DNA-dependent Protein Kinase-mediated Phosphorylation of Protein Kinase B Requires a Specific Recognition Sequence in the C-terminal Hydrophobic Motif

DNA-dependent protein kinase (DNA-PK) has been implicated in a variety of nuclear processes including DNA double strand break repair, V(D)J recombination, and transcription. A recent study showed that DNA-PK is responsible for Ser-473 phosphorylation in the hydrophobic motif of protein kinase B (PKB/Akt) in genotoxic-stressed cells, suggesting a novel role for DNA-PK in cell signaling. Here, we report that DNA-PK activity toward PKB peptides is impaired in DNA-PK knock-out mouse embryonic fibroblast cells when compared with wild type. In addition, human glioblastoma cells expressing a mutant form of DNA-PK (M059J) displayed a lower DNA-PK activity when compared with glioblastoma cells expressing wild-type DNA- PK (M059K) when PKB peptide substrates were tested. DNA- PK preferentially phosphorylated PKB on Ser-473 when compared with its known in vitro substrate, p53. A consensus hydrophobic amino acid surrounding the Ser-473 phospho-acceptor site in PKB containing amino acids Phe at position +1 and +4 and Tyr at position -1 are critical for DNA- PK activity. Thus, these data define the specificity of DNA- PK action as a Ser-473 kinase for PKB in DNA repair signaling.

Vision Modulates Corticospinal Suppression in a Functionally Specific Manner during Movement of the Opposite Limb

The effect of vision on the excitability of corticospinal projections to the flexor carpi radialis (FCR) and extensor carpi radialis (ECR) muscles of right human forearm was investigated before and during discrete movement of the opposite limb. An external force opposed the initial phase of the movement (wrist flexion) and assisted the reverse phase, so that recruitment of the wrist extensors was minimized. Three conditions were used as follows: viewing the inactive right limb (Vision), viewing the mirror image of the moving left limb (Mirror), and with vision of the right limb occluded (No Vision). Transcranial magnetic stimulation was delivered to the left motor cortex: before, at the onset of, or during the left limb movement to obtain motor evoked potentials (MEPs) in the muscles of the right forearm. At and following movement onset, MEPs obtained in the right FCR were smaller in the Vision condition than in the Mirror and No Vision conditions. A distinct pattern of variation was obtained for the ECR. In all conditions, MEPs in this muscle were elevated upon or following movement of the opposite limb. An additional analysis of ipsilateral silent periods indicated that interhemispheric inhibition plays a role in mediating these effects. Activity-dependent changes in corticospinal output to a resting limb during discrete actions of the opposite limb are thus directly contingent upon where one looks. Furthermore, the extent to which vision exerts an influence upon projections to specific muscles varies in accordance with the functional contribution of their homologs to the intended action.

Assessing the sensitivity and specificity of fish community indicators to management action

We assessed ten trophodynamic indicators of ecosystem status for their sensitivity and specificity to fishing management using a size-resolved multispecies fish community model. The responses of indicators to fishing depended on effort and the size selectivity (sigmoid or Gaussian) of fishing mortality. The highest specificity against sigmoid (trawl-like) size selection was seen from inverse fishing pressure and the large fish indicator, but for Gaussian size selection, the large species indicator was most specific. Biomass, mean trophic level of the community and of the catch, and fishing in balance had the lowest specificity against both size selectivities. Length-based indicators weighted by biomass, rather than abundance, were more sensitive and specific to fishing pressure. Most indicators showed a greater response to sigmoid than Gaussian size selection. Indicators were generally more sensitive at low levels of effort because of nonlinear sensitivity in trophic cascades to fishing mortality. No single indicator emerged as superior in all respects, so given available data, multiple complementary indicators are recommended for community monitoring in the ecosystem approach to fisheries management.

PROBLEM-SOLVING IN SCHIZOPHRENIA - A SPECIFIC DEFICIT IN PLANNING ABILITY

The study investigated problem solving ability in schizophrenia. Thirty DSM-IIIR schizophrenic patients and 27 matched normal controls were tested on the Three-Dimensional Computerised Tower of London Test (3-D CTL Test) (Morris et al., 1993). The patients took significantly more moves to solve a series of problems and solved significantly fewer problems in the predetermined minimum number of moves. The patients response times, as measured using a control version of the task (the 3-D CTL Control), were longer than those of the controls. However, when latencies were adjusted to take into account the slower responses overall, the patients planning times were not significantly increased. Inaccurate planning, as defined by taking more moves, did not correlate with either positive or negative symptoms, but the response times tended to be longer in patients who had more negative symptoms. The findings suggest that there is a deficit in problem solving activity in schizophrenia that may be associated with translating 'willed intentions' into action, independent of slower motor speed.

Steps to a better Belfast: physical activity assessment and promotion in primary care

BACKGROUND: Low physical activity (PA) levels which increase the risk of chronic disease are reported by two-thirds of the general UK population. Promotion of PA by primary healthcare professionals is advocated but more evidence is needed regarding effective ways of integrating this within everyday practice. This study aims to explore the feasibility of a randomised trial of a pedometer-based intervention, using step-count goals, recruiting patients from primary care. METHOD: Patients, aged 35-75, attending four practices in socioeconomically deprived areas, were invited to complete a General Practice PA Questionnaire during routine consultations. Health professionals invited 'inactive' individuals to a pedometer-based intervention and were randomly allocated to group 1 (prescribed a self-determined goal) or group 2 (prescribed a specific goal of 2500 steps/day above baseline). Both groups kept step-count diaries and received telephone follow-up at 1, 2, 6 and 11 weeks. Step counts were reassessed after 12 weeks. RESULTS: Of the 2154 patients attending, 192 questionnaires were completed (8.9%). Of these, 83 were classified as 'inactive'; 41(10 men; 31 women) completed baseline assessments, with the mean age of participants being 51 years. Mean baseline step counts were similar in group 1 (5685, SD 2945) and group 2 (6513, SD 3350). The mean increase in steps/day was greater in groups 1 than 2 ((2602, SD 1957) vs (748, SD 1997) p=0.005). CONCLUSIONS: A trial of a pedometer-based intervention using self-determined step counts appears feasible in primary care. Pedometers appear acceptable to women, particularly at a perimenopausal age, when it is important to engage in impact loading activities such as walking to maintain bone mineral density. An increase of 2500 steps/day is achievable for inactive patients, but the effectiveness of different approaches to realistic goal-setting warrants further study.

Establishing optimal quantitative-polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F-associated myeloproliferative neoplasms:a joint European LeukemiaNet/MPN&MPNr-EuroNet (COST action BM0902) study

Reliable detection of JAK2-V617F is critical for accurate diagnosis of myeloproliferative neoplasms (MPNs); in addition, sensitive mutation-specific assays can be applied to monitor disease response. However, there has been no consistent approach to JAK2-V617F detection, with assays varying markedly in performance, affecting clinical utility. Therefore, we established a network of 12 laboratories from seven countries to systematically evaluate nine different DNA-based quantitative PCR (qPCR) assays, including those in widespread clinical use. Seven quality control rounds involving over 21,500 qPCR reactions were undertaken using centrally distributed cell line dilutions and plasmid controls. The two best-performing assays were tested on normal blood samples (n=100) to evaluate assay specificity, followed by analysis of serial samples from 28 patients transplanted for JAK2-V617F-positive disease. The most sensitive assay, which performed consistently across a range of qPCR platforms, predicted outcome following transplant, with the mutant allele detected a median of 22 weeks (range 6-85 weeks) before relapse. Four of seven patients achieved molecular remission following donor lymphocyte infusion, indicative of a graft vs MPN effect. This study has established a robust, reliable assay for sensitive JAK2-V617F detection, suitable for assessing response in clinical trials, predicting outcome and guiding management of patients undergoing allogeneic transplant.

A highly potent and specific MET therapeutic protein antagonist with both ligand-dependent and ligand-independent activity

Activation of the MET oncogenic pathway has been implicated in the development of aggressive cancers that are difficult to treat with current chemotherapies. This has led to an increased interest in developing novel therapies that target the MET pathway. However, most existing drug modalities are confounded by their inability to specifically target and/or antagonize this pathway. Anticalins, a novel class of monovalent small biologics, are hypothesized to be "fit for purpose" for developing highly specific and potent antagonists of cancer pathways. Here, we describe a monovalent full MET antagonist, PRS-110, displaying efficacy in both ligand-dependent and ligand-independent cancer models. PRS-110 specifically binds to MET with high affinity and blocks hepatocyte growth factor (HGF) interaction. Phosphorylation assays show that PRS-110 efficiently inhibits HGF-mediated signaling of MET receptor and has no agonistic activity. Confocal microscopy shows that PRS-110 results in the trafficking of MET to late endosomal/lysosomal compartments in the absence of HGF. In vivo administration of PRS-110 resulted in significant, dose-dependent tumor growth inhibition in ligand-dependent (U87-MG) and ligand-independent (Caki-1) xenograft models. Analysis of MET protein levels on xenograft biopsy samples show a significant reduction in total MET following therapy with PRS-110 supporting its ligand-independent mechanism of action. Taken together, these data indicate that the MET inhibitor PRS-110 has potentially broad anticancer activity that warrants evaluation in patients.

Mobilizing against neoliberalism? Global affirmative action in context

This article uses the example of Northern Ireland to illustrate how political mobilization may
be deployed to challenge structural forms of inequality. The experience suggests that regulatory
models can be designed for particular contexts to shape approaches that present challenges to
dominant economic and political orthodoxies. The intention is not to overstate the significance
of this specific transitional context but simply to highlight elements that might feature in any
attempt to mobilize successfully around human rights and equality, and against aspects of neoliberal
thinking.

Gender Equality under the Charter of Fundamental Rights - a new lease for positive action?

This paper, which was published as a chapter of a Festskrift for Professor Ruth Nielsen, analyses Article 23 CFREU, the new provision on gender equality. It argues that Article 23 adds to the notion of gender equality in EU law, and not only allows, but also demands positive action measures if necessary to ensure equality between women and men. The provision also demands that positive action measures are suitable to achieve their aim. This implies that the EU legislator has to adapt positive action measure to the specific needs of the sector. The paper offers a critique of the proposal to introduce women quotas in board rooms, as proposed by the EU Commission in late 2012. It argues that the Commission unimaginatively copied rules developed for the German public service into a different sector, although these rules have not proven particularly efficient even in the public service. Consequently, a proposal that is demanding, but adapted to the sector should be developed.

Formação cívica e formação de professores: limites e potencialidades

A reorganização curricular do Ensino Básico de 2001 representou uma mudança de paradigma: uma aposta na flexibilidade, na integração e na autonomia. Uma das principais inovações desta reforma foi a criação de áreas curriculares não disciplinares, entre elas a Formação Cívica. Uma década volvida qual o impacto desta decisão? Estudos anteriores revelaram o uso administrativo e burocrático da Formação Cívica, ao serviço do Diretor de Turma e das suas responsabilidades de gestão da turma e resolução de conflitos. Num cenário de defraudamento das expetativas iniciais depositadas neste domínio não disciplinar e convencidos que estávamos da importância de potenciar espaços reais de promoção da Cidadania nas escolas desencadeámos um estudo de modelização dupla investigação-formação que nos permitisse compreender qual o papel da formação de professores neste domínio e os respetivos limites e potencialidades da Formação Cívica. Com base no pressuposto de que a ação de qualquer professor em contexto de sala de aula depende do conhecimento profissional que possui acerca de um determinado domínio curricular e reconhecendo a ausência de mecanismos de formação específica para a concretização dos tempos letivos de Formação Cívica, estruturámos o nosso trabalho com o intuito de contribuir para a construção de conhecimento sobre a natureza da formação de professores em Formação Cívica. Realizado no contexto da formação contínua de professores, o presente estudo desenvolveu-se em duas etapas fundamentais que se foram interligando entre momentos de investigação, reflexão, formação, recolha e análise de dados. A abordagem metodológica que se delineou para todas as fases assumiu um cariz qualitativo e crítico-interpretativo. Numa primeira fase, realizámos um estudo exploratório que nos permitiu identificar conceções, práticas e contextos no domínio da Formação Cívica. Selecionámos uma escola no distrito de Aveiro e entrevistámos 10 professores do 3.º CEB, coordenação e direção da escola, analisámos documentos oficiais e observámos aulas. Concluímos que a operacionalização da Formação Cívica está refém de um conjunto de condicionalismos, entre eles: a ausência de orientações curriculares; a sobreposição de funções do Diretor de Turma; o não reconhecimento das funções pedagógicas de uma área não disciplinar; a ausência de preparação científica e pedagógica dos professores responsáveis por este tempo letivo. A segunda fase do estudo contemplou a conceção, desenvolvimento, implementação e avaliação de um percurso formativo projetado com base nos resultados do estudo exploratório e da revisão da literatura realizada para o efeito. O percurso formativo, desenvolvido na modalidade de Oficina de Formação (OF), envolveu e alicerçou-se no potencial das tecnologias (participação numa comunidade online; criação de blogues; desenvolvimento de recursos digitais) e envolveu sete professores. Dos professores participantes na OF, foram analisados em detalhe o percurso formativo de três professoras de diferentes escolas do ensino básico e com diferentes níveis de experiência letiva em Formação Cívica. A realização deste percurso formativo permitiu-nos: i) identificar processos de reconstrução do conceito de cidadania e do lugar da Formação Cívica; ii) identificar uma visão mais alargada e transformadora da Formação Cívica ao invés de abordagens restritas de cariz administrativo; ii) analisar a importância da partilha de experiências, do feedback, do suporte e da reflexão no processo formativo para o desenvolvimento das práticas pedagógico-didáticas; iv) conceptualizar o conhecimento profissional docente associado à Formação Cívica. Consideramos que este estudo contribui para a necessária reflexão sobre o espaço e o papel da Educação para a Cidadania no sistema educativo Português, assim como para o debate da afirmação da Formação Cívica (com esta ou outra designação mas necessariamente com novos contornos), em particular no currículo do ensino básico. Sustentamos ainda a importância fundamental da formação contínua (de carácter ativo e reflexivo) e da construção partilhada do conhecimento para o desenvolvimento profissional dos professores de Formação Cívica, necessário para dar resposta aos desafios da Educação para a Cidadania.

Role of insulin and insulin-like peptides in bone formation: identification of bone-specific target genes and regulatory mechanisms, and characterization of the insulin-mimetic effect of vanadium

Tese de Doutoramento em Biologia, Especialidade em Biologia Molecular, Universidade do Algarve, 2008

A Benefit-Risk Assessment Framework for Development of Clinical Guidelines in Diagnostic Radiology

Thesis (Ph.D.)--University of Washington, 2015-12

Beach sand and the potential for infectious disease transmission: observations and recommendations

Recent studies suggest that sand can serve as a vehicle for exposure of humans to pathogens at beach sites, resulting in increased health risks. Sampling for microorganisms in sand should therefore be considered for inclusion in regulatory programmes aimed at protecting recreational beach users from infectious disease. Here, we review the literature on pathogen levels in beach sand, and their potential for affecting human health. In an effort to provide specific recommendations for sand sampling programmes, we outline published guidelines for beach monitoring programmes, which are currently focused exclusively on measuring microbial levels in water. We also provide background on spatial distribution and temporal characteristics of microbes in sand, as these factors influence sampling programmes. First steps toward establishing a sand sampling programme include identifying appropriate beach sites and use of initial sanitary assessments to refine site selection. A tiered approach is recommended for monitoring. This approach would include the analysis of samples from many sites for faecal indicator organisms and other conventional analytes, while testing for specific pathogens and unconventional indicators is reserved for high-risk sites. Given the diversity of microbes found in sand, studies are urgently needed to identify the most significant aetiological agent of disease and to relate microbial measurements in sand to human health risk.

Conceiving modern engineers within the framework of the sustainability action plan (PASUS) of ISEP: sustainability matters!

Engineering education practices have evolved not only due to the natural changes in the contents of the curricula and skills but also, and more recently, due to the requirements imposed by the Bologna revision process. In addition, industry is becoming more demanding, as society is becoming more and more aware of the global needs and consequences of industrial practices. Under this scope, higher education needs not only to follow but also to lead these trends. Therefore, the School of Engineering of the Polytechnic Institute of Porto (ISEP), a Global Reporting Initiative (GRI) training partner in Portugal, prepared and presented its Sustainability Action Plan (PASUS), with the main objective of creating a new kind of engineers, with Sustainable Development at the core of their graduation and MsC degrees. In this paper, the main strategies and activities of the referred plan along with the strategic approach, which guided its development and implementation, will be presented in detail. Additionally, a reflection about the above mentioned bridge between concept and application will be established and justified, in the framework of the action plan. Although in most of the situations, there was no prior discussion or specific request, many of the graduation and post-graduation programmes offered by ISEP already include courses that attend to PASUS philosophy. As a consequence, the number of Master thesis, Graduation projects and R&D projects that address sustainability problems has grown substantially, a proof that for ISEP community, sustainability really matters!

Reformulação de um Sistema de Gestão de Consumos de Energia

Atualmente a energia é considerada um vetor estratégico nas diversas organizações. Assim sendo, a gestão e a utilização racional da energia são consideradas instrumentos fundamentais para a redução dos consumos associados aos processos de produção do sector industrial. As ações de gestão energética não deverão ficar pela fase do projeto das instalações e dos meios de produção, mas sim acompanhar a atividade da Empresa. A gestão da energia deve ser sustentada com base na realização regular de diagnósticos energéticos às instalações consumidoras e concretizada através de planos de atuação e de investimento que apresentem como principal objetivo a promoção da eficiência energética, conduzindo assim à redução dos respetivos consumos e, consequentemente, à redução da fatura energética. Neste contexto, a utilização de ferramentas de apoio à gestão de energia promovem um consumo energético mais racional, ou seja, promovem a eficiência energética e é neste sentido que se insere este trabalho. O presente trabalho foi desenvolvido na Empresa RAR Açúcar e apresentou como principais objetivos: a reformulação do Sistema de Gestão de Consumos de Energia da Empresa, a criação de um modelo quantitativo que permitisse ao Gestor de Energia prever os consumos anuais de água, fuelóleo e eletricidade da Refinaria e a elaboração de um plano de consumos para o ano de 2014 a partir do modelo criado. A reformulação do respetivo Sistema de Gestão de Consumos resultou de um conjunto de etapas. Numa primeira fase foi necessário efetuar uma caraterização e uma análise do atual Sistema de Gestão de Consumos da Empresa, sistema composto por um conjunto de sete ficheiros de cálculo do programa Microsoft Excel©. Terminada a análise, selecionada a informação pertinente e propostas todas as melhorias a introduzir nos ficheiros, procedeu-se à reformulação do respetivo SGE, reduzindo-se o conjunto de ficheiros de cálculo para apenas dois ficheiros, um onde serão efetuados e visualizados todos os registos e outro onde serão realizados os cálculos necessários para o controlo energético da Empresa. O novo Sistema de Gestão de Consumos de Energia será implementado no início do ano de 2015. Relativamente às alterações propostas para as folhas de registos manuais, estas já foram implementadas pela Empresa. Esta aplicação prática mostrou-se bastante eficiente uma vez que permitiu grandes melhorias processuais nomeadamente, menores tempos de preenchimento das mesmas e um encurtamento das rotas efetuadas diariamente pelos operadores. Através do levantamento efetuado aos diversos contadores foi possível identificar todas as áreas onde será necessário a sua instalação e a substituição de todos os contadores avariados, permitindo deste modo uma contabilização mais precisa de todos os consumos da Empresa. Com esta reestruturação o Sistema de Gestão de Consumos tornou-se mais dinâmico, mais claro e, principalmente, mais eficiente. Para a criação do modelo de previsão de consumos da Empresa foi necessário efetuar-se um levantamento dos consumos históricos de água, eletricidade, fuelóleo e produção de açúcar de dois anos. Após este levantamento determinaram-se os consumos específicos de água, fuelóleo e eletricidade diários (para cada semana dos dois anos) e procedeu-se à caracterização destes consumos por tipo de dia. Efetuada a caracterização definiu-se para cada tipo de dia um consumo específico médio com base nos dois anos. O modelo de previsão de consumos foi criado com base nos consumos específicos médios dos dois anos correspondentes a cada tipo de dia. Procedeu-se por fim à verificação do modelo, comparando-se os consumos obtidos através do modelo (consumos previstos) com os consumos reais de cada ano. Para o ano de 2012 o modelo apresenta um desvio de 6% na previsão da água, 12% na previsão da eletricidade e de 6% na previsão do fuelóleo. Em relação ao ano de 2013, o modelo apresenta um erro de 1% para a previsão dos consumos de água, 8% para o fuelóleo e de 1% para a eletricidade. Este modelo permitirá efetuar contratos de aquisição de energia elétrica com maior rigor o que conduzirá a vantagens na sua negociação e consequentemente numa redução dos custos resultantes da aquisição da mesma. Permitirá também uma adequação dos fluxos de tesouraria à necessidade reais da Empresa, resultante de um modelo de previsão mais rigoroso e que se traduz numa mais-valia financeira para a mesma. Foi também proposto a elaboração de um plano de consumos para o ano de 2014 a partir do modelo criado em função da produção prevista para esse mesmo ano. O modelo apresenta um desvio de 24% na previsão da água, 0% na previsão da eletricidade e de 28% na previsão do fuelóleo.