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Processing of laser speckle contrast images : study of mathematical models and use of nonlinear analyses to investigate the impact of aging on microvascular blood flow / Traitement d’images de speckle laser : étude à partir de modèles mathématiques et utilisation d’analyses non linéaires pour appréhender l’effet de l’âge sur la microcirculation sanguineKhalil, Adil 05 April 2017 (has links)
Le vieillissement est un facteur de risque des maladies cardiovasculaires. Il est associé à des altérations fonctionnelles et structurelles du système vasculaire.Une étude approfondie du processus de vieillissement et le développement de systèmes d’imagerie et des traitements de données associés deviennent donc une priorité. Par l’analyse d’images de contraste par speckle laser (LSCI), l’objectif de cette thèse est d’étudier l’influence de l’âge sur la micro circulation.Pour ce faire, des données de LSCI ont été acquises sur l’avant-bras de sujets sains jeunes et âgés. A partir de modèles mathématiques, nous avons déterminé la vitesse des érythrocytes de la micro circulation chez les deux groupes de sujets. Par ailleurs, nous avons également mené une étude de la complexité de séries temporelles d’ LSCI s’appuyant sur des mesures d’entropie multi échelle. Nos résultats montrent que : 1) le groupe de sujets plus âgés présente des valeurs de vitesse des globules rouges significativement plus élevées que celles des sujets jeunes à l’hyperémie réactive post-occlusive; 2) les fluctuations des séries temporelles de LSCI dans le groupe des sujets jeunes ont une complexité supérieure à celles du groupe de sujets âgés. Ces modifications observées sur la micro circulation pourraient être attribuées à des modifications du système vasculaire dans son ensemble. La compréhension de ces altérations pourrait conduire à de nouvelles perspectives en matière de prévention et de traitement des pathologies liées à l’âge. / Aging is a primary risk factor for cardiovascular diseases. It is associated with functional and structural alterations in the vascular system. Therefore, a deep study of the aging process and the development of imaging systems and associated processing become of the utmost importance. By processing laser speckle contrast images (LSCI), this PhD work aims at studying the influence of age on microcirculation. In our work, LSCI data were acquired from the skin forearm of healthy subjects, subdivided into two age groups (younger and older). From mathematical models, we determined red blood cells velocity in microcirculation in the two groups of subjects. Moreover, we applied multiscale entropy-based algorithms to LSCI time series in order to study the complexity of microvascular signals. Our main findings are: 1) the older group has significantly higher velocity values than the younger group at post-occlusive reactive hyperaemia; 2) LSCI fluctuations in the younger group have significantly higher complexity than those of the older group. Age-related changes in skin microcirculation can be attributed to alterations in the vascular system as a whole. Understanding these changes in the microcirculatory system may give new insights for prevention and treatment of age-related diseases
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Control of scattered coherent light and photoacoustic imaging : toward light focusing in deep tissue and enhanced, sub-acoustic resolution photoacoustic imaging / Contrôle de la lumière cohérente diffusée et imagerie photoacoustique : focalisation de la lumière en profondeur dans les tissus biologiques et imagerie photoacoustique améliorée avec résolution sub-acoustiqueChaigne, Thomas 07 January 2016 (has links)
En microscopie, savoir focaliser la lumière à l’échelle micrométrique est déterminant. Dans les tissus biologiques néanmoins, les inhomogénéités du milieu diffusent la lumière, empêchant toute focalisation au-delà d’une profondeur de l’ordre du millimètre. Des techniques de façonnage de front d’onde ont été développées afin de pré-compenser la distorsion du faisceau lumineux induite par la propagation à travers un milieu diffusant. Pour parvenir à focaliser la lumière à l’intérieur même du milieu diffusant, l’enjeu est de mesurer l’intensité lumineuse en profondeur de manière non invasive. Nous proposons d’utiliser l’effet photoacoustique pour sonder cette intensité. Une structure optiquement absorbante éclairée par une impulsion lumineuse émet en effet un signal ultrasonore, dont l’amplitude est proportionnelle à l’intensité lumineuse. Ces ultrasons se propagent de façon quasi-balistique dans les tissus mous et peuvent donc être détectés à l’aide d’un transducteur acoustique externe. Cette mesure permet donc de déterminer l’intensité lumineuse éclairant l’absorbeur. Nous avons montré qu’il était possible d’utiliser l'imagerie photoacoustique pour mesurer la matrice de transmission d’un échantillon diffusant. Cette caractérisation nous permet de focaliser la lumière sur des structures absorbantes et de sonder des propriétés mésoscopiques du milieu diffusant. Nous avons montré que la large bande spectrale des signaux photoacoustiques permet d’améliorer la focalisation. Enfin, nous avons montré que l’utilisation d’une source de lumière cohérente permet de pallier certains artefacts de l’imagerie photoacoustique, ainsi que de franchir la limite de résolution acoustique. / Light focusing is a crucial requirement for high resolution optical imaging. In biological tissue though, refractive index inhomogeneities scatter light, preventing any focusing beyond one millimeter. Wavefront shaping techniques have been recently developed to partially compensate for light scattering after propagation through a scattering medium. These techniques require a measurement of the light intensity at the target point. These techniques hold much promise for performing wavefront correction in order to focus light deep inside scattering media. This would require a non-invasive measure of the light intensity at depth. In this PhD study, we propose to use the photoacoustic effect for such task. An optically absorbing structure under pulsed illumination indeed generates ultrasonic waves, whose amplitude is proportional to the absorbed light intensity. These ultrasounds mostly propagate in a ballistic way, and can therefore be detected with an external transducer. We have shown that photoacoustic imaging could be used to measure the transmission matrix of a scattering sample, enabling to focus light on absorbing structures as well as to retrieve mesoscopic properties of the medium. We have shown that the broadband spectral content of the photoacoustic signals can be harnessed to improve the focusing performances. Finally, we demonstrated that coherent illumination could be used to remove fundamentals artefacts, as well as to break the acoustic resolution limit of conventional deep tissue photoacoustic imaging.
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Approche multiparamètre de l'analyse du champ de speckle et applications / A multiparameter approach for the analysis of the speckle field and applicationsAbou Nader, Christelle 30 June 2016 (has links)
Quand une source de lumière cohérente éclaire un milieu présentant soit une rugosité de surface à l’échelle de la longueur d’onde de la lumière incidente, soit des variations d’indice de réfraction, les ondes diffusées se propagent suivant différents chemins optiques. Les ondelettes ainsi créées, toujours cohérentes, interfèrent entre elles dans un plan d’observation, créant un champ de speckle. Dans ce travail, nous couplons l’analyse du champ de speckle à l’analyse de la polarisation de la lumière en adoptant une approche multiparamètre regroupant des informations spatiales et temporelles. La capacité du speckle polarisé à caractériser des propriétés de milieux diffusants complexes est exploitée. Des informations liées à un changement de proportions de tailles de diffuseurs, de coefficient d’absorption, et de dynamique intrinsèque des diffuseurs sont extraites. Les méthodologies développées sont appliquées à la caractérisation d’échantillons biologiques, à la détection de phases préliminaires d’érosion dentaire, et au suivi de réactions de polymérisation. / When a coherent light source illuminates a random medium, having either surface roughness or refractive index variations, the scattered waves propagate following different optical paths. The coherent created wavelets interfere with each other in a certain observation plane, creating the speckle field. In this work, we simultaneously the analysis of the speckle field as well as the analysis of polarized light, in a multi-parameter approach combining spatial and temporal information. The capability of the polarized speckle field to characterize complex scattering media is investigated. Information related to changes in diffusers sizes and proportions, absorption coefficient, and intrinsic dynamics are extracted. The developed methodologies are applied to characterize biological samples, to diagnose early stages of dental erosion, and to monitor polymerization reactions.
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Dynamique des gaz quantiques ultrafroids dans des milieux aléatoires corrélés / Dynamics of ultracold quantum gases in correlated disordered potentialsAlamir, Ardavan 17 December 2013 (has links)
La problématique de cette thèse est l'étude de la localisation d'un condensat de Bose-Einstein confiné harmoniquement et quasi-1D à travers lequel différents potentiels désordonnés sont transportés. Cette problématique qui se veut pleinement pertinente pour les expérimentalistes est à priori difficile à traiter. Cela est dû au caractère non-linéaire, inhomogène et hors-équilibre du système. De ce fait, la plage des vitesses du désordre est limitée d'une part par la vitesse critique de superfluidité et d'autre part par la configuration inhomogène du système. Des notions habituelles de localisation telles que transmission ou exposant de Lyapunov ne sont plus applicables. Donc, il a fallu apporter une nouvelle mesure de localisation pour notre problématique: le ratio du déplacement du centre de masse du condensat au déplacement du désordre qu'on a identifié à la fraction d'atomes localisés. De plus, nous avons des corrélations dans le désordre qui introduisent l'effet d'un comportement non-monotone de l'efficacité de la localisation du potentiel désordonné en fonction de l'énergie. Ainsi, les corrélations peuvent être un moyen pour mettre en évidence la nature quantique de la localisation. Chose que nous avont fait dans un premier temps avec du désordre de type Modèle d'Edwards et dans une seconde partie avec du désordre de type speckle, qu'on nomme le Random Dimer speckle. Pour ce deuxième cas, nous avons proposé une procédure pour contrôler les corrélations et introduire un pic de localisation dans une certaine région énergétique. Cette configuration pourrait être vérifié dans les expériences à l'aide d'un modulateur spatial de lumière. / The topic of this thesis is the study of localization of a quasi-one-dimensional and harmonically trapped Bose-Einstein condensate through which various disordered potentials are transported. This problem, which wants itself to be fully relevant to experimenters, is a priori difficult to deal with. This is due to the non-linear, inhomhogeneous and out-of-equilibrium nature of the system. Because of this, the range of speeds of disorder is limited on one side by the critical speed of superfluidity and on the other side by the inhomogeneous setting of the system. Usual notions of localization like transmission and Lyapunov exponent are no longer applicable. Thus, we had to introduce a novel measure of localization for our problem: the ratio of the distance moved by the condensate center of mass to the distance moved by the disordered potential that we identify as the fraction of localized atoms. Furthermore, we have correlations in the disorder that introduce the effect of non-monotonic behavior of the localization efficiency of the disordered potential as a function of energy. A a result, correlations can be used as a tool to point the quantum nature of the localization. We did this in a first part with Edwards Model type disorders and in a second part with speckle type disorders, a new one that we call the Random Dimer speckle. For this second part, we propose a scheme to control the correlations and introduce a localization peak in a certain energy region. This device can be verified in experiments with the help of a Spatial Light Modulator.
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Etude expérimentale de l'instabilité de digitation visqueuse de fluides rhéofluidifiants modèles / Experimental study of viscous fingering instability of shear thinning fluid modelsChinaud, Maxime 17 December 2010 (has links)
Ce travail de thèse est consacré à l'étude des instabilités de Saffman-Taylor de fluides complexes modèles. Ces derniers sont des solutions de Xanthane dont le caractère rhéofluidifiant augmente avec la concentration en polymère. Dans un premier temps, nous avons étudié les propriétés rhéologiques de ces fluides modèles puis nous avons caractérisé leurs propriétés d'écoulement dans une cellule de Hele-Shaw de forte épaisseur. Dans un deuxième temps, nous avons mesuré par PIV (Particles Image Velocimetry) la distribution du champ de vitesse autour des doigts de Saffman-Taylor pour l'ensemble des fluides étudiés. Afin de valider les expériences de PIV, nous avons caractérisé les vitesses de sédimentation des traceurs, par la technique complémentaire de vélocimétrie par Speckle ultrasonore (Ultrasonic Speckle Velocimetry). Nous avons montré expérimentalement que le champ de vitesse pour tous les fluides est irrotationnel et que la forme du doigt peut être modélisé par un écoulement potentiel autour d'un solide de Rankine. Les expériences ont établi que le rayon de courbure à l'extrémité des doigts est l'unique paramètre qui dépend des propriétés rhéologiques des solutions de Xanthane. De plus, ce paramètre conditionne toute l'hydrodynamique autour des doigts de Saffman-Taylor. / This thesis is devoted to the study of Saffman-Taylor instabilities of complex fluids models. These are solutions of xanthan whose shear thinning behavior increases with polymer concentration.Initially, we studied the rheological properties of model fluids and then we characterized the properties of flow in a Hele-Shaw thick. In a second step, we measured by PIV (Particle Image Velocimetry) distribution of the velocity field around the fingers of Saffman-Taylor for all fluids studied. To validate the PIV experiments, we characterized the sedimentation rates of tracers, by the complementary technique of ultrasonic speckle velocimetry (Ultrasonic Speckle Velocimetry).We have shown experimentally that the velocity field for all fluids is irrotational and that the shape of the finger can be modeled by a potential flow around a Rankine solid. Experiments have shown that the radius of curvature at the top of the fingers is the only parameter which depends on the rheological properties of xanthan solutions.In addition, this parameter influences the whole hydrodynamic around Saffman-Taylor fingers.
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En jämförelsestudie mellan manuell, semiautomatisk och automatisk utlinjering av endokardiet vid bedömning av ejektionsfraktionen. / A comparative study between manual, s semiautomatic and automatic tracing of the endocardium the assessment of ejection fraction.Petersson, Ida January 2020 (has links)
Introduktion: Vid ekokardiografi används ultraljudsvågor med frekvens över 20 000Hz. Ultraljudssändare med piezoelektriska kristaller används för att generera ljudvågor som skickas ut i kroppen, reflekteras och sedan återvänder till kristallerna för att skapa en ultraljudsbild. Ejektionsfraktionen uttrycker hur stor del av den diastoliska volymen som pumpas ut i kroppen under systole och är en viktig parameter vid bedömning av den globala systoliska funktionen. Den rekommenderade metoden Simpsons biplan jämförs med Philips automatiska funktion a2Dq, som utgår ifrån speckle tracking principen, för bedömning av ejektionsfraktionen. Material och metod: 32 hjärtfriska testpersoner mellan 20–43 år inkluderades i tvärsnittsstudien. Apikala 4- och 2-kammarbilder insamlades ifrån testpersonerna och användes för beräkning av ejektionsfraktionen. Den manuella metoden Simpsons biplan grundas på operatörens utlinjering av endokardiet. Den automatiska funktionen a2Dq utgår ifrån 53 hjärtmodeller och placerar en region of interest automatiskt. Den semiautomatiska funktionen används genom att operatören modifierar region of interest men inte längden av vänsterkammaren. Resultat: Resultatet visade att det inte förelåg någon signifikant skillnad i mätvärdena av ejektionsfraktionen mellan de tre mätprinciperna. Den semiautomatiska och manuella metoden som är operatörsberoende hade likvärdiga mätresultat. Den automatiska funktionen erhöll totalt 6 mätvärden som hamnade utanför normalgränserna. Diskussion: Automatiska funktioner för bedömning av ejektionsfraktionen utvecklas snabbt och kräver utvärderingar. Automatiska mätningar har påvisats vara fördelaktiga då de inte är så tidskrävande som den manuella Simsons biplanmetoden. Tidsaspekten framhävs i de flesta studier dock på bekostnad av säkra mätresultat. Observationerna i denna studie påvisar att den automatiska funktionen inte är säker nog för användning. Konklusion: Enligt observationer ifrån denna studien rekommenderas att den automatiska funktionen ifrån Philips inte används. / Introduction: Ultrasound are soundwaves with frequencies above 20 000Hz. Piezoelectric crystals are used to generate sound waves that can be transmitted into the tissue and reflected back to the crystals to obtain an ultrasound image. The ejection fraction expresses how much blood the left ventricle pumps out with each contraction and is an important parameter for assessment of the hearts global systolic function. The recommended method Simpson´s biplane to assess ejection fraction is compared with Philips automatic function a2Dq, which is based on the principle of speckle tracking. Material and method: The study population included 32 tests subjects with age between 20-43 years old. The test subjects had no records of earlier heart pathologies. Apical 4- and 2-chamber images were collected for the purpose to measure the ejection fraction. The manual method Simpson´s biplane is based on the sonographers manual tracking of the endocardial border. The automatic function a2Dq is based on 53 heart models with different anatomy and physiology and applies a region of interest automatically along the endocardial border. The semi-automatic function allows the sonographer to adjust the region of interest but not the length of the chamber. Result: The result showed that there was no significant difference in the measured values of ejection fraction between the three different techniques. The semi-automatic and Simpson´s biplane method showed equivalent results. The automatic function showed a wide variance of measurements which resulted with a total of 6 measurement ended up below the normal values for ejection fraction. Discussion: Automatic functions for assessing the ejection fraction rapidly develops and requires validations. Automatic functions have shown to be advantageous as the are not as time-consuming as the manual Simpson´s biplane method. In most studies, however, the time aspect is emphasized at the expense of correct measurements results. The observations in this study show that the automatically function a2Dq from Philips are not providing correct measurements. Conclusion: According to observations in this study, it´s recommended that the automatic function from Philips shouldn’t be used.
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\"Estudo da evolução da reação de presa do cimento de fosfato de zinco através de luz laser, aplicando a técnica óptica de speckle dinâmico\" / Study of evolution of zinc phosphate cement´s setting reaction by laser, applying an optics technique of dynamic speckleKon, Dino Jeiset 09 October 2006 (has links)
A cimentação de próteses e/ou de outros elementos protéticos constituem-se em uma preocupação permanente, já que uma das grandes aplicações do cimento de fosfato de zinco é como agente cimentante. No entanto, a reação de presa do cimento de fosfato de zinco, assim como o tempo de sua presa final, não estão ainda completamente esclarecidos. O presente estudo visa analisar, microestruturalmente, a evolução da reação de presa do cimento de fosfato de zinco. Mais especificamente, buscou-se mensurar, baseado no fenômeno conhecido como speckle dinâmico, o tempo de duração da reação de presa do cimento de fosfato de zinco, desde o momento do término da espatulação até a sua presa completa, em ambiente com temperatura e umidade relativa do ar controlada. A metodologia utilizada consiste na análise do speckle dinâmico obtido através da luz laser, em um ensaio não-destrutivo. A partir dos dados coletados pela técnica óptica de speckle, usando algoritmos computacionais, foi possível estabelecer os coeficientes de correlação temporal e momentos de intensidade. O tratamento estatístico permitiu relacionar e comparar com outros parâmetros tradicionais, químicos e mecânicos macroestruturais já existentes e estudados por outros autores. Pode-se concluir que os resultados alcançados pela nova técnica e pelos métodos convencionais são convergentes. / The prosthesis cementation and/or other prosthetic elements, consists in permanent concern, since one of the major applications of zinc phosphate cement is as lutting agent. This study aims at the microstructural analyze to the evolution of the zinc phosphate cement´s setting reaction. More specifically, searched to verify, based on the known phenomenon as dynamic speckle, how long takes the setting reaction of zinc phosphate cement, since the moment of the ending of the mix until your complete setting, in environment with air relative humidity and temperature controlled. The used methodology consist in the analysis of dynamic speckle gotten through the laser, in a non-destructive assay. From the data supplied by the laser speckle, it was possible to establish the temporal correlation coefficients and intensity moments. The statistical treatment, allowed to compare and relate with other traditional, chemical and mechanical macrostructural parameters, already existing and studied by other authors. It can be conclude that outcomes achieved by the new technique and with the conventional methods are convergent.
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RhéoSpeckle : un nouvel outil d’étude du comportement multi-échelle des matériaux hétérogènes / RheoSpeckle : a new tool for studying the behavior of heterogeneous materials at multi-scalesAli, Nabil 12 February 2016 (has links)
Ce travail de thèse cherche à mieux caractériser le comportement des matériaux hétérogènes sous cisaillement avec une approche multi-échelles (macro-méso-microscopique).Cela est rendu possible en développant un montage innovant qui couple un rhéomètre à un système d’imagerie de speckle résolue spatialement et temporellement (RheoSpeckle). Nous montrons la validation de notre expérience en l’appliquant sur deux matériaux parfaits : un solide et un liquide. Sur le solide, on mesure le champ de déplacement sur les images de speckle avec une résolution meilleure que 1 µm. Puis on prouve l’élasticité du matériau à l’échelle microscopique. Sur le liquide, la taille des nanoparticules est déterminée avec un excellent accord avec la spécification du fabriquant. Le champ de vitesse dans l’entrefer du Couette est calculé avec une bonne précision sur un temps inférieur à 1 s et avec une résolution spatiale de 100 µm sur 5mm. La dynamique microscopique d’une solution brownienne est étudiée et l’influence du cisaillement sur la décorrélation est déterminée. Nous montrons les capacités de notre expérience à étudier des matériaux hétérogènes en l’appliquant sur une solution concentrée de micelles géantes. La rhéologie linéaire est étudiée en rhéometrie classique mais aussi en utilisant l’imagerie du speckle. La rhéologie non linéaire de ce matériau est déterminé en rhéometrie (macro) mais aussi en calculant le champ de vitesse et l’intensité des images de speckle (méso) ou on caractérise les bandes de cisaillement qui se forment à partir d’un cisaillement critique. En fin la relaxation spatio-temporelle des bandes de cisaillement (micro) est caractérisée. On observe pour la première fois l’existence de deux temps de relaxation après l’arrêt du cisaillement et que la relaxation des bandes est relativement lente. / This work tries to better characterize the behavior of homogeneous and heterogeneous materials under shear with a multi-scale approach (macro-meso-micro-scopic). To do that, we have developed an innovative setup by coupling a rheometer to a speckle imaging geometry witch is spatially and temporally resolved (RheoSpeckle). We validate our experience using two perfect materials: a solid and a pure viscous fluid. On a solid sample, we calculate the displacement field on the speckle images with a resolution better than 1 µm. we demonstrate than, the microscopic elasticity of this material. On a pure viscous fluid, we measured the nanoparticle’s size with excellent accuracy. When a constant shear rate is applied, the velocity profile is measured with a time less than 1 s with a spatial resolution of 100 µm over 5 mm. The microscopic dynamic of a Brownian solution under shear is probed and the shear induced on the decorrelation of the intensity correlation function is studied. We show the capabilities of our experience using a concentrated solution of wormlike micelles. The linear rheology is studied using rheometric measurements and our speckle imaging system. Nonlinear rheology is studied using rheometric measurements (macro), but also by calculating the velocity filed and the intensity of speckle images (meso). With mesocopics measurements, the formation of shear banding is proved and characterized. Finally, the spatio-temporal relaxation (micro) of shear bands of this material is studied. We show for the first time the existence of two relaxations times after shear and that the relaxation of bands is relatively slow.
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Avaliação de fibrose miocárdica pelo strain, comparado ao achado de realce tardio da ressonância magnética cardíaca, em pacientes portadores de cardiopatia chagásica crônica / Assessment of myocardial fibrosis by strain, compared with late enhancement by the cardiac magnetic resonance imaging in patients with chronic cardiopathy Chagas\' diseasePaladino Filho, Antonio Tito 07 June 2016 (has links)
Desde o primeiro relato a Doença de Chagas permanece endêmica na América Latina com 18 milhões de pessoas cronicamente infectadas e, aproximadamente, 200.000 novos casos por ano. Doença parasitária causada por Trypanosoma cruzi, tem como transmissor \"natural\" da doença um inseto hematófago, o Reduris (barbeiro). Este inseto se torna infectado ao ingerir o sangue de um animal ou indivíduo infectado com o Trypanosoma cruzi. O contágio ocorre basicamente nas áreas rurais nas quais o homem, frequentemente, está em contato com hospedeiros e vetores ao destruir a mata nativa. Apesar da infecção geralmente ocorrer nos primeiros anos de vida, os pacientes infectados podem manifestar os sinais e sintomas da cardiopatia chagásica até 20 anos mais tarde. O custo propriamente dito e o sofrimento humano representados pela Doença de Chagas são grandes. Medicamentos, internações hospitalares frequentes e tratamento com dispositivos de alto custo (ex: marcapasso/ cardiodesfibrilador). Com quadro clínico insidioso, os pacientes podem apresentar insuficiência cardíaca franca, eventos tromboembólicos, arritmias ventriculares, dor torácica atípica e morte súbita. Seu diagnóstico baseia-se em epidemiologia positiva, anamnese, exame físico, alterações eletrocardiográficas, radiológicas e testes sorológicos. O envolvimento cardíaco é a principal causa de morte, sendo que a fisiopatologia e a evolução clínica da doença não são completamente compreendidas e a estratificação de risco permanece um desafio. A presença de disfunção miocárdica associada ou não à doença arterial aterosclerótica vem acompanhada de áreas de fibrose miocárdica e tem se mostrado como importante fator de pior prognóstico. A Ressonância Magnética Cardíaca (RMC) é um método já consagrado na detecção de fibrose miocárdica pela técnica de realce tardio (RT). Os objetivos dessa tese são: 1-Correlacionar o strain miocárdico nos segmentos do ventrículo esquerdo com as áreas de realce tardio detectadas na Ressonância Magnética Cardíaca, 2- Correlacionar o strain global do ventrículo esquerdo avaliado pela técnica de Speckle Tracking com a extensão de fibrose (numero de segmentos) avaliada pela Ressonância Magnética Cardíaca; 3- Comparar a Fração de ejeção avaliada pelo Ecocardiograma bidimensional e a Ressonância Magnética Cardíaca em pacientes com cardiopatia chagásica crônica; 4- Avaliar se existe relação entre a fração de ejeção avaliada pelo ecocardiograma bidimensional e a extensão de fibrose (número de segmentos) pela Ressonância Cardíaca. Foram selecionados 31 pacientes com o diagnóstico confirmado de Doença de Chagas. 27 pacientes que respeitaram os critérios de inclusão e exclusão, e que assinaram o termo de consentimento livre e esclarecido realizaram Ressonância Magnética Cardíaca com a técnica de realce tardio. De todos os pacientes selecionados, 27 completaram o protocolo da RMC e realizaram em sequencia o Ecocardiograma transtorácico para avaliar o strain miocárdico (speckle tracking) em intervalo máximo de 5 dias. Todos que completaram o protocolo não apresentaram efeitos adversos e os exames foram considerados interpretáveis. Para calcular a amostra de pacientes necessária, os cálculos foram feitos pela estatística de teste t, com distribuição t de Student, para comparação entre duas médias, com poder de teste de 80% e nível de significância de 5%, o que resultou numa estimativa de tamanho mínimo de amostra de 22 lesões por grupo (44 lesões) para que seja possível identificar diferenças significativas em todas as regiões numa amostra com medidas de mesmo comportamento que as observadas em Yajima et al e Jitsuo Higaki et al. Com isso, a amostra mínima seria de 22 pacientes. Para avaliar a concordância entre variáveis quantitativas, utilizou-se o coeficiente de concordância de Lin com intervalo de confiança (IC) 95% estimado segundo método Bootstrap. O coeficiente de concordância de Lin (Rc) combinada precisão e acurácia para determinar se observações desviam-se significativamente da linha de perfeita concordância (linha de 45 graus com origem no 0 dos eixos x e y). O coeficiente de Kappa de Cohen com IC 95% foi utilizado em analises de concordância para variáveis categóricas. Valores de p foram calculados utilizando-se método exato. Os critérios de Landis & Koch foram utilizados na interpretação dos coeficientes de concordância, assim definidos: (a) quase-perfeita, para valores de 0,81 a 1,00; (b) substancial, para valores de 0,61 a 0,80; (c) moderada, para valores entre 0,41 e 0,60; (d) regular, para valores entre 0,21 e 0,40; (d) discreta, para valores de 0 a 0,20. As estimações de intervalos de confiança 95% via bootstrap basearam-se em 1000 replicações. Estimativas de sensibilidade, especificidade, valores preditivo positivo e negativo com IC95% foram determinados. Variáveis quantitativas foram comparadas entre 2 grupos independentes utilizando-se testes não-paramétrico de Mann-Whitney com métodos exatos de cálculo do valor-p. Variáveis quantitativas comparadas com Mann-Whitney foram descritas com mediana e intervalo interquartil. Dado o reduzido tamanho de amostra, analise de regressão logística binaria univariada foi conduzida utilizando método exato. Para covariável quantitativa foi testada, a suposição de linearidade com o log-odds no modelo de regressão logística através da construção de \"Smoothed Scatter Plots\". Quando suposição não foi satisfeita, covariável originalmente quantitativa foi dicotomizadas segundo a mediana da distribuição. Odds ratios e seus respectivos intervalos de confiança 95% foram estimados. Analises de correlação entre variáveis quantitativas foram conduzidas utilizando-se coeficiente de correlação de Spearman(rho)(com IC95%). Forte correlação foi definida como |rho| >=0.70; correlação moderada para 0.5 <= |rho| <0.7 ; correlação fraca a moderada para 0.3<= |rho| <0.5; e fraca para |rho| <0.3. Normalidade foi avaliada com a inspeção visual de histogramas e aplicação do teste de normalidade Shapiro-Wilks. Todos os valores de p apresentados são do tipo bilateral: p < 0.05 e 0.05<p<= 0.10 foram considerados significantes e marginalmente significantes respectivamente. O software R (R Foundation, Vienna, Austria) foi utilizado na análise estatística de dados. Resultados: Dos 27 pacientes do estudo temos a idade média dos pacientes de 53,1 +- 7,0 anos, com mediana de 54 anos, sendo 08 (29,7%) homens e 19 (70,3%) mulheres. A fração de ejeção média pelo ecocardiograma foi de 55,1 +- 14,7%, e pela Ressonância Magnética Cardíaca foi de 55,8 +- 13,4%. O número total de segmentos avaliados foi 453 (98,7%), com interpretação não realizada em apenas 6 segmentos (1,3%) pelo ecocardiograma, de um total de 459. Encontramos realce tardio em 61 segmentos analisados e do total, 86 segmentos apresentaram alteração do strain. Tendo como padrão ouro o realce tardio na RMC, comparamos segmento a segmento o resultado entre essa e o ecocardiograma (speckle tracking). Levamos em conta a literatura que descreve uma prevalência da fibrose em portadores de cardiopatia chagásica crônica de aproximadamente 40%. Com esses dados obtivemos uma sensibilidade de 95%, especificidade de 91%, Valor preditivo positivo de 88,6% e valor preditivo negativo de 96,6%. Correlacionando o Strain Global Longitudinal (SGL) e a extensão (número de segmentos com realce tardio pela RMC) da fibrose neste trabalho, não observou-se diferença nas medianas de número de segmentos na CMR comparadas entre os grupos Strain Global Longitudinal - normal x alterado (p=0.287 , teste exato de Mann-Whitney). Utilizando como limites de fração de ejeção do ventrículo esquerdo >= 55% como normal e <55% alterada, comparamos à extensão de fibrose- numero de segmentos- ( realce tardio) pela CMR. Observou-se uma diferença marginalmente significativa na comparação das medianas do numero de segmentos com realce tardio na CMR entre os 2 grupos (p=0.064). ¹based on exact Mann-Whitney test. Comparamos também a fração de ejeção do ventrículo esquerdo pela RMC e pelo Ecocardiograma transtorácico utilizando o método de Simpson. Ao nível de significância de 5% (coeficiente de Lin), observou-se uma quase perfeita concordância entre FEVE pela Ressonância Magnética Cardíaca e FEVE pelo Ecocardiograma transtorácico utilizando em ambos o método de Simpson. (Rc = 0.9335 IC95% 0.878-0.957; N=27). / Since the first report, Chaga\'s disease remains endemic in Latin America with 18 million chronically infected people and approximately 200,000 new cases per year. Parasitic disease caused by Trypanosoma cruzi, is \"natural\" transmitter disease of a hematophagous insect, the Reduris (barber). This insect becomes infected by ingesting blood of an animal or individual infected with Trypanosoma cruzi. The infection occurs primarily in rural areas where men often are in touch with hosts and vectors by destroying the native forest. Despite the infection usually occurs early in life, infected patients may exhibit signs and symptoms of Chagas disease 20 years later. The cost itself and human suffering represented by Chagas disease are a public issue. Medication, frequent hospitalization and treatment with costly devices (eg pacemaker / defibrillator) are frequently necessary. With insidious clinical signs and symptons, patients may present with heart failure, thromboembolic events, ventricular arrhythmias, atypical chest pain and sudden death. Its diagnosis is based on positive epidemiology, history, physical examination, electrocardiographic, radiological and serological changes. Cardiac involvement is the leading cause of death, and the pathophysiology and clinical course of the disease are not fully understood and the risk stratification remains a challenge. The presence of myocardial dysfunction with or without atherosclerotic arterial disease is accompanied by myocardial fibrosis areas and has been an important factor of poor prognosis. Cardiac Magnetic Resonance (CMR) is a method already established in the detection of myocardial fibrosis by delayed gadolinium enhancement technique (DGE). The objectives of this thesis are: 1-To correlate myocardial strain in left ventricular segments with areas of late enhancement detected in Cardiac Magnetic Resonance, 2- correlate the global left ventricular strain measured by Speckle Tracking technique with fibrosis extension (number of segments) evaluated by Cardiac Magnetic Resonance; 3- Compare the ejection fraction assessed by Two-dimensional echocardiography and cardiac Magnetic Resonance in patients with chronic cardiac Chagas\' disease; 4- To assess whether there is a relationship between the ejection fraction assessed by two-dimensional echocardiography and fibrosis extension (number of segments) by CMR. We selected 31 patients with confirmed diagnosis of Chagas\' disease. 27 patients who complied with the inclusion and exclusion criteria, and who signed the informed consent , performed Cardiac Magnetic Resonance with late gadolinium enhancement (DGE) technique. Of all the selected patients, 27 completed the RMC protocol and were in sequence directed to the transthoracic echocardiogram to assess myocardial strain (speckle tracking) in maximum interval of 5 days. Everyone who completed the protocol showed no adverse effects and the tests were considered interpretable. To calculate the sample required patients, we used the test statistic t, with t Student distribution for comparison between two averages, with the power to test 80% and 5% significance level, which resulted in an estimate minimum sample of 22 injuries per group size (44 injuries) so you can identify significant differences in all regions in a sample with the same measures of behavior than those observed in Yajima et al and Jitsuo Higaki et al. Thus, the minimum sample size would be 22 patients. To evaluate the correlation between quantitative variables, we used the Lin\'s concordance coefficient (1,2) with a confidence interval (CI) 95% estimated second method Bootstrap (4). The correlation coefficient Lin (Rc) combines precision and accuracy to determine if observations deviate significantly from perfect correlation line (45 degree line with origin 0 of the x and y axes). Cohen\'s kappa coefficient with 95% (3) was used for analysis of agreement for categorical variables. P values were calculated using the exact method. The Landis and Koch criteria (5) were used in the interpretation of correlation coefficients defined as follows: (a) quasi-perfect for values 0.81 to 1.00; (b) substantial amounts of 0.61 to the 0.80; (c) Moderate to values between 0.41 and 0.60; (d) regular, to between 0.21 and 0.40; (d) mild to values from 0 to 0.20. The estimation of 95% confidence intervals by bootstrap based on 1000 replicates. Quantitative variables were compared between two independent groups using non-parametric Mann-Whitney test with exact methods of calculating the p-value. (6-8) Quantitative variables compared with Mann-Whitney were described as median and interquartile range. Given the small sample size, regression analysis univariate binary logistic regression was conducted using exact method (9-11). For quantitative covariate was tested, the linearity assumption with the log-odds in the logistic regression model by building \"Smoothed Scatter Plots\". (9) When assumption was not met, originally quantitative covariate was dichotomized according to the median of the distribution. Odds ratios and their 95% confidence intervals were estimated. All significance probabilities (p values) presented are the bilateral type and values less than 0.05 considered statistically significant. The R (R Foundation, Vienna, Austria) software was used for statistical analysis of data. In order to measure the correlation between the results of both tests in the study were calculated sensitivity, specificity, positive predictive value and negative predictive value considering the CMR as the gold standard. Results: Of the 27 study patients have a average age of 53.1 + - 7.0 years and median of 54 years. 08 (29.7%) men and 19 (70.3%) women. The average ejection fraction by echocardiography was 55.1 + - 14.7%, and by Cardiac Magnetic Resonance was 55.8 + - 13.4%. The total number of evaluated segments was 453 (98.7%), with no interpretation performed in only 6 segments (1.3%) on echocardiography, a total of 459 segments. Delayed enhancement by Cardiac Magnetic Ressonance was found in 61 segments analyzed. In the order \"side\" 86 segmets registered an altered strain. As the gold standard the delayed enhancement in the MRC, compared to segment studied segment. We take into account the literature describing the prevalence of fibrosis in patients with chronic Chagas\' heart disease of approximately 40%. With this data we obtained a sensitivity of 95%, specificity 91%, positive predictive value of 88.6% and a negative predictive value of 96.6%. Correlating the Global Longitudinal Strain (SGL) and extent (number of segments with delayed enhancement by CMR) of fibrosis in this study, no difference was observed in the number of segments of medians in the CMR compared between Strain Global Longitudinal groups - Normal x changed (p = 0.287, exact Mann-Whitney). Using as an ejection fraction of the left ventricle limits > = 55% as normal and < 55% changed, compared to the extent of fibrose- number of segments- (delayed enhancement) by CMR. There was a marginally significant difference when comparing the median number of segments with delayed enhancement on CMR between the 2 groups (p = 0.064). ¹based on exact Mann-Whitney test. We also compared the ejection fraction of the left ventricle by CMR and transthoracic echocardiography using the Simpson method. At a significance level of 5% (Lin coefficient), there was an almost perfect correlation between LVEF by Cardiac Magnetic Resonance and LVEF by using transthoracic echocardiography in both the Simpson method. (Rc = 0.9335 95% CI 0878-0957; N = 27)
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Mesure des déplacements cellulaires dans les tissus non transparents : une application de la diffusion dynamique de la lumière / Measuring cell displacements inside non-transparent tissues : an application of dynamic light scatteringBrunel, Benjamin 29 October 2018 (has links)
Lorsqu'une tumeur grossit, elle exerce une pression sur les tissus environnants et est comprimée en retour. Des expériences sur un modèle de tumeur in vitro, appelé sphéroïde, ont montré que cette pression influence largement le devenir du tissu cancéreux, notamment en freinant sa croissance, mais aussi en le rendant plus invasif. Pour mieux comprendre ce dernier effet, nous avons cherché à étudier le comportement migratoire des cellules à l'intérieur d'un sphéroïde sous pression. Observer l'intérieur d'un sphéroïde pose cependant un problème technique car les méthodes usuelles d'imagerie ne sont pas utilisables dans des tissus épais (> 100 μm). L'imagerie classique étant limitée en profondeur à cause de la diffusion de la lumière, nous nous sommes tournés vers une méthode qui utilise justement celle-ci : la diffusion dynamique de lumière ou DLS (Dynamic Light Scattering). Nous avons développé son application à la migration cellulaire, afin d'obtenir la distribution des déplacements relatifs des cellules au cours du temps. Cette mesure est faite sans utiliser de marqueurs spécifiques et est applicable à des sphéroïdes allant jusqu'à 400 μm de diamètre. Nous avons ainsi mis en évidence une organisation radiale du sphéroïde en termes de mobilité, avec des cellules rapides en surface et plus lentes au centre. Nous avons aussi montré qu'en appliquant une contrainte au sphéroïde, la vitesse moyenne diminue jusqu'à être réduite de moitié pour des pressions supérieures à 15kPa. Une autre équipe a mesuré par ailleurs une augmentation de la vitesse des cellules en surface suite à une compression, ce qui indique que l'organisation radiale se retrouve dans la réponse à la pression. Nous avons montré que cette sensibilité à la pression est une propriété qui émerge de l'organisation 3D du tissu, dans laquelle la matrice extracellulaire joue un rôle primordial. Enfin, pour explorer les possibilités qu'offre notre technique, nous l'avons appliquée à une autre question : comment la migration des macrophages est-elle affectée par les signaux provenant de cellules apoptotiques ? Les résultats ont montré que les cellules apoptotiques précoces augmentent la vitesse des macrophages tandis que les cellules apoptotiques tardives la réduisent. D'un cas à l'autre, la longueur de persistance du mouvement est conservée. / As a tumor grows, it exerts a mechanical pressure on its surrounding tissue and is compressed back as a reaction. Recent experiments on an in vitro tumor model, called spheroid, have shown that this pressure is crucial for the fate of the cancerous tissue. In particular, the pressure slows down its growth, but makes it more invasive. To further understand the latter effect, we decided to study the migration of cells inside spheroids under pressure. However, imaging the inside of a spheroid is technically challenging as usual microscopy methods do not work on thick tissues (> 100 μm). Standard imaging methods are limited in terms of depth penetration because of light scattering. For this reason, we decided to take advantage of this scattered light with a method called Dynamic Light Scattering (DLS). We developed its application to cell migration in order to measure the distribution of cells displacements over time. The measurement is label-free and works with spheroids as thick as 400 μm in diameter. By this means, we revealed a radial organization inside the spheroid in terms of mobility, with fast cells at the surface and slower cells in the core. We also showed that applying a pressure onto spheroids decreases the average cell speed by a factor up to two for pressure greater than 15 kPa. Another team reported an increase in the speed of cells located at the surface of a compressed spheroid, which implies that the radial organization is also true for the impact of pressure. We demonstrated that this sensitivity to an external pressure is a 3D emergent property, in which the extracellular matrix plays an essential role. Finally, we explored the potential of our technique by addressing another question: how do apoptotic cells signals affect the migration of macrophages? We found that early apoptotic cells increase the speed of macrophages whereas late apoptotic cells decrease it. In both cases, the persistence length of the motion is the same.
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