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Many genes encoding synaptic proteins are associated with neurodevelopmental disorders (NDDs) such as autism spectrum disorders (ASDs), intellectual disability (ID), and epilepsy. Here we review recent studies on the synaptic effects of disease-associated rare variants identified in two families of synaptic proteins: NMDA receptors (NMDARs) and the postsynaptic adhesion molecules neuroligins (NLGNs). Many NMDAR subunit genes (GRINs) are highly intolerant to variation, and both gain-of-function (GOF) and loss-of-function (LOF) variants are implicated in disease. NLGN genes are also associated with ASDs, and in some cases, contribute to the male bias identified in these patients. Understanding the molecular basis of synaptic dysfunction of rare variants in these genes will help the design of new therapeutic approaches.

Comics are tools used to narrate stories through pictures and small texts in the form of balloons in a practical and fun way. In this perspective, this work has as main objective to report the practices of activities developed in a short course with Elementary School II students of a public school in the municipality of Santana de Araguaia - PA, online. We used a qualitative approach as a research methodology and, as descriptive analysis instruments, a video recording of the application of activities, a questionnaire and researchers' notes. The activities developed consisted of presenting and building comic books, involving mathematical content using StoryBoard That, as a digital tool. The results showed that the use of the technological resource provided students' curiosity and participation during the construction of comics, in addition to arousing interest in solving mathematical problems. The use of StoryBoard That generated autonomy and creativity in the students in developing their own stories and, at the same time, provided the opportunity for an exploratory and investigative learning spirit. As histórias em quadrinhos (HQs) são ferramentas utilizadas para narrar histórias por meio de figuras e pequenos textos em forma de balões de forma prática e divertida. Nessa perspectiva, este trabalho tem como objetivo principal relatar as práticas de atividades desenvolvidas em um minicurso com alunos do Ensino Fundamental de uma escola pública no município de Santana de Araguaia – PA, de forma online. Utilizamos como metodologia de pesquisa a abordagem qualitativa e, como instrumentos de análise descritiva, a vídeo gravação da aplicação das atividades, questionário e anotações dos pesquisadores. As atividades desenvolvidas consistiram em apresentar e construir histórias em quadrinhos, envolvendo conteúdos matemáticos com o uso do StoryBoard That, como ferramenta digital. Os resultados demonstraram que a utilização do recurso tecnológico proporcionou a curiosidade e participação dos alunos durante a construção de HQs, além de despertar o interesse em resolver problemas matemáticos. A utilização do StoryBoard That gerou nos alunos autonomia e criatividade em desenvolver suas próprias histórias e, ao mesmo tempo, oportunizou o espírito explorador e investigativo de aprendizagem.

This work aimed to investigate the potential of the use of digital tools in the teaching of cryptography and mathematics during the application of a Didactic Sequence (DS). The implementation of the SD was carried out with Mathematics students from a public university in the south of Pará, and the investigation was developed in a qualitative perspective. The instruments of analysis were video recordings of the DS implementation, direct observation, the questionnaire and the responses to the extra activities. The SD consisted of creating applications on the Mit App Inventor 2 platform for matrix calculations and the encoding and decoding of messages, making it unintelligible to students. In addition, problems related to the theme were presented in the format of digital comics (HQDs). The analysis consisted of planning, evaluation and reflection of the implemented SD. From the results, we showed that Mit App Inventor 2 and HQDs were technological tools suitable for the study of cryptography that aroused the interest and curiosity of academics, providing relevant knowledge for their future teaching practice, in addition to making the class more attractive, dynamic. and participatory. New studies on cryptography using digital technologies in Basic Education are indicated due to the lack of studies in this area. Este trabalho teve como objetivo investigar as potencialidades do uso de ferramentas digitais no ensino de criptografia e matemática durante aplicação de uma Sequência Didática (SD). A implementação da SD foi realizada com acadêmicas de Matemática de uma universidade pública do sul do Pará, sendo que a investigação foi desenvolvida numa perspectiva qualitativa. Os instrumentos de análise foram as gravações em vídeo da implementação da SD, a observação direta, o questionário e as respostas das atividades extras. A SD consistiu em criar aplicativos na plataforma do Mit App Inventor 2 para cálculos de matrizes e na codificação e decodificação de mensagens, tornando-a ininteligível aos estudantes. Além disso, foram apresentados problemas relacionados ao tema em formato de histórias em quadrinhos digitais (HQDs). A análise foi composta por planejamento, avaliação e reflexão da SD implementada. A partir dos resultados, evidenciamos que o Mit App Inventor 2 e HQDs foram ferramentas tecnológicas adequadas ao estudo de criptografia que despertaram o interesse e a curiosidade das acadêmicas, proporcionando saberes relevantes para sua futura prática docente, além de tornar a aula mais atrativa, dinâmica e participativa. Indicam-se novos estudos sobre criptografia com uso de tecnologias digitais na Educação Básica devido à carência de estudos nessa área.

The synaptic expression of glutamate receptors of the α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) type is dynamically controlled by interaction with binding partners and auxiliary proteins. These proteins can be regulated by posttranslational modifications, including ubiquitination. In this work, we investigated the regulation of glutamate receptor interacting protein-associated protein 1 (GRASP1) by ubiquitin-dependent mechanisms and its impact on surface expression and activity of synaptic AMPA receptors. Cotransfection of GFP-ubiquitin decreased myc-GRASP1 protein levels in HEK293T cells, and this effect was inhibited upon transfection of an ubiquitin mutant that cannot be ubiquitinated on Lys48. In addition, transfection of cultured hippocampal neurons with GFP-ubiquitin reduced the dendritic levels of endogenous GRASP1 and decreased the surface expression of GluA1 AMPA receptor subunits, an effect that was partly reversed by cotransfection with GRASP1. Similarly, transfection of hippocampal neurons with GFP-ubiquitin decreased the amplitude of miniature excitatory postsynaptic currents (mEPSCs) mediated by Ca2+ -impermeable AMPA receptors, and this effect was abrogated by cotransfection of GRASP1. Together, the results show a role for ubiquitination in the regulation of the postsynaptic protein GRASP1, which has an impact on the surface distribution of AMPA receptors and on their activity at the synapse.

As histórias em quadrinhos (HQs) são ferramentas utilizadas para narrar histórias por meio de figuras e pequenos textos em forma de balões de forma prática e divertida. Nessa perspectiva, este trabalho tem como objetivo principal relatar as práticas de atividades desenvolvidas em um minicurso com alunos do Ensino Fundamental de uma escola pública no município de Santana de Araguaia – PA, de forma online. Utilizamos como metodologia de pesquisa a abordagem qualitativa e, como instrumentos de análise descritiva, a vídeo gravação da aplicação das atividades, questionário e anotações dos pesquisadores. As atividades desenvolvidas consistiram em apresentar e construir histórias em quadrinhos, envolvendo conteúdos matemáticos com o uso do StoryBoard That, como ferramenta digital. Os resultados demonstraram que a utilização do recurso tecnológico proporcionou a curiosidade e participação dos alunos durante a construção de HQs, além de despertar o interesse em resolver problemas matemáticos. A utilização do StoryBoard That gerou nos alunos autonomia e criatividade em desenvolver suas próprias histórias e, ao mesmo tempo, oportunizou o espírito explorador e investigativo de aprendizagem.

Este trabalho teve como objetivo investigar as potencialidades do uso de ferramentas digitais no ensino de criptografia e matemática durante aplicação de uma Sequência Didática (SD). A implementação da SD foi realizada com acadêmicas de Matemática de uma universidade pública do sul do Pará, sendo que a investigação foi desenvolvida numa perspectiva qualitativa. Os instrumentos de análise foram as gravações em vídeo da implementação da SD, a observação direta, o questionário e as respostas das atividades extras. A SD consistiu em criar aplicativos na plataforma do Mit App Inventor 2 para cálculos de matrizes e na codificação e decodificação de mensagens, tornando-a ininteligível aos estudantes. Além disso, foram apresentados problemas relacionados ao tema em formato de histórias em quadrinhos digitais (HQDs). A análise foi composta por planejamento, avaliação e reflexão da SD implementada. A partir dos resultados, evidenciamos que o Mit App Inventor 2 e HQDs foram ferramentas tecnológicas adequadas ao estudo de criptografia que despertaram o interesse e a curiosidade das acadêmicas, proporcionando saberes relevantes para sua futura prática docente, além de tornar a aula mais atrativa, dinâmica e participativa. Indicam-se novos estudos sobre criptografia com uso de tecnologias digitais na Educação Básica devido à carência de estudos nessa área.

The real relationship and working alliance are considered sister concepts because even though they are separate constructs they are also interrelated. Both of them are considered to contribute significantly to the outcome of therapy, but there are still some questions regarding their relationship, like what contributes to their proximity? To examine the association of real relationship and working alliance across studies, and confirm their separation, we elaborated a systematic review of this two constructs. Based on the results from that study, we conducted an empirical investigation with 40 ongoing therapist-client dyads, where we evaluated how real relationship and working alliance contribute to the outcome of psychotherapy, and if that contribution was influenced by the rater’s perspective. The meta-analysis confirmed the theorized association between real relationship and working alliance, revealing an overall correlation of r = .66. The manner in which each of them contributes to outcome confirmed their differentiation. The empirical study also showed a significant correlation between real relationship and working alliance, and that the bond subscale of the Working Alliance Inventory (WAI) had an influence in this association. Overall, real relationship seemed to predict outcome beyond the working alliance, in both client and therapist’s perspectives. However, when we considered the subscales, the results depended on the perspective: for clients, genuineness was a better predictor; for therapists, realism was more important when combined with the task subscale of the WAI. The study emphasizes the need to keep the research on the concepts of real relationship and working alliance, in order to improve the therapeutic relationship and the outcome of psychotherapy. A relação terapêutica é tão importante para o sucesso da terapia como a própria intervenção psicoterapêutica. Assim, é importante que a primeira seja tão estudada como a segunda. O modelo tripartido da relação terapêutica, proposto por Gelso, afirma que existem três importantes componentes que são diferentes, mas interligados: a relação real, a aliança terapêutica e a configuração transferência-contratransferência. A relação real define-se como a relação pessoal existente entre duas ou mais pessoas, que se reflete no grau em que cada uma é genuína com a outra e percebe a outra de forma realista. Esta definição inclui duas dimensões importantes: a genuinidade e o realismo. A genuinidade é a capacidade de ser quem realmente se é, de ser autêntico no “aqui-e-agora”; o realismo é a experiência ou perceção do outro de formas que lhe são próprias, em vez de projeções do indivíduo baseadas nos seus medos ou desejos. A aliança terapêutica foi definida de várias maneiras ao longo dos anos, mas Gelso adota a definição de Bordin porque esta coloca o enfoque no trabalho terapêutico. Assim, consideramos a aliança terapêutica como a colaboração entre terapeuta e cliente que assenta em três componentes: acordo sobre objetivos terapêuticos, consenso sobre as tarefas que compõem a terapia, e um vínculo entre ambos os participantes. Embora a aliança terapêutica faça parte do tratamento, no sentido em que existe para realizar o trabalho da terapia, a relação real está presente sempre que duas ou mais pessoas se relacionam entre si. Por este motivo, o vínculo presente tanto na aliança como na relação real deve ser visto como um vínculo de trabalho e um vínculo pessoal, respetivamente. O facto de estes dois conceitos estarem tão interligados e, ainda assim, serem independentes, faz com que tenham sido nomeados “conceitos-irmãos”. A American Psychological Association (APA) organizou um grupo de trabalho cujo objetivo é identificar e estudar os elementos que compõem a relação terapêutica e contribuem para os resultados da terapia. Num conjunto de meta-análises reunido por Norcross e Lambert (2019), é demonstrado o contributo da relação real e da aliança terapêutica, de forma independente, para os resultados. No nosso estudo, que está dividido em duas partes, começámos por realizar uma revisão sistemática da literatura no que a estes dois construtos diz respeito. Pretendíamos examinar a sua associação e confirmar a sua diferenciação; e observar como estes construtos podem predizer o resultado da terapia. Tendo como base uma meta-análise realizada por Gelso et al. (2019), pesquisámos os conceitos “relação real” e “aliança terapêutica” em bases de dados científicas e definimos os critérios de inclusão e de exclusão, o que nos levou a um total de 23 artigos. Para a realização da meta-análise, tivemos de adotar critérios mais específicos, o que nos levou a excluir 7 artigos da amostra. Esta acusou uma correlação moderada entre relação real e aliança terapêutica de r = .66, confirmando a forte associação entre os construtos, mas sem revelar a que se devia esta associação. Na revisão sistemática, os resultados mostraram: que relação real e aliança terapêutica contribuem de maneiras diferentes para os resultados, confirmando que são conceitos diferentes; a relação real parece ser melhor preditora dos resultados da terapia; poderá haver influência do avaliador no valor preditivo de cada um dos construtos. Para tentar responder às questões levantadas neste estudo, realizámos um estudo empírico. Neste segundo estudo, propusemo-nos a responder se o fator vínculo do Inventário de Aliança Terapêutica (IAT) tinha influência na forte associação demonstrada entre relação real e aliança terapêutica; e se o avaliador (terapeuta ou cliente) dos construtos influencia o valor preditivo dos mesmos. Nesta investigação, reunimos informação de 40 díades terapeuta-cliente, constituídas por 6 terapeutas e 40 clientes. Todos os participantes completaram instrumentos de autorrelato sobre a relação real, a aliança terapêutica, os resultados de terapia e desejabilidade social. Os instrumentos foram aplicados à distância, através da plataforma Google Forms, devido à pandemia de Covid-19 que vigorava na altura em que o estudo foi conduzido. Foram realizadas correlações de Pearson na análise dos resultados e detetámos associações positivas e significativas entre a relação real e a aliança terapêutica. No caso dos clientes, o fator vínculo revelou uma correlação moderada e significativa com a relação real. Procedemos a uma análise posterior de significância de correlações, que demonstrou que o fator vínculo poderá estar a contribuir para as elevadas correlações entre relação real e aliança terapêutica. Tanto a relação real como a aliança terapêutica tiveram associações significativas com os resultados da terapia. Através de regressões lineares hierárquicas, observámos que, em ambas as perspetivas (terapeuta e cliente), a relação real era o melhor preditor de resultados. Quando observadas ao nível dos seus fatores, a subescala tarefas do IAT e a subescala genuinidade do Inventário de Relação Real (IRR) demonstraram ser as melhores preditoras dos resultados da terapia. No entanto, quando os fatores do IAT eram colocados no primeiro bloco de análise, havia diferenças ao nível dos avaliadores: na perspetiva dos clientes, a genuinidade roubava o valor preditivo das tarefas, tornando-se o melhor preditor; enquanto para os terapeutas o realismo e as tarefas, em conjunto, eram bons preditoresApesar de já estar estabelecido que a aliança terapêutica é um importante componente da relação terapêutica e um bom preditor dos resultados da terapia, neste estudo chegámos à conclusão de que a relação real também deve ser tida em consideração, pois mostrou ser um melhor preditor de resultados que o seu “conceito-irmão”. É necessário, contudo, algum cuidado a interpretar os nossos resultados, pois a amostra que apresentamos não é de grande dimensão, em particular no número de terapeutas, e os clientes apresentavam-se em diferentes fases da terapia, além de se encontrarem em psicoterapias diferentes. Além disso, apesar de uma tradução que consideramos adequada, o IRR ainda não foi validado para a população portuguesa. Este estudo vem acrescentar ao que tem sido feito no campo da relação terapêutica, não só ao nível da investigação, como ao nível clínico. Em termos de investigação, revela que o IAT talvez precise de ser revisto, porque o fator vínculo está a influenciar as correlações entre relação real e aliança terapêutica. Em termos clínicos, mostra que é necessário informar sobre a relação real e desenvolver estratégias que permitam aos terapeutas desenvolver as suas relações reais com os seus clientes e, consequentemente, melhorar os resultados da terapia..

NMDAR-dependent Ca2+ influx underpins multiple forms of synaptic plasticity. In the adult forebrain, the majority of synaptic NMDAR currents are mediated by GluN2A-containing NMDARs. These receptors are rapidly inserted into synapses during LTP; however, the underlying molecular mechanisms remain poorly understood. Here we show that GluN2A is phosphorylated at Ser-1459 by CaMKIIα in response to glycine stimulation that mimics LTP in primary neurons. Phosphorylation of Ser-1459 promotes GluN2A interaction with the SNX27-retromer complex, therefore enhancing the endosomal recycling of NMDARs. Loss of SNX27 or CaMKIIα function blocks the glycine-induced increase in GluN2A-NMDARs on the neuronal membrane. Interestingly, mutations of Ser-1459, including the rare S1459G human epilepsy variant, prolong decay times of NMDAR-mediated synaptic currents in heterosynapses by increasing the active duration of channel openings. Taken together, these findings not only identify a critical role of Ser-1459 phosphorylation in regulating the function of NMDARs, but also explain how the S1459G epilepsy variant dysregulates NMDAR function.

The N-methyl-D-aspartate receptors (NMDARs) are ionotropic glutamate receptors that mediate the flux of calcium (Ca(2+)) into the postsynaptic compartment. Calcium influx subsequently triggers the activation of various intracellular signaling cascades that underpin multiple forms of synaptic plasticity. Functional NMDARs are assembled as heterotetramers composed of two obligatory GluN1 subunits and two GluN2 or GluN3 subunits. Four different GluN2 subunits (GluN2A-D) are present throughout the central nervous system; however, they are differentially expressed, both developmentally and spatially, in a cell- and synapse-specific manner. Each GluN2 subunit confers NMDARs with distinct ion channel properties and intracellular trafficking pathways. Regulated membrane trafficking of NMDARs is a dynamic process that ultimately determines the number of NMDARs at synapses, and is controlled by subunit-specific interactions with various intracellular regulatory proteins. Here we review recent progress made towards understanding the molecular mechanisms that regulate the trafficking of GluN2-containing NMDARs, focusing on the roles of several key synaptic proteins that interact with NMDARs via their carboxyl termini.

Reducing emissions from deforestation and forest degradation and the role of conservation, sustainable management of forests and enhancement of forest carbon stocks in developing countries (REDD+), is still a promising mechanism of the UNFCCC for many tropical countries that would like to receive a fair financial compensation for their historical and current efforts to avoid forest conversion at the expense of more economically land uses. Brazil has a great opportunity to successfully participate in REDD+ not only because of its huge Amazon forest area (ca. 4 million km2) but also because of its advanced forest monitoring system “PRODES”. However, this opportunity could be threatened due to the current differentiated monitoring capacities of most Brazilian Amazon states, markedly in High-Forest and Low-Deforestation (HFLD) regions. This is evident in the State of Amapa, which despite its political will to support actions towards the design of its REDD+ strategy, is still struggling with key technical aspects of forest monitoring. To address this issue and to strengthen the on-going REDD+ design process we assessed a) land use and land cover (LULC) changes for period of 23 years (1985–2008); b) estimated CO2 emissions associated to these LULC changes; c) identified the main drivers and agents of deforestation, and d) discussed policy implications for REDD+ implementation in a HFLD area. We applied a methodology, which is capable of reducing cloud cover using temporal filters on the classified images, detecting deforestation (and forest degradation) in areas as small as 1 ha, and used the decision tree method to identify different LULC types. This methodology was able to demonstrate that forest cover in northern Amapa has remained almost untouched during the observed period of 23-years. As many other HFLD areas, this region has a great potential to receive financial benefits from the REDD+ mechanism, especially from voluntary markets that are largely interested in the conservation value of these areas. However, the use of high accuracy LULC classification approaches, with appropriate Measuring, Reporting and Verification systems should be part of the REDD+ implementation strategy of HFLD areas towards having high standards for certified carbon, and therefore improved chances to receive better prices for carbon offsets. The potential of REDD+ to be a fair and efficient mechanism will also depend on the recognition of the historical efforts to avoid deforestation in HFLD areas, mainly by Federal Governments, as an incentive for low-carbon development.

SUMMARY NMDA receptor (NMDAR)-dependent Ca2+ influx underpins multiple forms of synaptic plasticity. Most synaptic NMDAR currents in the adult forebrain are mediated by GluN2A-containing receptors, which are rapidly inserted into synapses during long-term potentiation (LTP); however, the underlying molecular mechanisms remain poorly understood. In this study, we show that GluN2A is phosphorylated at Ser-1459 by Ca2+/calmodulin-dependent kinase IIα (CaMKIIα) in response to glycine stimulation that mimics LTP in primary neurons. Phosphorylation of Ser-1459 promotes GluN2A interaction with the sorting nexin 27 (SNX27)-retromer complex, thereby enhancing the endosomal recycling of NMDARs. Loss of SNX27 or CaMKIIα function blocks the glycine-induced increase in GluN2A-NMDARs on the neuronal membrane. Interestingly, mutations of Ser-1459, including the rare S1459G human epilepsy variant, prolong the decay times of NMDAR-mediated synaptic currents in heterosynapses by increasing the duration of channel opening. These findings not only identify a critical role of Ser-1459 phosphorylation in regulating the function of NMDARs, but they also explain how the S1459G variant dysregulates NMDAR function., Graphical Abstract, In brief Yong et al. identify that activity-dependent phosphorylation of Ser-1459 in the GluN2A C-terminal domain by CaMKIIα promotes its interaction with the SNX27-retromer complex, thereby enhancing the surface expression of NMDARs during synaptic potentiation. Mutations of Ser-1459 prolong the decay times of NMDAR-mediated synaptic currents by increasing the duration of channel opening.

The present work is focused on the characterisation of the polyphenolic content of an Oxalis pes-caprae L. leaf extract and on the evaluation of its bioactivity with particular interest on its vascular activity and antioxidant potential. The polyphenolic content was characterised by HPLC-DAD and LC-MS/MS. The vascular activity was evaluated according to the influence on the serotonergic and adrenergic systems of the human internal mammary artery (HIMA). Antioxidant and neuroprotective studies were also conducted. Several luteolin and apigenin derivatives were identified as main constituents of the extract, which did not present any contractile effect nor had any effect on the serotonergic system of HIMA. However, it showed antagonistic effect on the adrenergic system, inhibiting the contraction to noradrenaline (reduction of 58.44% of maximum contraction). The extract showed antioxidant activity and standardised luteolin and apigenin derivatives showed neuroprotective potential, particularly homoorientin.

NMDA receptors (NMDARs) are ionotropic glutamate receptors that are crucial for neuronal development and higher cognitive processes. NMDAR dysfunction is involved in a variety of neurological and psychiatric diseases; however, the mechanistic link between the human pathology and NMDAR dysfunction is poorly understood. Rare missense variants within NMDAR subunits have been identified in numerous patients with mental or neurological disorders. We specifically focused on the GluN2B NMDAR subunit, which is highly expressed in the hippocampus and cortex throughout development. We analyzed several variants located in the GluN2B C terminus and found that three variants in patients with autism (S1415L) or schizophrenia (L1424F and S1452F) (S1413L, L1422F, and S1450F in rodents, respectively) displayed impaired binding to membrane-associated guanylate kinase (MAGUK) proteins. In addition, we observed a deficit in surface expression for GluN2B S1413L. Furthermore, there were fewer dendritic spines in GluN2B S1413L-expressing neurons. Importantly, synaptic NMDAR currents in neurons transfected with GluN2B S1413L in GluN2A/B-deficient mouse brain slices revealed only partial rescue of synaptic current amplitude. Functional properties of GluN2B S1413L in recombinant systems revealed no change in receptor properties, consistent with synaptic defects being the result of reduced trafficking and targeting of GluN2B S1413L to the synapse. Therefore, we find that GluN2B S1413L displays deficits in NMDAR trafficking, synaptic currents, and spine density, raising the possibility that this mutation may contribute to the phenotype in this autism patient. More broadly, our research demonstrates that the targeted study of certain residues in NMDARs based on rare variants identified in patients is a powerful approach to studying receptor function.SIGNIFICANCE STATEMENTWe have used a “bedside-to-bench” approach to investigate the functional regulation of NMDA receptors (NMDARs). Using information from deep sequencing of patients with neurological or psychiatric disorders, we investigated missense variants identified in the intracellular C-terminal domain of the GluN2B NMDAR subunit. We found several variants that displayed altered properties. In particular, one variant identified in a patient with autism, human GluN2B S1415L, displayed reduced surface expression and binding to PSD-95. Furthermore expression of GluN2B S1415L (S1413L in mouse) showed a deficit in rescue of synaptic NMDAR currents and fewer dendritic spines, consistent with other reports of spine abnormalities being associated with autism. More broadly, we demonstrate that using patient data is an effective approach to probing the structure/function relationship of NMDARs.

The five repetition sit-to-stand test (5STS) is a simple and validated functional test, suitable to be performed in different settings, that has been related with mortality in chronic obstructive pulmonary disease (COPD). Yet, its relationship with the need for hospitalisation is unknown. This study explored the 5STS ability to discriminate between hospitalised and non-hospitalised patients at the onset of an acute exacerbation of COPD (AECOPD). Patients with AECOPD visiting the urgency ward of 2 hospitals were recruited. Clinicians defined the need for hospitalisation based on their best judgment. The 5STS was completed within 48h of the AECOPD onset. Receiver operating characteristic analysis was performed and the area under the curve (AUC) was calculated. 47 patients (92% male, 69±7yrs, FEV150±20%predicted) participated. Age, gender, FEV1, body mass index and quadriceps muscle strength were not different between inpatients and outpatients. 5STS showed good ability to discriminate between inpatients and outpatients (AUC=0.73; 95%CI 0.59-0.88), with a sensitivity of 75% and specificity of 53% for a cut-off of 13.7s (Fig. 1). 5STS can accurately discriminate between inpatients and outpatients with AECOPD. A cut-off of 13.7s may be used to differentiate these patients. Results also emphasise the need to improve patients’ functionality during hospitalisation. Studies with larger samples are needed. published

Tese de mestrado, Psicologia (Área de Especialização em Psicologia Clínica e da Saúde - Psicologia Clínica Dinâmica), Universidade de Lisboa, Faculdade de Psicologia, 2019 Submitted by Biblioteca FPIE-ULisboa (bibliorul@fpie.ulisboa.pt) on 2020-01-31T14:31:04Z No. of bitstreams: 1 ulfpie053542_tm.pdf: 645160 bytes, checksum: cdc6f0ce4988743713f9033e66aa7eaf (MD5) Made available in DSpace on 2020-01-31T14:31:50Z (GMT). No. of bitstreams: 1 ulfpie053542_tm.pdf: 645160 bytes, checksum: cdc6f0ce4988743713f9033e66aa7eaf (MD5) Previous issue date: 2019

The originally published version of this Article contained errors in Figure 5, for which we apologise. In panel c, the scatter graph was inadvertently replaced with a scatter graph comprising a subset of data points from panel d. Furthermore, the legends to Figures 5c and 5d were inverted. These errors have now been corrected in both the PDF and HTML versions of the Article, and the incorrect version of Fig. 5c is presented in the Author Correction associated with this Article.

Transthyretin (TTR) is a protein whose function has been associated to binding and distribution of thyroid hormones in the body and brain. However, little is known regarding the downstream signaling pathways triggered by wild-type TTR in the CNS either in neuroprotection of cerebral ischemia or in physiological conditions. In this study, we investigated how TTR affects hippocampal neurons in physiologic/pathologic conditions. Recombinant TTR significantly boosted neurite outgrowth in mice hippocampal neurons, both in number and length, independently of its ligands. This TTR neuritogenic activity is mediated by the megalin receptor and is lost in megalin-deficient neurons. We also found that TTR activates the mitogen-activated protein kinase (MAPK) pathways (ERK1/2) and Akt through Src, leading to the phosphorylation of transcription factor CREB. In addition, TTR promoted a transient rise in intracellular calcium through NMDA receptors, in a Src/megalin-dependent manner. Moreover, under excitotoxic conditions, TTR stimulation rescued cell death and neurite loss in TTR KO hippocampal neurons, which are more sensitive to excitotoxic degeneration than WT neurons, in a megalin-dependent manner. CREB was also activated by TTR under excitotoxic conditions, contributing to changes in the balance between Bcl2 protein family members, toward anti-apoptotic proteins (Bcl2/BclXL versus Bax). Finally, we clarify that TTR KO mice subjected to pMCAO have larger infarcts than WT mice, because of TTR and megalin neuronal downregulation. Our results indicate that TTR might be regarded as a neurotrophic factor, because it stimulates neurite outgrowth under physiological conditions, and promotes neuroprotection in ischemic conditions.

Global cerebral ischemia induces selective degeneration of specific subsets of neurons throughout the brain, particularly in the hippocampus and cortex. One of the major hallmarks of cerebral ischemia is excitotoxicity, characterized by overactivation of glutamate receptors leading to intracellular Ca(2+) overload and ultimately neuronal demise. N-methyl-d-aspartate receptors (NMDARs) are considered to be largely responsible for excitotoxic injury due to their high Ca(2+) permeability. In the hippocampus and cortex, these receptors are most prominently composed of combinations of two GluN1 subunits and two GluN2A and/or GluN2B subunits. Due to the controversy regarding the differential role of GluN2A and GluN2B subunits in excitotoxic cell death, we investigated the role of GluN2B in the activation of pro-death signaling following an in vitro model of global ischemia, oxygen and glucose deprivation (OGD). For this purpose, we used GluN2B(-/-) mouse cortical cultures and observed that OGD-induced damage was reduced in these neurons, and partially prevented in wild-type rat neurons by a selective GluN2B antagonist. Notably, we found a crucial role of the C-terminal domain of the GluN2B subunit in triggering excitotoxic signaling. Indeed, expression of YFP-GluN2B C-terminus mutants for the binding sites to post-synaptic density protein 95 (PSD95), Ca(2+)-calmodulin kinase IIα (CaMKIIα) or clathrin adaptor protein 2 (AP2) failed to mediate neuronal death in OGD conditions. We focused on the GluN2B-CaMKIIα interaction and found a determinant role of this interaction in OGD-induced death. Inhibition or knock-down of CaMKIIα exerted a neuroprotective effect against OGD-induced death, whereas overexpression of this kinase had a detrimental effect. Importantly, in comparison with neurons overexpressing wild-type CaMKIIα, neurons overexpressing a mutant form of the kinase (CaMKII-I205K), unable to interact with GluN2B, were partially protected against OGD-induced damage. Taken together, our results identify crucial determinants in the C-terminal domain of GluN2B subunits in promoting neuronal death in ischemic conditions. These mechanisms underlie the divergent roles of the GluN2A- and GluN2B-NMDARs in determining neuronal fate in cerebral ischemia.

Transthyretin (TTR) has a neuroprotective role in the central nervous system (CNS) in Alzheimer's disease (AD) and cerebral ischemia. Increased levels of TTR and activated insulin-like growth factor I receptor (IGF-IR) are associated with reduced neurodegeneration in an AD mouse model. In the present study, we found that TTR and IGF-I have a synergistic effect on activation of one of the IGF-IR signaling pathways. Hippocampus of TTR null mice present decreased levels of phosphorylated IGF-IR and Akt when compared with TTR wild type littermate animals. Cell studies reveal the synergistic effect of TTR and IGF-I in promoting IGF-IR signaling even under glutamate induced toxicity. TTR:IGF-IR complexes are identified and a bio-layer interferometry assay demonstrated an interaction between TTR and IGF-IR with a KD ranging from 99 to 744nM. In summary, our results point to a new TTR role through the IGF-I axis, mediated through TTR-IGF-IR interactions.

Hepatitis C virus (HCV) is the cause of one of the most prevalent viral infections worldwide. Upon infection, the HCV genome activates the RIG‐I‐MAVS signalling pathway leading to the production of direct antiviral effectors which prevent important steps in viral propagation. MAVS localizes at peroxisomes and mitochondria and coordinate the activation of an effective antiviral response: peroxisomal MAVS is responsible for a rapid but short‐termed antiviral response, while the mitochondrial MAVS is associated with the activation of a stable response with delayed kinetics. The HCV NS3‐4A protease was shown to specifically cleave the mitochondrial MAVS, inhibiting the downstream response. In this study, we have analysed whether HCV NS3‐4A is also able to cleave the peroxisomal MAVS and whether this would have any effect on the cellular antiviral response. We show that NS3‐4A is indeed able to specifically cleave this protein and release it into the cytosol, a mechanism that seems to occur at a similar kinetic rate as the cleavage of the mitochondrial MAVS. Under these conditions, RIG‐I‐like receptor (RLR) signalling from peroxisomes is blocked and antiviral gene expression is inhibited. Our results also show that NS3‐4A is able to localize at peroxisomes in the absence of MAVS. However, mutation studies have shown that this localization pattern is preferred in the presence of a fully cleavable MAVS. These findings present evidence of a viral evasion strategy that disrupts RLR signalling on peroxisomes and provide an excellent example of how a single viral evasion strategy can block innate immune signalling from different organelles.

Rare variants in GRIN genes, which encode NMDAR subunits, are strongly associated with neurodevelopmental disorders. Among these, GRIN2A, which encodes the GluN2A subunit of NMDARs, is widely accepted as an epilepsy-causative gene. Here, we functionally characterize the de novo GluN2A-S1459G mutation identified in an epilepsy patient. We show that S1459 is a CaMKIIα phosphorylation site, and that endogenous phosphorylation is regulated during development and in response to synaptic activity in a dark rearing model. GluN2A-S1459 phosphorylation results in preferential binding of NMDARs to SNX27 and a corresponding decrease in PSD-95 binding, which consequently regulates NMDAR trafficking. Furthermore, the epilepsy-associated GluN2A-S1459G variant displays defects in interactions with both SNX27 and PSD-95, resulting in trafficking deficits, reduced spine density, and decreased excitatory synaptic transmission. These data demonstrate a role for CaMKIIα phosphorylation of GluN2A in receptor targeting and implicate NMDAR trafficking defects as a link to epilepsy.

As casas de fresco ressurgem, em território português, numa época em que se renova as preocupações com o conforto e recreio, em âmbito doméstico. À medida que nos aproximamos do século XVI, áreas exteriores como jardins, hortas ou pomares (que há muito integram as quintas e paços), tornam-se espaços cada vez mais cuidados e vocacionados para o lazer. É num ambiente de comunhão com a natureza que encontramos as casas de fresco, demonstrando uma vontade de expandir o espaço habitável para o exterior, sempre vocacionadas para a promoção de momentos de ócio prazeroso. Estas arquiteturas são geralmente de pequenas dimensões, proporcionando espaços de abrigo e de refrigério, que permitem momentos de contemplação e reflexão, em contacto direto com a natureza.Na primeira parte deste trabalho procurámos enquadrar o reaparecimento, estabelecer uma definição e enumerar algumas características destes dispositivos em território português. Na segunda parte procedemos à análise de quatro casos de estudo, situados no concelho de Évora e construídos no século XVI: A Fonte do Pomar do Divor, Casa da Água da Quinta de Valverde, Fonte do Arcediago e Casa de Fresco do Paço de S. Miguel. The casas de fresco [summer houses, coolhouses] reappear in Portuguese territory, at a time when concerns about comfort and recreation, in the domestic sphere, are being renewed. As we approach the 16th century, exterior areas such as gardens, vegetable gardens or orchards (which have long been a part of villas and palaces), start being planned as spaces of leisure and thus carefully designed. It's in an atmosphere of communion with nature that we find the casas de fresco, indicating a willingness to expand the habitable space for the outdoor, always geared toward the promotion of pleasurable moments of leisure. These architectures are generally small, providing spaces of shelter and refreshment, which allow moments of contemplation and reflection, in direct contact with nature.In the first part of this work we tried to frame the reappearance, establish a definition and enumerate some characteristics of these devices in Portuguese territory. In the second part, we analyze four case studies, located in the county of Évora and built in the 16th century: A Fonte do Pomar do Divor, Casa da Água da Quinta de Valverde, Fonte do Arcediago and Casa de Fresco do Paço de S. Miguel.

CaMKII is one of the most studied synaptic proteins, but many critical issues regarding its role in synaptic function remain unresolved. Using a CRISPR-based system to delete CaMKII and replace it with mutated forms in single neurons, we have rigorously addressed its various synaptic roles. In brief, basal AMPAR and NMDAR synaptic transmission both require CaMKIIα, but not CaMKIIβ, indicating that, even in the adult, synaptic transmission is determined by the ongoing action of CaMKIIα. While AMPAR transmission requires kinase activity, NMDAR transmission does not, implying a scaffolding role for the CaMKII protein instead. LTP is abolished in the absence of CaMKIIα and/or CaMKIIβ and with an autophosphorylation impaired CaMKIIα (T286A). With the exception of NMDAR synaptic currents, all aspects of CaMKIIα signaling examined require binding to the NMDAR, emphasizing the essential role of this receptor as a master synaptic signaling hub., Calcium-calmodulin-dependent protein kinase II (CaMKII) is well known for its roles in synaptic plasticity. Using a series of molecular replacement experiments, the authors show that the kinase function of CaMKII is required for long-term plasticity and basal AMPA receptor-mediated transmission.

Dissertação de Mestrado Integrado em Arquitetura, com a especialização em Arquitetura apresentada na Faculdade de Arquitetura da Universidade de Lisboa para obtenção do grau de Mestre. O Rio Mira é um sistema vivo que nasce em Santa Clara, passa por Odemira e desagua em Vila Nova de Milfontes. É parte integrante de uma paisagem muito característica. Entre as serras e a planície corre um troço de 30km onde as marés se fazem sentir. Este factor, aliado à produção de cereais no local, foi essencial para o desenvolvimento e construção de moinhos de maré neste território. Num total de 15 engenhos identificados são visíveis actualmente apenas cinco deles. Sendo este território um antigo porto fluvio-marítimo e zona de produção de alimento e de trabalho pretende-se aqui regenerar uma memória e uma história que se encontram hoje em dia num esquecimento quase completo. A existência naquele local de quatro moinhos abandonados e consequentemente em estado de ruína serviu de pretexto para uma intervenção. Este incide em particular nos do tipo moinho de maré, que são entre todos os mais ignorados. Em confronto com a situação deste local e do país desenvolveu-se uma proposta que ajudasse a dinamizar e a rehabitar este local através de uma estratégia inovadora, que actua num campo que se encontra hoje em grande desenvolvimento no país: o turismo. Para isso, este projecto integra três componentes - a criação de um percurso adaptado à paisagem, a reabilitação dos antigos moinhos de maré com funcionalidades que apoiem e constituam um hotel e a criação de unidades de alojamento complementares. O local, pelas suas características únicas, obriga à criação de uma arquitectura, que baseada na razão e na invenção humana, crie um diálogo com a natureza e a valorize. ABSTRACT:The Mira River is an ecosystem that stretches from Santa Clara, flowing through Odemira and into the Atlantic Ocean in Vila Nova de Milfontes. The river is a distinctive feature of this ecosystem and its landscape. Tucked between hills and plains is a 30km section of the river where the influence of the tide is present. This phenomenon, along with cereal production in the region, was the driving force behind the development of tide mills throughout this stretch of the river. Of a total of fifteen mills built, only five survived. This region was once a fluvial port and an important food production site which together employed many of the people living there. What is intended here is to bring back to life some of these memories and part of these mills history that has been forgotten. The existence in that place of four abandoned mills and consequently in a state of ruin, served as pretext for an intervention. This intervention occurs particularly on tide mills for being the ones that are of them all the most ignored. Taking into account the economical situation of the region, this project seeks to aid and invigorate this place through an innovating strategy, that takes into account tourism an ever growing sector in our country’s economy today. This project is composed of three main elements. The creation of a route integrated with the landscape, the creation of complementary accommodation lodging units and the rehabilitation of the old mills as support infrastructure with diverse functionalities. Due to the historical and ecological importance of the region, for its unique characteristics, the proposed intervention demands an architecture that is based in reason and human invention, creates a dialogue with nature, thus valuing it. N/A

Necroptosis is a form of regulated necrotic cell death, which is mediated by receptor-interacting protein 1 kinase (RIPK1) and RIPK3, and the downstream effector mixed lineage kinase domain-like (MLKL). Necroptotic signals induced by death receptors, such as TNF receptor 1, Toll-like receptors or interferon receptors lead to the formation of the necrosome, and result in cell death with morphological features of necrosis. The execution of necroptosis involves oligomerization of MLKL upon phosphorylation by RIPK3, its translocation to the plasma membrane and consequent plasma membrane permeabilization. Necroptosis participates in physiological functions but is also involved in cell death associated with several pathophysiological conditions. Here, we discuss key features of necroptosis, and evidence implicating necroptotic cell death in brain ischemia.

A presente investigação analisa as práticas interdisciplinares no sistema de proteção à criança em Portugal e identifica as dimensões de sustentabilidade que garantem os direitos das crianças. O campo empírico é constituído por profissionais que atuam na matéria de infância e juventude, ou sejam: assistentes sociais, juristas, advogados, psicólogos e educadores, que desempenham funções em entidades como as Comissões de Proteção de Crianças e Jovens em Risco e na Instituição de Apoio à Criança. Caracterizamos o sistema de proteção à criança em Portugal, as práticas interdisciplinares, dando particular enfase às práticas dos assistentes sociais e categoriza as dimensões de sustentabilidade na proteção da criança em Portugal. Utilizamos uma metodologia qualitativa, recorrendo ao método indutivo em conformidade com os objectivos definidos. Todos os dados foram recolhidos através de entrevistas semiestruturadas através de um guião previamente elaborado, onde posteriormente todas respostas obtidas foram analisadas, ficando a conhecer a realidade e a experiência de cada profissional. Concluímos que os profissionais sentem algumas dificuldades no momento de intervir derivado ao que teoricamente está escrito legalmente não se adequando ao que na prática necessitam de fazer, de acordo com os problemas contemporâneos. Estas dificuldades dizem respeito à forma como o sistema de proteção à criança em Portugal se encontra estruturado, necessitando ainda de algumas correcções legais. Conclui-se também que todos os profissionais trabalham no sentido de alcançarem uma resolução eficaz e eficiente nas suas intervenções. The present research aims to analyze the interdisciplinary practices in the child protection system in Portugal and to identify the sustainability dimensions that guarantee the rights of children. The empirical field is made up of professionals who work in the field of childhood and youth, namely: social workers, jurists, lawyers, psychologists and educators, who work in entities such as the Comissões de Proteção de Crianças e Jovens em Risco and the Instituição de Apoio à Criança. We characterize the child protection system in Portugal, to typify interdisciplinary practices, to identify and analyze professional practices, giving special attention to the practices of social workers and to categorize the dimensions of sustainability in child protection in Portugal. We use a qualitative methodology, applying the inductive method always focused on the defined objectives. All the data were collected through planned interviews through a previously elaborated script, Where later all answers obtained were analyzed, getting to know the reality and the experience of each professional. We conclude that the professionals feel some difficulties in the moment of intervening, since legally the theory is not adapted to the practice, within the contemporary problems. These difficulties are related to the way in which the child protection system in Portugal is structured, requiring some legal modifications. It is also concluded that, despite all the difficulties, all professionals are working towards an effective and efficient resolution of their interventions.

Mestrado em Engenharia Agronómica / Instituto Superior de Agronomia Para contribuir para um maior conhecimento das cultivares de oliveira portuguesas, avaliou-se a fenologia durante o ciclo em várias cultivares de oliveira, relacionando-a com outras áreas de interesse agronómico e económico. Para tal, analisou-se a qualidade da flor e o vingamento, relevantes para o potencial produtivo e avaliou-se a incidência de pragas e doenças importantes nesta cultura. Para a avaliação do Olho de Pavão (doença causada por Spilocaea oleagina), quantificou-se a proporção de área foliar com lesão visível. Para avaliar a Tuberculose (doença causada pela Pseudomonas savastanoi), prepararam-se suspensões com ramos, folhas e frutos que foram depois colocadas em meio favorável ao desenvolvimento da bactéria, seguindo-se uma avaliação qualitativa do crescimento das colónias. A mosca da azeitona, praga importante desta cultura, foi também avaliada em campo, observando-se semanalmente o número de indivíduos nas armadilhas. Para aferir a produtividade de cada cultivar, quantificaram-se a produção total de frutos e os teores de gordura e de humidade do fruto. O presente trabalho permitiu concluir que fenologicamente em 2016 a 'Verdeal de Serpa' apresentou o maior período de floração, contrariamente à 'Ocal' e 'Santulhana'. As cultivares com melhor qualidade da flor foram a 'Galega Vulgar' e a 'Lentrisca', tendo esta última registado também o maior vingamento. A 'Cobrançosa' foi identificada pelos ensaios como a mais sensível à mosca, por oposição à 'Quinta do Portado'. No que diz respeito às doenças, a 'Cordovil de Serpa', a 'Azeiteira' e a 'Cobrançosa' foram as mais sensíveis ao fungo Spilocaea oleagina enquanto a 'Lentrisca' e a 'Galega Vulgar' se mostraram mais tolerantes. Relativamente à Tuberculose, a 'Cornicabra' surpreendeu como sendo das cultivares mais sensíveis juntamente com a 'Cordovil de Castelo Branco', mostrando-se mais tolerantes a 'Cobrançosa' e a 'Quinta do Portado' (descrita como sensível). A 'Azeiteira' revelou-se a cultivar mais produtiva, ao contrário da 'Quinta do Portado N/A

Objectives:Presentation of the study design u201cCharacterization of risk of depression and suicide in a sample of elderly people living in the municipality of Beja u2013 an exploratory studyu201d.Background:Suicide has a strong impact on public health, not only because it is generally associated with treatable pathology like major depressive disorder, but also because it has a dramatic impact on survivors. Suicide is a growing problem in our country, and Beja, a district in the interior south of Portugal, is among the regions with the highest suicide rates. The issues of rurality, isolation and old age are known suiciderisk factors that are important in Beja, where 25% of the population is over 65 years old.Materials and Methods:Three groups of elderly people were constituted: Group A - isolated elderly, identified by the national police

Global cerebral ischemia induces selective acute neuronal injury of the CA1 region of the hippocampus. The type of cell death that ensues may include different programmed cell death mechanisms namely apoptosis and necroptosis, a recently described type of programmed necrosis. We investigated whether necroptosis contributes to hippocampal neuronal death following oxygen-glucose deprivation (OGD), an in vitro model of global ischemia. We observed that OGD induced a death receptor (DR)-dependent component of necroptotic cell death in primary cultures of hippocampal neurons. Additionally, we found that this ischemic challenge upregulated the receptor-interacting protein kinase 3 (RIP3) mRNA and protein levels, with a concomitant increase of the RIP1 protein. Together, these two related proteins form the necrosome, the complex responsible for induction of necroptotic cell death. Interestingly, we found that caspase-8 mRNA, a known negative regulator of necroptosis, was transiently decreased following OGD. Importantly, we observed that the OGD-induced increase in the RIP3 protein was paralleled in an in vivo model of transient global cerebral ischemia, specifically in the CA1 area of the hippocampus. Moreover, we show that the induction of endogenous RIP3 protein levels influenced neuronal toxicity since we found that RIP3 knock-down (KD) abrogated the component of OGD-induced necrotic neuronal death while RIP3 overexpression exacerbated neuronal death following OGD. Overexpression of RIP1 also had deleterious effects following the OGD challenge. Taken together, our results highlight that cerebral ischemia activates transcriptional changes that lead to an increase in the endogenous RIP3 protein level which might contribute to the formation of the necrosome complex and to the subsequent component of necroptotic neuronal death that follows ischemic injury.

Transthyretin is the carrier protein of thyroxine and retinol in plasma and cerebrospinal fluid and has been described also as a neuroprotective molecule. 14-3-3 Proteins are very important in many cellular processes, being their absence related with deficits in memory and learning. The analysis of the relationship between these two proteins is the main objective of this work. We found that hippocampi of young TTR null mice presented lower levels of 14-3-3ζ protein, but no changes in gene expression when compared to TTR wild type littermates were noted. Cellular studies ascribed this finding to increased degradation of 14-3-3ζ in lysosomes in the absence of TTR, increasing autophagy.