data, and consensus decision-making. A key recommendation from this study is for small business leaders to use voice-of-the-customer data to create new revenue streams and improve product and service quality. Implications for positive business and social change include the potential for other small business leaders to use CSR strategies to manage financial risk. Because stakeholders use CSR initiatives to hold business leaders accountable for their influence on society, this study demonstrated how CSR helped create social infrastructure. Viewing risk from a stakeholder perspective might help other small business leaders balance stakeholder interests leading to collaborative, loyal relationships with their stakeholders. Therefore, small business leaders could manage financial risk by incorporating CSR initiatives based on stakeholder interests into their organization

Acute effects of flavonoid-rich blueberry on cognitive and vascular function in healthy older adults

BACKGROUND Consumption of flavonoids, natural compounds found in foods such as berries and cocoa, have been shown to be beneficial for cognitive function. However, less is known about potential mechanisms and acute benefits for the older population. OBJECTIVE To determine whether acute intake of flavonoid-rich blueberry could have beneficial effects on cognitive function in a sample of healthy older adults and identify possible mechanisms. METHODS A cross-over randomised controlled trial (RCT) was conducted (N = 18) with volunteers receiving a flavonoid-rich blueberry beverage (579mg of antho- and pro-cyanidins) on one visit and a sugar-matched control on another. Cognitive function was measured at baseline, 2 and 5 hours post consumption of the intervention and blood pressure (BP), arterial stiffness and plasma brain-derived neurotrophic factor (BDNF) concentration at baseline and an hour post-intervention. RESULTS Whilst there was no significant effect of the intervention on global cognitive function, performance was significantly different at 2 compared to 5 hours following the control beverage (p   0.05). CONCLUSIONS A single dose of flavonoid-rich blueberry could have a potentially protective effect on cognitive function in healthy older adults, possibly due to improved cerebrovascular function and positive interactions with cell signalling pathways involved in cognitive processes

Contribuição para uma apreciação jus-internacional da integração regional africana

Tese de doutoramento, Direito (Ciências Jurídico-Políticas), Universidade de Lisboa, Faculdade de Direito, 2016A partir da pretensão inicial, expressa no título e tema, este trabalho acaba por resultar numa reflexão aturada e solitária (como aliás muitos autores o destacam no caso das suas teses de doutoramento) que ultrapassou o mero âmbito da “apreciação jus-internacional da integração regional africana”, para desembocar num levantamento (pelo menos) de pistas sobre o fenómeno e as concepções do Direito, em geral, e particularmente na interacção historicamente inevitável entre os direitos públicos africano e euro-ocidental. Diferentemente da maioria dos trabalhos académicos sobre a África dita moderna (tanto por africanos como por outras entidades, que não precisam sequer de exibir qualquer tipo de ostensivo eurocentrismo, já minimamente extirpado do mainstream formal hodierno), que partem, geralmente, dos pressupostos teorético-formais euro-ocidentais, transportados, quase sempre de forma acrítica para o Continente, em processos legislativos e noutros aspectos hermenêutico-aplicativos do Direito, nós seguimos uma metodologia que julgamos singular. Singularidade que não reside sequer ou simplesmente na ideia de partirmos de uma pretensa rede de sistemas teorético-formais “genuinamente africanos”, o que, desde logo, conflituaria com a ideia apurada de que essa “África moderna” é uma “invenção” tão recente e prenhe de tensões que mal a deixariam criar tais sistemas com alguma consistência e coerência. Entendemos que a singularidade metodológica deste empreendimento reside, essencialmente, no facto de que tentamos partir da realidade nua e crua da factologia histórica, antropológica e cultural múltipla e diversa dessa África moderna, que resultou da traumática experiência colonial e pós-colonial, sem no entanto deixarmos de relacioná-la com o seu passado mais remoto (perante si e perante o resto da Humanidade). Ora, ao confrontarmos essa pura realidade africana com as metodologias aplicativas do direito moderno (estamos essencialmente no domínio da criação e “efetivização” do direito público, base do nosso objeto de dissertação) de matriz, necessariamente, euro-ocidental, embembido no seu extasiante perfume formal-positivista que lhe vem de Roma, do Renascimento europeu e do Século das Luzes, deparamo-nos com um enorme “buraco negro”, devorador atroz do conteúdo e do sentido desse tipo de direito, no Continente, e desintegrador, ao mesmo tempo, de sistemas tradicionais positivos, no sentido da dignidade e da dignificação humana. Concluímos, pois, que o problema não reside nem na realidade, que é e será sempre inelutável como a força do vento, nem propriamente na formulação da proposição jurídica de cariz euro-ocidental que acompanha essa infiltração, inicialmente exógena, mas que se tornaria ela própria irreversível e integradora necessária da realidade africana hodierna. Propomos, assim, uma nova metodologia de aplicação do direito: olhar para as atitudes dos agentes políticos e de outros aplicadores do Direito, acima das estritas normas jurídicas formais, perante os imperativos ético-morais que a situação do Continente exige. Quanto ao que ao “direito de integração regional e continental” diz respeito, nem sequer é o seu conteúdo e sentido que são absorvidos pelo aludido “buraco negro”, mas antes os das próprias regras adjectivas programadas para a sua criação, autonomizando-o de um direito internacional africano, ele próprio vogando em atribulados mares e agitados ”grandes lagos”. Na África Austral, particularmente na África do Sul, com base na presumível sublimação da filosofia tradicional Ubuntu, tentando sustentar uma convivência na multirracialidade e multiculturalidade, não obstante a pressão a que está sujeita pelo mainstream “afrocentrista”, encontramos o modelo em que tal “buraco negro” é, aparentemente, diminuto, tendo em conta a preservação dos dois pilares (europeu e nativo) em que assenta a sociedade, com reflexos positivos no funcionamento e aplicação do Direito, na base de um casamento, aparentemente ideal, entre a tradição e a modernidade. Enquanto isso, em Angola, estudada como caso médio da África Negra, encontramos o protótipo de uma sociedade em que o “buraco” devorador de conteúdos e sentidos jurídicos é avassalador, como resultado tanto de uma política assimilacionista-integracionista colonial, seguida de outra, a do racionalismo marxista-leninista, que contribuiu para o acanhamento e desvirtuamento da axiologia nativa ou tradicional, completando-se o drama com a diluição dos valores positivos da modernidade ocidental, com as prolongadas guerras civis que determinaram, inicialmente, um êxodo considerável de populações de origem europeia, como suportes deste outro pilar da construção da nação africana moderna, em Angola. De todo o modo, podemos nos aperceber de alguns traços de um direito de integração regional africana, reflectidos e sustentados, ao mesmo tempo, no e pela emergência de uma frágil jurisprudência. Fica a dúvida de se saber se esses diáfanos traços poderão sobreviver à virulência das “próximas chuvas” e preencher-se de vez o vazio lamentado pelos últimos pan-africanistas como Ki-Zerbo e Édem KodjoStarting from the initial intention set out in its title and theme, this work has ultimately resulted in a thorough and solitary reflection (a point that many authors emphasise regarding their doctoral theses), which went beyond the "assessment based in international law of African regional integration", and ended up outlining, at least, the phenomenon and the conceptions of law in general, and particularly the historically inevitable interaction between African and Euro-Western public law. Most academic work on so-called modern Africa tends to start from Euro-Western theoretical and formal assumptions that are transported almost always in an uncritical manner to the African continent in the legislative process and in other hermeneutic-applicative aspects of the law. This is true for work both by Africans and by others, even if they do not display the slightest signs of Eurocentrism, something that has already started to be removed from the mainstream of today’s thinking. Instead, we follow a methodology that we believe to be unique. This uniqueness does not reside simply in the idea that we are starting from a supposed network of “genuinely African” theoretical-formal systems; such an approach would soon come into conflict with the established idea that this “modern Africa” is an “invention” so recent and so riven with tensions that would barely allow it to create such systems in a coherent and consistent manner. Instead, we believe that the methodological uniqueness of this project resides essentially in the fact that we try to start from the bare and crude reality of the multiple and diverse historical, anthropological and cultural facts of this modern Africa, which result from the traumatic colonial and postcolonial experience, but which we also relate to its more distant past (both within itself and in relation to the rest of humanity). Now when we confront this purely African reality with the applicative methodologies of modern law (we are essentially in the domain of the creation and implementation of public law, the basis of our object of study) that is necessarily within a Euro-Western matrix, suffused with the heady formal-positivist perfume that comes from Rome, from the European Renaissance and from the Enlightenment, we find ourselves with a huge “black hole” that swallows up the content and meaning of this type of law on the African continent, and at the same time destroys traditional systems that are positive in the sense of conferring human dignity. We conclude, therefore, that the problem lies neither in the reality, which is and will always be as elusive as the wind, nor exactly in the formulation of the Euro-Western version of juridical proposition that comes with this infiltration, at first from the outside, but which would become irreversible and which would necessarily merge into contemporary African reality. Thus we propose a new methodology for the application of the law: to study the attitudes of political actors and others who apply the law outside of the strict formal legal norms when faced with the ethical and moral imperatives that the continent’s situation demands. As for law governing “continental and regional integration”, it is not so much its content and meaning that are swallowed up by the supposed “black hole”, but rather the very procedural rules that were meant to allow the creation of such a law and to empower it as African international law, are themselves drifting in stormy seas. Within Southern Africa, it is in South Africa that we find a model in which the “black hole” is reduced and which takes into account the two pillars (European and indigenous) on which the society is constructed. This is presumably on the basis of South Africa having enshrined the traditional Ubuntu philosophy in an attempt to sustain multiracial and multicultural coexistence despite coming under pressure from the “Afrocentric” mainstream, and this decision has had positive consequences in the functioning and application of the law based on an apparently ideal marriage between tradition and modernity. Meanwhile, in Angola, taken as a more typical African case, we find the prototype of a society in which the “black hole” that swallows legal content and meanings is overwhelming, the result of the assimilationist / integrationist colonial policy followed by one of Marxist-Leninist rationalism, which contributed to the weakening and distortion of indigenous or traditional axiology. The drama was completed with the erosion of the positive aspects of western modernity with the long civil wars, which caused the exodus of a large part of the population of European origin who could have been a pillar for the construction of a modern African nation in Angola. Notwithstanding, we could catch a glimpse of some footsteps of an african modern communitarian law, reflected in and sustained by a few and fragile number of justice cases. What we can not predict is wether this faint footsteps are capable to resist to the turbulent rains coming up, so that the regretted void by the last pan-africanists like Ki-Zerbo and Edem Kodjo can be filled up.Fundação Calouste Gulbenkia

Medium- and high-cell-density production of Adeno-Associated virus serotype 6 and the mitigation of cell density effect via medium supplementation

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Metabolomics for bioactivity assessment of natural products.

Natural products historically have been a rich source of lead molecules in drug discovery, based on their capability to create unique and diverse chemical structures. However, it is also true that the vast number of metabolites typically present in natural products and their huge dynamic range results in the loss of many possibly bioactive natural compounds, becoming an inextricable obstacle for drug development. Recently, new strategies which favour a holistic approach as opposed to the traditional reductionist methods used previously, have been introduced with the purpose of overcoming the bottlenecks in natural product research. This approach is based on the application of new technologies, including metabolomics, for example. Metabolomics allows a systematic study of a complex mixture such as a phytochemical preparation, which can be linked to observations obtained through biological testing systems without the need for isolating active principles. This may put drug discovery from natural products back in the limelight again. In this review paper, the description of some examples of successful metabolomics applications in several important fields related to drug discovery from natural sources aims at raising the potential of metabolomics in reducing the gap between natural products (NP) and modern drug discovery demand