Estudos longitudinais sobre o crescimento somático e desempenho motor: delineamentos, desafios, necessidades

O objetivo principal desta revisão é fornecer uma visão geral dos principais estudos longitudinais e longitudinais-mistos que se centraram sobre o crescimento somático, maturação biológica e, mais recentemente, também no desempenho físico. Somente foram considerados os estudos realizados na América do Norte, Europa e países de língua portuguesa. Em primeiro lugar, são apresentadas as principais considerações teóricas, características gerais, o delineamento do estudo e análise estatística multivariada dos dados. Na segunda parte, é edificado o panorama geral sobre os estudos emblemáticos de natureza longitudinal e longitudinal-mista. Finalmente, foram considerados alguns dos principais desafios que se colocam à pesquisa longitudinal.

Microfluidic platforms for the characterization of in meso membrane protein crystallization

Membrane proteins, which reside in the membranes of cells, play a critical role in many important biological processes including cellular signaling, immune response, and material and energy transduction. Because of their key role in maintaining the environment within cells and facilitating intercellular interactions, understanding the function of these proteins is of tremendous medical and biochemical significance. Indeed, the malfunction of membrane proteins has been linked to numerous diseases including diabetes, cirrhosis of the liver, cystic fibrosis, cancer, Alzheimer's disease, hypertension, epilepsy, cataracts, tubulopathy, leukodystrophy, Leigh syndrome, anemia, sensorineural deafness, and hypertrophic cardiomyopathy.1-3 However, the structure of many of these proteins and the changes in their structure that lead to disease-related malfunctions are not well understood. Additionally, at least 60% of the pharmaceuticals currently available are thought to target membrane proteins, despite the fact that their exact mode of operation is not known.4-6 Developing a detailed understanding of the function of a protein is achieved by coupling biochemical experiments with knowledge of the structure of the protein. Currently the most common method for obtaining three-dimensional structure information is X-ray crystallography. However, no a priori methods are currently available to predict crystallization conditions for a given protein.7-14 This limitation is currently overcome by screening a large number of possible combinations of precipitants, buffer, salt, and pH conditions to identify conditions that are conducive to crystal nucleation and growth.7,9,11,15-24 Unfortunately, these screening efforts are often limited by difficulties associated with quantity and purity of available protein samples. While the two most significant bottlenecks for protein structure determination in general are the (i) obtaining sufficient quantities of high quality protein samples and (ii) growing high quality protein crystals that are suitable for X-ray structure determination,7,20,21,23,25-47 membrane proteins present additional challenges. For crystallization it is necessary to extract the membrane proteins from the cellular membrane. However, this process often leads to denaturation. In fact, membrane proteins have proven to be so difficult to crystallize that of the more than 66,000 structures deposited in the Protein Data Bank,48 less than 1% are for membrane proteins, with even fewer present at high resolution (< 2Å)4,6,49 and only a handful are human membrane proteins.49 A variety of strategies including detergent solubilization50-53 and the use of artificial membrane-like environments have been developed to circumvent this challenge.43,53-55 In recent years, the use of a lipidic mesophase as a medium for crystallizing membrane proteins has been demonstrated to increase success for a wide range of membrane proteins, including human receptor proteins.54,56-62 This in meso method for membrane protein crystallization, however, is still by no means routine due to challenges related to sample preparation at sub-microliter volumes and to crystal harvesting and X-ray data collection. This dissertation presents various aspects of the development of a microfluidic platform to enable high throughput in meso membrane protein crystallization at a level beyond the capabilities of current technologies. Microfluidic platforms for protein crystallization and other lab-on-a-chip applications have been well demonstrated.9,63-66 These integrated chips provide fine control over transport phenomena and the ability to perform high throughput analyses via highly integrated fluid networks. However, the development of microfluidic platforms for in meso protein crystallization required the development of strategies to cope with extremely viscous and non-Newtonian fluids. A theoretical treatment of highly viscous fluids in microfluidic devices is presented in Chapter 3, followed by the application of these strategies for the development of a microfluidic mixer capable of preparing a mesophase sample for in meso crystallization at a scale of less than 20 nL in Chapter 4. This approach was validated with the successful on chip in meso crystallization of the membrane protein bacteriorhodopsin. In summary, this is the first report of a microfluidic platform capable of performing in meso crystallization on-chip, representing a 1000x reduction in the scale at which mesophase trials can be prepared. Once protein crystals have formed, they are typically harvested from the droplet they were grown in and mounted for crystallographic analysis. Despite the high throughput automation present in nearly all other aspects of protein structure determination, the harvesting and mounting of crystals is still largely a manual process. Furthermore, during mounting the fragile protein crystals can potentially be damaged, both from physical and environmental shock. To circumvent these challenges an X-ray transparent microfluidic device architecture was developed to couple the benefits of scale, integration, and precise fluid control with the ability to perform in situ X-ray analysis (Chapter 5). This approach was validated successfully by crystallization and subsequent on-chip analysis of the soluble proteins lysozyme, thaumatin, and ribonuclease A and will be extended to microfluidic platforms for in meso membrane protein crystallization. The ability to perform in situ X-ray analysis was shown to provide extremely high quality diffraction data, in part as a result of not being affected by damage due to physical handling of the crystals. As part of the work described in this thesis, a variety of data collection strategies for in situ data analysis were also tested, including merging of small slices of data from a large number of crystals grown on a single chip, to allow for diffraction analysis at biologically relevant temperatures. While such strategies have been applied previously,57,59,61,67 they are potentially challenging when applied via traditional methods due to the need to grow and then mount a large number of crystals with minimal crystal-to-crystal variability. The integrated nature of microfluidic platforms easily enables the generation of a large number of reproducible crystallization trials. This, coupled with in situ analysis capabilities has the potential of being able to acquire high resolution structural data of proteins at biologically relevant conditions for which only small crystals, or crystals which are adversely affected by standard cryocooling techniques, could be obtained (Chapters 5 and 6). While the main focus of protein crystallography is to obtain three-dimensional protein structures, the results of typical experiments provide only a static picture of the protein. The use of polychromatic or Laue X-ray diffraction methods enables the collection of time resolved structural information. These experiments are very sensitive to crystal quality, however, and often suffer from severe radiation damage due to the intense polychromatic X-ray beams. Here, as before, the ability to perform in situ X-ray analysis on many small protein crystals within a microfluidic crystallization platform has the potential to overcome these challenges. An automated method for collecting a "single-shot" of data from a large number of crystals was developed in collaboration with the BioCARS team at the Advanced Photon Source at Argonne National Laboratory (Chapter 6). The work described in this thesis shows that, even more so than for traditional structure determination efforts, the ability to grow and analyze a large number of high quality crystals is critical to enable time resolved structural studies of novel proteins. In addition to enabling X-ray crystallography experiments, the development of X-ray transparent microfluidic platforms also has tremendous potential to answer other scientific questions, such as unraveling the mechanism of in meso crystallization. For instance, the lipidic mesophases utilized during in meso membrane protein crystallization can be characterized by small angle X-ray diffraction analysis. Coupling in situ analysis with microfluidic platforms capable of preparing these difficult mesophase samples at very small volumes has tremendous potential to enable the high throughput analysis of these systems on a scale that is not reasonably achievable using conventional sample preparation strategies (Chapter 7). In collaboration with the LS-CAT team at the Advanced Photon Source, an experimental station for small angle X-ray analysis coupled with the high quality visualization capabilities needed to target specific microfluidic samples on a highly integrated chip is under development. Characterizing the phase behavior of these mesophase systems and the effects of various additives present in crystallization trials is key for developing an understanding of how in meso crystallization occurs. A long term goal of these studies is to enable the rational design of in meso crystallization experiments so as to avoid or limit the need for high throughput screening efforts. In summary, this thesis describes the development of microfluidic platforms for protein crystallization with in situ analysis capabilities. Coupling the ability to perform in situ analysis with the small scale, fine control, and the high throughput nature of microfluidic platforms has tremendous potential to enable a new generation of crystallographic studies and facilitate the structure determination of important biological targets. The development of platforms for in meso membrane protein crystallization is particularly significant because they enable the preparation of highly viscous mixtures at a previously unachievable scale. Work in these areas is ongoing and has tremendous potential to improve not only current the methods of protein crystallization and crystallography, but also to enhance our knowledge of the structure and function of proteins which could have a significant scientific and medical impact on society as a whole. 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Estudos longitudinais sobre o crescimento somático e desempenho motor: delineamentos, desafios, necessidades

O objetivo principal desta revisão é fornecer uma visão geral dos principais estudos longitudinais e longitudinais-mistos que se centraram sobre o crescimento somático, maturação biológica e, mais recentemente, também no desempenho físico. Somente foram considerados os estudos realizados na América do Norte, Europa e países de língua portuguesa. Em primeiro lugar, são apresentadas as principais considerações teóricas, características gerais, o delineamento do estudo e análise estatística multivariada dos dados. Na segunda parte, é edificado o panorama geral sobre os estudos emblemáticos de natureza longitudinal e longitudinal-mista. Finalmente, foram considerados alguns dos principais desafios que se colocam à pesquisa longitudinal.

Inequities in energy-balance related behaviours and family environmental determinants in European children: baseline results of the prospective EPHE evaluation study

BACKGROUND:Tackling inequalities in overweight, obesity and related determinants has become a top priority for the European research and policy agendas. Although it has been established that such inequalities accumulate from early childhood onward, they have not been studied extensively in children. The current article discusses the results of an explorative analysis for the identification of inequalities in behaviours and their determinants between groups with high and low socio-economic status. METHODS: This study is part of the Epode for the Promotion of Health Equity (EPHE) evaluation study, the overall aim of which is to assess the impact and sustainability of EPODE methodology to diminish inequalities in childhood obesity and overweight. Seven community-based programmes from different European countries (Belgium, Bulgaria, France, Greece, Portugal, Romania, The Netherlands) participate in the EPHE study. In each of the communities, children aged 6-8 years participated, resulting in a total sample of 1266 children and their families. A parental self-administrated questionnaire was disseminated in order to assess the socio-economic status of the household, selected energy balance-related behaviours (1. fruit and vegetable consumption; 2. soft drink/ fruit juices and water consumption; 3. screen time and 4. sleep duration) of the children and associated family environmental determinants. The Mann-Whitney U test and Pearson's chi-square test were used to test differences between the low and high education groups. The country-specific median was chosen as the cut-off point to determine the educational level, given the different average educational level in every country. RESULTS: Children with mothers of relatively high educational level consumed fruits and vegetables more frequently than their peers of low socio-economic status. The latter group of children had a higher intake of fruit juices and/or soft drinks and had higher screen time. Parental rules and home availability were consistently different between the two socio-economic groups in our study in all countries. However we did not find a common pattern for all behaviours and the variability across the countries was large. CONCLUSIONS: Our findings are indicative of socio-economic inequalities in our samples, although the variability across the countries was large. The effectiveness of interventions aimed at chancing parental rules and behaviour on health inequalities should be studied.

Prevalência de sobrepeso e de obesidade em crianças e adolescentes na Região Autónoma da Madeira, Portugal

O objectivo foi estimar a prevalência de sobrepeso e obesi dade em crianças e adolescentes da Região Autónoma da Madeira, Portugal. Um procedimento estratificado proporcional foi usado para obter uma amostra representativa de crianças e adolescentes madeirenses dos 7 aos 18 anos. No total, 2503 sujeitos, 1266 rapazes e 1237 raparigas, participaram no estudo. A prevalência de sobrepeso e obesidade foi definida a partir do índice de massa corporal e de acordo com os pontos de corte propostos pela ‘International Obesity TaskForce’. A prevalência de sobrepeso foi de 14.22% e 10.99% para os rapazes e raparigas dos 7-18 anos, respectivamente. Os valores correspondentes para a obesidade foram 2.61% e 1.86%. Na maioria dos grupos etários, os elementos do sexo mas culino apresentaram uma prevalência mais elevada de sobrepeso e obesidade do que o sexo feminino. Percentagens mais baixas ou ausência de sobrepeso e obesidade foram obser vadas aos 16-17 anos. A prevalência de sobrepeso e obesid ade para as crianças e adolescentes madeirenses foi similar ou inferior a pesquisas desenvolvidas em Portugal e em outros países europeus. A prevalência de sobrepeso e obesidade, embora baixa, requer prevenção adequada.

Prevalência de sobrepeso e de obesidade em crianças e adolescentes na Região Autónoma da Madeira, Portugal

O objectivo foi estimar a prevalência de sobrepeso e obesi dade em crianças e adolescentes da Região Autónoma da Madeira, Portugal. Um procedimento estratificado proporcional foi usado para obter uma amostra representativa de crianças e adolescentes madeirenses dos 7 aos 18 anos. No total, 2503 sujeitos, 1266 rapazes e 1237 raparigas, participaram no estudo. A prevalência de sobrepeso e obesidade foi definida a partir do índice de massa corporal e de acordo com os pontos de corte propostos pela ‘International Obesity TaskForce’. A prevalência de sobrepeso foi de 14.22% e 10.99% para os rapazes e raparigas dos 7-18 anos, respectivamente. Os valores correspondentes para a obesidade foram 2.61% e 1.86%. Na maioria dos grupos etários, os elementos do sexo mas culino apresentaram uma prevalência mais elevada de sobrepeso e obesidade do que o sexo feminino. Percentagens mais baixas ou ausência de sobrepeso e obesidade foram obser vadas aos 16-17 anos. A prevalência de sobrepeso e obesid ade para as crianças e adolescentes madeirenses foi similar ou inferior a pesquisas desenvolvidas em Portugal e em outros países europeus. A prevalência de sobrepeso e obesidade, embora baixa, requer prevenção adequada.

Congruencia entre los valores personales y organizacionales y su influencia con el engagement en el trabajo : estudio de caso en una organización del sector salud

La capacidad de gestión del personal se ha convertido en un imperativo para las organizaciones modernas. Por tanto se vienen introduciendo temas como la administración en valores y el engagement de los trabajadores. Sin embargo, la relación entre estos dos aún no ha sido estudiada. El presente estudio tiene como objetivo analizar el efecto que tiene el grado de articulación y reconocimiento de los valores organizacionales y personales y organizacionales sobre los niveles de engagement de los empleados. Para esta investigación se utilizó una muestra constituida por 54 trabajadores de una organización del sector salud de la ciudad de Bogotá a quienes les fueron aplicadas dos escalas: el Inventario para Medir la Articulación entre la Persona y la Organización (Inventario APO) y Utrech Work Engagement Scale (UWES). Como principal resultado se obtuvo que de las tres dimensiones consideradas predictoras del engagement, solamente el reconocimiento de los valores organizacionales tuvo un efecto estadísticamente significativo.

Tratamiento quirúrgico del síndrome de apnea hipopnea obstructiva del sueño en la Clínica Rivas, Bogotá – Colombia

Introducción: El Síndrome de Apnea Hipopnea Obstructiva del Sueño es un trastorno respiratorio del sueño mayor ampliamente conocido, con importantes implicaciones para los pacientes y cuya incidencia ha venido en aumento durante los últimos años; comprende diversas manifestaciones clínicas que varían desde el ronquido hasta consecuencias cardiovasculares importantes. Objetivo: Describir la experiencia de los procedimientos quirúrgicos más utilizados para el tratamiento de pacientes con Trastornos Respiratorios del Sueño en la Clínica Rivas. Diseño: Estudio observacional descriptivo. Métodos: Revisión de 366 historias clínicas de pacientes con diagnóstico clínico y Polisomnográfico de SAHOS intervenidos quirúrgicamente debido al Trastorno Respiratorio del Sueño por rechazo de terapia de presión positiva en 3 años de observación. Resultados: Se evaluaron diferencias en medianas de los cambios del IAH, índice de Saturación de oxigeno basal y mínima, y el índice de microdespertares nocturnos tanto prequirúrgica como postquirúrgicamente. Como medida de evaluación secundaria se evaluaron las complicaciones quirúrgicas. Conclusión: En nuestra institución, como centro de referencia en apnea del sueño, la cirugía ha demostrado que disminuye de forma significativa gravedad del SAHOS y disminuye el riesgo de los pacientes con trastornos respiratorios del sueño que han rechazado el dispositivo de presión positiva.

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Pharmacological and Toxicological Studies of Essential Oil of Lavandula stoechas subsp. luisieri.

The present study was carried out to evaluate the chemical and pharmacological properties of essential oil (EO) of Lavandula stoechas L. subsp. luisieri that is a spontaneous shrub widespread in Alentejo (Portugal). Oxygenated monoterpenes, as 1,8-cineole, lavandulol and necrodane derivatives are the main components of essential oil. It revealed important antioxidant activity with high ability to inhibit the lipid peroxidation and showed an outstanding effect against a wide spectrum of microorganisms, such as Gram-positive and Gram-negative bacteria and pathogenic yeasts. The analgesic effect studied in rats was dose dependent, reaching a maximum of 67 % at 60 min. with the dose of 200 mg/kg and the anti-inflammatory activity with this dose caused an inhibition in carrageenan-induced rat paw oedema (83 %) that is higher than dexamethasone 1 mg/Kg (69 %). Besides, animals exhibited a normal behaviour after EO administration revealing low toxicity. Essential oil of L. luisieri from Alentejo that presents important pharmacological properties and low toxicity is a promised candidate to be used as food supplement or in pharmaceutical applications.

Expression of rice genes homologous of Arabidopsis genes previously related to drought tolerance.

The identification and validation of candidate genes related to traits of interest is a time consuming and expensive process and the homology among genes from different species can facilitate the identification of genes of the target species from the genomic information of a model species. This study aimed to quantify the expression of homologous rice genes previously related to drought tolerance in Arabidopsis. Five genes (CPK6, PLDa, GluR2, CesA8, and EIN2) were identified in rice by the homology of the amino acid sequence between rice and Arabidopsis. The genotypes Douradão (drought tolerant) and Primavera (drought susceptible) were subjected to a water deficit experiment, and subsequently evaluated for gene expression by qPCR for the five homologous and Lsi1 genes. The qPCR analysis clearly showed that the five homologous genes were expressed in rice, which is an indication that these genes could preserve their function in rice as a response to drought. In Douradão, of the five homologous genes, all but OsGluR2 displayed an increase in the average expression in drought treatment when compared to the control, while in Primavera, the average expression of the five genes did not differ between the control and drought treatment. In Douradão, the OsPLDa1, which showed the higher expression level in drought in relation to the control (10.82), significantly increased the gene expression in the leaf and root tissues as a response to drought, in both vegetative and reproductive stages, whereas in Primavera, this gene was suppressed in both tissues and stages under drought. Therefore, the OsPLDa1 gene was the most important in relation to drought response and is an interesting candidate for further studies in developing rice cultivars that are more tolerant to this stress.