Fisiopatologia da Dor

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Lupus Eritematoso Sistémico

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Novos sistemas terapêuticos para administração nasal de fármacos

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Synthesis of new Aβ-ligands useful in diagnosis of Alzheimer's disease

Alzheimer’s disease is a chronic progressive neurodegenerative disease and is the most common form of dementia (estimated 50−60% of all cases), associated with loss of memory (in particular episodic memory), cognitive decline, and behavioural and physical disability, ultimately leading to death. Alzheimer’s disease is a complex disease, mostly occurring sporadically with no apparent inheritance and being the age the main risk factor. The production and accumulation of amyloid-beta peptide in the central nervous system is a key event in the development of Alzheimer’s disease. This project is devoted to the synthesis of amyloid-beta ligands, fluorophores and blood brain barrier-transporters for diagnosis and therapy of Alzheimer’s disease. Different amyloid-beta ligands will be synthesized and their ability to interact with amyloid-beta plaques will be studied with nuclear magnetic resonance techniques and a process of lead optimization will be performed. Many natural and synthetic compounds able to interact as amyloid-beta ligands have been identified. Among them, a set of small molecules in which aromatic moieties seem to play a key role to inhibit amyloid-beta aggregation, in particular heteroaromatic polycyclic compounds such as tetracyclines. Nevertheless tetracyclines suffer from chemical instability, low water solubility and possess, in this contest, undesired anti-bacterial activity. In order to overcome these limitations, one of our goals is to synthesize tetracyclines analogues bearing a polycyclic structure with improved chemical stability and water solubility, possibly lacking antibacterial activity but conserving the ability to interact with amyloid-beta peptides. Known tetracyclines have in common a fourth cycle without an aromatic character and with different functionalisations. We aim to synthesize derivatives in which this cycle is represented by a sugar moiety, thus bearing different derivatisable positions or create derivatives in which we will increase or decrease the number of fused rings. In order to generate a potential drug-tool candidate, these molecules should also possess the correct chemical-physical characteristics. The glycidic moiety, not being directly involved in the binding, it assures further possible derivatizations, such as conjugation to others molecular entities (nanoparticles, polymeric supports, etc.), and functionalization with chemical groups able to modulate the hydro/lipophilicity. In order to be useful such compounds should perform their action within the brain, therefore they have to be able to cross the blood brain barrier, and to be somehow detected for diagnostic purposes.

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.

The role of lymphotoxin-B receptor signaling in t-cell acute lymphoblastic leukemia development

Dissertação de mest., Ciências Biomédicas, Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2010

Differential regulation of hippocampal neurogenesis by nitric oxide following seizures

The fact that the adult brain is able to produce new neurons or glial cells from neural stem cells (NSC) became one of the most interesting and challenging fields of research in neuroscience. Endogenous adult neurogenesis occurs in two main regions of the brain: the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone (SGZ) in the dentate gyrus. Brain injury may be accompanied by increased neurogenesis, although neuroinflammation promotes the activation of microglial cells that can be detrimental to the neurogenic process. Nitric oxide (NO) is one of the factors released by microglia that can be proneurogenic. The mechanism by which NO promotes the proliferation of NSCs has been intensively studied. However, little is known about the role of NO in migration, survival and differentiation of the newborn cells. The aim of this work was to investigate the role of NO from inflammatory origin in proliferation, migration, differentiation and survival of NSCs from the dentate gyrus in a mouse model of status epilepticus. We also assessed neuroinflammation in the same injury model. Our work showed that NO increased proliferation of the early-born cells after seizures, but is detrimental for their survival. NO also increased migration of neuroblasts. Moreover, NO was important to maintain long-term neuroinflammation. Taken together, these results show that NO may be a good target to promote proliferation and migration of NSCs following seizures, but compromises survival of early-born cells.

Captura-recaptura como método epidemiológico a aplicarà esclerose múltipla

Tese de Doutoramento, Neurologia, Faculdade de Medicina, Universidade de Lisboa, 2014

Exploring the modulatory role of A2A receptors in oligodendrogenesis derived from neural stem/progenitor cells of the subventricular zone

Tese de mestrado, Neurociências, Faculdade de Medicina, Universidade de Lisboa, 2015

Characterization of adenosinergic system in Rett syndrome

Tese de mestrado, Neurociências, Faculdade de Medicina, Universidade de Lisboa, 2015

Development of a new cellular model for the visualization and study of GFAP oligomerization in living cells

Tese de mestrado, Neurociências, Faculdade de Medicina, Universidade de Lisboa, 2016

Deciphering the mechanisms underlying the loss of BDNF neuroprotection in an Alzheimer’s disease model

Tese de mestrado, Neurociências, Faculdade de Medicina, Universidade de Lisboa, 2016

Programa de intervenção segundo o modelo de auto-regulação na esclerose múltipla

A esclerose múltipla é um doença crónica do sistema nervoso central, que afecta mais frequentemente adultos jovens, no auge da sua carreira profissional e desenvolvimento pessoal, sem cura e de causas desconhecidas. Os sintomas e sinais mais comuns são a fadiga, fraqueza muscular, alterações da sensibilidade, ataxia, alterações do equilíbrio, dificuldades na marcha, dificuldades de memória, alterações cognitivas e dificuldades na resolução de problemas. A esclerose múltipla é uma doença progressiva e imprevisível, resultando, nalguns casos, em incapacidades e limitações de actividade de vida diária, causando danos irreparáveis para os indivíduos. Esta doença pode surgir através de surtos ou de uma forma progressiva.

Variabilidade dos limites de estabilidade na posição ortostática em utentes com Acidente Vascular Encefálico

Introdução: O controlo postural (CP), intimamente ligado aos limites de estabilidade, é fundamental nas atividades da vida diária e encontra-se comprometido após Acidente Vascular Encefálico (AVE). Objectivo: Descrever as alterações nos limites de estabilidade, na posição ortostática, em utentes com AVE, face à aplicação de um programa de intervenção baseado num processo de raciocínio clínico. Métodos: Estudo de série de casos, de três indivíduos com alterações do CP do tronco e/ou das grandes articulações, decorrentes de AVE, mas capazes de assumir e manter a posição ortostática. Para avaliar os limites de estabilidade foram utilizados a plataforma de pressões Emed (coordenadas do centro de pressão, COPx e COPy) e o MultidirectionalReachTest (MDRT). Foram também aplicadas, a Fugl-Meyer Assessmentof Sensoriomotor Recovery After Stroke (FMA) e a Berg Balance Scale (BBS). Os instrumentos de avaliação foram aplicados antes e após a intervenção (M0 e M1), de 8 semanas, baseada nos princípios do conceito de Bobath. Resultados: Os valores dos testes de alcance no MDRT aumentaram em todos os utentes. Os limites e os deslocamentos médio-lateral (COPx) e, ântero-posterior (COPy) dos valores do CPr aumentaram, entre M0 e M1. No que se refere à FMA e a BBS, verificou-se que todos apresentaram um aumento dos scores. Conclusão: Os utentes modificaram os seus limites de estabilidade no sentido do aumento, assim como, da função motora e do equilíbrio. Deste modo, a intervenção de acordo com os princípios do conceito de Bobath aparenta introduzir os estímulos necessários à reorganização funcional do sistema nervoso central lesado.

Reorganização do controlo postural do tronco e a sua influência na distribuição de carga na posição ortostática em indivíduos com Acidente Vascular Encefálico

Introdução: O Acidente Vascular Encefálico (AVE) consiste numa das primeiras causas de mortalidade e morbilidade em Portugal. Esta lesão do Sistema Nervoso Central (SNC) desencadeia alterações ao nível do controlo postural (CP), que interferem com a recuperação funcional dos indivíduos. Objetivo: Deste modo, torna-se premente descrever as alterações do CP do tronco através da análise dos alinhamentos dos segmentos corporais do tronco no grupo de indivíduos selecionados, face à aplicação de um programa de intervenção baseado nos princípios do Conceito de Bobath. Metodologia: Estudo de série de casos, em seis indivíduos com alterações neuromotoras decorrentes de AVE, os quais foram avaliados antes e após o plano de intervenção segundo a abordagem baseada nos princípios do Conceito de Bobath, através do registo observacional, da Classificação Internacional de Funcionalidade Incapacidade e Saúde (CIF), da utilização do Software de Avaliação Postural (SAPO) e da Plataforma de Pressões da Emed (PPE), modelo AT. Os dados recolhidos foram trabalhados em função do valor médio através do software Excel. Resultados: A análise do SAPO, na posição ortostática observam-se mudanças quer na vista posterior quer nas laterais, indicando uma maior simetria entre hemitroncos, e mudanças nos alinhamentos verticais indicando uma maior aproximação dos 180º. Na PPE observam-se os valores da área plantar, da pressão plantar média e do centro de pressão, tendem globalmente a uma maior semelhança e simetria. Quanto à CIF também se verificou uma diminuição da restrição na participação e limitação na atividade. Conclusão: A intervenção baseada no processo de raciocínio clínico aparenta introduzir os estímulos necessários à reorganização funcional do SNC lesado, produzindo melhorias ao nível dos alinhamentos dos segmentos corporais e desta forma melhorar a atividade muscular.