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

Search for new modulators of Phe508del-CFTR retention at the plasma membrane of epithelial cells

Tese de doutoramento, Biologia (Biologia de Sistemas), Universidade de Lisboa, Faculdade de Ciências, 2018Cystic fibrosis (CF) is a complex inherited disorder caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Around 2000 disease causing mutations are known for this gene, which encodes a Chloride (Cl−) channel expressed at the plasma membrane (PM) of epithelial cells. The most frequent CFTR mutation, the deletion of phenylalanine 508 (Phe508del), causes the protein to misfold and be prematurely degraded. Low temperature or pharmacological “correctors” can partly rescue Phe508del-CFTR processing defect and enhance the channel traffic to the cell surface. Nevertheless, the rescued channels show partial channel function and a highly decreased PM half-life, due to accelerated endocytosis and fast turnover. Given this accelerated endocytic rate, new strategies aiming to retain rescued Phe508del-CFTR at the cell surface could be relevant as to enhance the efficacy of currently available pharmacological correctors. For that reason, the major objective of this dissertation is to identify novel cellular pathways or key interactors for the modulation of CFTR surface retention. Previous results from the host laboratory had showed that stimulation of endogenous RAC1 by Hepatocyte Growth Factor (HGF) signaling potentiated the retention of rescued Phe508del-CFTR at the PM by promoting an interaction between the actin-binding adaptor ezrin (EZR) and the Na+/H+ exchange regulatory factor-1 (NHERF1), enhancing CFTR anchoring to the actin cytoskeleton. In chapter 2 we showed that the mechanism behind this stabilization lies on a conformational change in NHERF1, triggered by EZR activation upon RAC1 signaling, which is then able to bind and stabilize misfolded CFTR at the PM. However, HGF/RAC1 signaling pathway is known to have proliferative and pleiotropic biological functions, which limit its application for therapeutic intervention. Therefore, in chapter 3, we investigated the effect of HGF treatment in epithelium-like cellular models, in combination with the most common administrated drugs. Contrary to what would be commonly assumed, we found that prolonged co-administration of HGF actually prevented previously unrecognized epithelial dedifferentiation effects of prolonged exposure to the FDA-approved Phe508del-CFTR corrector VX-809. It also significantly increased the Phe508del-CFTR functional rescue by the FDA- and EMA-approved VX-809/VX-770 drug combination, preventing the destabilization of the PM rescued channels by prolonged exposure to the VX-770 potentiator drug. These results suggest that HGF co-administration could indeed be beneficial for CF patients and should be further clinically explored. Lastly, since we showed that the type of protein interactions that wt- and rescued Phe508del-CFTR establish at the cell surface can be major determinants of their different PM stabilities, in chapter 4 we identified, for the first time, the core components of the macromolecular complexes assembled around wt- and rescued Phe508del-CFTR proteins at the PM. By identifying exclusive PM interactions between rescued Phe508del-CFTR, NHERF1 and EZR, we were able to recognize Calpain 1 as a key contributor for the decreased surface stability of pharmacologically rescued Phe508del-CFTR, probably acting through the disruption of the EZR-actin cytoskeleton binding.Fundação para a Ciência e a Tecnologia (PD/BD/52490/2014

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 multimorbidity of asthma and rhinitis: from epidemiologic data to molecular traits

RESUMO: Introdução e Objetivos: A asma afeta a vida de várias centenas de milhões de pessoas de todas as idades, em todo o mundo. Apesar dos avanços nas últimas décadas, a asma e a sua inerente multimorbilidade permanecem um ónus significativo para as pessoas com a doença, para as suas famílias e para a sociedade e economia da saúde. Um número elevado de questões permanece por responder, abrangendo vários aspetos da doença, relacionados com lacunas no conhecimento científico atual, desde a epidemiologia à fisiopatologia e aos cuidados prestados à pessoa com asma. O objetivo principal desta dissertação foi contribuir para a abordagem de algumas destas questões relativas à asma e a sua associação com a rinite. Em particular, os trabalhos originais visavam: (1) estimar a prevalência de asma em Portugal e analisar a sua associação com a rinite em grupos populacionais particularmente vulneráveis e sobre os quais há carência de dados a nível internacional - crianças e idosos; (2) identificar características para um reconhecimento precoce de asma, através de fenótipos clínicos multidimensionais de sibilância recorrente em idade pré-escolar, estabelecidos “sem hipóteses pré-definidas" e relacionados com a persistência de asma na adolescência; (3) analisar a associação entre parâmetros funcionais respiratórios das vias aéreas superiores e inferiores, em conjunto com a avaliação subjetiva do controlo da rinite alérgica e da asma, em crianças em idade escolar; (4) explorar estratégias inovadoras para identificar características metabólicas associadas ao fenótipo de asma e rinite alérgica em crianças, em amostras colhidas de forma não invasiva. Métodos: Esta dissertação baseou-se em três tipos de estudos: 1. Estudos transversais, baseados na população nacional, de cidadãos que viviam em Portugal, tendo sido aplicados questionários por entrevista, usando procedimentos padronizados, para a obtenção de dados epidemiológicos relativos à asma e à rinite. Para o estudo pediátrico, foram analisados os dados de todos os indivíduos com idade inferior a 18 anos que participaram no estudo INAsma (estudo por entrevista telefónica, de base populacional, nacional, para estimar a prevalência de asma em Portugal). O estudo dirigido aos idosos foi desenhado para estimar a prevalência de rinite em adultos com 65 anos ou mais, residentes em Portugal continental, tendo sido os dados colhidos por entrevista direta; 2. Estudo prospetivo de coorte de crianças com idade inferior a 7 anos com sibilância recorrente, avaliadas sistematicamente em pontos de tempo específicos, até 13 anos de seguimento. Foram desenvolvidos modelos de regressão logística multivariável para persistência de asma na adolescência, com base em respostas a questionários e resultados de testes cutâneos por picada. Os fenótipos clínicos foram identificados por análise de agrupamento das variáveis (cluster), selecionadas com base na análise de regressão logística, e comparados a respeito da persistência de asma, uso de tratamentos de controlo e avaliação funcional respiratória em idade escolar e na adolescência; 3. Estudo transversal, caso-controlo, de crianças em idade escolar, com rinite alérgica e asma, e crianças saudáveis (amostra emparelhada para a idade e género), avaliadas no que diz respeito a: - Análise laboratorial funcional respiratória, i.e., avaliações sequenciais do débito inspiratório máximo nasal (PNIF) antes e após a aplicação de vasoconstritor tópico nasal, e espirometria com prova de broncodilatação. O teste de controlo da rinite alérgica e da asma para crianças (CARATkids) foi utilizado para a avaliação subjetiva do controlo destas doenças. As associações entre os parâmetros de avaliação funcional respiratória e de controlo subjetivo foram analisadas usando modelos de regressão linear múltipla. - Análise laboratorial analítica por espectroscopia de ressonância magnética nuclear (NMR) para análise metabolómica não dirigida das amostras de urina e saliva de cada criança. Os dados espectroscópicos e clínicos foram analisados estatisticamente, incluindo abordagens multivariável e univariável. Adicionalmente foram colhidas amostras de condensado de ar exalado (EBC) de voluntários, em conjunto com amostras de ar ambiente da sala de colheitas. As amostras colhidas foram analisadas por NMR, com comparação dos espectros resultantes.Resultados: A prevalência estimada de asma ativa em crianças foi de 8,4% (intervalo de confiança a 95% (95%CI) 6,6%-10,7%). As prevalências estimadas de rinite e de asma diagnosticada por médico em idosos foram 29,8% (95CI% 28,4%-31,3%) e 10,9% (95%CI 9,9%-11,9%), respetivamente. Foi encontrada uma associação forte entre asma e rinite a nível populacional, tanto nas crianças (odds-ratio (OR) 5,2, 95%CI 3,1-8,9), como nos idosos (OR variando de 8,3 95%CI 6,1-11,4 na rinite intermitente ligeira, a 39,9 95%CI 27,5-58,0 na rinite persistente moderada-grave). No estudo de coorte, a presença de atopia e de rinite em idade pré-escolar foram fatores de risco independentes para a persistência de asma na adolescência (OR 11,8 95%CI 4,0-34,6 e OR 10,4 95%CI 3,7-29,1, respetivamente). Foram identificados três fenótipos de sibilância em idade pré-escolar, que foram preditivos para a persistência de asma, uso de medicamentos de controlo e parâmetros funcionais respiratórios em idade escolar e na adolescência. A multimorbilidade, em particular a presença de rinite, com ou sem atopia, associou-se a um pior prognóstico. Na avaliação funcional nasal e pulmonar, observaram-se correlações entre os valores de PNIF pré e pós-vasoconstritor e do débito expiratório máximo instantâneo (PEF) e volume expiratório forçado no primeiro segundo (FEV1), pré e pós-broncodilatação, observado independentemente da presença de rinite e asma. O melhor modelo de regressão linear múltipla para o PNIF incluiu as variáveis PEF, idade e género. Em crianças com rinite alérgica e asma, não foi encontrada associação entre o PNIF e a pontuação no questionário CARATkids, exceto no que diz respeito à obstrução nasal auto-reportada. A análise metabolómica não dirigida em amostras de saliva e urina mostrou um subconjunto de áreas do espetro de NMR significativamente diferente nas crianças com rinite alérgica e asma, em comparação com crianças saudáveis. Alguns metabolitos que contribuíram para estas áreas do espetro foram identificados: arginina, taurina, citrato e aspartato (na saliva), e quinolinato, butirato, pantotenato, gluconato, pseudouridina e lisina (na urina). Observou-se uma correlação entre parâmetros espirométricos e a concentração urinária dos metabolitos quinolinato, butirato e pantotenato, enquanto a dos metabolitos quinolinato, gluconato e pseudouridina estava correlacionada com os níveis de óxido nítrico no ar exalado (FeNO). Observou-se uma associação entre a presença de sensibilização alergénica múltipla e as concentrações urinárias de quinolinato e salivares de citrato e aspartato. O perfil metabólico do EBC foi semelhante à composição espectral do ar ambiente. Discussão e Conclusões: Estes estudos epidemiológicos foram os primeiros de base populacional nacional que reportaram a prevalência de sintomas de asma em todas as idades pediátricas, bem como de sintomas de rinite, sua classificação e associação com asma em idosos. Os resultados reforçaram a asma como uma doença comum em crianças e em idosos, frequentemente associada a rinite. Em crianças com sibilância recorrente em idade pré-escolar, a presença de multimorbilidade, particularmente rinite com ou sem atopia associada, tende a prever um pior prognóstico no que respeita à persistência de asma e compromisso da função respiratória em idade escolar e na adolescência. Estes resultados apoiam a necessidade de uma abordagem integrada da rinite e da asma, desde idades precoces. Para a avaliação funcional respiratória global, o PNIF pode constituir uma medida objetiva complementar à avaliação subjetiva do controlo da rinite alérgica e da asma, em crianças em idade escolar. Os resultados sugerem que na interpretação dos valores do PNIF nesta faixa etária, os valores do PEF devem idealmente ser considerados, para além da idade e do género. A análise metabolómica exploratória de amostras de urina e saliva revelou subconjuntos de áreas do espectro de NMR associadas à rinite alérgica e asma em crianças, gerando novas hipóteses que necessitam de análises suplementares. Os resultados obtidos na análise do perfil metabólico do EBC reforçaram a importância do controlo do ar ambiente durante a colheita de amostras e a necessidade de procedimentos analíticos que permitam distinguir a presença de compostos exógenos nas amostras de EBC. Em resumo, os resultados apresentados nesta dissertação adicionam evidência para uma avaliação global integrada da asma em conjunto com a rinite, tanto na prática clínica, como na investigação. Prevemos que a avaliação funcional nasal possa ser generalizada na prática clínica, numa abordagem global funcional das vias aéreas. O conjunto de metabolitos identificados na análise exploratória metabolómica estimula a continuação dos estudos nesta área para validação dos resultados, seguida da identificação das moléculas/vias metabólicas responsáveis pelas diferenças encontradas, o seu papel na fisiopatologia da rinite alérgica e asma e, por fim, como potenciais (novos) alvos terapêuticos.ABSTRACT: Introduction and Aims: Worldwide and across all age groups, asthma affects the lives of several hundred million people. In spite of the advances over the last decades, asthma and its multimorbidity continue to impart a significant onus on individuals with the disease, their families and society and also on health economies. A high number of unmet needs remain to be resolved, related to gaps in current scientific knowledge covering many aspects of asthma, from epidemiology and pathophysiology to patient care. The main objective of this dissertation was to contribute to address some of these existing unmet needs in asthma and its link with rhinitis. In particular, the original work aimed to (1) estimate nationwide asthma prevalence and analyze its association with rhinitis in particularly vulnerable and internationally data-lacking population groups – the children and the elderly; (2) unveil features for an early recognition of asthma, identifying multidimensional “hypothesis-free” early childhood wheezing clinical phenotypes related to asthma persistence in adolescence; (3) analyze the association between nasal and lower airway function, together with the subjective evaluation of allergic rhinitis and asthma concurrent control in children; (4) explore innovative strategies to uncover “unbiased” differentiating metabolic features of childhood allergic rhinitis and asthma multimorbidity in non-invasively collected samples. Methods: This dissertation was based on three types of studies: 1. Cross-sectional, population-based, nationwide surveys of citizens living in Portugal, applied by interview using standardized procedures, to collect epidemiological data related to asthma and rhinitis and to analyze the association between these two conditions. For the pediatric study, data from all individuals aged below 18 years who participated in the INAsma study (population-based, all-age, nationwide telephone interview study to estimate asthma prevalence in Portugal) was analyzed. The elderly-targeted study was originally designed to estimate rhinitis prevalence in individuals aged 65 years or above living in mainland Portugal and the data was collected by direct face-to-face interview; 2. Prospective cohort study of children aged below 7 years with recurrent wheezing, systematically evaluated at specific time-points, up to 13 years of follow-up. Multivariable logistic regression models for persistent asthma in adolescence were developed based on questionnaires and skin prick tests data. Clinical phenotypes were identified by cluster analysis of variables selected with the logistic regression analysis, and compared for predicting asthma prevalence, use of control treatments and lung function in childhood and adolescence; 3. Cross-sectional, case-control study of school-aged children with allergic rhinitis and asthma multimorbidity and healthy children (matched for age and gender), evaluated with respect to: a. Respiratory functional laboratorial assessments, i.e., sequential assessments of peak nasal inspiratory flow (PNIF) before and after nasal decongestion and spirometry with bronchodilation test. The Control of Allergic Rhinitis and Asthma Test for children (CARATkids) was used for these diseases concurrent subjective control evaluation. Associations between objective and subjective scores were investigated by multiple linear regression models. b. Analytical laboratorial study using untargeted metabolomics analysis by nuclear magnetic resonance (NMR) spectroscopy of urine and saliva samples collected from each child. Spectroscopic and clinical data were subjected to statistical analysis including multivariable and univariable approaches. Additionally, exhaled breath condensate (EBC) samples were collected from volunteers, together with room air samples, which were analyzed by NMR spectroscopy. The resulting spectra were compared.Results: The estimated prevalence of current asthma in children was 8.4% (95% confidence interval (CI) 6.6%-10.7%). The prevalence of rhinitis and of physician-diagnosed asthma in the elderly were estimated to be 29.8% (95%CI 28.4%-31.3%) and 10.9% (95%CI 9.9%-11.9%), respectively. A strong association between asthma and rhinitis at the population-level was found both in children (odds-ratio (OR) 5.2, 95%CI 3.1-8.9) and in the elderly (OR varying from 8.3, 95%CI 6.1-11.4 in mild intermittent rhinitis to 39.9, 95%CI 27.5-58.0 in moderate-severe persistent rhinitis). In the cohort study, atopy and rhinitis at preschool-age were independent risk factors for asthma persistence in adolescence (OR 11.8, 95%CI 4.0-34.6, and OR 10.4, 95%CI 3.7-29.1, respectively). Three distinct early childhood wheezing phenotypes were identified, which were predictive of asthma persistence, use of control treatments and lung function in school-age and adolescence. Multimorbidity, particularly rhinitis, with or without associated atopy, tended to predict a worse prognosis. In the nasal and lung function study, baseline and decongested PNIF correlated with baseline and post-bronchodilation peak expiratory flow (PEF) and forced expiratory volume in one second (FEV1) in school-aged children, observed independently of rhinitis and asthma diagnosis. The best linear regression model for PNIF included the variables PEF, age and gender. In children with allergic rhinitis and asthma, no association was found between PNIF and CARATkids scores, except for nasal obstruction self-report. Untargeted metabolomics analysis of saliva and urine samples revealed a subset of the spectral areas significantly different in the children with allergic rhinitis and asthma, compared to healthy controls. Some metabolites contributing to these variables were identified: arginine, taurine, citrate and aspartate (in saliva), and quinolinate, butyrate, pantothenate, gluconate, pseudouridine and lysine (in urine). Urinary quinolinate, butyrate and pantothenate concentrations correlated with spirometric parameters, while quinolinate, gluconate and pseudouridine concentrations correlated with exhaled nitric oxide (FeNO) levels. Urinary quinolinate and salivary citrate and aspartate were associated with multiple allergenic sensitization. The EBC metabolic profile was found to be highly comparable to the ambient air spectral composition. Discussion and Conclusions: These were the first population-based nationwide epidemiologic studies reporting asthma symptoms prevalence among all pediatric ages, and rhinitis prevalence, its classification and association with asthma in the elderly. Our results further support that asthma is a common disease in children and the elderly, frequently associated with rhinitis. In early childhood, the presence of multimorbidity, particularly rhinitis with or without associated atopy, tended to predict a worse prognosis of recurrent wheezing regarding asthma persistence and impaired lung function in later childhood and adolescence. These results reinforce the need for a global, integrated care pathway in asthma and rhinitis, since early ages. In this integrated assessment, PNIF may provide complementary objective information to subjective concurrent control assessment of allergic rhinitis and asthma in school-aged children. The results suggested that PEF values should ideally be considered, besides age and gender, when interpreting PNIF values in this age group. Exploratory metabolomics revealed differentiating subsets of NMR spectral features in saliva and urine associated with allergic rhinitis and asthma multimorbidity in children, generating hypotheses to be further analyzed. The results obtained in the EBC metabolic profile analysis reinforced the importance of ambient air controls during samples collection and the need for analytical procedures to distinguish exogenously originated metabolites in EBC. In summary, the results presented in this dissertation added compelling information for an integrated, global assessment of asthma together with rhinitis, in clinical practice and in research. We foresee the clinical general application of nasal and lung function evaluation in a global airways assessment strategy. The differentiating subsets of metabolites found in exploratory metabolomics analysis stimulate further studies in order to validate our findings, followed by the identification of molecules/metabolic pathways involved, its role in allergic rhinitis and asthma pathophysiology and ultimately the potential as (novel) therapeutic targets

Trajectory and determinants of sexual well-being in the transition to parenthood: a couple-centered approach

Cerca de 90% dos casais numa relação de compromisso fará a transição para a parentalidade em algum momento da sua relação. A gravidez e o pós-parto são considerados períodos de vulnerabilidade para o bem-estar sexual dos casais, sendo frequente surgirem dificuldades sexuais (e.g., distress sexual, baixo desejo, dor genital) que podem iniciar-se na gravidez e persistir até 12 meses pós-parto. O bem-estar sexual é uma dimensão crítica para a qualidade e longevidade das relações de casal e está associado a melhores índices de saúde global e qualidade de vida. Contudo, o conhecimento acerca de fatores associados com o início e com a persistência de problemas sexuais perinatais é pouco robusto e, atualmente, não existem intervenções baseadas em evidência focadas no bem-estar sexual desta população. O presente trabalho visa compreender processos psicológicos intra- e interpessoais envolvidos no bem-estar sexual dos casais durante a transição para a parentalidade. Com base em modelos de ajustamento diádico e modelos cognitivo-afetivos de resposta sexual, apresentamos cinco estudos que investigam de que forma padrões de interdependência entre as experiências de casal contribuem para o bem-estar sexual durante este período. Recolhemos dados de amostras de casais durante a gravidez e após o parto (total n = 512 casais) usando desenhos diádicos transversais e longitudinais. Os resultados destes estudos indicam a presença de vários mecanismos interpessoais (e.g., padrões de [des]semelhança cognitivo-afetiva intradíade, bidirecionalidade entre dimensões de bem-estar sexual e relacional) que contributem para índices de bem-estar sexual (e.g., função sexual, satisfação sexual, distress sexual) durante a gravidez e no pós-parto, de forma transversal e longitudinal. Adicionalmente, identificamos classes de casais com trajetórias de bem-estar sexual distintas ao longo do tempo e examinamos o contributo de determinantes biológicos, psicológicos e relacionais para a pertença a estas trajetórias. Os resultados desta tese contribuem para uma maior compreensão dos processos interpessoais envolvidos no bem-estar sexual em períodos críticos de vida, bem como para o desenvolvimento de intervenções baseadas em evidência para promover o bem-estar sexual desta população

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