(Re)configuração das práticas educativas na prevenção do stresse na infância

O presente trabalho resulta do estudo que procurou analisar a reconfiguração das práticas educativas na prevenção do stresse na infância. O principal objetivo foi o de analisar de que forma os educadores podem desenvolver práticas educativas, estruturadoras e estruturantes do bem-estar da criança, investindo intencionalmente no desenvolvimento de atividades educativas, de modo a prevenir os índices de stresse das crianças, na educação pré-escolar. Foram realizados cinco estudos complementares, de natureza quantitativa e qualitativa. O estudo 1: contextualização e caracterização de indicadores sociofamiliares de crianças que frequentam a educação pré-escolar; Estudo 2: adaptação/avaliação do PKBS-2 de Merrell, para identificação das aptidões sociais e os problemas de comportamento em crianças dos 3 aos 6 anos numa amostra portuguesa (N=150) e estudos comparativos Portugal/Brasil (N=300) e Portugal/Cabo-Verde (N=150); Estudo 3: validação da Escala Comportamental para crianças em idade Pré-Escolar – PKBSpt, versão portuguesa do PKBS-2 em crianças dos 2 aos 7 anos (N=581); Estudo 4: contributos para a reconfiguração das práticas pedagógicas e o stresse na infância sob o olhar dos educadores. No âmbito deste estudo 4, desenvolvemos e validamos o Protocolo para a Avaliação do Stresse na Infância–PASI, constituído por três subescalas que aplicamos a educadores ou equiparados (N=188): ESISI, EPELSI e ECPLSI; Estudo 5: análise das competências dos profissionais da educação ao nível da ansiedade (IAB de Beck) e estratégias de coping; (EC de Gomes & Pereira). Para análise estatística dos dados utilizamos o programa SPSS e o Excel. Os resultados evidenciaram a validade e fidelidade dos instrumentos: ESISI, EPELSI, ECPLSI e PKBSpt, bem como a validação para a língua portuguesa deste último. A variável género apresentou diferenças estatisticamente significativas nas aptidões sociais e problemas de comportamento, no entanto não estão correlacionadas com os problemas de ansiedade. Os Educadores focalizam a sua praxis educativa na observação, identificação e definição de estratégias orientadas para o stresse na infância e, ainda, na cooperação escola/família e na prevenção das situações indutoras de stresse. Contudo apontam algumas fragilidades na formação básica e contínua, para lidarem com o stresse na primeira infância. As causas psicossociais são preditoras das situações indutoras de stresse na infância e são explicadas pelas causas externas de componente escolar, ou seja, as perturbações de ansiedade na infância poderão ter repercussões no contexto escolar. Importa que a comunidade educativa perspetive particular atenção às crianças que estão em situação de maior vulnerabilidade, de modo a prevenir e a intervir nas situações de stresse na educação pré-escolar. São referidas implicações psicopedagógicas deste estudo.

Pregnancy and newborns disorders followed by urine metabolomics

Chapter 1 introduces the scope of the work by identifying the clinically relevant prenatal disorders and presently available diagnostic methods. The methodology followed in this work is presented, along with a brief account of the principles of the analytical and statistical tools employed. A thorough description of the state of the art of metabolomics in prenatal research concludes the chapter, highlighting the merit of this novel strategy to identify robust disease biomarkers. The scarce use of maternal and newborn urine in previous reports enlightens the relevance of this work. Chapter 2 presents a description of all the experimental details involved in the work performed, comprising sampling, sample collection and preparation issues, data acquisition protocols and data analysis procedures. The proton Nuclear Magnetic Resonance (NMR) characterization of maternal urine composition in healthy pregnancies is presented in Chapter 3. The urinary metabolic profile characteristic of each pregnancy trimester was defined and a 21-metabolite signature found descriptive of the metabolic adaptations occurring throughout pregnancy. 8 metabolites were found, for the first time to our knowledge, to vary in connection to pregnancy, while known metabolic effects were confirmed. This chapter includes a study of the effects of non-fasting (used in this work) as a possible confounder. Chapter 4 describes the metabolomic study of 2nd trimester maternal urine for the diagnosis of fetal disorders and prediction of later-developing complications. This was achieved by applying a novel variable selection method developed in the context of this work. It was found that fetal malformations (FM) (and, specifically those of the central nervous system, CNS) and chromosomal disorders (CD) (and, specifically, trisomy 21, T21) are accompanied by changes in energy, amino acids, lipids and nucleotides metabolic pathways, with CD causing a further deregulation in sugars metabolism, urea cycle and/or creatinine biosynthesis. Multivariate analysis models´ validation revealed classification rates (CR) of 84% for FM (87%, CNS) and 85% for CD (94%, T21). For later-diagnosed preterm delivery (PTD), preeclampsia (PE) and intrauterine growth restriction (IUGR), it is found that urinary NMR profiles have early predictive value, with CRs ranging from 84% for PTD (11-20 gestational weeks, g.w., prior to diagnosis), 94% for PE (18-24 g.w. pre-diagnosis) and 94% for IUGR (2-22 g.w. pre-diagnosis). This chapter includes results obtained for an ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) study of pre-PTD samples and correlation with NMR data. One possible marker was detected, although its identification was not possible. Chapter 5 relates to the NMR metabolomic study of gestational diabetes mellitus (GDM), establishing a potentially predictive urinary metabolic profile for GDM, 2-21 g.w. prior to diagnosis (CR 83%). Furthermore, the NMR spectrum was shown to carry information on individual phenotypes, able to predict future insulin treatment requirement (CR 94%). Chapter 6 describes results that demonstrate the impact of delivery mode (CR 88%) and gender (CR 76%) on newborn urinary profile. It was also found that newborn prematurity, respiratory depression, large for gestational age growth and malformations induce relevant metabolic perturbations (CR 82-92%), as well as maternal conditions, namely GDM (CR 82%) and maternal psychiatric disorders (CR 91%). Finally, the main conclusions of this thesis are presented in Chapter 7, highlighting the value of maternal or newborn urine metabolomics for pregnancy monitoring and disease prediction, towards the development of new early and non-invasive diagnostic methods.

Mitochondrial dysfunction in fatty acid β-oxidation disorders

Mitochondria are central organelles for cell survival with particular relevance in energy production and signalling, being mitochondrial fatty acid β–oxidation (FAO) one of the metabolic pathways harboured in this organelle. FAO disorders (FAOD) are among the most well studied inborn errors of metabolism, mainly due to their impact in health. Nevertheless, some questions remain unsolved, as their prevalence in certain European regions and how pathophysiological determinants combine towards the phenotype. Analysis of data from newborn screening programs from Portugal and Spain allowed the estimation of the birth prevalence of FAOD revealing that this group of disorders presents in Iberia (and particularly in Portugal) one of the highest European birth prevalence, mainly due to the high birth prevalence of medium chain acyl-CoA dehydrogenase deficiency. These results highlight the impact of this group of genetic disorders in this European region. The characterization of mitochondrial proteome, from patients fibroblasts with FAOD, namely multiple acyl-CoA dehydrogenase deficiency (MADD) and long chain acyl-CoA dehydrogenase deficiency (LCHADD), provided a global perspective of the mitochondrial proteome plasticity in these disorders and highlights the main molecular pathways involved in their pathogenesis. Severe MADD forms show an overexpression of chaperones, antioxidant enzymes (MnSOD), and apoptotic proteins. An overexpression of glycolytic enzymes, which reflects cellular adaptation to energy deficiency due to FAO blockage, was also observed. When LCHADD fibroblasts were analysed a metabolic switching to glycolysis was also observed with overexpression of apoptotic proteins and modulation of the antioxidant defence system. Severe LCHADD present increased ROS alongside with up regulation of MnSOD while moderate forms have lower ROS and down-regulation of MnSOD. This probably reflects the role of MnSOD in buffering cellular ROS, maintain them at levels that allow cells to avoid damage and start a cellular response towards survival. When ROS levels are very high cells have to overexpress MnSOD for detoxifying proposes. When severe forms of MADD were compared to moderate forms no major differences were noticed, most probably because ROS levels in moderate MADD are high enough to trigger a response similar to that observed in severe forms. Our data highlights, for the first time, the differences in the modulation of antioxidant defence among FAOD spectrum. Overall, the data reveals the main pathways modulated in FAOD and the importance of ROS levels and antioxidant defence system modulation for disease severity. These results highlight the complex interaction between phenotypic determinants in FAOD that include genetic, epigenetic and environmental factors. The development of future better treatment approaches is dependent on the knowledge on how all these determinants interact towards phenotype.!