Expression of the functional carbohydrate-binding module (CBM) of human laforin

Laforin is a human protein associated with the glycogen metabolism, composed of two structurally and functionally independent domains: a phosphatase catalytic domain and a substrate-binding module with glycogen and starch affinity. The main goal of this work is the development of a methodology for the expression of the so far poorly characterized carbohydrate-binding module (CBM) of laforin, allowing its study and development of biomedical applications. The laforin’s CBM sequence was originally cloned by PCR from a human muscle cDNA library. The recombinant protein, containing laforin’s CBM fused to an Arg-Gly-Asp sequence (RGD), was cloned and expressed using vector pET29a and recovered as inclusion bodies (IBs). Refolding of the IBs allowed the purification of soluble, dimeric and functional protein, according to adsorption assays using starch and glycogen. Several other experimental approaches, using both bacteria and yeast, were unsuccessfully tested, pointing towards the difficulties in producing the heterologous protein. Indeed, this is the first work reporting the production of the functional CBM from human laforin.Fundação para a Ciência e a Tecnologia (FCT, Portugal)

ADRITEM: a diferença no panorama do desenvolvimento rural

Mestrado em Administração e Gestão PúblicaO presente trabalho pretende, no âmbito do estágio divulgar as atividades do Grupo de Ação Local - ADRITEM (Associação de Desenvolvimento Rural Integrado das Terras de Santa Maria), tendo em conta todos os atores envolvidos, hierarquicamente ou ao nível das atividades desenvolvidas, nomeadamente ao nível de projetos de cooperação e candidaturas ao Eixo 3 do Programa ProDeR. O objetivo principal e final é a consciência da influência que a ADRITEM tem, com a análise de alguns indicadores, tais como a criação de emprego e o investimento feito na sua zona de intervenção.The present work intends, within the framework of the training course to disclose the activities of the Local Action Group - ADRITEM (Integrated Rural Development Association of the Lands of Santa Maria), taking into account all actors involved, either hierarchically or at the level of activities undertaken, namely cooperation projects and applications to the axis 3 of the ProDeR Program. The main and final goal is the awareness of the influence that ADRITEM has regarding to the analysis of some indicators such as the creation of employment and investment made in their intervention zone

Ideação suicida nos adolescentes da Ilha de São Miguel (Açores): análise dos fatores de risco e de proteção

Os comportamentos suicidários e autolesivos constituem um problema de saúde pública, sendo o suicídio uma das 10 maiores causas de morte mundial e a segunda maior em jovens. Destaca-se a Região Autónoma dos Açores (RAA) onde o suicídio jovem é preocupante e a ideação suicida é alarmante. Sabendo-se que para uma predição adequada do risco suicidário é fundamental a compreensão dos fatores de risco e proteção, a presente investigação incluiu dois estudos. O primeiro estudo objetiva a exploração das características psicométricas do Inventário de Atitudes Face à Procura de Serviços de Saúde Mental (IAPSSM), numa amostra de 750 adolescentes, com idades compreendidas entre os 12 e os 19 anos, na ilha de São Miguel (Açores). Os resultados apontam que a estrutura trifatorial proposta é adequada, tendo o IAPSSM sido reduzido a 20 itens. O instrumento revela boas propriedades ao nível da consistência interna, validade convergente, divergente e estabilidade temporal. O segundo estudo visa analisar a ideação suicida e os fatores de risco e proteção nos adolescentes, recorrendo à mesma amostra do primeiro estudo. Os participantes preencheram instrumentos que avaliavam diversas variáveis: ideação suicida; satisfação com o suporte social; atitudes e intenções face à procura de ajuda; acontecimentos de vida negativos; razões para viver; sintomatologia depressiva e desesperança Observou-se que apesar de os adolescentes apresentarem valores mais inferiores nos fatores de risco e superiores nos fatores de proteção, uma porção significativa revela algum grau de ideação suicida e 38.3% enquadram-se como população de risco. Verificam-se associações entre os fatores, salientando-se que a depressão é o melhor preditor da ideação suicida, porém, a relação entre ambas não é direta. A presente investigação constituiu-se como um contributo para o conhecimento do suicídio, complementando a informação existente na RAA, promovendo a abertura para investigações futuras e revelando implicações práticas.Suicidal and self-harm behaviors are a public health problem, being that suicide is one of the 10 leading causes of death worldwide and the second leading cause of death for young people. In the Autonomous Region of the Azores (ARA) suicide in younger age groups is worrisome and suicidal ideation is alarming. Knowing that to achieve an adequate prediction of suicidal risk it is essential to understand risk and protective factors, this investigation included two studies. The first study aims to explore the psychometric characteristics of the Inventory of Attitudes toward Seeking Mental Health Services (IASMHS), in a sample comprised of 750 adolescents, with ages ranging from 12 to 19 years, in São Miguel island (Azores). Results suggest that the proposed three-factor structure was adequate, with the IASMHS being shortened to 20 items. This inventory revealed satisfactory psychometric properties in terms of internal consistency, convergent and divergent validity, and temporal stability. The second study intends to analyze suicidal ideation and its associated risk and protective factors, in the same adolescent sample as the first study. The participants filled in several tools to assess different variables: suicidal ideation; social support satisfaction; help-seeking attitudes and intentions; negative life events; reasons for living; depressive symptoms and hopelessness. It was indicated that even though the adolescents displayed less risk factors and more protective factors, a significant portion revealed suicidal ideation and 38.3% were considered a risk group. Furthermore, associations between the different factors were exhibited, highlighting that depression is the best predictor of suicidal ideation, however, this relationship was mediated. The present investigation constituted itself as a contribution to the knowledge on suicide, supplementing the already existing information in the ARA, promoting the openness to future studies and presenting practical implications

Escherichia coli expression, refolding and characterization of human laforin

Laforin is a unique human dual-specificity phosphatase as it contains an amino terminal carbohydrate binding module (CBM). Laforin gene mutations lead to Lafora disease, a progressive myoclonus epilepsy with an early fatal issue. Previous attempts to produce recombinant laforin faced various difficulties, namely the appearance of protein inclusion bodies, the contamination with bacterial proteins and a high tendency of the protein to aggregate, despite the use of fusion tags to improve solubility and ease the purification process. In this work, we have expressed human laforin in Escherichia coli in the form of inclusion bodies devoid of any fusion tags. After a rapid dilution refolding step, the protein was purified by two chromatographic steps, yielding 5–7 mg of purified protein per liter of bacterial culture. The purified protein was shown to have the kinetic characteristics of a dual-specificity phosphatase, and a functional carbohydrate binding module. With this protocol, we were able for the first time, to produce and purify laforin without fusion tags in the amounts traditionally needed for the crystallographic structural studies paving the way to the understanding of the molecular mechanisms of laforin activity and to the development of novel therapies for Lafora disease.Fundação para a Ciência e a Tecnologia (FCT) – Programa Operacional “Ciência, Tecnologia, Inovação” (POCTI

Workaholism and work-family interaction among emergency and critical care nurses

Objectives: Investigating the prevalence of workaholism as well as the relationship between work-family interaction among emergency and critical care nurses. Research methodology/design: A quantitative cross-sectional study. Setting: A total of 219 nurses took part in the Dutch Work Addiction Scale (DUWAS-10) and the Survey WorkHome Interaction Nijmegen (SWING), which included socio–demographic and occupational question. Data was gathered in Spain between June and September 2019. Results: Workaholism was found to be prevalent in 28.3% of the participants. In all four categories, workaholism was statistically connected to work-home interaction, with workaholics having higher means than nonworkaholics. Perceived work stress was related to workaholism (p =.036). In the Work Excessively dimension, female nurses had significantly higher mean scores (M = 2.26) than their male counterparts (M = 1,88). In addition, in the Negative Work-Home Interaction (M = 2.04), the global mean scores were higher than in the Negative Home-Work Interaction (M = 1.34), indicating conflict and a negative impact of work on the family. Conclusion: The findings of this study demonstrated the necessity of taking into account demands and resources from both the work and home domains since it has been shown that both have an impact on one other. Furthermore, given the vital responsibilities that emergency and critical care nurses play in the health care system, our findings suggest that occupational health treatments should be used to identify those working profiles that are particularly at risk.info:eu-repo/semantics/publishedVersio

Intracellular Trafficking Mechanisms of Synaptic Dysfunction in Alzheimer’s Disease

The Almeida lab has been supported by FCTJPCOFUND/0004/2015; Alzheimer’s Association Research Grant (AARG-19-618007); Maratona da Saúde; H2020 Spreading Excellence and Widening Participation, H2020-WIDESPREAD01-2016-2017-TeamingPhase2-GA739572; iNOVA4Health (UID/Multi/04462/2019), a program financially supported by Fundação para a Ciencia e Tecnologia (FCT)/Ministério da Educação e Ciencia, through national funds and co-funded by FEDER under the PT2020 Partnership Agreement. CG’s salary is supported by FCT-CEECIND/00410/2017. FM has been the recipient of an FCT doctoral fellowship (PD/BD/128344/2017). CP has been the recipient of an FCT doctoral fellowship (SFRH/BD/128374/2017).Alzheimer’s disease (AD) is the most common neurodegenerative disease characterized by progressive memory loss. Although AD neuropathological hallmarks are extracellular amyloid plaques and intracellular tau tangles, the best correlate of disease progression is synapse loss. What causes synapse loss has been the focus of several researchers in the AD field. Synapses become dysfunctional before plaques and tangles form. Studies based on early-onset familial AD (eFAD) models have supported that synaptic transmission is depressed by β-amyloid (Aβ) triggered mechanisms. Since eFAD is rare, affecting only 1% of patients, research has shifted to the study of the most common late-onset AD (LOAD). Intracellular trafficking has emerged as one of the pathways of LOAD genes. Few studies have assessed the impact of trafficking LOAD genes on synapse dysfunction. Since endocytic traffic is essential for synaptic function, we reviewed Aβ-dependent and independent mechanisms of the earliest synaptic dysfunction in AD. We have focused on the role of intraneuronal and secreted Aβ oligomers, highlighting the dysfunction of endocytic trafficking as an Aβ-dependent mechanism of synapse dysfunction in AD. Here, we reviewed the LOAD trafficking genes APOE4, ABCA7, BIN1, CD2AP, PICALM, EPH1A, and SORL1, for which there is a synaptic link. We conclude that in eFAD and LOAD, the earliest synaptic dysfunctions are characterized by disruptions of the presynaptic vesicle exo- and endocytosis and of postsynaptic glutamate receptor endocytosis. While in eFAD synapse dysfunction seems to be triggered by Aβ, in LOAD, there might be a direct synaptic disruption by LOAD trafficking genes. To identify promising therapeutic targets and biomarkers of the earliest synaptic dysfunction in AD, it will be necessary to join efforts in further dissecting the mechanisms used by Aβ and by LOAD genes to disrupt synapses.publishersversionpublishe

Life-cycle assessment of microalgae biodiesel: a review

Microalgae are an attractive way to produce biofuels due to the ability to accumulate lipids and very high photosynthetic yields. This article presents a review of life-cycle assessment studies of microalgae biodiesel production, including an analysis of modeling choices and assumptions. A high variation in GHG emissions (between -0.75 and 2.9 kg CO2eq MJ-1) was found and the main causes were investigated, namely modeling choices (e.g. the approach used to deal with multifunctionality), and a high parameter uncertainty in microalgae cultivation, harvesting and oil extraction processes

Bioclimatic modeling in the Last Glacial Maximum, Mid-Holocene and facing future climatic changes in the strawberry tree (Arbutus unedo L.)

Research ArticleIncreasing forest wildfires in Portugal remain a growing concern since forests in the Mediterranean region are vulnerable to recent global warming and reduction of precipitation. Therefore, a long-term negative effect is expected on the vegetation, with increasing drought and areas burnt by fires. The strawberry tree (Arbutus unedo L.) is particularly used in Portugal to produce a spirit by processing its fruits and is the main income for forestry owners. Other applications are possible due to the fruit and leaves’ anti-oxidant properties and bioactive compounds production, with a potential for clinical and food uses. It is a sclerophyllous plant, dry-adapted and fire resistant, enduring the Mediterranean climate, and recently considered as a possibility for afforestation, to intensify forest discontinuity where pines and eucalypts monoculture dominate the region. To improve our knowledge about the species’ spatial distribution we used 318 plots (the centroid of a 1 km2 square grid) measuring the species presence and nine environmental attributes. The seven bioclimatic variables most impacting on the species distribution and two topographic features, slope and altitude, were used. The past, current and future climate data were obtained through WorldClim. Finally, the vulnerability of the strawberry tree to the effects of global climate change was examined in the face of two emission scenarios (RCP 4.5 and 8.5), to predict distribution changes in the years 2050 and 2070, using a species distribution models (MaxEnt). The reduction of suitable habitat for this species is significant in the southern regions, considering the future scenarios of global warming. Central and northern mountainous regions are putative predicted refuges for this species. Forest policy and management should reflect the impact of climate change on the usable areas for forestry, particularly considering species adapted to the Mediterranean regions and wildfires, such as the strawberry tree. The distribution of the species in the Last Glacial Maximum (LGM) and Mid-Holocene (MH) agrees with previous genetic and paleontological studies in the region, which support putative refuges for the species. Two in the southern and coastal-central regions, since the LGM, and one in the eastcentral mountainous region, considered as cryptic refugiainfo:eu-repo/semantics/publishedVersio