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Additional file 1: Figure S1. Gating strategy to detect CD69+ or CD25+ cells following sAg-mediated activation. E6.1-CD6WT and E6.1-CD6Dcyt cells were cultured for 24 h in the presence of unloaded or sAg-loaded Raji cells, and expression of CD69 and CD25 was assessed by flow cytometry. (a) The gate was defined based on unstained E6.1 cells, and positivity is represented in the right half of dot plots of FSC vs. marker expression. (b) Representative dot plots for each condition analyzing CD69 (left panels) or CD25 (right panels) expression. FSC: Forward scatter. Figure S2. Activity of the HRas biosensor MRS2 in unstimulated and activated E6.1 cells. (a) MRS2- and MRS2-S17N-expressing E6.1 single cells were filmed for 5 min at 37 ºC (1 min frames). At the 5 min mark, medium was added and cells were filmed for another 30 min. FRET/Clover ratios were assessed for each time-point. (b) Biosensor-expressing cells were initially filmed for 5 min alone. Then, unloaded or sAg-loaded Raji cells were added and allowed to form conjugates with the E6.1 cells for 30 min. FRET/Clover ratios were calculated for cells expressing MRS2 that interacted with Raji or Raji + sAg, and the same was performed for E6.1-MRS2-S17N:Raji pairs. (c) E6.1-CD6WT or E6.1-CD6∆cyt cells expressing either MRS2 or MRS2-S17N were filmed during conjugate formation with sAg-primed Raji cells. Biosensor-expressing E6.1 cells were filmed alone for 5 min, and for 30 min after sAg-Raji cells were added to the preparation. FRET/Clover ratios were calculated for each of the four experimental conditions. (a-c) Dots and lines represent mean ± SD of 5 independent experiments, with 10 individual cells (in a) or 9–10 cell pairs (in b and c) assessed for each condition in each experiment.

The development of autonomy is influenced by different individual and contextual variables, that include socio-demographic and family/institutional (in cases where children live in residential care) background. Hence, the role residential care play in the process of providing children and youth at risk/danger with the adequate skills to prepare them to have an independent adult life is fundamental. A quantitative exploratory study was thus developed to characterize adolescents/young people autonomy (68.8% living in residential care) and to explore individual variables relevance in this area. Most of the participants lived in coastal areas of the country (69%), were female (76.2%) and age ranged between 13 and 24 (17.38±2.33). They completed the Autonomy Questionnaire for Adolescents. SPSS IBM 24 was used for data analysis and statistically significative differences were found (p˂.05): i) between males (with higher results) and females; ii) in functional autonomy according to age (higher scores among the 13-17 year-old group vs. 18- 24); iii) in functional and total autonomy, better in those living in residential care. These results stress the importance of investing in parental education initiatives that may foster the construction of youths’ autonomy in their own families considering the role played by gender and age in intervention. info:eu-repo/semantics/publishedVersion

Mestrado em Educação Pré-Escolar e Ensino no 1º Ciclo do Ensino Básico Neste trabalho apresenta-se o estudo realizado durante um estágio curricular do Mestrado em Educação Pré-Escolar e Ensino do 1.º Ciclo do Ensino Básico, da Universidade de Aveiro, num contexto de Pré-escolar, numa sala constituída por 21 crianças, com idades compreendidas entre os 3 e os 6 anos. Neste âmbito, assumindo a importância do direito de participação das crianças, procurámos promover competências sociais e capacidade de questionamento e reflexão crítica nas crianças, baseando-nos numa abordagem tipo “filosofia para crianças”. Em particular, pretendeu-se estimular nas crianças o desenvolvimento de atitudes positivas na relação entre pares, capacidade de resolução de conflitos, de entreajuda, bem como o estímulo à reflexão no sentido de desenvolver o seu sentido crítico, de escuta e respeito pela opinião do outro. A avaliação das sessões de filosofia baseou-se na análise dos níveis de implicação das crianças e no registo dos comentários e atividades realizadas pelas crianças, daí se inferindo acerca dos seus níveis de participação e interesse das crianças pelos temas de cariz social. Ainda, procurámos perceber até que ponto as sessões contribuíam para uma melhoria das relações entre pares no quotidiano. Concluímos que, apesar dos níveis de implicação das crianças não serem, globalmente, muito elevados, o balanço geral foi positivo, na medida em que as crianças foram adotando novas formas de resolver os seus conflitos, passaram a falar mais sobre os seus sentimentos, a emitir a sua opinião e a aceitar melhor o ponto de vista do outro. Atendendo aos resultados, muito haverá a melhorar, algo que só poderá ser alcançado com um investimento e melhoria continuada na abordagem filosofia para crianças, visando educar para o direito à participação. In this paper it’s presented a study done during an Infant Education and Basic Compulsory Education Masters Degree’s traineeship from the University of Aveiro. It was done in a context of a preschoolers’ class of twentyone children, between the ages of three and six years old. In this sphere of action, and taking in consideration the importance of the children’s participation we tried to develop their ability to have a critical reflection, based on an attitude of “philosophy for kids”. It was made the attempt, in particular, to stimulate the development of positive attitudes between peers, the capacity to resolve conflicts, the development of mutual help felling and to stimulate, also, the reflection so it’s possible to the children to develop their critical sense, listening capacity and respect of other’s opinion. The evaluation of the philosophy sessions was based on the analysis of the levels of implication presented by the children and on the records of commentaries and activities done by them. From this is possible to imply the children’s levels of participation and interest when it comes to social themes. We also tried to understand the importance of this sessions in the improvement of the peers’ relationships. We concluded that, despite the not very high levels of implication presented by the children, the overall result was positive, since the children acquired new ways to resolve conflicts, they also started to talk more about their feelings and opinions and they learn to embrace other’s opinions. There’s still a lot to improve, but this is only possible if the investment and improvement of the approach of the “philosophy for kids” can be done, always considering a children’s participation based education.

A autonomia traduz-se numa habilidade relacional cujo desenvolvimento ocorre ao longo de toda a vida sendo influenciada por variáveis individuais, contextuais e familiares diversas (Barbosa & Wagner, 2015). O estudo apresentado procura compreender a autonomia, percecionada por 42 adolescentes/jovens e por 66 monitores/animadores, e o contributo da participação em Colónias de Férias neste âmbito, atendendo a variáveis sociodemográficas e institucionais. A amostra de adolescentes/jovens foi sobretudo do litoral (69%), género feminino (76.2%) e 68.8% que ainda se encontravam institucionalizados. Quanto aos monitores, a maioria era do litoral (77.3%), encontrava-se a trabalhar (84.8%), 63.6% do género masculino e apresentava idade média de 28 anos. Foi aplicado aos adolescentes o Questionário de Autonomia dos Adolescentes e um instrumento construído especificamente para o estudo aos monitores. Recorreu-se ao SPSS IBM 23 para a análise de dados. Relativamente à autonomia dos adolescentes/jovens percecionada pelos próprios verificaram-se diferenças estatisticamente significativas entre: 1) rapazes (melhores resultados) e raparigas na independência e autonomia cognitiva; 2) grupos etários (13-17 vs. 18-24) na auto-determinação (resultados superiores dos 13 aos 17); 3) os que residiam ainda em instituição ou não (médias mais elevadas) na independência; 4) e adolescentes e jovens institucionalizados (resultados mais favoráveis) e não institucionalizados na auto-determinação e autonomia total. Em relação à contribuição das Colónias de Férias para a autonomia dos adolescentes e jovens, percecionada pelos próprios e pelos monitores/animadores, não se verificaram resultados estatisticamente significativos. Será fundamental continuar a explorar estas questões inerentes à compreensão da autonomia, nomeadamente atendendo a variáveis de natureza diversa, incluindo aspetos pessoais e contextuais. The autonomy refers to a relational hability that grows lifelong influenced by individual, contextual and familiar variables (Barbosa & Wagner, 2015). The following study aims to understand the autonomy perceived by 42 teenagers/young people and 66 monitors/animators as well as the contribution of participations in Holidays Camps in this sense, considering sociodemografic and institucional. The teenagers/youngs were in large number from coast (69%), females (76.2%) and people who still are institutionalized (68.8%). As for the monitors, the majority lived in the coast (77.3%), most of them had a job (84.8%) and there were more males (63.6%), with the medium age of 28 years. The Adolescents Autonomy Questionnaire was applied to adolescents and an instrument built specifically for the study of the monitors. SPSS IBM23 was used for data analysis. For the teenagers and young's autonomy, there was some statistically significant differences between: 1- boys (better results) and girls on independence and cognitive autonomy; 2- age groups (13-17 vs 18-24) on self-determination (higher levels on 13-17 group); 3- those who still live in institution and those who don’t (higher results), on independence; 4- institutionalized and non-institutionalized individuals (better results) on self-determination and total autonomy. In relation to the contribution of Holidays Camps for the teenagers and youngs autonomy, there were not statistically significant results. Its fundamental that we continue to explore this questions about autonomy to have a better knowledge, particularly about the contribution of the personal and contextual variables.