Caractérisation des ataxies hérédodégénératives par spectroscopie de résonance magnétique nucléaire in vivo

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal

Synthèse et caractérisation physicochimique de peptides de polyglutamines

Neuf maladies neurodégénératives sont le produit de l’expression de gènes mutés, dans lesquels le codon CAG est répété au-delà d’un seuil pathologique. Ceci produit des protéines mutantes dans lesquelles sont insérés des segments de polyglutamines (polyGln), qui perdent leur activité et acquièrent une nouvelle fonction, ce qui est toxique pour le neurone. Ces altérations sont attribuables aux propriétés particulières de la polyGln. En effet, ces dernières possèdent la capacité de s’assembler pour former des corps d’inclusion intracellulaires. Cette propension à l’agrégation de la polyGln rend difficile l’étude de ces pathologies. C’est ainsi que l’utilisation de peptides peut s’avérer une approche avantageuse. Toutefois, la synthèse de polyGln est associée à de nombreuses délétions et nécessite l’ajout de groupements chargés afin de permettre leur purification. Cependant, ce prérequis donne lieu à des interactions électrostatiques qui biaisent la structure et la cinétique d’agrégation de ces peptides, en plus d’interférer avec l’évaluation d’éventuels agents thérapeutiques. L’objectif du projet est de développer un système permettant l’étude de la polyGln en s’affranchissant des effets de charges. Pour ce faire, deux approches ont été explorées, la première utilise la polyGln non chargée et la seconde utilise une structure polyGln-morpholine ayant des charges labiles en fonction du pH. Ces peptides ont été produits en utilisant une approche linéaire de synthèse peptidique sur support solide avec protection maximale des chaînes latérales. La purification a été effectuée par chromatographie de haute performance en phase inverse en milieu acide. Ces stratégies ont permis de produire des peptides de polyGln de grande pureté avec des rendements acceptables. Une procédure de solubilisation des peptides alliant sonication et lyophilisation a été développée afin d’étudier chacun de ces peptides à l’aide de diverses techniques physicochimiques, telles que la diffusion de la lumière, la spectroscopie de résonance magnétique nucléaire, Raman et UV-visible, le dichroïsme circulaire et la microscopie optique polarisée. La polyGln non chargée solubilisée dans le trifluoroéthanol-eau a montré que la taille des particules et la vitesse d’agrégation sont proportionnelles à la fraction volumique en eau. De plus, la structure secondaire en solution est à prédominance alpha et semble être peu sensible à la fraction d’eau jusqu’à un certain seuil (25%) après lequel la structure aléatoire prédomine. L’analyse des agrégats à l’état solide montre des structures hélicoïdales > aléatoires et ont les caractéristiques des fibrilles amyloïdes. Le peptide de polyGln-morpholines a un pKa de 7,3 en milieu aqueux. Il demeure en solution lorsque le pH bêta > alpha. De plus, en RMN, l’élargissement des signaux du 1Hγ en cours d’agrégation suggère une interaction via les chaînes latérales. Les analyses en phase solide ont plutôt montré une prédominance de structures bêta et alpha. L’inhibition de l’agrégation à pH 8 varie selon rouge de Congo > tréhalose, alors que le peptide liant la polyGln 1 et la thioflavine T ne semble pas avoir d’effet. Ces approches ont donc permis pour la première fois de s’affranchir des effets de charges auparavant inhérents à l’étude de la polyGln en solution et par conséquent d’obtenir des informations inédites quant à la solubilité, la structure et la cinétique d’agrégation. Enfin, le dispositif à charges labiles permet d’évaluer l’efficacité d’éventuels agents thérapeutiques à pH quasi physiologique.Nine neurodegenerative diseases come from mutated genes expression in which the CAG codon is repeated above a pathological threshold. This is producing mutant proteins, in which are inserted polyglutamine (polyGln) segments, which lose their activity and acquire a new function that is toxic for the neuron. These alterations are related to the peculiar properties of the polyGln. Indeed, these polypeptides have the capacity of autoassemble to form intracellular inclusion bodies. This aggregation tendency of polyGln makes difficult the study of these pathologies. Thus, the use of peptides could constitute an advantageous approach. However, the synthesis of polyGln is associated with numerous deletions and necessitates the addition of charged moieties to achieve purification. Unfortunately, this requirement creates electrostatic interactions that modify the structure and aggregation kinetics of these peptides, in addition to interfering with the evaluation of potential therapeutical agents. The aim of this project is to develop a system to study polyGln without the charge effects. To do so, two approaches were explored, the first used uncharged polyGln and the second used a polyGln-morpholine structure bearing pH-dependent labile charges. These peptides were produced by solid-support synthesis using a linear and maximal protection approach. Purification was performed by reverse phase high-performance liquid chromatography. These strategies allowed the production of peptides of high purity in good yields. A solubilization procedure combining sonication and lyophilization was developed to study each of these peptides by physicochemical techniques such as light scattering, magnetic resonance, Raman and UV-visible spectroscopies, circular dichroism and polarized optical microscopy. The uncharged polyGln solubilized in trifluoroethanol-water showed that particle size and aggregation kinetics are proportional to volumetric water fraction. Furthermore, the secondary structure in solution is alpha-predominant and seems rather insensitive to water fraction up to a threshold (25%) above which random coil structure predominates. The analysis of solid-state aggregates showed that helicoidal structures are more abundant than random structures and have the characteristics of amyloid fibrils. The polyGln-morpholines peptide has a pKa of 7.3 in aqueous media. It is soluble when pH beta > alpha. Furthermore, the broadening of 1Hγ NMR signals while the peptide is aggregating suggests an interaction between side-chains. Solid-phase studies showed predominant beta and alpha structures. The aggregation inhibition at pH 8.0 was higher for Congo red than for trehalose, while polyglutamine binding peptide 1 and thioflavine T did not seem to be effective. These approaches permitted for the first time to overcome the charge effects that were previously inherent to polyGln solution studies and to obtain new information about solubility, structure and aggregation kinetics. Finally, the labile charge groups allow the evaluation of the efficiency of potential therapeutic agents at near physiological pH

Reaching within a dynamic virtual environment

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A kinematic analysis of a haptic handheld stylus in a virtual environment: a study in healthy subjects

<p>Abstract</p> <p>Background</p> <p>Virtual Reality provides new options for conducting motor assessment and training within computer-generated 3 dimensional environments. To date very little has been reported about normal performance in virtual environments. The objective of this study was to evaluate the test-retest reliability of a clinical procedure measuring trajectories with a haptic handheld stylus in a virtual environment and to establish normative data in healthy subjects using this haptic device.</p> <p>Methods</p> <p>Fifty-eight normal subjects; aged from 20 to 69, performed 3 dimensional hand movements in a virtual environment using a haptic device on three occasions within one week. Test-retest stability and standardized normative data were obtained for all subjects.</p> <p>Results</p> <p>No difference was found between test and retest. The limits of agreement revealed that changes in an individual's performance could not be detected. There was a training effect between the first test occasion and the third test occasion. Normative data are presented.</p> <p>Conclusion</p> <p>A new test was developed for recording the kinematics of the handheld haptic stylus in a virtual environment. The normative data will be used for purposes of comparison in future assessments, such as before and after training of persons with neurological deficits.</p

Evaluation of a modern-analogue methodology for reconstructing Australian palaeoclimate from pollen

Quantitative palaeoclimate reconstructions are widely used to evaluate climatemodel performance. Here, as part of an effort to provide such a data set for Australia, we examine the impact of analytical decisions and sampling assumptions on modern-analogue reconstructions using a continent-wide pollen data set. There is a high degree of correlation between temperature variables in the modern climate of Australia, but there is sufficient orthogonality in the variations of precipitation, summer and winter temperature and plant–available moisture to allow independent reconstructions of these four variables to be made. The method of analogue selection does not affect the reconstructions, although bootstrap resampling provides a more reliable technique for obtaining robust measures of uncertainty. The number of analogues used affects the quality of the reconstructions: the most robust reconstructions are obtained using 5 analogues. The quality of reconstructions based on post-1850 CE pollen samples differ little from those using samples from between 1450 and 1849 CE, showing that European post settlement modification of vegetation has no impact on the fidelity of the reconstructions although it substantially increases the availability of potential analogues. Reconstructions based on core top samples are more realistic than those using surface samples, but only using core top samples would substantially reduce the number of available analogues and therefore increases the uncertainty of the reconstructions. Spatial and/or temporal averaging of pollen assemblages prior to analysis negatively affects the subsequent reconstructions for some variables and increases the associated uncertainties. In addition, the quality of the reconstructions is affected by the degree of spatial smoothing of the original climate data, with the best reconstructions obtained using climate data froma 0.5° resolution grid, which corresponds to the typical size of the pollen catchment. This study provides a methodology that can be used to provide reliable palaeoclimate reconstructions for Australia, which will fill in a major gap in the data sets used to evaluate climate models

The intramembrane protease SPPL2a promotes B cell development and controls endosomal traffic by cleavage of the invariant chain

Peer reviewe

Computational models and motor learning paradigms: Could they provide insights for neuroplasticity after stroke? An overview

Computational approaches for modelling the central nervous system (CNS) aim to develop theories on processes occurring in the brain that allow the transformation of all information needed for the execution of motor acts. Computational models have been proposed in several fields, to interpret not only the CNS functioning, but also its efferent behaviour. Computational model theories can provide insights into neuromuscular and brain function allowing us to reach a deeper understanding of neuroplasticity. Neuroplasticity is the process occurring in the CNS that is able to permanently change both structure and function due to interaction with the external environment. To understand such a complex process several paradigms related to motor learning and computational modeling have been put forward. These paradigms have been explained through several internal model concepts, and supported by neurophysiological and neuroimaging studies. Therefore, it has been possible to make theories about the basis of different learning paradigms according to known computational models. Here we review the computational models and motor learning paradigms used to describe the CNS and neuromuscular functions, as well as their role in the recovery process. These theories have the potential to provide a way to rigorously explain all the potential of CNS learning, providing a basis for future clinical studies

Children reading to dogs: a systematic review of the literature

Background Despite growing interest in the value of human-animal interactions (HAI) to human mental and physical health the quality of the evidence on which postulated benefits from animals to human psychological health are based is often unclear. To date there exist no systematic reviews on the effects of HAI in educational settings specifically focussing on the perceived benefits to children of reading to dogs. With rising popularity and implementation of these programmes in schools, it is essential that the evidence base exploring the pedagogic value of these initiatives is well documented. Methods Using PRISMA guidelines we systematically investigated the literature reporting the pedagogic effects of reading to dogs. Because research in this area is in the early stages of scientific enquiry we adopted broad inclusion criteria, accepting all reports which discussed measurable effects related to the topic that were written in English. Multiple online databases were searched during January-March 2015; grey literature searches were also conducted. The search results which met the inclusion criteria were evaluated, and discussed, in relation to the Oxford Centre for Evidence Based Medicine levels of evidence; 27 papers were classified as Level 5, 13 as Level 4, 7 as Level 2c and 1 as Level 2b. Conclusion The evidence suggests that reading to a dog may have a beneficial effect on a number of behavioural processes which contribute to a positive effect on the environment in which reading is practiced, leading to improved reading performance. However, the evidence base on which these inferences are made is of low quality. There is a clear need for the use of higher quality research methodologies and the inclusion of appropriate controls in order to draw causal inferences on whether or how reading to dogs may benefit children’s reading practices. The mechanisms for any effect remain a matter of conjectur

A global multiproxy database for temperature reconstructions of the Common Era

Source at https://doi.org/10.1038/sdata.2017.88 .Reproducible climate reconstructions of the Common Era (1 CE to present) are key to placing industrial-era warming into the context of natural climatic variability. Here we present a community-sourced database of temperature-sensitive proxy records from the PAGES2k initiative. The database gathers 692 records from 648 locations, including all continental regions and major ocean basins. The records are from trees, ice, sediment, corals, speleothems, documentary evidence, and other archives. They range in length from 50 to 2000 years, with a median of 547 years, while temporal resolution ranges from biweekly to centennial. Nearly half of the proxy time series are significantly correlated with HadCRUT4.2 surface temperature over the period 1850–2014. Global temperature composites show a remarkable degree of coherence between high- and low-resolution archives, with broadly similar patterns across archive types, terrestrial versus marine locations, and screening criteria. The database is suited to investigations of global and regional temperature variability over the Common Era, and is shared in the Linked Paleo Data (LiPD) format, including serializations in Matlab, R and Python

Anisotropic nanomaterials: structure, growth, assembly, and functions

Comprehensive knowledge over the shape of nanomaterials is a critical factor in designing devices with desired functions. Due to this reason, systematic efforts have been made to synthesize materials of diverse shape in the nanoscale regime. Anisotropic nanomaterials are a class of materials in which their properties are direction-dependent and more than one structural parameter is needed to describe them. Their unique and fine-tuned physical and chemical properties make them ideal candidates for devising new applications. In addition, the assembly of ordered one-dimensional (1D), two-dimensional (2D), and three-dimensional (3D) arrays of anisotropic nanoparticles brings novel properties into the resulting system, which would be entirely different from the properties of individual nanoparticles. This review presents an overview of current research in the area of anisotropic nanomaterials in general and noble metal nanoparticles in particular. We begin with an introduction to the advancements in this area followed by general aspects of the growth of anisotropic nanoparticles. Then we describe several important synthetic protocols for making anisotropic nanomaterials, followed by a summary of their assemblies, and conclude with major applications