Plasma membrane-specific interactome analysis reveals calpain 1 as a druggable modulator of rescued Phe508del-CFTR cell surface stability

Cystic fibrosis (CF) is a genetic disease caused by mutations in the gene encoding CF transmembrane conductance regulator (CFTR), a chloride channel normally expressed at the surface of epithelial cells. The most frequent mutation, resulting in Phe-508 deletion, causes CFTR misfolding and its premature degradation. Low temperature or pharmacological correctors can partly rescue the Phe508del-CFTR processing defect and enhance trafficking of this channel variant to the plasma membrane (PM). Nevertheless, the rescued channels have an increased endocytosis rate, being quickly removed from the PM by the peripheral protein quality-control pathway. We previously reported that rescued Phe508del-CFTR (rPhe508del) can be retained at the cell surface by stimulating signaling pathways that coax the adaptor molecule ezrin (EZR) to tether rPhe508del–Na+/H+-exchange regulatory factor-1 (NHERF1) complexes to the actin cytoskeleton, thereby averting the rapid internalization of this channel variant. However, the molecular basis for why rPhe508del fails to recruit active EZR to the PM remains elusive. Here, using a proteomics approach, we characterized and compared the core components of wt-CFTR– or rPhe508del–containing macromolecular complexes at the surface of human bronchial epithelial cells. We identified calpain 1 (CAPN1) as an exclusive rPhe508del interactor that prevents active EZR recruitment, impairs rPhe508del anchoring to actin, and reduces its stability in the PM. We show that either CAPN1 downregulation or its chemical inhibition dramatically improves the functional rescue of Phe508del-CFTR in airway cells. These observations suggest that CAPN1 constitutes an attractive target for pharmacological intervention, as part of CF combination therapies restoring Phe508del-CFTR function.This work was supported by a center grant UID/MULTI/04046/2019 to BioISI and project PTDC/BIA-CEL/28408/2017 and IF2012 to PM, both from FCT, Portugal. AMM was recipient of fellowship SFRH/BD/52490/2014 from BioSYS PhD programme PD65-2012, and PB of fellowship SFRH/BPD/94322/2013.N/

From Fun to Nightmare. Measuring Students’ Senior Trip Experiences Through a Mixed-method Approach

From Fun to Nightmare. Measuring Students’ Senior Trip Experiences Through a Mixed-method Approac

Factorial validity and group invariance of the Portuguese short version of the Social Physique Anxiety Scale in adolescents

Social physique anxiety (SPA) has been associated with a range of psychosocial and health-related variables, thus it can be considered is an indicator of social-psychological adjustment. The purpose of this paper is to determine factorial validity and group invariance of Motl and Conroy (2000) translated 7-item SPA among Portuguese adolescents. A nationally representative sample of 3330 8th and 10th grade students (15.07 1.34 years old; 47.5% males and 52.5% females) answered a survey as a part of a larger collaborative cross-national survey, the Health Behaviour in School-Aged Children (HBSC) 2006 study. Exploratory and confirmatory factor analyses resulted on a uni-dimensional factor structure of 6 items [Satorra-Bentler c2 = 30.85, df = 8, p<.01; CFI = .996; NNFI = .992; RMSEA = .038 (90% C.I.: .024 - .052); SRMR = .010]. Further analises, confirmed configurational (all CFI and RMSEA > .99) and metric invariances (CFI difference between restricted and unrestricted models were lower than .01 for all groups) across gender, grade level, diet beliefs, physical activity, perception of body, and BMI. The present version can be used confidently by researchers in analysing and interpreting scores of SPA across a variety of samples in Portuguese adolescents, and that this instrument can be used in cross-cultural research

Life satisfaction in adolescents:the role of individual and social health assets

The aim of this study is to explore the relationship between adolescents’ life satisfaction and individual and social health assets. A nationally representative sample of 3,494 Portuguese adolescents (mean age = 14.94 ± 1.30 years; 53.6% girls) completed the Health Behavior in School-aged Children survey measuring a variety of health behaviors and beliefs. A sequential regression analysis was conducted with gender, individual assets (academic achievement, social competence, self-regulation and life objectives) and social assets (family support, peer support, parental monitoring and school connectedness) entered in separate steps. A second regression analysis was conducted with social assets entered before individual assets. The final model explained 18.3% of life satisfaction. School connectedness (β = .198, p < .001) and family support (β = .154, p < .001) were the strongest predictors of adolescents’ life satisfaction followed by social competence (β = .152, p < .001), academic achievement (β = .116, p < .001) and self-regulation (β = .064, p < .001). Social assets explained a larger variance of life satisfaction than individual assets when entered first in the regression (r2 = .134 and r2 = .119, respectively, p < .001). When entered last step in the regression analysis, social assets added more to life satisfaction’s variance than when individual assets were added in the last step (r2 = .060 and r2 = .045, respectively, p < .001). These results reinforce the role social interaction and social capital models in the promotion of well-being

Efectos del programa de gestión personal en la promoción de la actividad física en adolescentes

Las enfermedades cardiovasculares son la principal causa de muerte prematura en todo el mundo. La prevención del sedentarismo es pues una preocupación de la salud pública. El desarollo de hábitos de actividad física debe ser uno de los principales objetivos de las políticas de educación para la salud en las que la Educación Física en las escuelas tiene un papel fundamental. El objetivo del presente estudio es verificar la eficacia de un programa de promoción de la actividad física en adolescentes en el medio escolar. El programa se ha desarollado en ocho sesiones (una a la semana) en las cuales se han enseñado estrategias cognoscitivas y de comportamiento que facilitaran la implicación en actividades físicas a lo largo de la vida. Se evaluaron las intenciones de comportamiento, en cuatro momentos distintos a lo largo del año lectivo: evaluación de control, pre-test, post-test y evaluación de seguimiento. Estos momentos delimitan tres fases del estudio: periodo de control (cuatro semanas), periodo experimental (nueve semanas) y periodo de seguimiento (ocho semanas). Los niveles de actividad física han disminuido de una forma significativa durante los periodos de control y de seguimientos y han aumentado durante el periodo experimental, aunque no de forma significativa. Las intenciones de comportamiento han aumentado significativamente durante la intervención y disminuido durante el periodo de seguimiento. Se sugiere que una intervención más prolongada podría ser necesaria para alterar de un forma consistente los hábitos de los individuos.Cradiovascular diseases actually are the main cause of death all around the world. Sedentarism prevention is a first line preocupation for the public health authorities. The development of physical activity habits must be one the main aims for the eduaction on the youth. The objective of the present study, is to verify the efficacy of a program of physical activity promotion of teenagers in the school. This program was developed by the physical education teachers, and directed to the different groups of young schoolars. The duration of the program was a total of eight sessions. Each session was performed one time at week. The content of each session are programed in order to provide strategies both cognoscitive and behavioural to the students, which facilitate their adherence to the physical activities lifelong. We evaluated the behavior intentions, at four different moments during the school periode: control, pre-test, post-test, and follow-up. These moments are the three phases of the study: control (four weeks); experimental phase (nine weeks); and follow-up (eight weeks). The results shown that the physical activity levels decreased during the phases number one and three, and increased during the experimental phase, but not significatively. However, the behaviour intentions increased significatively during the intervention phase, and decreased at the follow-uperiod. We suggest that one intervention more extented could be necessary in order to obtanin a singificatively change over the individuals behavior

The information professionals in the Algarve region: a study of the users perspective

In this article it is presented the research carried out in the South of Portugal (region of Algarve) with the general objective to know the users perspective about the information services (public libraries and archives) and their professionals and afterwards define a marketing strategy to improve the image of these services. After the literature review it was decided to apply a questionnaire (quantitative approach) to the users and non-users of archives and libraries of the region. The data collected were analysed and although the perspectives of the users were very good there is the need to develop a marketing campaign to show the non-user the information services potentiality.CIDHEUS-UE/FC

Proof in classroom social practice

How does the construction of proof relate to the social practice developed in the mathematics classroom? This report addresses the role of diagrams in order to focus the complementarity of participation and reification in the process of constructing a proof and negotiating its meaning. The discussion is based on the analysis of the mathematical practice developed by a group of four 9th grade students and is inspired by the social theory of learnin

Os estilos de vida dos jovens adultos com diabetes tipo 1.

A maioria dos jovens apresenta boas competências pessoais e sociais, considera ter bom suporte social (família, colegas, amigos, equipa de saúde) e refere os benefícios das atividades em grupo com outros jovens com diabetes. Média de satisfação com a vida 6,6 ±1,7 (escala 0 -10). Os jovens evidenciam bom fracionamento alimentar (83%), um terço pratica a atividade física recomendada. A adesão à insulinoterapia (77% administram ≥ 4 vezes dia, 6% utilizam “bomba” de insulina) e à vigilância glicemica (74% pesquisam ≥ 3 vezes dia) é satisfatória. A média de HbA1c é 8,7 ±1,6%. Quanto mais elevada a HbA1c: menor adesão global ao tratamento da diabetes (r=-0,153*), menor perceção de saúde (r=0,377***) menor satisfação com a vida (r=-0,185**), menor adaptação psicológica à diabetes (r=- 0,167*). 25% dos jovens apresentam complicações da diabetes A maioria dos jovens manifestou concordar com as representações positivas sobre a diabetes e demonstrou boa adaptação psicológica. As melhores representações sobre a diabetes estão positivamente relacionadas com a adesão à alimentação (r=0,190**) e à insulinoterapia (r=0,247***). A melhor adaptação psicológica à diabetes está positivamente relacionada com a adesão à alimentação (r=0,181**), à vigilância glicemica (r=0,179**) e à insulinoterapia (r=0,241***)

Role of oxidative stress in ERK and p38 MAPK activation induced by the chemical sensitizer DNFB in a fetal skin dendritic cell line

The intracellular mechanisms involved in the early phase of dendritic cell (DC) activation upon contact with chemical sensitizers are not well known. The strong skin sensitizer 2,4-dinitrofluorobenzene (DNFB) was shown to induce the activation of mitogen-activated protein kinases (MAPK) in DC. In the present study, we investigated a putative role for oxidative stress in DNFB-induced MAPK activation and upregulation of the costimulatory molecule CD40. In a DC line generated from fetal mouse skin, DNFB induced a significant increase in protein oxidation, measured by the formation of carbonyl groups, while it had almost no effect on lipid peroxidation. The antioxidants glutathione and vitamin E, which inhibit protein and lipid oxidation, respectively, were used to assess the role of oxidative stress in DNFB-induced MAPK activation. Glutathione, but not vitamin E, inhibited DNFB-induced p38 MAPK and ERK1/2 phosphorylation, whereas none of the antioxidants interfered significantly with the DNFB-induced upregulation of CD40 protein levels. Taken together, these results indicate that DNFB activates p38 MAPK and ERK1/2 via production of reactive oxygen species, and that protein oxidation plays an important role in MAPK activation