Molecular Interactions Underlying Neuronal Ceroid Lipofuscinoses CLN1 and CLN5

Disorders resulting from degenerative changes in the nervous system are progressive and incurable. Both environmental and inherited factors affect neuron function, and neurodegenerative diseases are often the sum of both factors. The cellular events leading to neuronal death are still mostly unknown. Monogenic diseases can offer a model for studying the mechanisms of neurodegeneration. Neuronal ceroid lipofuscinoses, or NCLs, are a group of monogenic, recessively inherited diseases affecting mostly children. NCLs cause severe and specific loss of neurons in the central nervous system, resulting in the deterioration of motor and mental skills and leading to premature death. In this thesis, the focus has been on two forms of NCL, the infantile NCL (INCL, CLN1) and the Finnish variant of late infantile NCL (vLINCLFin, CLN5). INCL is caused by mutations in the CLN1 gene encoding for the PPT1 (palmitoyl protein thioesterase 1) enzyme. PPT1 removes a palmitate moiety from proteins in experimental conditions, but its substrates in vivo are not known. In the Finnish variant of late infantile NCL (vLINCLFin), the CLN5 gene is defective, but the function of the encoded CLN5 has remained unknown. The aim of this thesis was to elucidate the disease mechanisms of these two NCL diseases by focusing on the molecular interactions of the defective proteins. In this work, the first interaction partner for PPT1, the mitochondrial F1-ATP synthase, was described. This protein has been linked to HDL metabolism in addition to its well-known role in the mitochondrial energy production. The connection between PPT1 and the F1-ATP synthase was studied utilizing the INCL-disease model, the genetically modified Ppt1-deficient mice. The levels of F1-ATP synthase subunits were increased on the surface of Ppt1-deficient neurons when compared to controls. We also detected several changes in lipid metabolism both at the cellular and systemic levels in Ppt1-deficient mice when compared to controls. The interactions between different NCL proteins were also elucidated. We were able to detect novel interactions between CLN5 and other NCL proteins, and to replicate the previously reported interactions. Some of the novel interactions influenced the intracellular trafficking of the proteins. The multiple interactions between CLN5 and other NCL proteins suggest a connection between the NCL subtypes at the cellular level. The main results of this thesis elicit information about the neuronal function of PPT1. The connection between INCL and neuronal lipid metabolism introduces a new perspective to this rather poorly characterized subject. The evidence of the interactions between NCL proteins provides the basis for future research trying to untangle the NCL disease mechanisms and to develop strategies for therapies.

Development of a Rapid Multiplex PCR Assay To Genotype Pasteurella multocida Strains by Use of the Lipopolysaccharide Outer Core Biosynthesis Locus

Pasteurella multocida is a Gram-negative bacterial pathogen that is the causative agent of a wide range of diseases in many animal species, including humans. A widely used method for differentiation of P. multocida strains involves the Heddleston serotyping scheme. This scheme was developed in the early 1970s and classifies P. multocida strains into 16 somatic or lipopolysaccharide (LPS) serovars using an agar gel diffusion precipitin test. However, this gel diffusion assay is problematic, with difficulties reported in accuracy, reproducibility, and the sourcing of quality serovar-specific antisera. Using our knowledge of the genetics of LPS biosynthesis in P. multocida, we have developed a multiplex PCR (mPCR) that is able to differentiate strains based on the genetic organization of the LPS outer core biosynthesis loci. The accuracy of the LPS-mPCR was compared with classical Heddleston serotyping using LPS compositional data as the "gold standard." The LPS-mPCR correctly typed 57 of 58 isolates; Heddleston serotyping was able to correctly and unambiguously type only 20 of the 58 isolates. We conclude that our LPS-mPCR is a highly accurate LPS genotyping method that should replace the Heddleston serotyping scheme for the classification of P. multocida strains.

The Molecular Basis of Hydrolethalus Syndrome

Hydrolethalus syndrome (HLS) is a severe fetal malformation syndrome that is inherited by an autosomal recessive manner. HLS belongs to the Finnish disease heritage, an entity of rare diseases that are more prevalent in Finland than in other parts of the world. The phenotypic spectrum of the syndrome is wide and it is characterized by several developmental abnormalities, including hydrocephalus and absent midline structures in the brain, abnormal lobation of the lungs, polydactyly as well as micrognathia and other craniofacial anomalies. Polyhydramnios are relatively frequent during pregnancy. HLS can nowadays be effectively identified by ultrasound scan already at the end of the first trimester of pregnancy. One of the main goals in this study was to identify and characterize the gene defect underlying HLS. The defect was found from a previously unknown gene that was named HYLS1. Identification of the gene defect made it possible to confirm the HLS diagnosis genetically, an aspect that provides valuable information for the families in which a fetus is suspected to have HLS. Neuropathological findings of mutation confirmed HLS cases were described for the first time in detail in this study. Also, detailed general pathological findings were described. Since HYLS1 was an unknown gene with no relatives in the known gene families, many functional studies were performed in order to unravel the function of the gene and of the protein it codes for. Studies showed, for example, that the subcellular localization of the HYLS1 protein was different when the normal and the defective forms were compared. In addition, HYLS1 was shown to possess transactivation potential which was significantly diminished in the defective form. According to the results of this study it can be stated that HYLS1 most likely participates in transcriptional regulation and also in the regulation of cholesterol metabolism and that the function of HYLS1 is critical for normal fetal development.

Aspectos antropológicos de las terapias con células madre

Resumen: Las células madre tienen dos características naturales únicas, que son su capacidad de división celular indefinida, y de regeneración y reparación de tejidos. Esto genera un gran interés en las áreas de investigación y tratamiento de enfermedades que aún no tienen cura y son motivadoras de una importante fuente de esperanza para los pacientes. Surge, entonces, el dilema ético de definir cuáles serán los recursos para obtenerlas, procesarlas y determinar la seguridad y eficacia de su uso en la aplicación clínica. Un enfoque personalista ontológico considerará la protección de todos los participantes en los procesos de investigación: los donantes de células, los participantes de la investigación y los pacientes que serán beneficiarios de estos tratamientos, de manera de proteger a todos los involucrados sin exponer a riesgos a unos para salvar a otros.

Fitoterapia aplicada ao tratamento da osteoartrite

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

Towards An Acoustic Model-Based Poroelastic Imaging Method: I. Theoretical Foundation

The ultrasonic measurement and imaging of tissue elasticity is currently under wide investigation and development as a clinical tool for the assessment of a broad range of diseases, but little account in this field has yet been taken of the fact that soft tissue is porous and contains mobile fluid. The ability to squeeze fluid out of tissue may have implications for conventional elasticity imaging, and may present opportunities for new investigative tools. When a homogeneous, isotropic, fluid-saturated poroelastic material with a linearly elastic solid phase and incompressible solid and fluid constituents is subjected to stress, the behaviour of the induced internal strain field is influenced by three material constants: the Young's modulus (E(s)) and Poisson's ratio (nu(s)) of the solid matrix and the permeability (k) of the solid matrix to the pore fluid. New analytical expressions were derived and used to model the time-dependent behaviour of the strain field inside simulated homogeneous cylindrical samples of such a poroelastic material undergoing sustained unconfined compression. A model-based reconstruction technique was developed to produce images of parameters related to the poroelastic material constants (E(s), nu(s), k) from a comparison of the measured and predicted time-dependent spatially varying radial strain. Tests of the method using simulated noisy strain data showed that it is capable of producing three unique parametric images: an image of the Poisson's ratio of the solid matrix, an image of the axial strain (which was not time-dependent subsequent to the application of the compression) and an image representing the product of the aggregate modulus E(s)(1-nu(s))/(1+nu(s))(1-2nu(s)) of the solid matrix and the permeability of the solid matrix to the pore fluid. The analytical expressions were further used to numerically validate a finite element model and to clarify previous work on poroelastography.

Online health information - what the newspapers tell their readers: a systematic content analysis

Background: This study investigated the nature of newspaper reporting about online health information in the UK and US. Internet users frequently search for health information online, although the accuracy of the information retrieved varies greatly and can be misleading. Newspapers have the potential to influence public health behaviours, but information has been lacking in relation to how newspapers portray online health information to their readers.

Methods: The newspaper database Nexis (R) UK was searched for articles published from 2003 - 2012 relating to online health information. Systematic content analysis of articles published in the highest circulation newspapers in the UK and US was performed. A second researcher coded a 10% sample to establish inter-rater reliability of coding.

Results: In total, 161 newspaper articles were included in the analysis. Publication was most frequent in 2003, 2008 and 2009, which coincided with global threats to public health. UK broadsheet newspapers were significantly more likely to cover online health information than UK tabloid newspapers (p = 0.04) and only one article was identified in US tabloid newspapers. Articles most frequently appeared in health sections. Among the 79 articles that linked online health information to specific diseases or health topics, diabetes was the most frequently mentioned disease, cancer the commonest group of diseases and sexual health the most frequent health topic. Articles portrayed benefits of obtaining online health information more frequently than risks. Quotations from health professionals portrayed mixed opinions regarding public access to online health information. 108 (67.1%) articles directed readers to specific health-related web sites. 135 (83.9%) articles were rated as having balanced judgement and 76 (47.2%) were judged as having excellent quality reporting. No difference was found in the quality of reporting between UK and US articles.

Conclusions: Newspaper coverage of online health information was low during the 10-year period 2003 to 2012. Journalists tended to emphasise the benefits and understate the risks of online health information and the quality of reporting varied considerably. Newspapers directed readers to sources of online health information during global epidemics although, as most articles appeared in the health sections of broadsheet newspapers, coverage was limited to a relatively small readership.

Factores sociodemográficos na saúde oral : influências nos comportamentos de rotinas de saúde oral, frequências de idas a consultas, auto avaliação do estado de saúde oral e presença de dor em jovens de 12 anos

RESUMO - Introdução: A saúde oral é uma componente essencial na saúde geral e no bem-estar dos indivíduos. Sabe-se que os problemas de saúde oral afectam predominantemente os elementos de níveis socioeconómicos mais baixos, evidenciando a influência dos determinantes sociais da saúde na saúde oral das populações. Os objectivos deste estudo são caracterizar os comportamentos de rotinas diárias de higiene oral, frequências de idas a consultas de saúde oral, auto-avaliação do estado de saúde oral e percepção de dor na cavidade oral em crianças de 12 anos em Portugal e analisar a associação entre estes e os factores sociodemográficos. Métodos: Foi realizado um estudo observacional, transversal e analítico, abrangendo 1309 jovens e baseado em informação recolhida no III Estudo Nacional de Prevalência de Doenças Orais (ENPDO). Para além das estatísticas descritivas usuais, as estatísticas inferenciais basearam-se predominantemente em modelos de regressão logística binária. Resultados: Dos participantes, 70.6% (n=924) escova “duas ou mais vezes por dia” com associação com todas as variáveis sociodemográficas. Na análise multivariada, o género masculino (OR=2.088; IC95%: 1.574-2.770, em relação ao género feminino), a área de residência predominantemente rural ou mediamente urbana (OR= 1.800; IC95%: 2.587; OR=1.516; IC95%: 1.093-2.103, em relação a zonas predominantemente urbanas), a escolaridade da mãe ser o ensino básico (OR= 2.112; IC95%: 1.408-3.168, em relação ao ensino superior) e a actividade laboral do pai ser desempregado (OR= 1.938; IC95%: 1.280-2.934, em relação a ser trabalhador) foram as variáveis com mais impacto para a adopção de comportamentos de escovagem potencialmente inadequados (p<0.05). A maioria dos inquiridos (94.2%; n=1247) já tinham ido a uma consulta de saúde oral e 74.5% (n=860) nos últimos 12 meses, 95.5% (n=1250) encontram-se satisfeitos com a saúde oral e 44.5% (n=578) afirma ter tido algum tipo de dor na cavidade oral nos últimos 12 meses. Conclusão: Os resultados obtidos estão de acordo com a literatura em termos de factores de associação. Desta forma, a saúde oral nos jovens de 12 anos em Portugal, nos diversos contextos aqui analisados, pode ser considerada como satisfatória. A única excepção relevante é a componente da dor, com valores alarmantes embora de natureza mais subjectiva. A influência dos factores sociodemográficos sugere que futuras abordagens para a promoção da saúde oral tenham em conta os determinantes de saúde no delineamento de estratégias quer a nível individual quer a nível comunitário.

Facteurs associés à l'alimentation traditionnelle au sein de trois communautés cries du Nord du Québec

Cette étude propose d’identifier les facteurs affectant la consommation d’aliments traditionnels à travers une perspective écologique, afin de réduire les taux de prévalence élevés de maladies chroniques et ralentir la forte diminution de consommation d’aliments traditionnels chez les Cris du nord québécois. Pour ce faire, une méthode mixte « sequential explanatory », fut utilisée, combinant quatre groupes focus (n=23) et une régression logistique (n=374) à partir de données secondaires issues de trois études transversales. Selon les résultats de la régression logistique: l’âge, chasser, marcher, le niveau d’éducation et la communauté de résidence étaient associées à une consommation d’aliments traditionnelle trois fois/semaine (p<0,05). Subséquemment, des groupes focus vinrent enrichir et contredire ces résultats. Par exemple : les participants étaient en désaccord avec le fait qu’il n’y avait aucune association entre les aliments traditionnels et l’emploi. Ils croyaient que les personnes sans emploi ont plus d’opportunités pour aller chasser mais peu d’argent pour couvrir les dépenses et inversement pour ceux avec emploi. Ce double effet aurait possiblement fait disparaître l’association dans la régression logistique. Suite aux groupes focus, plusieurs facteurs furent identifiés et distribués dans un modèle écologique suggérant que la consommation d’aliments traditionnels est principalement influencée par des facteurs sociaux, communautaires et environnementaux et ne se limite pas aux facteurs individuels. En conclusion, afin de promouvoir l’alimentation traditionnelle, quatre suggestions de priorités d’action sont proposées. L’alimentation traditionnelle doit faire partie des stratégies de santé publique pour réduire les taux de maladies chroniques et améliorer le bien-être des populations autochtones.

La stabilité des réponses physiologiques face au stress psychologique telle qu’étudiée sur une période de trois ans

Le stress augmente le risque de développer des maladies cardiovasculaires (CV) ainsi que de mourir de ces maladies. Selon certaines hypothèses, ce phénomène se produirait par le biais de réponses répétés de réactivité physiologique élevée ou de récupération physiologique prolongée, suite à un épisode de stress. La stabilité à long terme des réponses physiologiques face au stress a reçu peu d’attention. Objectifs: (1) Évaluer la stabilité temporelle de la réactivité et de la récupération physiologique suite au stress, à travers l’évaluation des systèmes cardiovasculaires et nerveux autonome, et ce sur un intervalle de 3 ans. (2) Déterminer si le sexe et l'âge agissent comme des variables modératrices. Méthodologie: Un total de 134 hommes et femmes en santé ont été recrutés au sein de la communauté et ont pris part à 2 séances en laboratoire. Quatre tâches, d’une durée de 5 minutes chacune et composée d’un élément de stress interpersonnel différent, ont été administrées. Chaque tâche était suivie d’une période de récupération de 5 min. Des mesures de la fréquence cardiaque (FC), de la pression artérielle (PA) et de la variabilité de la fréquence cardiaque (VFC : HF, LF et VLF) ont été obtenues. Des corrélations de Spearman et des régressions linéaires ont été effectuées. Résultats: Des corrélations test-retest significatives ont été obtenues pour toutes les mesures physiologiques, sauf pour la PA diastolique et la VLF, lors de la période de récupération. Aucune différence significative quant à la stabilité des réponses face au stress en fonction du sexe ou de l’âge des participants, n’a été trouvée. Conclusion: Les réponses physiologiques face au stress représentent des caractéristiques individuelles stables sur trois ans, peu affectées par le sexe et l’âge.

Exámenes de laboratorio clínico en menores de cinco años atendidos en sala ERA (enfermedad respiratoria aguda) en una empresa social del estado de primer nivel de atención

Las Salas ERA (Enfermedad Respiratoria Aguda) son áreas definidas en las Unidades Primarias de Atención (UPAs), Centros de Atención Médica Inmediata (CAMIs) y de algunas instituciones de segundo nivel, para la atención de pacientes con ERA, que puedan ser manejados con esquemas terapéuticos básicos que en muchas oportunidades se ocupan las salas de observación de urgencias por casos que podrían manejarse en un nivel de complejidad más bajo, pero tampoco pueden ser manejados a nivel domiciliario. La enfermedad respiratoria aguda (ERA) es un conjunto de patologías que afectan el sistema respiratorio, siendo esta una causa muy frecuente de morbilidad y mortalidad en los niños y niñas menores de 5 años, en especial por infección respiratoria aguda (IRA). El pago de la atención integral en las salas ERA a los niños menores de cinco años (población vinculada) de edad corresponde al Fondo Financiero Distrital el cual ha establecido un paquete que incluye valoración médica, valoración con la terapeuta respiratoria, analgésicos, corticosteroides e inhaladores. En el momento en que el médico ordena un examen de laboratorio clínico como ayuda diagnóstica el Fondo Financiero Distrital deja de pagar el paquete completo de prestación de servicios en Salas ERA y paga los medicamentos y dispositivos médicos empleados en la atención de pacientes de manera individual, con esta segunda manera de pago tiene menos ganancias el hospital ya que este debe asumir los costos de la valoración por parte del médico y la terapeuta respiratoria. El objetivo de esta investigación es conocer la magnitud en la solicitud de exámenes de Laboratorio Clínico a los niños menores de cinco años que recibieron atención por parte de la Sala ERA de una Empresa Social del Estado de primer nivel de atención en la ciudad de Bogotá.

Evaluación a la implementación de un modelo de atención para riesgo biológico en una administradora de riesgos profesionales en Colombia en el periodo de enero a octubre de 2009

Debido al alto riesgo que presentan los trabajadores que están expuestos a adquirir enfermedades infecto contagiosas en nuestro medio, se determina la necesidad de evaluar el modelo administrativo de atención para riesgo biológico que se implemento en una Administradora de Riesgos Profesionales (ARP) en Colombia, con el fin de establecer medidas para el manejo pertinente y oportuno del evento, así como el seguimiento de los trabajadores, buscando disminuir la incidencia y prevalencia de enfermedades de alto costo como VIH, Hepatitis B y Hepatitis C, y de esta manera generar un impacto positivo en la atención y administración de los recursos. Para ello se realiza un análisis de datos de la población afiliada a la ARP que reportaron un accidente de riesgo biológico durante el 2009, y se realizan comparaciones de variables entre los casos positivos del primer período, comprendido entre los meses de enero y junio de 2009, y los que se presentaron posterior a la implementación del nuevo modelo de atención, período comprendido desde julio hasta octubre del 2009. Se encontró que el género masculino fue el más afectado, los departamentos con mayor reporte de casos fueron Atlántico y Bolívar, el sector económico más afectado fue el de recolección de basura y las lesiones fueron provocadas en su mayoría por objeto cortopunzantes. Con el nuevo modelo se evidenció un cambio positivo en términos de pertinencia, oportunidad de aviso y calificación del evento, en cuanto a costos se encontró que, por el momento, los períodos no son comparables por la variabilidad en tiempo, sin embargo, se proyecta un cambio positivo, no solo por los costos cuantificables, sino por los costos indirectos.

Pedometer: intervention strategy and physical activity promotion

World Health Organization (WHO) has prioritized physical activity (PA) as one of the mostrelevant strategies leading the decreasing prevalence of noncommunicable chronic diseases. Pedometer has emerged as one of the valid intervention programs, reliable and useful to assess,measure and promote the physical activity practice, through counts the number of steps perday. One of the aims is to establish the goals based on steps per day made by a person and thepositive feedback, which can generate behavior changes and adoption of healthy habits, from a regular physical activity practice perspective. This review attends to enhance the current state ofpedometer program, as an intervention one, in all kind of population; its health impact and theapplication methodologies, using the pedometer as a steps quantifier device, with feasible access,use and management. Additionally, the review will be useful as a framework to design futureresearch projects, aim to develop, adapt and apply evidence based pedometer protocols, insideclinical, academic and community context.

A geostatistical analysis of soil, vegetation, and image data characterizing land surface variation

The elucidation of spatial variation in the landscape can indicate potential wildlife habitats or breeding sites for vectors, such as ticks or mosquitoes, which cause a range of diseases. Information from remotely sensed data could aid the delineation of vegetation distribution on the ground in areas where local knowledge is limited. The data from digital images are often difficult to interpret because of pixel-to-pixel variation, that is, noise, and complex variation at more than one spatial scale. Landsat Thematic Mapper Plus (ETM+) and Satellite Pour l'Observation de La Terre (SPOT) image data were analyzed for an area close to Douna in Mali, West Africa. The variograms of the normalized difference vegetation index (NDVI) from both types of image data were nested. The parameters of the nested variogram function from the Landsat ETM+ data were used to design the sampling for a ground survey of soil and vegetation data. Variograms of the soil and vegetation data showed that their variation was anisotropic and their scales of variation were similar to those of NDVI from the SPOT data. The short- and long-range components of variation in the SPOT data were filtered out separately by factorial kriging. The map of the short-range component appears to represent the patterns of vegetation and associated shallow slopes and drainage channels of the tiger bush system. The map of the long-range component also appeared to relate to broader patterns in the tiger bush and to gentle undulations in the topography. The results suggest that the types of image data analyzed in this study could be used to identify areas with more moisture in semiarid regions that could support wildlife and also be potential vector breeding sites.

Absorption of isoflavones in humans: effects of food matrix and processing

If soy isoflavones are to be effective in preventing or treating a range of diseases, they must be bioavailable, and thus understanding factors which may alter their bioavailability needs to be elucidated. However, to date there is little information on whether the pharmacokinetic profile following ingestion of a defined dose is influenced by the food matrix in which the isoflavone is given or by the processing method used. Three different foods (cookies, chocolate bars and juice) were prepared, and their isoflavone contents were determined. We compared the urinary and serum concentrations of daidzein, genistein and equol following the consumption of three different foods, each of which contained 50 mg of isoflavones. After the technological processing of the different test foods, differences in aglycone levels were observed. The plasma levels of the isoflavone precursor daidzein were not altered by food matrix. Urinary daidzein recovery was similar for all three foods ingested with total urinary output of 33-34% of ingested dose. Peak genistein concentrations were attained in serum earlier following consumption of a liquid matrix rather than a solid matrix, although there was a lower total urinary recovery of genistein following ingestion of juice than that of the two other foods. (c) 2006 Elsevier Inc. All rights reserved.