Barnacle Allergy: Allergen Characterization and Cross-Reactivity with Mites

Background: Barnacles are a type of seafood with worldwide distribution and abundant along the shores of temperate seas. They are particularly appreciated and regularly consumed in Portugal as well as in Spain, France and South America, but barnacle allergy is a rare condition of which there is only one reference in the indexed literature. The molecular allergens and possible cross-reactivity phenomena implicated (namely with mites) have not been established. Objective: To demonstrate the IgE-mediated allergy to barnacle and to identify the proteins implicated as well as possible cross-reactivity phenomena with mites. Methods: We report the clinical and laboratory data of five patients with documented IgE-mediated allergy to barnacle. The diagnosis was based on a suggestive clinical history combined with positive skin prick tests (SPT) to barnacle – prick to prick method. Two barnacle extracts were prepared (raw and cooked barnacle) and sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) and IgE-immunoblotting were performed. An immunoblotting inhibition assay with Dermatophagoides pteronyssinus was also done in order to evaluate cross-reactivity. Results: All patients had mite-related asthma and the allergic rhinoconjunctivitis; they all experienced mucocutaneous symptoms. All of them had positive SPT to barnacle, and the immunoblotting showed several allergenic fractions with a wide molecular weight range (19 – 94 kDa). The D. pteronyssinus extract inhibited several IgE-binding protein fractions in the barnacle extract. Conclusions: We describe five patients with IgE-mediated barnacle allergy. We also describe a group of IgEbinding+ proteins between 30 and 75 kDa as the allergenic fractions of this type of Crustacea. Cross-reactivity with D. pteronyssinus was demonstrated in two cases.

Adhesive selection for single lap bonded joints: Experimentation and advanced techniques for strength prediction

The integrity of multi-component structures is usually determined by their unions. Adhesive-bonding is often used over traditional methods because of the reduction of stress concentrations, reduced weight penalty, and easy manufacturing. Commercial adhesives range from strong and brittle (e.g., Araldite® AV138) to less strong and ductile (e.g., Araldite® 2015). A new family of polyurethane adhesives combines high strength and ductility (e.g., Sikaforce® 7888). In this work, the performance of the three above-mentioned adhesives was tested in single lap joints with varying values of overlap length (LO). The experimental work carried out is accompanied by a detailed numerical analysis by finite elements, either based on cohesive zone models (CZM) or the extended finite element method (XFEM). This procedure enabled detailing the performance of these predictive techniques applied to bonded joints. Moreover, it was possible to evaluate which family of adhesives is more suited for each joint geometry. CZM revealed to be highly accurate, except for largely ductile adhesives, although this could be circumvented with a different cohesive law. XFEM is not the most suited technique for mixed-mode damage growth, but a rough prediction was achieved.

Tensile fracture characterization of adhesive joints by standard and optical techniques

The use of adhesive joints has increased in recent decades due to its competitive features compared with traditional methods. This work aims to estimate the tensile critical strain energy release rate (GIC) of adhesive joints by the Double-Cantilever Beam (DCB) test. The J-integral is used since it enables obtaining the tensile Cohesive Zone Model (CZM) law. An optical measuring method was developed for assessing the crack tip opening (δn) and adherends rotation (θo). The proposed CZM laws were best approximated by a triangular shape for the brittle adhesive and a trapezoidal shape for the two ductile adhesives.

Characterization of an antioxidant surfactant-free topical formulation containing Castanea sativa leaf extract

Context: Inclusion of antioxidants in topical formulations can contribute to minimize oxidative stress in the skin, which has been associated with photoaging, several dermatosis and cancer. Objective: A Castanea sativa leaf extract with established antioxidant activity was incorporated into a semisolid surfactant-free formulation. The objective of this study was to perform a comprehensive characterization of this formulation. Materials and methods: Physical, microbiological and functional stability were evaluated during 6 months storage at 20 °C and 40 °C. Microstructure elucidation (cryo-SEM), in vitro release and in vivo moisturizing effect (Corneometer® CM 825) were also assessed. Results and discussion: Minor changes were observed in the textural and rheological properties of the formulation when stored at 20 °C for 6 months and the antioxidant activity of the plant extract remained constant throughout the storage period. Microbiological quality was confirmed at the end of the study. Under accelerated conditions, higher modifications of the evaluated parameters were observed. Cryo-SEM analysis revealed the presence of oil droplets dispersed into a gelified external phase. The release rate of the antioxidant compounds (610 ± 70 µgh−0.5) followed Higuchi model. A significant in vivo moisturizing effect was demonstrated, that lasted at least 4 h after product’s application. Conclusion: The physical, functional and microbiological stability of the antioxidant formulation was established. Specific storage conditions should be recommended considering the influence of temperature on the stability. A skin hydration effect and good skin tolerance were also found which suggests that this preparation can be useful in the prevention or treatment of oxidative stress-mediated dysfunctions.

Comparing the performance of geostatistical models with additional information from covariates for sewage plume characterization

In this work, kriging with covariates is used to model and map the spatial distribution of salinity measurements gathered by an autonomous underwater vehicle in a sea outfall monitoring campaign aiming to distinguish the effluent plume from the receiving waters and characterize its spatial variability in the vicinity of the discharge. Four different geostatistical linear models for salinity were assumed, where the distance to diffuser, the west-east positioning, and the south-north positioning were used as covariates. Sample variograms were fitted by the Mat`ern models using weighted least squares and maximum likelihood estimation methods as a way to detect eventual discrepancies. Typically, the maximum likelihood method estimated very low ranges which have limited the kriging process. So, at least for these data sets, weighted least squares showed to be the most appropriate estimation method for variogram fitting. The kriged maps show clearly the spatial variation of salinity, and it is possible to identify the effluent plume in the area studied. The results obtained show some guidelines for sewage monitoring if a geostatistical analysis of the data is in mind. It is important to treat properly the existence of anomalous values and to adopt a sampling strategy that includes transects parallel and perpendicular to the effluent dispersion.

Ankylosing spondylitis: genomic and functional characterization of candidate genes and their repercussion in clinical practice

RESUMO: Introdução: A espondilite anquilosante (EA) é uma doença inflamatória crónica caracterizada pela inflamação das articulações sacroilíacas e da coluna. A anquilose progressiva motiva uma deterioração gradual da função física e da qualidade de vida. O diagnóstico e o tratamento precoces podem contribuir para um melhor prognóstico. Neste contexto, a identificação de biomarcadores, assume-se como sendo muito útil para a prática clínica e representa hoje um grande desafio para a comunidade científica. Objetivos: Este estudo teve como objetivos: 1 - caracterizar a EA em Portugal; 2 - investigar possíveis associações entre genes, MHC e não-MHC, com a suscetibilidade e as características fenotípicas da EA; 3 - identificar genes candidatos associados a EA através da tecnologia de microarray. Material e Métodos: Foram recrutados doentes com EA, de acordo com os critérios modificados de Nova Iorque, nas consultas de Reumatologia dos diferentes hospitais participantes. Colecionaram-se dados demográficos, clínicos e radiológicos e colhidas amostras de sangue periférico. Selecionaram-se de forma aleatória, doentes HLA-B27 positivos, os quais foram tipados em termos de HLA classe I e II por PCR-rSSOP. Os haplótipos HLA estendidos foram estimados pelo algoritmo Expectation Maximization com recurso ao software Arlequin v3.11. As variantes alélicas dos genes IL23R, ERAP1 e ANKH foram estudadas através de ensaios de discriminação alélica TaqMan. A análise de associação foi realizada utilizando testes da Cochrane-Armitage e de regressão linear, tal como implementado pelo PLINK, para variáveis qualitativas e quantitativas, respetivamente. O estudo de expressão génica foi realizado por Illumina HT-12 Whole-Genome Expression BeadChips. Os genes candidatos foram validados usando qPCR-based TaqMan Low Density Arrays (TLDAs). Resultados: Foram incluídos 369 doentes (62,3% do sexo masculino, com idade média de 45,4 ± 13,2 anos, duração média da doença de 11,4 ± 10,5 anos). No momento da avaliação, 49,9% tinham doença axial, 2,4% periférica, 40,9% mista e 7,1% entesopática. A uveíte anterior aguda (33,6%) foi a manifestação extra-articular mais comum. Foram positivos para o HLA-B27, 80,3% dos doentes. Os haplótipo A*02/B*27/Cw*02/DRB1*01/DQB1*05 parece conferir suscetibilidade para a EA, e o A*02/B*27/Cw*01/DRB1*08/DQB1*04 parece conferir proteção em termos de atividade, repercussão funcional e radiológica da doença. Três variantes (2 para IL23R e 1 para ERAP1) mostraram significativa associação com a doença, confirmando a associação destes genes com a EA na população Portuguesa. O mesmo não se verificou com as variantes estudadas do ANKH. Não se verificou associação entre as variantes génicas não-MHC e as manifestações clínicas da EA. Foi identificado um perfil de expressão génica para a EA, tendo sido validados catorze genes - alguns têm um papel bem documentado em termos de inflamação, outros no metabolismo da cartilagem e do osso. Conclusões: Foi estabelecido um perfil demográfico e clínico dos doentes com EA em Portugal. A identificação de variantes génicas e de um perfil de expressão contribuem para uma melhor compreensão da sua fisiopatologia e podem ser úteis para estabelecer modelos com relevância em termos de diagnóstico, prognóstico e orientação terapêutica dos doentes. -----------ABSTRACT: Background: Ankylosing Spondylitis (AS) is a chronic inflammatory disorder characterized by inflammation in the spine and sacroiliac joints leading to progressive joint ankylosis and in progressive deterioration of physical function and quality of life. An early diagnosis and early therapy may contribute to a better prognosis. The identification of biomarkers would be helpful and represents a great challenge for the scientific community. Objectives: The present study had the following aims: 1- to characterize the pattern of AS in Portuguese patients; 2- to investigate MHC and non-MHC gene associations with susceptibility and phenotypic features of AS and; 3- to identify candidate genes associated with AS by means of whole-genome microarray. Material and Methods: AS was defined in accordance to the modified New York criteria and AS cases were recruited from hospital outcares patient clinics. Demographic and clinical data were recorded and blood samples collected. A random group of HLA-B27 positive patients and controls were selected and typed for HLA class I and II by PCR-rSSOP. The extended HLA haplotypes were estimated by Expectation Maximization Algorithm using Arlequin v3.11 software. Genotyping of IL23R, ERAP1 and ANKH allelic variants was carried out with TaqMan allelic discrimination assays. Association analysis was performed using the Cochrane-Armitage and linear regression tests as implemented in PLINK, for dichotomous and quantitative variables, respectively. Gene expression profile was carried out using Illumina HT-12 Whole-Genome Expression BeadChips and candidate genes were validated using qPCR-based TaqMan Low Density Arrays (TLDAs). Results: A total of 369 patients (62.3% male; mean age 45.4±13.2 years; mean disease duration 11.4±10.5 years), were included. Regarding clinical disease pattern, at the time of assessment, 49.9% had axial disease, 2.4% peripheral disease, 40.9% mixed disease and 7.1% isolated enthesopathic disease. Acute anterior uveitis (33.6%) was the most common extra-articular manifestation. 80.3% of AS patients were HLA-B27 positive. The haplotype A*02/B*27/Cw*02/DRB1*01/DQB1*05 seems to confer susceptibility to AS, whereas A*02/B*27/Cw*01/DRB1*08/DQB1*04 seems to provide protection in terms of disease activity, functional and radiological repercussion. Three markers (two for IL23R and one for ERAP1) showed significant single-locus disease associations. Association of these genes with AS in the Portuguese population was confirmed, whereas ANKH markers studied did not show an association with AS. No association was seen between non-MHC genes and clinical manifestations of AS. A gene expression signature for AS was established; among the fourteen validated genes, a number of them have a well-documented inflammatory role or in modulation of cartilage and bone metabolism. Conclusions: A demographic and clinical profile of patients with AS in Portugal was established. Identification of genetic variants of target genes as well as gene expression signatures could provide a better understanding of AS pathophysiology and could be useful to establish models with relevance in terms of susceptibility, prognosis, and potential therapeutic guidance.

Avaliação de Segurança de um Edifício Antigo na Baixa do Porto

Com o presente trabalho pretende-se avaliar a segurança de um elemento estrutural de um edifício antigo (tradicional) da baixa da Porto edificado nos finais do última quartel do século XVIII. Após uma pequena introdução onde será exposta a estrutura do trabalho, capítulo a capítulo, será feita uma explanação histórica sobre o meio urbano onde o edifício está construído, os métodos construtivos adotados desde 1755 até aos nossos dias, passando pelo historial do próprio edificado. Passar-se-á de seguida à análise do edifício, apresentando evidências do que eventualmente terá sido construído de base até aos dias de hoje. Será feita uma apresentação dos elementos construtivos que compõe actualmente o edificio, apresentando evidências do estado de cada um, ou seja, será feita um levantamento das patologias evidentes e visíveis a olho nu, recorrendo a métodos expeditos de análise. Também será feito um levantamento dimensional da volumetria , planimétria do edifício e da compartimentação, registando as dimensões dos elementos estruturais e arquitectónicos relevantes para a realização dos respectivos projectos. Posteriormente passar-se-á para uma exposição dos diversos tipos de ensaios, começando pela apresentação dos métodos disponíveis atualmente para a caracterização física e mecânica dos materiais, seguida da realização dos ensaios, recolha de dados no edificado e proposta de definição dos materiais. Após a recolha de todos estes elementos, estão reunidas as condições para se passar a uma análise estrutural, onde para estes objectivos, serão aplicados os mais avançados programas de cálculo estrutural existentes na actualidade, suportados em metodologias que analisam não só o elemento mas também a sua interação com os que lhe servem de fronteira acabando-se num todo que é o edifício. Pretende-se com este estudo estrutural avaliar a segurança de uma parte significativa e representativa do edifico baseado na regulamentação em vigor. A parte a analisar representa o pavimento aplicado na construção. Finalizar-se-á com a apresentação das conclusões, propondo eventuais ações futuras.

Algorithm for the Parkinson’s disease behavioural models characterization using a biosensor

Dissertação para obtenção do Grau de Mestre em Engenharia Biomédica

GENOTYPE CHARACTERIZATION OF Leishmania (Viannia) braziliensis ISOLATED FROM HUMAN AND CANINE BIOPSIES WITH AMERICAN CUTANEOUS LEISHMANIASIS

Introduction:American tegumentary leishmaniasis (ATL) can be caused by Leishmania (Viannia) braziliensis complex. The evolution of ATL initially results in lesions and can develop into disseminated or diffuse forms. The genetic diversity of L. (V.) braziliensis in some endemic areas of Brazil has been poorly studied, such as in the state of São Paulo. This study analyzed the genetic diversity of L. (V.) braziliensis isolates collected from patients and dogs with LTA from the state of São Paulo.Methods:Leishmaniasis diagnosis was determined by PCR. The 132 biopsies were collected in different regions of Sao Paulo State, Brazil (36 municipalities). The genetic characterization of L. (V.) braziliensis isolates was tested by RFLP-PCR using DNA extracted from biopsies. The primer set amplified a specific region of Leishmania internal transcribed spacers of the ribosomal DNA locus.Results:Of the 132 samples, 52 (40%) were completely genotyped by RFLP-PCR (44 from human patients and eight from dogs). The results showed nine distinct patterns. The majority of the genotyped samples were from Sorocaba (30), and the others were distributed among 14 other municipalities. The first pattern was more frequent (29 samples), followed by pattern 2 (nine samples) and pattern 3 (three samples). Patterns 4, 6, 7, 8 and 9 were composed of two samples each and pattern 5 of one sample.Conclusion:These results suggest that polymorphic strains of L. (V.) braziliensis circulate in the state of São Paulo. These data agree with studies from other regions of Brazil, showing great variability among the natural populations of endemic foci.

Optical clearing mechanisms characterization in muscle

Optical immersion clearing is a technique that has been widely studied for more than two decades and that is used to originate a temporary transparency effect in biological tissues. If applied in cooperation with clinical methods it provides optimization of diagnosis and treatment procedures. This technique turns biological tissues more transparent through two main mechanisms — tissue dehydration and refractive index (RI) matching between tissue components. Such matching is obtained by partial replacement of interstitial water by a biocompatible agent that presents higher RI and it can be completely reversible by natural rehydration in vivo or by assisted rehydration in ex vivo tissues. Experimental data to characterize and discriminate between the two mechanisms and to find new ones are necessary. Using a simple method, based on collimated transmittance and thickness measurements made from muscle samples under treatment, we have estimated the diffusion properties of glucose, ethylene glycol (EG) and water that were used to perform such characterization and discrimination. Comparing these properties with data from literature that characterize their diffusion in water we have observed that muscle cell membrane permeability limits agent and water diffusion in the muscle. The same experimental data has allowed to calculate the optical clearing (OC) efficiency and make an interpretation of the internal changes that occurred in muscle during the treatments. The same methodology can now be used to perform similar studies with other agents and in other tissues in order to solve engineering problems at design of inexpensive and robust technologies for a considerable improvement of optical tomographic techniques with better contrast and in-depth imaging.

Advanced instrumentation for superheated liquid detectors in dark matter searches

Dissertação para obtenção do Grau de Doutor em Engenharia Física

Synthesis, characterization and applications of new schiff base fluorescent chemosensors for metal and DNA interactions: conventional and "green" approaches

Dissertação apresentada para a obtenção do Grau de Doutor em Química Sustentável, especialidade de Química-Física Inorgânica, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Establishment and characterization of a human model of the blood-brain barrier

Dissertação apresentada para a obtenção do Grau de Mestre em Genética Molecular e Biomedicina, pela Universidade N ova de Lisboa, Faculdade de Ciências e Tecnologia

Avaliação clínica na disfunção fronto-estriatal : movimento e cognição

RESUMO: Os circuitos fronto-estriatais constituem um sistema em ansa fechada que une diversas regiões do lobo frontal aos gânglios da base, participando, com outras áreas cerebrais, no controlo do movimento, cognição e comportamento. As Distonias Primárias, a Doença de Parkinson e a Hidrocefalia de Pressão Normal, são doenças do movimento caracterizadas por disfunção do circuito fronto-estriatal motor. A conectividade funcional entre as diversas ansas do sistema fronto-estriatal, permite prever que as doenças do movimento possam também acompanhar-se de sintomas da esfera cognitiva e comportamental, cuja avaliação seria importante no manejo diagnóstico e terapêutico dos doentes. Objectivos Os nossos objectivos foram avaliar, por estudos clínicos, a relação entre sintomas motores, cognitivos e comportamentais em três doenças do movimento com fisiopatologias diversas - distonias Primárias, Doença de Parkinson e Hidrocefalia de Pressão Normal - analisando os dados sob a perspectiva teórica fornecida pelo conhecimentos dos vários circuitos frontoestriatais. Os nossos objectivos específicos para cada doença foram: a) Distonias Primárias: avaliação de disfunção executiva em doentes com Distonia Primária e relação com a gravidade dos sintomas motores b) Doença de Parkinson: 1. avaliação breve das funções mentais nas fases iniciais da doença, incluindo análise longitudinal para determinação de factores preditivos para declínio cognitivo; 2. relação entre a função motora e cognitiva e a Perturbação do Comportamento do sono REM, incluindo análise longitudinal; 3.avaliação de sintomas psiquiátricos, de um ponto de vista global e especificamente com incidência sobre as Perturbações do Controlo do Impulso (PCI). c) Hidrocefalia de Pressão Normal: 1. caracterização das alterações da marcha, incluindo comparação com a Doença de Parkinson; 2. caracterização das alterações cognitivas e da relação entre estas e a disfunção da marcha; 3. estudo evolutivo das alterações da marcha e cognitiva em doentes submetido a cirurgia e doentes não submetidos a cirurgia. Métodos: A Distonia Primária, a Doença de Parkinson e a Hidrocefalia de Pressão Normal foram diagnosticadas segundo critérios clínicos validados. Sempre que justificado, foram recrutados grupos de controlo, com indivíduos sem doença, emparelhados para idade, sexo e grau de escolaridade. Os doentes foram avaliados com instrumentos de aplicação clinica directa, incluindo escalas de função motora, testes neuropsicológicos globais e dirigidos às funções executivas e escalas de avaliação psiquiátrica. Testes aplicados nas Distonias Primárias: Unified Dystonia Rating Scale, Wisconsin Card Sorting Test, teste de Stroop, teste de cubos da WAIS, Teste de Retenção Visual de Benton; na Doença de Parkinson: Unified Parkinson's Disease Rating Scale, Frontal Assessment Battery (FAB), Mini-Mental State Examination (MMSE), REM-sleep behaviour disorder Questionnaire; Symptom Chek-list 90-R, Brief Psychiatric Rating Scale, FAS (fluência verbal lexical) Nomeação de Animais (Fluência verbal semântica), prova de repetição de dígitos (WAIS), Rey auditory verbal learning test, teste de Stroop, matrizes progressivas de Raven, Questionnaire for Impulsive-Compulsive Disorders; na HPN: prova cronometrada de marcha,MMSE, prova de memória imediata da WAIS, prova de repetição de dígitos (WAIS), FAB, desenho complexo de Rey, teste de Stroop, cancelamento de letras, teste Grooved Pegboard. Os doentes com HPN foram também submetidos a estudo imagiológico. A avaliação estatística foi adaptada às características de cada um dos estudos.Resultados Distonias Primárias: encontrámos défices de função executiva, envolvendo dificuldade na mudança entre sets cognitivos, bem como correlação significativa entre as pontuações nos testes cronometrados e a gravidade dos sintomas motores. Doença de Parkinson: os doentes com DP obtiveram pontuações significativamente inferiores na FAB e em sub-testes do MMSE (memória e função visuo-espacial). A pontuação no MMSE encontrava-se significativamente correlacionada com itens da função motora não relacionados com o tremor. A disfunção da marcha, a disartria, o fenótipo não tremorígeno, a presença de alucinações e pontuação abaixo do ponto de corte na MMSE, foram factores preditivos de demência na avaliação longitudinal. A rigidez e a disartria foram factores preditivos de declínio nas funções frontais. A disfunção frontal foi factor preditivo de declínio na pontuação do MMSE. Encontrámos uma prevalência elevada de RBD nas fases iniciais da DP, que o estudo longitudinal mostrou ser factor preditivo de declínio motor, nomeadamente por agravamento da bradicinésia. Encontrámos também uma prevalência elevada de sintomas psiquiátricos, nomeadamente psicose, depressão, ansiedade, somatização e sintomas obsessivo-compulsivos. As PCI não se encontravam relacionadas com o fenótipo motor, com as complicações motoras do tratamento dopaminérgico ou com a disfunção cognitiva. HPN: os doentes com HPN e os DP apresentaram um padrão disfunção da marcha semelhante, caraterizado por passos curtos, lentidão e dificuldades de equilíbrio, sendo os sintomas mais graves na HPN. Os doentes de Parkinson com maior duração de doença, maior dose de dopaminérgicos e fenótipo motor acinético-rígido apresentaram um padrão de disfunção da marcha de gravidade semelhante ao encontrado na HPN. As alterações vasculares da substância branca, em particular as encontradas na região frontal, encontravam-se negativamente correlacionadas com a melhoria da marcha após PL. O estudo das funções cognitivas mostrou um padrão de atingimento global, com valores mais baixos na cópia do desenho complexo de Rey. Os resultados nas provas de função cognitiva não se encontravam significativamente correlacionados com os resultados na prova da marcha. A progressão na disfunção da marcha encontrava-se relacionada com o tratamento não cirúrgico, idade superior na primeira avaliação, presença de lesões da substância branca, e presença de factores de risco vascular, ao passo que não foram encontrados factores que predissessem de modo significativo o agravamento da função cognitiva. Conclusões: Os resultados dos diversos estudos, evidenciam a presença de alterações cognitivas e comportamentais nas três doenças de movimento. O padrão destas alterações e o modo como estas se relacionaram com os sintomas motores variou de doença para doença. Nas Distonias primárias, a perseveração cognitiva poderá ser o sintoma correspondente à perseveração motora própria da doença, sugerindo disfunção no circuito dorso-lateral frontoestriatal. A correlação entre a gravidade motora da doença e o resultado nos testes cognitivos cronometrados, poderá ser o efeito da relação entre bradicinésia e bradifrenia. Na Doença de Parkinson, o espectro de alterações é mais acentuado, espelhando a disseminação do processo degenerativo no SNC. Para além dos sintomas de disfunção executiva, sugerindo disfunção das tês ansas não motoras, existem sinais de disfunção cognitiva global, estas com uma influência mais significativa no desenvolvimento da demência. A relação entre os diferentes sintomas motores e cognitivos é também complexa, embora se evidencie uma dissociação significativa entre o tremor, sem relação com os sintomas não motores, e os sintomas motores não tremorígenos, relacionados com o declínio cognitivo. Enquanto que a presença de RBD parece ser um factor preditivo de agravamento motor, os sintomas psiquiátricos, também muito frequentes, apresentam uma relação menos clara com a função motora. Destes, os sintomas obsessivo-compulsivos são aqueles que com mais frequência se atribuem a disfunção do sistema fronto-estriatal, nomeadamente da ansa orbito-frontal. As PCI também não mostraram ter relação com os sintomas motores ou cognitivos. Na HPN, é patente o carácter fronto-estriatal das alterações da marcha, demonstrado tanto na sua caracterização quanto no efeito deletério das lesões vasculares da substância branca do lobo frontal na recuperação da marcha após PL. As alterações cognitivas parecem ter um padrão mais difuso, o que talvez explique a falta de correlação com os sintomas motores - esta dissociação pode ser causada quer por diferença nos mecanismos fisiopatológicos quer por presença de comorbilidades cognitivas. --------- ABSTRACT: Fronto-striatal circuits constitute a closed loop system which connects different parts of the frontal lobes to the basal ganglia. They are engaged in motor, cognitive and behavioural control. Primary Dystonia, Parkinson's Disease and Normal-Pressure Hydrocephalus are movement disorders caused by disturbance of the motor fronto-striatal circuit. The existence of cognitive and behavioural dysfunction in these movement disorders is predictable, given the functional connectivity between the several distinct loops of the circuit. Evaluation of cognitive and behavioural dysfunction in these three disorders is thus both of clinical and theoretical relevance. Objectives Our objectives were to evaluate, by clinical means, the relation between motor, cognitive and behavioural symptoms in three movement disorders with different pathophysiological backgrounds - Primary Dystonia, Parkinson's Disease and Normal-Pressure Hydrocephalus - and to analyse the study results under the theoretical framework formed by present knowledge of the fronto-estriatal system. Specific objectives: a) Primary Dystonia: executive dysfunction assessment and correlation analysis with motor dysfunction severity; b) Parkinson's Disease: 1. brief cognitive assessment in the early stages of disease, including a longitudinal analysis for determination of predictive factors for cognitive decline; 2. to investigate the relation between RBD and cognitive and motor dysfunction, including a longitudinal analysis; 3. psychiatric symptom assessment, with particular incidence on Impulse Control Disorders; c) Normal-Pressure Hydrocephalus: 1. gait dysfunction characterization and comparison with Parkinson's Disease patients; 2. determination of cognitive dysfunction profile and its relation with gait dysfunction; 3. follow-up study of cognitive and motor outcome in patients submitted and not submitted to shunt surgery. Methods: Primary Dystonia, Parkinson's Disease and Normal Pressure Hydrocephalus were diagnosed according to clinically validate criteria. Where warranted, we recruited control groups formed by healthy individuals, matched for age, sex and educational level. Patients were evaluated with instruments of direct clinical application, including motor function scales, neuropsychological tests aimed at global and executive functions and psychiatric rating scales. Tests used in Primary Dystonia: Unified Dystonia Rating Scale, Wisconsin Card Sorting Test, Stroop Test, Cube Assembly test (WAIS), Benton’s Visual Retention Test; in Parkinson's Disease: Unified Parkinson's Disease Rating Scale, Frontal Assessment Battery (FAB) , Mini-mental State Examination (MMSE), REM-sleep behavior disorder Questionnaire, Symptom Check-list 90- R, Brief Psychiatric Rating Scale, FAS (phonetic verbal fluency), semantic verbal fluency test, digit span test (WAIS), auditory verbal learning test,Stroop test, Raven's progressive Matrices, Questionnaire for Impulsive-Compulsive Disorders; in NPH: timed walking test, MMSE, immediate memory task (WAIS), digit span test (WAIS), FAB, Rey’s Complex Figure test, Stroop test, letter cancellation test, Perdue Pegboard test. NPH patients were also subjected to an imaging study. Statistics were adapted to the characteristics of each study.Results: Primary Dystonia: we found set-shifting deficits as well as significant correlation between timed neuropsychological tests and dystonia severity. Parkinson's Disease: PD patients had significantly lower scores on the FAB and on the memory and visuo-spatial tests of the MMSE; MMSE scores were significantly correlated to non-tremor motor scores; gait dysfunction and speech scores, non-tremor motor phenotype, hallucinations and scores bellow cut-off on the MMSE were predictive of dementia at follow-up; speech and rigidity scores were predictive of frontal type decline; frontal dysfunction was predictivy of decline in MMSE scores; RBD bradykinesia worsening; psychiatric symptoms were prevalent, particularly Psychosis, Depression, Anxiety, Somatisation and Obsessive-Compulsive Symptoms; Impulse Control Disorders were unrelated to motor phenotype,motor side effects of dopamine treatment and executive function; NPH: gait dysfunction was worse in NPH when compared to PD patients, although the pattern was similarly characterized by slowness, short steps and disequilibrium; PD patients whose gait disturbance was as severe as that of NPH patients were characterized by longer disease duration, predominance of non-tremor motor scores, more advanced disease stage and higher dopamine dose; frontal white matter lesions correlated negatively with improvement after LP; cognitive function assessment revealed wide spread deficits, with lower results on the drawing of the complex figure of Rey, which were not significantly correlated to gait dysfunction; older age, white matter lesions and the presence of vascular risk factors were predictive factors for motor but not cognitive function worsening. Conclusion: Results from our studies highlight the presence of cognitive and behavioural dysfunction in all three movement disorders. Symptom pattern and the relation with ovement derangement varied according to the disease. In Primary Dystonia, set-shifting difficulties could be the cognitive counterpart of motor perseveration characteristic of this disorder, suggesting dysfunction of the dorso-lateral circuit. The relation between timed tests and dystonia severity could suggest a relation between bradyphrenia and bradykinesia in Primary Dystonia. In Parkinson's Disease patients, the spectrum of non-motor symptoms is wider, probably reflecting the spread of neurodegeneration beyond the fronto-striatal circuits. While frontal type deficits predominate, suggestive of dorso-lateral and orbito-frontal dysfunction, non-frontal deficits were also apparent in the initial stages of disease, and were predictive of dementia at follow-up. The relationship between cognitive and motor symptoms is complex, although the results strongly suggest a dissociation between tremor symptoms, which bore no relation with non-motor symptoms, and non-tremor symptoms,whichwas frequent, and a predictive factor for which were related with cognitive decline. While RBD was found to be a predictive factor for bradykinesia worsening, psychiatric symptoms, which were also frequent, showed no apparent relation with motor dysfunction. Relevant to our theoretical consideration was the high prevalence of OCS, which have been attributed to orbito-frontal dysfunction. As to the particular case of ICD, we found no relation either with motor or cognitive dysfunction. The fronto-striatal nature of gait dysfunction in NPH is suggest by the clinical characterization study and by the effects of frontal white matter lesions on gait recovery after LP, whereas cognitive dysfunction presented a more diffuse pattern, which could explain the lack or relation with gait assessment results and also the different outcome on the longitudinal study - this dissociation could be caused by a real difference in pathophysiological mechanisms or, in alternative, be due to the existence of cognitive comorbidities.

Spectroscopic characterization of natural dyes by their non-invasive identification on pre-Columbian codices: the Maya yellow

Mesoamerican cultures had a strong tradition of written and pictorial manuscripts, called the codices. In studies already performed it was found the use of Maya Blue, made from a mixture of indigo and a clay called palygorskite, forming an incredibly stable material where the dye is trapped inside the nanotubes of the clay, after heating. However, a bigger challenge lies in the study of the yellows used, for these civilizations might have used this clay-dye mixture to produce their yellow colorants. As a first step, it was possible to provide identification, by non-invasive methods, of two colorants (a flavonoid and a carotenoid). While the flavonoid absorbed between 368-379 nm, the carotenoid would absorb around 455 nm. A temperature study also conducted allowed to set 140ºC as the desirable temperature to heat the samples without degrading them. FT-IR, conventional Raman and SERS allowed us to understand the existence of a reaction between the dyes and the clays (palygorskite and kaolinite), however it is difficult to understand it in a molecular point of view. As a second step, five species of Mexican dyes were selected on the basis of historical sources. The Maya yellow samples were produced adapting the recipe proposed by Reyes-Valerio, supporting the yellow dyes extracted from the dried plants on the clays, with addition of water, and then heated at 140ºC. It was found that the addition of water in palygorskite would increase the pH, hence deprotonating the molecules having a clear negative effect in the color. A second recipe was developed, without the addition of water; however, it was found that the use of water based binders would still alter the color of the samples with palygorskite. In this case, kaolinite without heating yield better results as a Maya yellow hybrid. It was found that the Maya chemistry might not have been the same for all the colors. The Mesoamericans might have found that different dyes could work better to their desires if matched with different clays. It was noticeable that for a clear distinction between flavonoids and carotenoids the reflectance and emission studies suffice, but when clay is added, Raman techniques will perform better. For this reason, conventional Raman and SERS were employed in order to create a database for the Mesoamerican dyestuffs for a future identification.