Dissertationen zum Thema „Functional UltraSound“

L'objectif de cette thèse était d'explorer le potentiel de l’imagerie du cerveau humain par ultrasons. L'anatomie, le flux sanguin et la rigidité des tissus mous ont déjà été étudiés avec l'imagerie ultrasonore ultrarapide chez l'homme et validés sur plusieurs organes, tels que le sein et le foie, mais pas encore sur le cerveau adulte. La principale limitation de l'imagerie échographique transcrânienne est aujourd'hui le très fort artefact d'aberration induit par le crâne. En effet, l’os, de par sa composition ne permet pas la propagation des ultrasons comme ailleurs dans le corps humain. Dans cette thèse, nous avons utilisé l'imagerie ultrasonore ultrarapide pour l'évaluation de la rigidité des tissus mous et l'imagerie neurofonctionnelle dans le cerveau humain adulte, lors de chirurgies du cerveau afin de contourner dans un premier temps le problème des aberrations induites par le crâne. La dernière partie de cette thèse était axée sur la correction d’aberration pour l’échographie quantitative et l’imagerie ultrasonore transcrânienne. Nous avons tout d’abord fourni plusieurs preuves de l'intérêt d'utiliser l’élastographie par onde de cisaillement pendant la chirurgie du cerveau. Nous avons également présenté notre nouvelle technique d’élastographie par onde de cisaillement en 3D à l'aide d'une sonde matricielle dans le but de pouvoir dépasser les limitations du 2D et notamment être moins dépendant de l’opérateur.Dans un second volet, nous avons démontré la capacité des ultrasons ultrarapides à identifier, cartographier et différencier en profondeur les régions d'activation corticales en réponse à un stimulus, à la fois chez les patients éveillés et chez les patients anesthésiés. Nous avons démontré que l'imagerie neurofonctionnelle par ultrasons a le potentiel de devenir une modalité complète de neuroimagerie avec des avantages majeurs pour une utilisation peropératoire.Dans un troisième volet, nous avons utilisé une technique en trois étapes pour calculer précisément la vitesse du son (SSE) dans un milieu. Cette technique a été testée dans des fantômes ultrasonores et in vivo dans les foies de patients. Dans les deux cas, notre méthode a été capable de trouver la vitesse du son correspondant au milieu. Nous avons démontré que la SSE était liée à la fraction de graisse. Cette analyse a permis de conclure que la SSE était en mesure de distinguer un foie sain et d’un foie malade aussi bien avec la biopsie qu’avec l’IRM comme méthode de référence. Combiné à l'utilisation de la formule de Wood, nous avons même pu avoir accès à une fraction de graisse mesurée par ultrasons de manière non invasive. Puis nous avons combiné la correction d’aberration de phase, d'amplitude et de vitesse du son pour faire de l’imagerie transcrânienne en simulation numérique. Nous avons atteint notre objectif en obtenant des images représentant fidèlement le milieu (position latérale et profondeur) et caractérisées par une résolution et un contraste similaires à ceux obtenus avec une source ponctuelle dans le milieu
The objective of this thesis was to explore the potential of human brain ultrasound imaging. Anatomy, blood flow and soft tissue stiffness have already been studied with ultrafast ultrasound imaging in humans and validated in several organs, such as, the breast and liver but not yet on the adult brain. The main limitation of transcranial ultrasound imaging is today the very strong skull-induced aberration artefact. Indeed, the bone, due to its composition, does not allow for ultrasound propagation as elsewhere in the human body. Therefore, this thesis was focused on the development of ultrafast ultrasound imaging for the evaluation of soft tissue stiffness and neurofunctional imaging in the adult human brain, during brain surgery to bypass the problem of skull aberration, and on an aberration correction technique for transcranial ultrasound imaging.We first provided several evidence of the benefit of using shear wave elastography during brain surgery. We also presented our new technique for 3D shear wave elastography using a matrix array in order to be able to overcome the limitations of 2D imaging and in particular to reduce the operator dependence.In a second phase, we demonstrated the capability of ultrasound to identify, map and differentiate in depth cortical regions of activation in response to a stimulus, both in awake patients and in anaesthetized patients. We have demonstrated that ultrasound neurofunctional imaging has the potential to become a comprehensive modality of neuroimaging with major benefits for intraoperative use. In a third part, we developed a new sound speed estimation (SSE) technique, based on a three-step technique that estimates the sound speed accurately corresponding to the illuminated medium. This technique was tested in ultrasound phantoms and in vivo in patient’s liver. In both cases, our method was able to find the sound speed corresponding to the medium. We demonstrated that SSE was related to the fat fraction. This analysis led to the conclusion that SSE was able to distinguish a healthy liver from a diseased liver with both biopsy and MRI as gold standard. Combined with the use of the Wood’s formula, we were even able to access a fat fraction measured by non-invasive ultrasound. Finally, by combining the phase, the amplitude and the sound speed estimation, we have developed a new aberration correction algorithm to perform transcranial ultrasound imaging. By performing numerical simulations, we obtained images that faithfully represented the medium (lateral position and depth) and characterized by one resolution and one contrast similar to those obtained with a punctual source in the medium

Depuis le milieu du XXème siècle, les techniques d’imagerie fonctionnelles ont un rôle grandissant dans notre compréhension sur les fonctions du cerveau en conditions physiologique et pathologique. Bien que l’IRMf fasse partie des techniques les plus communément utilisées pour l’imagerie du cerveau complet lors d’études préclinique et clinique, cette modalité souffre de sa résolution spatiotemporelle et sa sensibilité pour enregistrer finement les fonctions et activités cérébrales. Récemment l’imagerie fonctionnelle par ultrason (ifUS) a subi des développements permettant d’être complémentaires à l’IRMf ainsi qu’aux autres techniques d’imagerie cérébrales classiquement employées. Contrairement aux ultrasons focalisés conventionnels, l’imagerie hémodynamique proposé par l’ifUS repose sur une illumination ultrasonore plane permettant la détection des globules rouges en mouvement et la mesure de leur vitesse dans les micro-vaisseaux cérébraux. De ce fait, l’ifUS est indirectement lié à l’activité cérébrale d’où l’importance d’une meilleure compréhension des mécanismes du couplage neuro-vasculaire liant l’activité neuronale et les variations cérébrales d’apport en sang. De plus, cette technique a le potentiel pour fournir des informations précises sur les processus de certaines pathologies à la fois sur des modèles précliniques et chez l’homme. Dans un premier temps, j’exposerais mes travaux sur les récents développements techniques permettant l’ifUS in vivo (i) en condition chronique, (ii) sur l’animal éveillé, libre de mouvement et effectuant une tache comportementale et (iii) des vaisseaux capillaires chez le rongeur et l’homme. Dans un second temps, je démontrerais que l’ifUS in vivo peut fournir des informations nouvelles sur des pathologies telles que l’accident vasculaire cérébrale
Since the middle of the 20th century, functional imaging technologies are making an increasing impact on our understanding on brain functions in both physiological and pathological conditions. Even if fMRI is nowadays one of the most used tool for whole brain imaging in pre-clinical and clinical studies, it lacks sufficient spatiotemporal resolution and sensitivity to assess fine brain function and activity. Functional ultrasound imaging (fUSi) has been recently developed and presents a potential to complement fMRI and other existing brain imaging modalities. Contrary to conventional ultrasound using focus beams, fUSi relies on hemodynamic imaging based on ultrasound plane-wave illumination to detect red blood cells movement and velocity in brain micro-vessels. Consequently, the fUSi signal is indirectly related to brain activity and it is therefore important to better understand the mechanisms of the neurovascular coupling linking neural activity and cerebral blood changes. Here again, fUSi may provide relevant information about disease processes in preclinical models but also in humans. First, I will present recent technical developments allowing in vivo fUSi (i) in chronic condition, (ii) in freely moving and behaving rats and (iii) in rodents and human brain capillaries. Second, I will demonstrate how fUSi could provide new insights in brain pathologies such as stroke

Ces travaux de thèse portent sur les récents progrès de l’imagerie fonctionnelle par ultrasons et ses nouvelles applications en ophtalmologie. Dans le cadre d’un projet mêlant physique des ondes, imagerie, neurosciences et ophtalmologie, nous avons appliqué cette technologie à l’imagerie du système visuel et à l’étude préclinique de thérapies le ciblant. Au cours de ce projet, nous avons accompagné nos études précliniques de progrès constants dans notre imagerie.Un dispositif permettant l’imagerie du cerveau en 3 dimensions a été conçu. Cette imagerie a été réalisée en temps réel, ou à une fréquence ultrasonore de 30 MHz grâce au procédé d’entrelacement. Grâce à une connaissance a priori de l’architecture vasculaire cérébrale et de l’effet Doppler, il est possible de réaliser une décomposition spectrale des écoulements sanguins cérébraux selon leurs vitesses et de leurs orientations.Ceci a permis une étude des fonctions visuelles du rat et du primate non-humain. Nous avons imagé la rétine du primate en Doppler de puissance, mais sa forte mobilité en fait un organe délicat à imager en imagerie fonctionnelle. En revanche, nous avons réalisé une imagerie fonctionnelle de la rétine de rat à 30 MHz. Nous avons caractérisé en détail le système visuel cérébral de ce rongeur. Nous avons mis en évidence ses principales structures et redémontré leurs caractéristiques les plus connues, comme leur organisation rétinotopique ou leur différence de temps de réponse neurovasculaire à un stimulus. Des animaux traités par des thérapies de restauration visuelle a été imagée. La première imagerie de primates non-humains anesthésiés puis éveillés et exécutant une tâche comportementale, et la détection de variations de flux sanguins dues à des erreurs uniques ont été réalisées. Enfin, une étude préclinique aiguë et une étude chronique de traitements ayant des effets neurovasculaires ont été menées grâce au suivi du flux sanguin par nos procédés
This thesis focuses on recent improvements in the functional ultrasound imaging (fUS) technique and their applications in the field of ophthalmology. Within the framework of a synergetic project blending waves physics, medical imaging, neuroscience and ophthalmology, fUS was shown to be capable of imaging and studying the visual system of healthy and diseased animals for the purpose of preclinical studies. To tackle these issues, constant upgrades in the fUS technique had to support the preclinical studies.An experimental set-up was built to image the visual pathway in three dimensions with fUS. Using a new imaging facility, fUS was proven to be feasible in real time and at high ultrasound frequencies such as 30 MHz. Interleaved sampling had to be implemented in that case. Furthermore, the a priori knowledge of the vascular cerebral architecture and the Doppler Effect were exploited to spectrally decompose cerebral blood flux and vessels according to their velocities and orientations.Leveraging these improvements, functional ultrasound imaging of rats and non-human primates was performed. Primate retina was imaged with Power Doppler, but proved to be too mobile to be functionally imaged. However, fUS has been performed on rat retina after 30 MHz fUS imaging had been implemented. The rat visual pathway has then been characterised with fUS. Some of its known features where highlighted such as its retinotopic organisation or the time response differences between some of its structures. The same set-up has been leveraged to map the cerebral activity of animal that underwent visual restauration therapies. These tools were then used to map cerebral activity in anesthetized and awake and behaving monkeys. Unique blood volume variations due to unique mistakes were detected. These tools were finally applied to two preclinical trials on a depressive state of the brain vascular contractility. Blood volume and blood velocity changes were highlighted throughout an acute and a chronical study

The hip joint center (HJC) is needed for calculation of hip kinematics in various applications. In the functional method, the center is determined by moving femur with respect to acetabulum. A popular way for measuring this movement is through an optical motion capture system. This method is fast and economical for most applications where we require an instant HJC even though the reconstruction error in bone position calculation exists due to skin artifact. This error is caused by movement of markers placed on skin rather than on actual bone. Here we introduce ultrasound imaging as an additional modality to measure the change in soft tissue thickness above bone while hip is flexed. We use this information on the tissue thickness change to recalculate position of markers placed on skin to match the movement of bone. A good advantage of using ultrasound machine is its non-invasiveness. We calculated HJC using a symmetric center of rotation estimation (SCoRE) algorithm, which uses the concept of coordinate transformation on 3D marker position data. The algorithm gives the 3D position of two centers, one for each hip bone. The distance between these two centers (SCoRE residual) gives us a hint on the accuracy of the HJC calculation and has been proved to be proportional to the error with respect to actual center in previous studies. These two centers should ideally coincide as they collectively form a spherical joint. Our new algorithm for HJC calculation with tissue thickness compensation, measured using ultrasound imaging shows the error has been reduced from 9.13 mm to 4.87 mm

The buckwheat is a naturally gluten-free pseudocereal, rich in phytochemicals, phenols and functional proteins, however, the functional characteristics of its flour in baking process are not as performing as those of wheat flour. The aim of this thesis was to analyse how the functional, viscosimetric and antioxidant properties of two variety of buckwheat (Polish and Italian) flours change in response to an ultrasound (US) pre-treatment carried out on different solid:liquid ratio. For this, in the differently treated Polish buckwheat samples some functional properties (water absorption index, water solubility index, swelling power, water holding capacity, water absorption capacity, foaming capacity and stability, emulsifying activity and emulsion stability) were analysed. Moreover, the thermoviscous test, antioxidant capacity and polyphenol content were quantified. Finally, bread samples, obtained with treated and untreated Polish buckwheat flour were analysed for baking loss, specific volume, colour analysis of crust, porosity and hardness. The obtained results showed that the US pre-treatment increased the water holding capacity, improved the foam stability, but decreased the ability to form foams and emulsions as well as the viscosity of the treated flour samples. The breads formulated with US treated flours had a more uniform pore distribution in the crumb and were softer than the breads formulated with untreated flour. Moreover, US pre-treatment had no effects on surface colour and phenolic content of obtained bread. In conclusion, the application of US on buckwheat flour can be a good alternative to the use of additives in formulation in order to obtain gluten-free and nutritionally rich final products.

Le monde extérieur regorge de sons complexes, que chaque animal doit interpréter afin de survivre. Pour ce faire, leur cerveau se doit de représenter toute la richesse de la structure acoustique de ces sons, jusque dans leurs propriétés les plus complexes. Dans cette thèse, cette question est explorée à travers un nouveau prisme, l'imagerie fonctionnelle ultrasonore (fUS). Dans un premier temps, l'imagerie fUS est utilisée pour étudier avec une haute fidélité l'organisation topographique du système auditif, ainsi que ses connexions avec d'autres aires cérébrales. Dans un deuxième temps, elle permet d'explorer des aspects fondamentaux de la façon dont le cortex auditif encode les sons naturels, ainsi que les spécificités humaines pour le traitement du langage. Enfin, elle révèle des formes topographiques mais non continues d'encodage, avec l'exemple de la localisation spatiale des sons. À travers ces trois aspects sont révélés les différents modules de traitement de l'information auditive, spatialement organisés, qui se superposent au sein d'une aire cérébrale unique
The world teems with complex sounds that animals have to interpret in order to survive. To do so, their brain must represent the richness of the sounds' acoustic structure, from simple to high-order features. Understanding how it does it, however, remains filled with challenges. In this thesis, these questions were explored through a new technical prism, namely functional UltraSound imaging (fUSi). First, fUSi was used to investigate with a high fidelity the topographical organization of the auditory system, as well as its connectivity with other brain areas. Second, it provided fundamental clues for our understanding of how natural sounds are encoded in the auditory cortex, and hints at the human particularities for speech processing. Last, it gave us access to non-continuous topographical encoding, with the example of spatial localization. Through these three aspects, we exposed the different spatially organized modules of processing that overlap within a single brain area

This thesis aims to advance the understanding of ultrasonic processing for the alteration of food microstructures. It considers the impact of ultrasonic processing upon protein structure and for emulsification. It was shown that ultrasound treatment of proteins reduced the hydrodynamic volume of protein aggregates by ultrasonic cavitations. Insufficient acoustic energy was provided to achieve proteolysis. Emulsions prepared with ultrasound treated milk protein isolate, pea protein isolate and bovine gelatin yielded smaller, stable emulsion droplets in comparison to their untreated counterparts. This behaviour is ascribed to more rapid adsorption of protein at the oil-water interface and improved interfacial packing, due to reduction in protein aggregate size. The droplet size of emulsions with sufficient emulsifier (> 0.5 wt. %) emulsion droplet size can be predicted from a mathematical relation between emulsion droplet size (d3,2) and energy density (Ev), an inverse power law. Droplet size predictions were unattainable at low emulsifier concentrations (≤ 0.5 wt. %) due to re-coalescence behaviour attributed to insufficiency of emulsifier and droplet collisions within the acoustic field. Continuous processing yields more efficient utilisation of acoustic energy in comparison to batch configurations due to the intense transmission of acoustic energy within the smaller processing volumes.

Cette thèse porte sur deux branches majeures de l'utilisation d'agents sensibles aux ultrasons: l'échographie ultrarapide du cerveau assistée par microbulles et la délivrance par ultrasons de médicaments pour la thérapie du cancer. Dans la première approche, des microbulles remplies de gaz fluoré ont été utilisés pour observer l'activation du cerveau à travers le crâne des rongeurs. Nous avons été en mesure de reconstituer de manière non invasive le réseau vasculaire du cerveau, puis de récupérer sa réponse hémodynamique avec une résolution spatio-temporelle élevée. La validation de cette approche d'imagerie fonctionnelle par échographie (FUS) a été facilitée par la grande sensibilité de la technique du Doppler ultrarapide ultrasensible. En effet, cette modalité d'imagerie permet de détecter les changements hémodynamiques dus au couplage neurovasculaire avec une grande résolution (1ms, 100pm). Ces résultats suggèrent que la combinaison des agents de contraste et l'imagerie ultrarapide peut aider à compenser entièrement l'atténuation par le crâne, et ce en préservant la résolution et en augmentant la profondeur de pénétration. L'injection d'agents de contraste ultrasonore a également conduit à des résultats remarquables en imagerie ultrasonore ultrarapide. La barrière de la diffraction a été contournée pour aller au-delà de la limite de demi-longueur d'onde de résolution. Nous avons démontré que des microvaisseaux cérébraux de 9pm de diamètre peuvent être distingués par microscopie échographie ultrarapide de localisation (uULM). Des millions de sources «clignotantes» sont localisées dans l'espace et dans le temps, conduisant à des images super-résolues (cartographie de densité de microbulles) de l'ensemble du réseau vasculaire du cerveau du rat avec une résolution spatiale de À / 10. En outre, les trajets des microbulles au cours du temps ont pu être relevés et ainsi permettre d'extraire les vitesses des flux sanguins avec une grande dynamique. Dans la seconde approche, nous avons exploité la manière dont nous pouvons contrôler, spatialement et temporellement, la vaporisation de micro gouttes composites de perfluorocarbone (PFC) lorsque leur activation est déclenchée par de courtes impulsions ultrasonore. Le concept de "chimie in-situ" est introduit dès lors que nous avons été en mesure de contrôler une réaction chimique spontanée in vitro. En outre, dans le cadre des applications in vivo de la chimie in situ, un nouveau dispositif microfluidique en verre a été proposé afin de permettre une production stable et rapide de gouttes monodisperses. Ce nouveau dispositif présente 128 générateurs en parallèles avec deux canaux sous pression. Finalement, de nouvelles séquences d'échographie de contrôle ultra-rapides ont été développées dans le but de contrôler et de surveiller la libération des gouttelettes composites
This thesis focuses on two main branches of the application of ultrasound contrast agents: microbubbles-aided ultrafast ultrasound imaging of the brain and ultrasound-triggered drug delivery for cancer therapy. At first, gas-filled microbubbles have been used to retrieve the brain activation through the skull in large animais. With this approach we have been able to non-invasively reconstruct the cerebral network of the brain, as well as retrieve its hemodynamic response to specific evoked tasks with high spatiotemporal resolution. The validation of this novel functional ultrasound (fUS) imaging approach was facilitated by the high sensitivity of the ultrasensitive Doppler technique able to detect subtle hemodynamic changes due to the neurovascular coupling. These resuits suggested that combining microbubbles injections with ultrafast imaging may help to fully compensate for the attenuation from the skull. Indeed, by combining both, we preserved resolution and increased penetration depth. The injection of ultrasound contrast agents has also lead to outstanding resuits in ultrafast ultrasound imaging by breaking the diffraction barrier and move beyond the half-wavelength limit in resolution. We have demonstrated that cerebral microvessels of 9pm in diameter can me distinguished via ultrafast ultrasound localization microscopy (uULM). Millions of blinking sources were localized in space and in time in few seconds in a higher dimensional space, leading to super-resolved images (microbubble density map) of the whole rat brain with a spatial resolution of À/10. Moreover, a displacement vector allowed microbubbles-tracking within frames yielding to in-plane velocity measurements retrieving a large dynamic of cerebral blood velocities. Next, we have exploited how we can spatiotemporally control the vaporization of composite perfluorocarbon (PFC) microdroplets when their activation is triggered by short ultrasound pulses. The concept 'chemistry in-situ' is introduced as we have been able to control a spontaneous chemical reaction in-vitro. Moreover, a new microfluidic device in glass has been proposed to robustly produce monodisperse droplets for future in-vivo applications of the chemistry in situ. This new device presents 128-parallel generators with two pressurized rivers. Eventually, new ultrafast ultrasound monitoring sequences have been developed in order to control and monitor the release of composite droplets

The effects of high intensity ultrasound (HIU) on the crystallization behavior and functional properties of shortenings were evaluated. HIU was applied to different shortenings such as interesterified soybean oil (IESBO), multi-purpose commercial shortening, palm oil, and palm stearin. The functional properties measured include crystal morphology, solid fat content, melting profile, viscoelastic properties, hardness, and polymorphism. Different experimental set-ups were evaluated including a static batch system, a temperature cycling design, and flow cell system. Results showed that HIU generated harder material on IESBO, commercial shortening, and palm oil with more uniform and smaller crystal size, sharper melting profile, and higher elasticity. No chemical changes on triacylglycerol (TAG) and fatty acids were observed on IESBO under the sonication conditions used in this dissertation. Application of HIU maintained the texture of the commercial shortening that was subjected to temperature fluctuations, especially when HIU was applied before changes in temperature occurred. When sonication was applied in a flow-cell system lower power levels applied in a continuous manner was proved to be the most effective at inducing crystallization of palm oil. Research also showed that pulse irradiation of sonication and higher flow rates could be used to decrease the thermal effects generated by higher power levels of HIU. In addition, HIU was used in a highly saturated fat (palm stearin) at low power levels with long durations to delay lipid crystallization and generate a softer material. All the research findings suggest the great potential use of HIU in shortening production and food processing to improve the texture and its stability, as well as other functional properties.

In obstetrics, the application of computer-based image analysis to provide deeper insight into pathology in early pregnancy is highly desirable but underdeveloped. One such pathology, fetal growth restriction (FGR) is a leading cause of mortality and morbidity in pregnancy. FGR affects approximately 3-10% of pregnancies in the western world leading to increased risk of stillbirth and health problems in later life. Morphometric or functional measurement of the placenta in pregnancy in utero may aid diagnosis of this pathology as the interface between placenta and mother is the site where the pathology manifests itself. Detection of growth restriction is yet to be resolved as poor, unreliable biochemical and image-based biomarkers have made it hard to detect and manage these pregnancies effectively. By the provision and development of tools for quantification of the placenta by three dimensional (3D) ultrasound (US) using image segmentation and mesh pro- cessing, this thesis aims to better facilitate clinical investigation of this major problem in obstetric healthcare. In a first contribution, 3D placental volume measurement using 3D US is used to classify the difference between normal and FGR pregnancies. Volume was estim- ated using the semi-automated random walker (RW) algorithm. The repeatability and reliability of the method was tested between three observers and showed the new method to be equivalent to manual segmentation. In a study of 143 women performed by our clinical partners, significantly smaller placental volume was found in pregnancies defined as small-for-gestational age (SGA). Expanding on volumetry, the utero-placental interface (UPI) is the location where the pathology that leads to FGR occurs. Manual manipulation of the volume is requiredtovisualisetheinterface, soweinvestigatedapplyinga“meshflattening” process to convert the contorted UPI into a disc to provide a standardised way to view the interface between acquistions and subjects. Finally, an existing, two dimensional (2D) Doppler standardisation technique was extended into 3D to provide standardisation of values of Doppler vascularity. This technique was then applied to measure the vascularity of volumes of interest relat- ive to the interface between placenta and mother. This test was then applied clin- ically in 143 women and found that the vascularity of the small-for-gestational age (SGA) pregnancies was significantly smaller than that of the population who produced appropriately sized babies. These three tools each provide augmentation of our understanding of placental health and function in pregnancy. From measuring the gross volume to estimat- ing the blood flow we show the potential clinical application for image analysis performed on 3D power Doppler (PD) ultrasound volumes.

The Achilles tendon is the largest tendon in the human body, which elastically stores and releases energy to facilitate walking and running. Tendons can suffer from a range of pathologies, most notably that of complete rupture, which affects athletes, physically active workers and the aged. There is a growing demand for in vivo methods of objectively measuring tendon health for aiding diagnosis, monitoring therapy and for assessment of new treatments. Knowledge of the changes in mechanical properties during the healing process is also limited and new methods to accurately and consistently estimate these could provide insights into the healing process and guide future research efforts. This thesis presents the development and use of 2D ultrasound elastography, a quantitative strain estimation imaging technique, as a tool to measure changes in the tensile mechanical properties of the Achilles tendon. This technique performs frame-to-frame block matching of image texture to track motion in an ultrasound signal sequence and create a strain estimation field from the spatial derivative of the motion. Elastography in the image-lateral direction of sagittal plane scans is of particular interest as this is in line with the longitudinal axis of the tendon, but presents extra accuracy issues from out of plane motion and lower image spatial resolution. Tendon rupture also presents unique problems to image acquisition and analysis- patient pain and safety are important considerations and disruption of the ultrasound texture can make 2D motion tracking more difficult. A new 2D elastography block matching algorithm, named `AutoQual', was developed to enable accurate tracking of motion in the image-lateral direction and reduce the impact of artefacts and errors common with damaged Achilles tendons image sequences. It was shown to outperform a multiscale block matching method when tested using ultrasound sequences from in vivo and gelatine phantom experiments. The input parameters of this algorithm were then optimised using the phantom data for benchmarking. The AutoQual algorithm was then used to analyse ultrasound sequences from a 24-week longitudinal study of 21 subjects with ruptured Achilles tendons to assess lateral, axial and principal strains during controlled passive motion of the foot or axial palpation of the ultrasound probe. Lateral and principal strains from controlled dorsiflexion were shown to be more repeatable and more sensitive to change than axial strains with manual palpation. This experience with lateral strain imaging from ruptured Achilles tendons gave an increased knowledge of the strain imaging artefacts and features that can occur. These are described in detail in order that they may be further mitigated in quantitative analysis by optimising acquisition protocols, further amendment of the block tracking algorithm, or exclusion of erroneous areas when selecting regions of interest. Regularisation is a potential solution to some common artefacts such as discontinuities from poor tracking in shadow regions. Regularisation of the lateral displacement fields is investigated using 2D bicubic smoothing splines. The regularisation parameters used are shown to have minimal effect on quantitative analysis and can aid visual clarity or reduce artefacts within certain settings. However, regularisation was also shown to cause large errors when parameters were set more aggressively. Finally, it is identified that cumulative lateral strain measurement of the Achilles and other tendons is feasible but that future work is needed to further improve the quality of force and cross sectional area measurements in order to infer mechanical properties accurately. Repeatable high force motion protocols also need to be developed to measure healthy tendons and to ensure comparable results between different patients and research groups.

Contactless ultrasonic power transfer (UPT), using piezoelectric transducers, is based on transferring energy using acoustic waves, in which the waves are generated by an acoustic source or transmitter and then transferred through an acoustic medium such as water or human tissue to a sensor or receiver. The receiver then converts the mechanical strain induced by the incident acoustic waves to electricity and delivers to an electrical load, in which the electrical power output of the system can be determined. The execution and efficiency of this technology can be significantly enhanced through patterning, focusing, and localization of the transmitted acoustic energy in space to simultaneously power pre-determined distributed sensors or devices. A passive 3D-printed acoustic hologram plate alongside a single transducer can generate arbitrary and pre-designed ultrasound fields in a particular distance from the hologram mounted on the transmitter, i.e., a target plane. This dissertation presents the use of these simple, cost-effective, and high-fidelity acoustic holograms in UPT systems to selectively enhance and pattern the electrical power output from the receivers. Different holograms are numerically designed to create single and multi-focal pressure patterns in a target plane where an array of receivers are placed. The incident sound wave from a transmitter, after passing through the hologram, is manipulated, hence, the output field is the desired pressure field, which excites the receivers located at the pre-determined focal points more significantly. Furthermore, multi-functional holograms are designed to generate multiple images at different target planes and driving frequencies, called, respectively, multi-image-plane and multi-frequency patterning holograms. The multiple desired pressure distributions are encoded on the single hologram plate and each is reconstructed by changing the axial distance and by switching the frequency. Several proof-of-concept experiments are performed to verify the functionality of the computationally designed holograms, which are fabricated using modern 3D-printers, i.e., the desired wavefronts are encoded in the hologram plates' thickness profile, being input to the 3D-printer. The experiments include measurement of output pressure fields in water using needle hydrophones and acquisition of receivers' voltage output in UPT systems. Another technique investigated in this dissertation is the implementation of acoustic impedance matching layers deposited on the front leading surface of the transmitter and receiver transducers. Current UPT systems suffer from significant acoustic losses through the transmission line from a piezoelectric transmitter to an acoustic medium and then to a piezoelectric receiver. This is due to the unfavorable acoustic impedance mismatch between the transducers and the medium, which causes a narrow transducer bandwidth and a considerable reflection of the acoustic pressure waves at the boundary layers. Using matching layers enhance the acoustic power transmission into the medium and then reinforce the input as an excitation into the receiver. Experiments are performed to identify the input acoustic pressure from a cylindrical transmitter to a receiver disk operating in the 33-mode of piezoelectricity. Significant enhancements are obtained in terms of the receiver's electrical power output when implementing a two-layer matching structure. A design platform is also developed that can facilitate the construction of high-fidelity acoustically matched transducers, that is, the material layers' selection and determination of their thicknesses. Furthermore, this dissertation presents a numerical analysis for the dynamical motions of a high-intensity focused ultrasound (HIFU)-excited microbubble or stable acoustic cavitation, which includes the effects of acoustic nonlinearity, diffraction, and absorption of the medium, and entails the problem of several biomedical ultrasound applications. Finally, the design and use of acoustic holograms in microfluidic channels are addressed which opens the door of acoustic patterning in particle and cell sorting for medical ultrasound systems.
Doctor of Philosophy
This dissertation presents several techniques to enhance the wireless transfer of ultrasonic energy in which the sound wave is generated by an acoustic source or transmitter, transferred through an acoustic medium such as water or human tissue to a sensor or receiver. The receiver transducer then converts the vibrational energy into electricity and delivers to an electrical load in which the electrical power output from the system can be determined. The first enhancement technique presented in this dissertation is using a pre-designed and simple structured plate called an acoustic hologram in conjunction with a transmitter transducer to arbitrarily pattern and shape ultrasound fields at a particular distance from the hologram mounted on the transmitter. The desired wavefront such as single or multi-focal pressure fields or an arbitrary image such as a VT image pattern can simply be encoded in the thickness profile of this hologram plate by removing some of the hologram material based on the desired shape. When the sound wave from the transmitter passes this structured plate, it is locally delayed in proportion to the hologram thickness due to the different speed of sound in the hologram material compared to water. In this dissertation, various hologram types are designed numerically to implement in the ultrasonic power transfer (UPT) systems for powering receivers located at the predetermined focal points more significantly and finally, their functionality and performances are verified in several experiments. Current UPT systems suffer from significant acoustic losses through the transmission from a transmitter to an acoustic medium and then to a receiver due to the different acoustic impedance (defined as the product of density and sound speed) between the medium and transducers material, which reflects most of the incident pressure wave at the boundary layers. The second enhancement technology addressed in this dissertation is using intermediate materials, called acoustic impedance matching layers, bonded to the front side of the transmitter and receiver face to alleviate the acoustic impedance mismatch. Experiments are performed to identify the input acoustic pressure from a transmitter to a receiver. Using a two-layer matching structure, significant enhancements are observed in terms of the receiver's electrical power output. A design platform is also developed that can facilitate the construction of high-fidelity acoustically matched transducers, that is, the material layers' selection and determination of their thicknesses. Furthermore, this dissertation presents a numerical analysis for the dynamical motions of a microbubble exposed to a high-intensity focused ultrasound (HIFU) field, which entails the problem of several biomedical ultrasound applications such as microbubble-mediated ultrasound therapy or targeted drug delivery. Finally, an enhancement technique involving the design and use of acoustic holograms in microfluidic channels is addressed which opens the door of acoustic patterning in particle and cell sorting for medical ultrasound systems.

La douleur neuropathique est une sensation de douleur anormale qui persiste au-delà du cours temporel de la guérison naturelle. Elle interfère avec la qualité de vie du patient et est associée à plusieurs comorbidités telles que l'anxiété et la dépression. Des études antérieures ont suggéré que la douleur chronique pourrait résulter d’une plasticité neuronale anormale et inadaptée dans les structures connues pour être impliquées dans la perception de la douleur (Bliss et al. 2016). Cela signifie qu'une lésion nerveuse déclencherait une potentialisation à long terme de la transmission synaptique dans les aires cérébrales liées à la douleur (Zhuo et al. 2014). Comme ces régions sont également impliquées dans les aspects émotionnels de la douleur, notre hypothèse est que la plasticité inadaptée susmentionnée dans ces zones cérébrales pourrait constituer un mécanisme clé pour le développement de comorbidités, telles que l'anxiété et la dépression.Au cours de ma thèse, nous avons choisi de tester cette hypothèse de travail par l’étude des altérations de la connectivité fonctionnelle (CF) intrinsèque des réseaux cérébraux par imagerie fonctionnelle ultrasonore (fUS) dans un modèle murin de douleur neuropathique. Cette technique de neuro-imagerie relativement récente a permis de nombreuses avancées en neurosciences, grâce à sa haute résolution spatio-temporelle, à sa sensibilité, mais aussi son adaptabilité, permettant des études chez l’animal anesthésié ou éveillé.Dans une première étude, j’ai mis au point un protocole expérimental permettant d’imager le cerveau des souris éveillées de façon reproductible et avec un minimum de stress et d artefacts de mouvements et ai également été impliquée dans le développement d’un nouvel algorithme d’analyse des signaux générées par ces acquisitions. Cette première approche étant réalisée avec une sonde linéaire en mouvement qui ne permet pas de visualiser l’entièreté du cerveau, dans une seconde étude, j’ai participe au développement d’une nouvelle technologie de sonde compilées et motorisée.Fort de ces développements technologiques, j’ai alors utilisé ces nouvelles approches pour tester mon hypothèse neurobiologique. J’ai entrepris deux études en parallèle chez des animaux anesthésiés pour l’une et éveillés pour la seconde, chez lesquelles nous avons étudié le lien temporel entre les altérations de la CF cérébrale et le développement de la douleur neuropathique et/ou des comorbidités associées. Pour cela, nous avons mesuré la CF (en période de repos) chez des souris atteintes de douleur neuropathique, à trois moments différents : I) 2 semaines après l’induction de la douleur neuropathique (manchon autour du nerf sciatique) II) à 8 semaines post-induction, lorsque l'anxiété émerge et III) à 12 semaines post-induction, lorsque la dépression apparait (12W). Ce suivi longitudinal a également été réalisé en parallèle sur un groupe d’animaux contrôles.Nos résultats indiquent des changements significatifs de la CF dans les principales régions cérébrales impliquées dans la transmission ou la modulation de la sensibilité ou de la douleur, suggérant la mise en place d’une plasticité inadaptée du réseau de la douleur, suite à la lésion nerveuse. De plus, nous observons une évolution temporelle de ces altérations, potentiellement corrélée à l'apparition des comorbidités associées. Ainsi, ces mécanismes pourraient participer à la chronicisation de la douleur
Neuropathic pain is an abnormal pain sensation that persists longer than the temporal course of natural healing. It interferes with the patient’s quality of life and leads to several comorbidities, such as anxiety and depression. It has been suggested that chronic pain may result from abnormal and maladaptive neuronal plasticity in the structures known to be involved in pain perception (Bliss et al. 2016). This means that nerve injury would trigger long-term potentiation of synaptic transmission in pain-related areas (Zhuo et al. 2014). Since these regions are also involved in the emotional aspects of pain, our hypothesis is that the aforementioned maladaptive plasticity in these brain areas could constitute a key mechanism for the development of comorbidities such as anxiety and depression.My PhD aimed at testing this working hypothesis, through the study of brain resting state functional connectivity (FC) using functional ultrasound imaging (fUS) in a mouse model of neuropathic pain. FUS is a relatively recent neuroimaging technique that enabled numerous advances in neuroscience, thanks to its high spatio-temporal resolution, its sensitivity, but also its adaptability, allowing studies in anesthetized or awake animals.In a first study, I developed an experimental protocol allowing the brains of awake mice to be imaged in a reproducible manner and with minimal stress and movement artifacts and was also involved in the development of a new algorithm for the analysis of the signals generated by these acquisitions. As this first approach was carried out with a moving linear probe which does not allow the entire brain to be visualized, in a second study, I participated in the development of a new compiled and motorized probe technology.Building on these technological developments, I then used these new approaches to test my neurobiological hypothesis. I undertook two parallel studies in animals anesthetized for one and awake for the second, in which we studied the temporal link between alterations in cerebral FC and the development of neuropathic pain and/or associated comorbidities. To do this, we measured the resting-state functional connectivity (FC) in anesthetized and in awake head-fixed mice, at three time points: I) 2 weeks after induction of neuropathic pain (cuff around the sciatic nerve), II) at 8 weeks post-induction during the emergence of anxiety (8W) and III) at 12 weeks post-induction during the emergence of depression. This longitudinal follow-up has been conducted concurrently on a control group.Our results show significant changes in FC in major pain-related brain regions in accordance with the development of neuropathic pain symptoms. These findings suggest that the pain network undergoes maladaptive plasticity following nerve injury which could contribute to pain chronification. Moreover, the time course of these connectivity alterations between regions of the pain network could be correlated with the subsequent apparition of associated comorbidities

L’échographie ultrarapide est une nouvelle méthode d'imagerie basée sur la transmission des ondes planes et qui permet d'imager les tissus biologiques à très haute cadence et avec une excellente résolution spatiale. Lorsqu'elle est appliqué à l'imagerie Doppler, elle permet d'améliorer considérablement la détection du flux sanguin dans les vaisseaux avec une sensibilité considérablement augmentée par rapport à l'imagerie conventionnelle basée sur la transmission des ondes focalisées. Ainsi le Doppler ultrarapide a ouvert d'autres champs d'applications à l'échographie faisant d’elle une nouvelle modalité précieuse de neuro-imagerie fonctionnelle capable de reconstruire la micro-vascularisation cérébrale mais aussi de mesurer indirectement l’activité neuronale en se basant sur le couplage neuro-vasculaire. Cependant l’expansion de l’imagerie fonctionnelle ultrasonore (fUS) est limitée par la difficulté à se repérer dans le réseau vasculaire complexe du cerveau, ce qui rend complexe le positionnement de la sonde échographique et l’analyse des données. Il est donc crucial de mettre en place des outils de neuro-informatique dédiés à l’imagerie fonctionnelle ultrasonore au même titre que l’imagerie fonctionnelle par IRM qui est la modalité de référence.Les travaux de cette thèse portent donc sur le développement et la validation du GPS cérébral, un outil de neuro-navigation automatique à partir des empreintes vasculaires Doppler ultrasensible de la souris et du rat. Dans un premier temps un template anatomique vasculaire a été construit puis recalé sur des atlas familiers (Atlas Allen pour la souris et l’atlas SIGMA pour le rat) permettant ainsi la création d’un atlas vasculaire pouvant servir de référence lors des sessions d’imagerie fUS pour recaler des données expérimentales et leur fournir un contexte anatomique. La précision du recalage vasculaire a été quantifié à partir des images vasculaires super-résolues obtenues par microscopie de localisation ultrasonore.Par la suite après avoir développé la chaîne neuro-informatique intégrée à une nouvelle génération de neuro-imageurs, nous avons montré la capacité du GPS cérébral à guider le positionnement d’une sonde linéaire sur des zones fonctionnelles choisies afin d’obtenir des cartes d’activation fonctionnelle même sur des plans obliques complexes. Nous avons aussi montré que le GPS cérébral peut être utilisé pour calculer automatiquement la matrice de connectivité en fournissant un contexte anatomique et une segmentation automatique des zones fonctionnelles.Pour aller plus loin nous avons exploré le potentiel du GPS vasculaire à guider automatiquement les injections intra-cérébrales dans des structures profondes. Des résultats encourageants confirmés par des images de microscopie de fluorescence ont pu être obtenus après injection d’un traceur neuronal dans le noyau thalamique. Toujours à titre exploratoire nous nous sommes intéressés à la reconstruction des images anatomiques structurelles du cerveau en plus des images vasculaires généralement générées afin d’évaluer l’échogénicité de certaines zones cérébrales identifiées à l’aide du GPS vasculaire. Enfin nous avons conduit une étude de quantification longitudinale dans le cadre du processus de consolidation du trace mnésique chez la souris. Ces travaux permettent d’apporter des nouveaux outils de neuro-imagerie pour renforcer le potentiel de l’imagerie fonctionnelle ultrasonore et permettre aux experts et aux non-experts neuroanatomistes de réaliser des protocoles standardisés, reproductibles, avec plus de précision et impliquant des études sur de grosses cohortes
Ultrafast ultrasound imaging is a recent method based on transmission of plane waves which enables the visualization of biological media with high spatio-temporal resolution. When applied to Doppler imaging, it enables detection of blood flow with very high sensitivity compared to conventional ultrasound Doppler usually limited to high blood flow imaging in cardiology. This advances have rendered functional ultrasound (fUS) imaging a valuable neuroimaging modality capable of mapping cerebral vascular networks, but also to indirectly capture neuronal activity with high sensitivity thanks to the neurovascular coupling. However, the expansion of fUS imaging is still limited by the difficulty to identify cerebral structures during experiments based solely on the Doppler images and the shape of the vessels, which complicates the positioning of the ultrasound probe and the data analysis of the data. It is therefore crucial to set up tools dedicated to functional ultrasound imaging in the same way as functional imaging by MRI, which remains the reference modality.This thesis focuses on the development and validation of cerebral GPS, an automatic neuro-navigation tool based on ultrasensitive Doppler vascular footprint of mice and rats. Initially, a vascular anatomical template was built and then registered on familiar atlases (Allen Atlas for the mouse and the SIGMA atlas for the rat) thus allowing the creation of a vascular atlas that can serve as a reference during fUS imaging sessions to align experimental data and provide an anatomical context. The accuracy of the vascular registration was quantified from the super-resolved vascular images obtained with ultrasound localization microscopy.Secondly, the neuro-informatic pipeline has been developed and integrated into a new generation of neuroimaging devices to perform online navigation. We then demonstrated the capability of the system to position itself automatically over chosen anatomical structures and to obtain corresponding functional activation maps even in complex oblique planes. Additionally, we show that the system can be used to acquire and estimate functional connectivity matrices automatically.To go further we explored the potential of vascular GPS to automatically guide intracerebral injections into deep structures. Encouraging results confirmed by fluorescence microscopy images were obtained after injection of a neuronal tracer into the thalamic nucleus. Still on an exploratory basis, we were interested in the reconstruction of structural anatomical images of the brain in addition to the vascular images generally reconstructed in order to assess the echogenicity of a few brain areas identified using vascular GPS. Finally we carried out a longitudinal study in the framework of memory consolidation in rats.This work provides new neuroimaging tools to strengthen the potential of functional ultrasound imaging and allow neuroanatomists experts and non-expert to carry out standardized, reproducible protocols with more accuracy and involving studies on large cohorts

Les cultures de racines et tubercules jouent un rôle important, diversifié et complexe dans l'alimentation des populations. Ces cultures ont le potentiel d'éradiquer la pauvreté et d'améliorer la sécurité alimentaire. L'amidon est le principal composant des racines et des tubercules. L'importance de l'amidon en tant que matière première dans les industries alimentaires et non alimentaires augmente. Dans la présente étude, l'efficacité de l'extraction assistée par ultrasons (EAU) pour augmenter le rendement d’extraction de l’amidon et des polyphénols à partir des légumes lontan (Canna et manioc) a été analysée ainsi que son influence sur les propriétés physico-chimiques des composés extraits. Les paramètres d'extraction (température, temps, rapport solide : liquide) ont été optimisés grâce à la conception de surface de réponse Box Behnken (BBD). Les propriétés physicochimiques et fonctionnelles de l'amidon et des polyphénols ont été étudiées par SEM, indice de gonflement et de solubilité, indice d'absorption d'huile et d'eau, rendement total en polyphénols, dosage DPPH et ORAC. Les rendements en amidon obtenu à partir du canna et du manioc aux conditions d'extraction optimales (Canna : température 40 ° C, durée 10 min, rapport solide : liquide 1:30 g / ml ; Manioc : température 40 ° C, durée 10 min, rapport solide : liquide 1:25 g / ml) étaient respectivement de 19,81% et 16,51%. Nous avons mis en évidence que le rendement en amidon obtenu était significativement plus élevé que le rendement atteint par la méthode d'extraction conventionnelle sans aucun changement significatif ou défavorable de ses propriétés physico-chimiques et fonctionnelles. Les rendements d’extraction des polyphénols du canna et du manioc étaient de 1061,72 mg GAE / & 100g et 7644,46 mg GAE / & 100g respectivement. Le rendement total en polyphénols obtenu par EAU s'est révélé significativement plus élevé que pour la méthode conventionnelle. L'activité antioxydante des composés bioactifs analysés par DPPH et ORAC s'est avérée être directement proportionnelle au rendement en polyphénol. Les résultats que nous avons obtenus suggèrent que les EAU augmenter les rendements d’extraction en amidon et en polyphénols sans altérer les propriétés des composés isolés
Roots and Tuber crops plays an eminent, countless and complex part in feeding the world. These crops have the potential to eradicate poverty and improve food security. Starch is the principal component of roots and tuber crops. The significance of starch as a raw material in both food and non-food industries are increasing. In present study efficiency of ultrasound assisted extraction (UAE) to increase the yield of starch and polyphenols from lontan (Canna and Cassava) crops were analysed along with its influence on the physiochemical properties of extracted compounds. Extraction parameters (Temperature, time, solid: liquid ratio) were optimized through Box Behnken response surface design (BBD). Physiochemical and functional properties of starch and polyphenols were investigated through SEM, swelling and solubility index, oil and water absorption index, Total polyphenol yield, DPPH and ORAC assay. Starch yield obtained from canna and cassava at the optimum extraction conditions (Canna: temperature 40°C, time 10 min, solid: liquid ratio 1:30 g/ml; Cassava: temperature 40°C, time 10 min, solid: liquid ratio 1:25 g/ml) were 19.81% and 16.51% respectively. Obtained starch yield was found to be significantly higher than the yield attained through conventional extraction method without any significant or adverse changes in the physicochemical and functional properties. Total polyphenol yield from canna and cassava rhizome was 1061.72 mg GAE/&100g and 7644.46 mg GAE/&100g respectively. Total polyphenol yield obtained through UAE was found to be significantly higher than conventional method. Antioxidant activity of bioactive compounds analysed through DPPH and ORAC was found to be directly proportional to the attained polyphenol yield. From the obtained results it can be observed that UAE has the efficiency to increase the yield of starch and polyphenols without altering the properties of isolated compounds

Le rythme thêta est un rythme cérébral associé à l’activité locomotrice et au sommeil paradoxal. Bien que son implication dans la communication entre régions du cerveau et processus mnésiques ait largement été démontrée, il persiste un manque de données extensives dû à la difficulté d’imager l’ensemble de l’activité cérébrale dans des conditions naturelles de locomotion et d’exploration. Dans cette thèse, j’ai développé une approche qui combine l’enregistrement des potentiels de champs locaux à l’imagerie ultrasonore fonctionnelle (fUS) sur l’animal en mouvement libre. Pour la première fois, j’ai pu révéler les réponses hémodynamiques associées au rythme thêta dans la plupart des structures du système nerveux central avec de bonnes résolutions spatiale (100 x 100 x 400 μm) et temporelle (200 ms). Pendant la locomotion et le sommeil, les variations hémodynamiques de l’hippocampe, du thalamus dorsal et du cortex (rétrosplenial, somatosensoriel) corrèlent fortement avec la puissance instantanée du signal thêta hippocampique, avec un décalage temporel variant de 0.7 s à 2.0 s selon les structures. De manière intéressante, les rythmes gamma hippocampiques moyen (55-95 Hz) et rapide (100-150 Hz) expliquent la variance des signaux hémodynamiques mieux que le seul rythme thêta, alors que le rythme gamma lent (30-50 Hz) est non pertinent. L’hyperémie fonctionnelle de l’hippocampe suit séquentiellement la boucle tri-synaptique (gyrus denté - région CA3 - région CA1) et se renforce considérablement à mesure que la tâche progresse. Lors du sommeil paradoxal, j’ai observé une hyperémie tonique globale ainsi que des activations phasiques de grande amplitude initiées dans le thalamus et terminant dans les aires corticales, que nous avons appelées “poussées vasculaires”. De fortes bouffées d’activité gamma rapide (100-150 Hz) précèdent de manière robuste ces poussées vasculaires, l’inverse n’étant pas vrai. Dans l’ensemble, ces résultats révèlent la dynamique spatio-temporelle des signaux hémodynamiques associés à la locomotion et au sommeil paradoxal et suggèrent un lien fort entre rythmes thêta, gamma rapide et activité vasculaire globale
Theta rhythm is a prominent oscillatory pattern of EEG strongly associated with active locomotion and REM sleep. While it has been shown to play a crucial role in communication between brain areas and memory processes, there is a lack of extensive data due to the difficulty to image global brain activity during locomotion behavior. In this thesis, I developed an approach that combines local field potential recordings (LFP) and functional ultrasound imaging (fUS) to unrestrained rats. For the first time, I could image the hemodynamic responses associated with theta rhythm in most central nervous system (CNS) structures, with high spatial (100 x 100 x 400 μm) and temporal (200 ms) resolutions. During running and REM sleep, hemodynamic variations in the hippocampus, dorsal thalamus and cortices (S1BF, retrosplenial) correlated strongly with instantaneous theta power, with a delay ranging from 0.7 to 2.0 s after theta peak. Interestingly, mid (55-95 Hz) and high gamma (100-150 Hz) instantaneous power better explained hemodynamic variations than mere theta activity, while low-gamma (30-50 Hz) did not. Hippocampal hyperaemia followed sequentially the trisynaptic circuit (dentate gyrus - CA3 region - CA1 region) and was considerably strengthened as the task progressed. REM sleep revealed brain-wide tonic hyperaemia, together with phasic high-amplitude vascular activation starting in the dorsal thalamus and fading in cortical areas, which we referred to as “vascular surges”. Strong bursts of hippocampal high gamma (100-150 Hz) robustly preceded these surges, while the opposite was not true. Taken together, these results reveals the spatio-temporal dynamics of hemodynamics associated with locomotion and REM sleep and suggest a strong link between theta, high-gamma rhythms and brain-wide vascular activity

京都大学
新制・課程博士
博士(人間健康科学)
甲第23426号
人健博第93号
新制||人健||6(附属図書館)
京都大学大学院医学研究科人間健康科学系専攻
(主査)教授 山田 重人, 教授 林 悠, 教授 森本 尚樹
学位規則第4条第1項該当
Doctor of Human Health Sciences
Kyoto University
DFAM

Les épilepsies sont des hyperactivités neuronales pathologiques largement distribuées au sein du système nerveux. Aborder la question de l'organisation spatiotemporelle de ces crises est un premier pas crucial vers la dissection des mécanismes qui les sous-tendent. Alors qu'il existe de nombreux modèles d'hyperactivité épileptiforme, il est plus difficile d'étudier les crises spontanées, qui sont altérées par la sédation. Dans cette thèse, j'ai développé une approche combinant l'électroencéphalogramme (EEG) avec l'imagerie fonctionnelle ultrasonore (fUS), sur le rat mobile. Ainsi, sur un modèle d'épilepsie absence, j'ai pu enregistrer simultanément la survenue des crises et les variations hémodynamiques, marqueurs du métabolisme cellulaire. Le suivi additionnel de l'activité hémodynamique n'avait pas d'effet en soi sur l'occurrence et la durée des crises épileptiques. L'analyse des enregistrements a permis de mettre en évidence des corrélations entre les activités électriques et vasculaires durant les crises. Tandis que le thalamus présentait des zones d'hyperperfusion pendant les crises, le cortex présentait des corrélats variables suivant les aires, avec une hyperémie des aires somato-sensorielles accompagnée parfois d'une baisse de perfusion des tissus adjacents. La sensibilité du fUS a révélé, à partir d'événements uniques, qu'une série de pointe-ondes observée par une électrode EEG ne s'accompagne pas toujours d'une hyperactivité vasculaire au même endroit. Ainsi, cette approche permet de délimiter le contour des aires présentant une activité vasculaire pendant les crises et montre une dichotomie partielle entre les composantes électriques et vasculaires des crises
Epilepsies consist in neuronal hyperactivities distributed across the nervous system that need first to be located in order to later decipher the mechanisms of these pathologies. While there are many models of epileptiform hyperactivity, it is more difficult to study spontaneous seizures, which are altered by sedation. In this thesis, I developed an approach that combines electroencephalography (EEG) and functional ultrasound imaging (fUS), on the mobile rat. Thus, on a model of absence epilepsy, I could record simultaneously the occurrence of seizures and the hemodynamic variations, which reflect cellular metabolism. Seizures were unaltered by the recording protocol, compared to rats with EEG alone. Correlations were observed between electric and vascular activities. The thalamus showed areas of hyperperfusion during seizures. The cortex exhibited different correlates in distinct areas, with hyperaemia in somato-sensory areas, occasionally associated with a decrease in perfusion in adjacent tissue. The sensitivity of fUS, which could resolve blood changes from single occurrences, revealed that series of spike-wave discharges recorded from an EEG electrode were not always associated with vascular hyperactivity in the same region. Thus, this approach can delimit the contour of areas presenting vascular activity during seizures and shows a partial dichotomy between the electric and vascular components of seizures

Stato dell’arte: I microrganismi probiotici negli alimenti potrebbero causare cambiamenti significativi nel sapore e nella reologia, a causa del loro metabolismo attivo. È importante, quindi, controllare il metabolismo dei microrganismi probiotici negli alimenti, senza influenzare negativamente la vitalità e le caratteristiche funzionali. Un modo per superare questo problema è l'utilizzo di colture starter attenuate attraverso l’utilizzo di metodi fisici o chimici. Alcuni autori hanno studiato l'omogeneizzazione come metodo per attenuare/modulare il metabolismo delle colture starter nei prodotti caseari (Lanciotti et al., 2004, 2006, 2007); in questo progetto di tesi ho utilizzato una nuova tecnologia emergente, gli ultrasuoni. (i) È stato effettuato uno screening delle combinazioni possibili di ultrasuoni (US) (potenza/durata) su diversi microrganismi probiotici e sono state studiate le caratteristiche probiotiche e tecnologiche dopo l'esposizione agli US. (ii) Sono stati studiati gli effetti degli US sul rilascio di componenti intracellulari. (iii) È stata valutata l'interazione dei ceppi attenuati con il microbiota intestinale utilizzando la fermentazione in vitro (batch culture fermentation). Problematiche: (i) Non si conoscono gli effetti dell’attenuazione mediante US sulle caratteristiche tecnologiche/probiotiche dei microrganismi; (ii) non è chiara la risposta delle cellule microbiche al trattamento con gli US e il danno che ne deriva, in quanto in letteratura sono disponibili pochi dati; (iii) non sono disponibili dati sull'interazione di ceppi attenuati con il microbiota intestinale. Obiettivi: (i) Scelta della migliore combinazione di ultrasuoni (potenza/durata), in grado di evitare la post-acidificazione senza compromettere la vitalità dei ceppi, studio dei cambiamenti delle caratteristiche probiotiche e tecnologiche che i microrganismi potrebbero subire a seguito dell’attenuazione; (ii) valutazione del rilascio di componenti intracellulari (acidi nucleici e proteine) dopo l'applicazione degli ultrasuoni; e (iii) studio dell’effetto di ceppi attenuati sul microbiota intestinale. Pianificazione della ricerca: Nella prima parte sono stati usati tre diversi generi di microrganismi probiotici: Lactobacillus, Bifidobacterium, Propionibacterium. I ceppi sono stati trattati con ultrasuoni e sono state studiate le caratteristiche tecnologiche e probiotiche. Nella seconda parte, sono stati studiati gli effetti degli US sul rilascio di componenti intracellulari. I ceppi sono stati studiati a seguito del trattamento fisico per valutare il rilascio di costituenti intracellulari (acidi nucleici, proteine) ed eventuali lesioni della membrana cellulare. Nell'ultima parte, è stata studiata l'interazione di ceppi attenuati con il microbiota intestinale. Questo studio è stato condotto presso l'Università di Roehampton (Regno Unito). Materiali e metodi: (i) Caratteristiche tecnologiche: Prove di acidificazione in mezzo di laboratorio, crescita a diverse temperature, pH e contenuto di sale; caratteristiche probiotiche: antibiotico resistenza, sopravvivenza a pH 2,5 e in presenza di sali biliari (0,3%), idrofobicità e formazione di biofilm. (ii) La valutazione del danno sulle cellule microbiche è stato determinato attraverso letture spettrofotometriche a 260 e 280 nm per quantificare il rilascio di DNA e proteine. (iii) L'interazione dei ceppi attenuati con il microbiota intestinale è stata valutata utilizzando la fermentazione in vitro (batch culture fermentation). Risultati: (i) Le migliori combinazioni in grado di evitare la post-acidificazione sono state le seguenti: potenza, 60%; tempo, 6 minuti; impulsi, 2 s per Lactobacillus e Bifidobacterium, 40%, 8 min P. jensenii; 60%, 4 min P. freudenreichii subsp. freudenreichii. Gli US non hanno influenzato la vitalità a 45 °C e a pH 9, ma hanno determinato una diminuzione della crescita microbica a pH 4 (lattobacilli e bifidobatteri). Tuttavia, gli US non hanno influenzato il GI dei propionibatteri. L'effetto dell'attenuazione potrebbe essere migliorato con la refrigerazione. Il trattamento con gli US non ha influenzato la maggior parte dei tratti tecnologici, ma ha causato un aumento della suscettibilità ad alcuni antibiotici. Per quanto riguarda i tratti probiotici, gli US hanno causato un aumento di idrofobicità per L. reuteri e P. freudenreichii spp. freudenreichii. Questi risultati sono stati confermati con l'adesione alle cellule Caco-2 per L. reuteri. L. reuteri attenuato ha registrato un significativo aumento di idrofobicità (dal 3 al 25%) e una maggiore adesione alle cellule Caco-2. Inoltre, gli US hanno migliorato la stabilità dei biofilm nel tempo e questo risultato ha confermato i dati ottenuti con l'idrofobicità. (ii) Il rilascio di acidi nucleici e proteine, evidenzia che la membrana cellulare potrebbe essere un altro target del trattamento fisico. (iii) Per quanto riguarda gli effetti degli US sul microbiota intestinale: i ceppi attenuati non hanno influenzato il microbiota intestinale, ma in alcuni casi sono stati evidenziati effetti positivi. Importanza e impatto della ricerca di dottorato: Il metabolismo attivo dei microrganismi probiotici potrebbe costituire un problema quando questi ultimi vengono aggiunti agli alimenti. Infatti alcuni ceppi di batteri lattici continuano a produrre acido lattico e causano post-acidificazione (diminuzione del pH durante la conservazione). Pertanto, è importante controllare il loro metabolismo. Un modo possibile per controllare il metabolismo dei probiotici negli alimenti è l'attenuazione attraverso metodi fisici o chimici. Una delle tecnologie emergenti sono gli ultrasuoni (US). Questo approccio è stato utilizzato per evitare la post-acidificazione in una bevanda commerciale di riso (Bevilacqua et al., 2016). La presente tesi di dottorato ha contribuito a valutare gli effetti dell'attenuazione mediante ultrasuoni, su alcune caratteristiche tecnologiche e probiotiche, testando tre diversi generi di ceppi probiotici. Inoltre, questo progetto di dottorato ha studiato i cambiamenti in termini di caratteristiche tecnologiche e probiotiche che i microrganismi probiotici potrebbero subire a seguito dell'attenuazione; il rilascio di proteine, acidi nucleici. Infine, la novità di questa tesi di dottorato è stata lo studio degli effetti di ceppi attenuati sul microbiota intestinale. Proposte future: una prospettiva futura potrebbe essere focalizzata sull'uso degli US per migliorare o modulare l'adesione dei ceppi probiotici, considerando l'aumento di idrofobicità e la maggiore adesione alle cellule Caco-2. È importante studiare gli effetti di altri ceppi attenuati sul microbiota intestinale modulando le variabili del trattamento.
Scientific background: Probiotics in foods could lead to significant changes in food flavor and rheology, due to their active metabolism. A possible way to overcome this problem is the attenuation of probiotics through a physical or chemical method. Some authors studied homogenization as a way to attenuate/modulate the metabolism of starter cultures in dairy products (Lanciotti et al., 2004, 2006, 2007); in this project I have used a new emerging technology, the ultrasounds. (i) A screening of the ultrasound (US) (power/duration) on different probiotic microorganisms was perfomed, and were studied the probiotic and technological characteristics after US-exposure. (ii) The effects of US on the release of intracellular components, was investigate. (iii) The interaction of attenuated strains with gut microbiota was evaluated, using in vitro batch culture fermentation. Open questions: (i) Few data are available on effects of US on probiotics and technological characteristics of probiotic strains; (ii) few data are available on the effect of attenuation with ultrasound on the sub-lethal injury; (iii) no data are available on the interaction of attenuated strains with gut microbiota. Aims: (i) Choice of the best combination of ultrasound to avoid post-acidification without affecting the viability of the strains, and study of the probiotic and technological characteristics to evaluate if attenuation could change them; (ii) study the release of intracellular components (nucleic acids and proteins) after the application of ultrasound; and (iii) evaluation the effects of attenuated strains on gut microbiota. Planning of the research: In the first part three different genera of probiotics were used: Lactobacillus, Bifidobacterium, Propionibacterium. The strains were treated with ultrasound and studied for technological and probiotic characteristics. In the second part, the effects of US on the release of intracellular components, was investigated. The strains were studied after physical treatment to assess the release of intra-cellular constituents (nucleic acids, proteins) and injury of the membrane. In the last part, the interaction of attenuated strains with gut microbiota, was studied. This study was carried out at University of Roehampton (UK). Materials and Methods: (i) Technological traits: acidification in lab medium, growth at different temperatures, pHs and salt content; probiotic traits: antibiotic-resistance, survival at pH 2.5 and in the presence of 0.3% bile salt, hydrophobicity, and biofilm formation. (ii) Injury characterization was evaluated by leakage of UV-absorbing substances. (iii) The interaction of attenuated strains with gut microbiota was evaluated, using in vitro batch culture fermentation. Results: (i) The best combinations to avoid post-acidification were the following: power, 60%; time, 6 min; pulse, 2 s for Lactobacillus and Bifidobacterium, 40%, 8 min P. jensenii; 60%, 4 min P. freudenreichii subsp. freudenreichii. US did not affect viability at 45 °C or at pH 9, but it determined a decrease of microbial growth to pH 4 (lactobacilli and bifidobacteria). However, the US did not affect the GI of propionibacteria. The effect of attenuation could be enhanced by the storage under refrigeration. US-treatment did not affect most of the technological traits, but generally caused an increase of susceptibility to some antibiotics. Concerning probiotic traits, US caused an increase of hydrophobicity for L. reuteri and P. freudenreichii spp. freudenreichii, after US-exposure. These results were confirmed with adhesion to Caco-2 cells for L. reuteri. US-attenuated L. reuteri experienced a significant increase of hydrophobicity (from 3 to 25%) and a higher adhesion to Caco-2 cells. Moreover, US improved the stability of the biofilm over the time, and this result confirmed the data obtained with hydrophobicity. (ii) The release of nucleic acids and proteins was found, highlighting that cell membrane could be another target physical treatments. (iii) Concerning the effects of US on gut microbiota, the ultrasound didn’t affect the gut microbiota, but in some cases, it could have a positive effect. Significance and Impact of PhD research: A main drawback of probiotics in foods can relate to their active metabolism, some strains of lactic acid bacteria continue to produce lactic acid and cause post-acidification (the decrease of pH within the storage). Therefore, it is important to control their metabolism. A possible way to control the metabolism of probiotic in foods is the attenuation through physical or chemical methods. One of the emerging technologies is ultrasound (US). This approach was used to avoid post-acidification in a commercial rice drink (Bevilacqua et al., 2016). The present PhD thesis contributed to evaluate the effects of attenuation with ultrasound, on some technological and probiotic strains, testing three different genera of probiotic strains. Moreover, this PhD project has investigate the changes that may affect probiotic strains after attenuation; the release of proteins, nucleic acids. Finally, the novelty of this PhD thesis was the study of the effects of attenuated strains on gut microbiota. Future trends: A future perspective could be a focus on the use of US to improve or modulate the adhesion of probiotic strains, considering the increase of hydrophobicity and the higher adhesion to Caco-2-cells. It is important to investigate the effects of other attenuated strains on gut microbiota by modulating the variables of the treatment. Key words: Hydrophobicity, acidification, growth, attenuation, gut-microbiota, proteins, nucleic acid, ultrasound, attenuated microorganisms, probiotics, lactic acid bacteria.

Orientadores: Maria de Lourdes Setsuko Ayrizono, Raquel Franco Leal
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: Introdução: A incidência da doença de Crohn Perineal (DCP) varia entre 20% a 25% nos portadores de doença de Crohn . O padrão-ouro para a investigação da DCP é a avaliação clínica clássica seguida do exame sob anestesia, mas a ultrassonografia endoanal (USEA) e a ressonância nuclear magnética (RNM) da pelve podem ser úteis na pesquisa de abscessos, fistulas e lesões esfincterianas que podem levar à incontinência fecal e perda da qualidade de vida. Objetivo: Correlacionar a avaliação clínica clássica da função esfincteriana por meio do escore de Jorge-Wexner nos pacientes com DCP, comparando com os achados de USEA, através do escore de Starck. Casuística e Método: Vinte e quatro pacientes participaram do estudo, sendo 14 do sexo feminino e média de idade de 40,54 anos. Todos os doentes tinham diagnóstico confirmado de DCP, sendo que 7 (29,16%) também apresentavam acometimento do intestino delgado; 7 (29,16%) do cólon e reto, 2 (8,33%) do intestino delgado, do cólon e reto e 8 pacientes (33,33%) apresentavam DCP como manifestação única da doença. Resultados: Os dois escores apresentaram distribuição normal, com escore de Jorge-Wexner médio de 3,8333 (DP 4,52689) e escore de Starck médio de 9,7500 (DP 2,54097). A análise estatística mostrou não haver correlação entre os dois escores, com um intervalo de confiança de 95%. Conclusão: O escore de Jorge- Wexner apresentou utilidade limitada nessa categoria de pacientes, enquanto a USEA foi ferramenta útil para alcançar melhor correlação com deficiência muscular esfincteriana. No entanto, mais estudos com propostas similares são necessários para aumentar a validade desses achados e para estabelecer se a USEA pode predizer a função esfincteriana e a incontinência fecal antes de procedimento cirúrgico em pacientes com DCP
Abstract: Introduction: The incidence of perianal CD (PCD) is variable between 20% and 25% of patients with CD. The gold standard for assessment of PCD is still the classic clinical examination followed eventually by examination under anesthesia (EUA) but complementary examinations like endoanal ultrasound (EAUS) and magnetic resonance imaging of the pelvis (MRI) were introduced as useful to completely assess the anatomical lesions of the sphincteric muscles caused by fistulas and abscesses. Objective: Verify the adequacy of the classical clinical evaluation to which most of the patients are submitted, in particular regarding the adequacy of assessing the sphincteric function through the Jorge- Wexner score in patients with PCD, comparing it with the findings of EAUS trough a score published by Stark and colleagues. Patients and Methods: Twenty four patients participated to the study, being 16 females and the mean age is 40.54 years old. All patients have an established diagnosis of PCD, being 7 (29,16%) with a diagnosis of CD involving the small bowel, 7 (29,16%) CD involving the colon and rectum, 2 (8,33%) CD involving the small bowel and the colon-rectum, and 8 (33,33%) have a diagnosis of PCD as the only clinical manifestation of CD. Results: The two scores has a normal distribution, with a mean Wexner score of 3.8333 (SD 4,52689) and a mean Starck score of 9,7500 (SD 2,54097). The statistical analysis showed that there is no correlation between the two scores with a confidence interval of 95%. Conclusion: The Jorge-Wexner score had a limited usefulness in this category of patients, whereas EAUS was an useful tool for achieving better correlation with muscle deficiency. This study investigated CD, a very particular and still not fully understood disease, being the perianal disease important feature of CD. However, more studies with similar purpose are needed, in order to improve the validity of these findings, and establish if the EAUS can predict the sphincter function and fecal continence before perianal surgery for CD
Mestrado
Fisiopatologia Cirúrgica
Mestre em Ciências

L’Accident Vasculaire Cérébral (AVC) ischémique, causé par l’obstruction d’une artère cérébrale, est l’une des principales causes de mortalité et d’invalidité dans le monde. Bien que des traitements comme le rtPA et la thrombectomie mécanique existent, seul un faible pourcentage de patients y a accès. Par ailleurs, malgré l’investissement de la recherche, les difficultés à transposer les résultats des modèles animaux aux essais cliniques humains limitent le développement de nouvelles thérapies. Cette thèse présente un modèle d’AVC chez la souris éveillée visant à améliorer la transposabilité des études précliniques. Grâce à l’imagerie ultrasonore fonctionnelle (fUS) et à l’IRM, ce modèle permet une évaluation en temps réel des paramètres hémodynamiques ainsi qu’une analyse de la récupération fonctionnelle après l’ischémie. Nous proposons également un nouvel outil d’évaluation de la connectivité cérébrale post-AVC, apportant des informations sur la récupération cérébrale et la réponse au traitement. L’efficacité du traitement thrombolytique standard, le rtPA, a été testée, et une approche innovante de traitement ciblé a été explorée. Les résultats soulignent le potentiel de ces approches multimodales et thérapeutiques ciblées pour améliorer la prise en charge des AVC ischémiques, ouvrant la voie à de nouvelles perspectives de recherche translationnelle
Ischemic stroke, caused by the obstruction of a cerebral artery, is one of the leading causes of mortality and disability worldwide. Despite the availability of treatments such as rtPA and endovascular thrombectomy, only a small percentage of patients have access to these. Moreover, despite substantial research efforts, challenges in translating results from animal models to human clinical trials limit the development of new therapies. This thesis presents a novel stroke model in awake mice to improve the translatability of preclinical studies. Through the use of functional ultrasound (fUS) and MRI, this model enables real-time assessment of hemodynamic parameters and functional recovery following ischemia. Additionally, we propose a new tool for evaluating post-stroke brain connectivity, providing insights into brain recovery and treatment response. The efficacy of the stantard thrombolytic treatment, rtPA, was evaluated, and an innovative targeted treatment approach was explored. The results underscore the potential of these multimodal imaging approaches and targeted therapies to enhance ischemic stroke management, opening new avenues for translational research

Avec l’avènement de l’imagerie ultrarapide 4D au laboratoire Physics for Medecine Inserm 1273, la possibilité d’acquérir dans les trois dimensions et avec une haute résolution spatio-temporelle a été démontrée. Plusieurs des modalités phares de l’imagerie 2D ultrarapide ont pu être étendues à l’imagerie volumique (Doppler de puissance ultrasensible 3D, élastographie 3D …). Leur dissémination en clinique bénéficierait grandement aux praticiens. Cependant les moyens nécessaires à leur mise en œuvre sont encore trop importants pour espérer une transposition rapide en clinique. Le développant de stratégies intelligentes de réduction de voies est donc devenu un enjeu central. Une stratégie originale basée sur l’architecture de la sonde consiste en l’adressage orthogonal, par lignes et colonnes, du réseau de sonde matricielle, ou Row and Column Adressing RCA. Elle propose une solution transducteur tout à fait adaptée à l’imagerie ultrarapide par ondes planes. Grâce à cette solution, la sonde peut être pilotée par un seul échographe standard, tout en conservant une ouverture d’imagerie étendue. La grille matricielle 2D organisée selon N+N voies orthogonales, représentant ainsi un facteur de réduction de N/2. Cette stratégie présente un changement de paradigme important d’imagerie par dissociation des voies de focalisation en émission et réception et pose un nouveau compromis en termes de résolution spatio-temporelle. Au cours de ces travaux de thèse, les performances de la RCA associées à l’imagerie ultrarapide 4D sont étudiées pour divers cas. Le principe d’imagerie 4D ultrarapide RCA avec sommation orthogonale OPW sont étudié. L’imagerie vectorielle 3D pour la RCA est développée. Un nouveau prototype de sonde RCA haute fréquence (15MHz) est présenté et testé sur un protocole d’imagerie fonctionnelle cérébral en 3D. Enfin une nouvelle modalité d’imagerie 3D d’intensité du flux est présentée offrant un nouveau champ d’exploitation de la sonde RCA
With the advent of 4D ultrafast imaging at the Physics for Medicine Inserm 1273 laboratory, the ability to acquire in all three dimensions and with a high spatio-temporal resolution has been demonstrated. Several of the most effective 2D ultrafast imaging modalities have been extended to volume imaging (3D ultrasensitive power Doppler, 3D elastography ...). Their dissemination in clinic would greatly benefit to physicians. However the necessary means to implement ultrafast 4D are still too heavy and costly to hope for a transposition in the short or mid-term to the radiology departments. Developing smart strategies to reduce channel number has become a central issue. An original strategy based on the probe architecture consists of orthogonal row and column addressing of the Matrix Probe array, the Row and Column Adressing RCA. It offers a transducer solution perfectly adapted with ultra-fast plane waves imaging. With this approach, the probe can be driven by a single standard ultrasound unit, while maintaining a large aperture. The 2D matrix grid is organized according to N + N orthogonal channels, thus representing a reduction factor of N / 2. This strategy presents an important paradigm shift of imaging by dissociation of the focus pathways in transmission and reception and offers a new compromise in terms of spatio-temporal resolution. During this thesis work, the performances of the RCA associated with the ultra fast 4D imaging are studied for various cases. The principle of 4D ultrafast RCA imaging with orthogonal summation OPW are studied. 3D vector imagery for RCA is developed. A new high frequency RCA probe prototype (15MHz) is presented and tested on a 3D functional brain imaging protocol. Finally, a new modality of 3D imaging of the flux intensity is presented offering a new way of exploitation for the RCA probe

L'objectif était de quantifier les modifications de la contractilité cardiaque au cours d’un bedrest de 21 jours (HDBR) par la methode speckle tracking 4D et de déterminer si la contre-mesure « exercice » était capable de préserver cette contractilité cardiaque. Les méthodes traditionnelles 2D ne mesurent qu'une variation de la taille du ventricule de la systole à la diastole et ne suivent pas la cinetique des mouvements de contractilité à l’interieur de la paroi. De plus l’echographie 2D ne permet d’acceder qu’a la contractilite longitudinale globale par la mesure de la distance Apex-Valve mitrale. Le suivi des mouvements des speckles (points singuliers a l’interieur du myocarde) en 4D montre que la contractilité radiale diminuée signifcativement pendant le HDTBR chez les sujets témoins alors qu’elle est conservée dans le groupe exercice. Par contre la contractilité longitudinale n’est pas affectée dans aucun des 2 groupes. Bien sûr, l'IRM pourrait fournir des données similaires, mais il n’est pas possible à ce jour d’imaginer avoir une IRM a bord de l’ISS dans un futur proche. A la suite de ces resultats nous avons mis au point un algorithme de traitement des video cardiaque qui permet d’acceder a la contractilite radiale et longitudinale en vol (prog Vasc-Aging en cours) .L’objectif de l’experimentation était de montrer que quatre jours en immersion seche suffisaient pour realiser un transfert liquidien important vers les régions céphaliques comparable à celui observé en vol spatial. Notre protocole consistait donc a mesurer les volume veineux au niveau cervical cerebral et porte au debut de la periode en immersion (a 2h) puis a 4 jours d’immersion. Compte tenu des problemes engendrés par ce transfert liquidien en vol nous avions proposé de tester la capacité des brassarts de cuisse a reduire l’amplitude du transfert veineux. Les résultats montrent que l’immersion provoque un transfert de sang veineux au niveau cervical maximal à 2H et que les Brassarts de cuisse reduisent signifcativement ce transfert dans cette phase precoce. Par contre a 4 jours d’immersion l’amplitude du transfert sanguin est considerablement diminuée (bien que toujours presente) et les brassarts de cuisse n’ont pas d’effet visible à ce moment-là. En fait le volume plasmatique decroit significativement à la fin du premier jour (env 20%) des lors la masse de sang deplacées vers la tete par l’immersion est insuffisante pour generer une stase importante au niveau cervical comme à 2h d’immersion. Pour cette meme raison la vitesse dans les veines cerebrales n’est pas augmentée à 4 jours d’immersion contrairement à ce qu’on avait observé à 2h d’immersion lors d’une precedente etude. Donc le modele immersion seche est un modele pour etudier les transferts liquidiens en microgravité mais seulement en debut de phase d’immersion. Sur cette période de temps, les Brassarts de cuisse ont bien un effet protecteur pour les organes de la zone cephalique
Objective: to evaluate functional myocardial contractility after 21 days of head-down bed rest (HDTBR) in sedentary control (CON) or with a resistive vibration exercise (RVE) countermeasure (CM) applied, by using 4D echocardiographic (4D Echo) imaging and speckle tracking strain quantification.Methods: Twelve volunteers were enrolled in a crossover HDTBR design, and 4D Echo was performed in supine position (REST) at BDC-2 and at R+2, and in -6° HDTBR (on day 18), and also during the first and the last minute of the 80° head-up step of Standard Measures tilt test, performed at both BDC-2 and R+2. Radial (Rad-Str), longitudinal (Lg-Str) and twist (Tw-Str) strain were measured by 4D speckle tracking, as well as left ventricle diastolic volume (LVDV) and mass (LVmass).Results: On day 18: in the CON group, LVDV and LVmass were reduced (p<0.05), the Rad-Str decreased (p<0.05) and Tw-Str showed a tendency to increase (p< 0.11), with no changes in Lg-Str. In RVE group, LVDV and LV mass, as well as all the strain parameters remained unchanged.On R+2: in the CON group, LVDV and LVmass were not recovered in all subjects compared to pre-HDTBR (p<0.08), Rad-Str was still decreased (p<0.05), while Tw-Str tended to increase (p<0.09). These parameters remained unchanged in the RVE group.Tilt 80°: Rad-Str and Lg-Str values at 80° tilt were similar post HDT in both groups.Conclusion: 4D Echo and speckle tracking analysis showed that in the CON group, Rad-Str decreased concomitant with LVmass and LVDV with HDTBR, but this observation did not support the hypothesis that this HDTBR induced remodelling or a muscle atrophy. RVE acted to preserve both LVmass, LVDV and contractility during HDTBR, thus proving its effectiveness to this aim. Nevertheless, the significant HDTBR-induced changes observed in the CON group had only a limited effect on the cardiac contractile response as observed during post HDTBR tilt test. The level of contractility at 80° Tilt position was not affected neither by HDTBR nor by RVE CM.Purpose: The objective was to quantify the venous redistribution during a 4-day dry immersion (DI) and evaluate the effect of thigh cuffs.Methods: The study included 9 control (Co) and 9 subjects wearing thigh cuffs during daytime hours (CU). Ultrasound images were collected Pre DI, on the fourth day in the morning (D4 AM) and on the fourth day in the afternoon (D4 PM), to assess the following outcome variables: left ventricle dimension, stroke volume, and ejection fraction (LVD, SV, EF), jugular vein volume (JV), portal vein dimension (PV), middle cerebral vein velocity (MCVv). An additional measure of JV dimension was performed on the first day after having worn the cuffs for two hours (D1 2H).Results: The JV volume increased significantly from Pre to D1 2H in both groups, but increased more in the Co compare to the CU subjects (Co: 0,27+/0.15cm3 to 0.94+/-0;22 cm3;P<0.01 CU: 0,32+/-0.13 cm3 to 0.64+/-0.32 cm3 P<0.042).At D4 AM no difference was found between the two treatment groups for any of the parameters listed above.Stroke volume and EF decreased from Pre (SV:111+/-23cm3 to 93+/-24 cm3 p<0.05; EF:0.66+/-0.07 to 0.62+/-0.07 p<0.05). JV volume was slightly, but significantly increased (Co: 0.47+/-0.22cm3 CU:0.35+/-014cm3 P<0.05), while MCVv and PV remained unchanged from Pre DI. From D4 AM to PM these parameters did not show any significant change.Conclusion: The results confirm that DI induces, during the first 2-3 h, a significant cephalic fluid shift as observed in spaceflight. During this early phase the thigh cuffs reduced the amplitude of the fluid shift towards the head, but after 4 days in DI there was only a slight memory (residual) effect of DI on the jugular volume and no residual effect of thigh cuffs

Le problème d'estimer une fonction de plusieurs variables à partir d'une observation corrompue surgit dans de nombreux domaines d'ingénierie. Par exemple, en imagerie médicale cette tâche a attiré une attention particulière et a, même, motivé l'introduction de nouveaux concepts qui ont trouvé des applications dans de nombreux autres domaines. Cet intérêt est principalement du au fait que l'analyse des données médicales est souvent effectuée dans des conditions difficiles car on doit faire face au bruit, au faible contraste et aux transformations indésirables inhérents aux systèmes d'acquisition. D'autre part , le concept de parcimonie a eu un fort impact sur la reconstruction et la restauration d'images au cours des deux dernières décennies. La parcimonie stipule que certains signaux et images ont des représentations impliquant seulement quelques coefficients non nuls. Cela est avéré être vérifiable dans de nombreux problèmes pratiques. La thèse introduit de nouvelles constructions d'a priori de parcimonie dans le cas des ondelettes et de la variation totale. Ces constructions utilisent une notion d'anisotopie généralisée qui permet de regrouper des variables ayant des comportements similaires : ces comportement peuvent peut être liée à la régularité de la fonction, au sens physique des variables ou bien au modèle d'observation. Nous utilisons ces constructions pour l'estimation non-paramétriques de fonctions. Dans le cas des ondelettes, nous montrons l'optimalité de l'approche sur les espaces fonctionnelles habituels avant de présenter quelques exemples d’applications en débruitage de séquences d'images, de données spectrales et hyper-spectrales, écoulements incompressibles ou encore des images ultrasonores. En suite, nous modélisons un problème déconvolution de données d'imagerie par résonance magnétique fonctionnelle comme un problème de minimisation faisant apparaître un a priori de variation totale structuré en espace-temps. Nous adaptons une généralisation de l'éclatement explicite-implicite pour trouver une solution au problème de minimisation
The problem of estimating a multivariate function from corrupted observations arises throughout many areas of engineering. For instance, in the particular field of medical signal and image processing, this task has attracted special attention and even triggered new concepts and notions that have found applications in many other fields. This interest is mainly due to the fact that the medical data analysis pipeline is often carried out in challenging conditions, since one has to deal with noise, low contrast and undesirable transformations operated by acquisition systems. On the other hand, the concept of sparsity had a tremendous impact on data reconstruction and restoration in the last two decades. Sparsity stipulates that some signals and images have representations involving only a few non-zero coefficients. The present PhD dissertation introduces new constructions of sparsity priors for wavelets and total variation. These construction harness notions of generalized anisotropy that enables grouping variables into sub-sets having similar behaviour; this behaviour can be related to the regularity of the unknown function, the physical meaning of the variables or the observation model. We use these constructions for non-parametric estimation of multivariate functions. In the case of wavelet thresholding, we show the optimality of the procedure over usual functional spaces before presenting some applications on denoising of image sequence, spectral and hyperspectral data, incompressible flows and ultrasound images. Afterwards, we study the problem of retrieving activity patterns from functional Magnetic Resonance Imaging data without incorporating priors on the timing, durations and atlas-based spatial structure of the activation. We model this challenge as a spatio-temporal deconvolution problem. We propose the corresponding variational formulation and we adapt the generalized forward-backward splitting algorithm to solve it

O ultra-som é uma das formas de tratamento terapêutico, não-invasivo, que pode auxiliar na regeneração nervosa periférica. Dentre todas estas técnicas não-invasivas de regeneração nervosa periférica, pouco se sabe sobre a influência do ultra-som na regeneração desse tecido. Para tanto foi realizado um trabalho experimental com o objetivo de avaliar o efeito da terapia do ultra-som pulsado de baixa intensidade após lesão por esmagamento do nervo ciático de rato. Foram utilizados 26 ratos Wistar machos, com massa corporal média de 289,61g divididos em 3 grupos: normal (N), lesado (L) e lesado+ultra-som (L+US). A aplicação do ultra-som pulsado (1 MHz, 40 mW/\'CM POT.2\', 8 minutos de duração) iniciou-se 1 dia pós-lesão e repetido por 4 semanas em dias alternados. A análise funcional do nervo ciático (análise de marcha em pista) foi avaliada semanalmente durante todo o período experimental. Para verificar a eficácia do ultra-som sobre a regeneração nervosa foram analisados índice funcional do ciático, morfologia do nervo ciático e morfologia e morfometria muscular dos músculos tibial anterior (TA) e sóleo (SOL). Concluiu-se que o tratamento com o ultra-som pulsado de baixa intensidade na regeneração nervosa periférica não obteve resultados significativos quando se comparou os grupos lesado e lesado+ultra-som, apenas foi observada uma manutenção da área de secção transversa do grupo lesado+ultra-som do músculo TA quando comparado ao grupo normal, levando a indícios de que o ultra-som estimulou o nervo para tal manutenção.
The ultrasound is one of the forms therapeutic treatment, no-invasive, that it assists in regeneration peripheral nerve. For in such a way an experimental work was carried through with the objective to verify the effect of the therapy of low intensity pulsed ultrasound after crush of sciatic nerve. Amongst all these no-invasive techniques of regeneration peripheral nerve, little known on the influence of the ultrasound in the regeneration of this tissue. Twenty six male Wistar rats weighing 289,61 g on average were used and divided into three groups: normal (N), injury (L) and injury+ultrasound (L+US). The application of the pulsed ultrasound (1 Mhz, 40 mW/\'CM POT.2\' and 8 minutes duration) was started one day after lesion and repeated for 4 week in alternate day. The functional analysis of the sciatic nerve (analysis of the march in track) was evaluated at weekly during all the experimental period. To verify the effectiveness of the ultrasound on nerve regeneration they had been analyzed sciatic functional index, morphology of the sciatic nerve and muscle morphology and morfometry of the muscles tibial anterior (TA) and sóleo (SOL). It was concluded that the treatment with the device of ultrasound of low intensity in regeneration peripheral nerve did not get resulted significant when compared the groups injured and injury+ultrasound, was only observed a maintenance of the cross-sectional area of the group injury+ultrasound of the muscle TA when compared with the normal group, taking the indications of that the ultrasound stimulated the nerve for such maintenance.

There is a large need for accurate methods detecting cardiovascular diseases, since they are one of the leading causes of mortality in the world, accounting for 29.3% of all deaths. Due to the complexity of the cardiovascular system, it is very challenging to develop methods for quantification of its function in order to diagnose, prevent and treat cardiovascular diseases. Ultrasound is a technique allowing for inexpensive, noninvasive imaging, but requires an experienced echocardiographer. Nowadays, methods like Tissue Doppler imaging (TDI) and Speckle tracking imaging (STI), measuring motion and deformation in the myocardium and the vessel walls, are getting more common in routine clinical practice, but without a proper visualization of the data provided by these methods, they are time-consuming and difficult to interpret. Thus, the general aim of this thesis was to develop novel ultrasound-based methods for accurate quantification and easily interpretable visualization of cardiovascular function. Five methods based on TDI and STI were developed in the present studies. The first study comprised development of a method for generation of bull’s-eye plots providing a color-coded two-dimensional visualization of myocardial longitudinal velocities. The second study proposed the state diagram of the heart as a new circular visualization tool for cardiac mechanics, including segmental color-coding of cardiac time intervals. The third study included development of a method describing the rotation pattern of the left ventricle by calculating rotation axes at different levels of the left ventricle throughout the cardiac cycle. In the fourth study, deformation data from the artery wall were tested as input to wave intensity analysis providing information of the ventricular – arterial interaction. The fifth study included an in-silico feasibility study to test the assessment of both radial and longitudinal strain in a kinematic model of the carotid artery. The studies showed promising results indicating that the methods have potential for the detection of different cardiovascular diseases and are feasible for use in the clinical setting. However, further development of the methods and both quantitative comparison of user dependency, accuracy and ease of use with other established methods evaluating cardiovascular function, as well as additional testing of the clinical potential in larger study populations, are needed.
QC 20100727

Nowadays, most of the medical technology manufacturers try to meet the high demands of quality and design of ultrasound equipment, including transducers: easy to use, flexible image handling, multi frequencies operation use of various applications, good resolution and penetration. It is known that this kind of medical devices is extremely sensitive and should be maintained in compliance with the necessary physical and functional requirements, via regular tests (Moore, 2006). This master thesis has two purposes. The major one is to examine the physical and functional status of every single transducer in Gävleborg hospitals. It allows defining a possible problem before it has influenced formation image and diagnosis. The second purpose is to find out the advantages and disadvantages of using the ultrasound evaluation test FirstCall aPerio Testing Accreditation Program, manufactured by Sonora Medical System in Longmont, USA (www.4sonora.com).The scientific method of this evaluation test is based on three steps which are as follows: testing of ultrasound transducers, classified data collection and analysis. The evaluation test used in this report is a highly qualified system which has been designed to avoid and minimize diagnostic risks, and at the same time, maximize patient’s safety. It allows distinguishing any hidden manufacturing defects with new devices or those appearing during the operation period. For this evaluation test, 73 transducers were controlled, belonging to clinics of four different hospitals within the Gävleborg area. Tested transducers were manufactured by Siemens, GE Healthcare and Aloka. A total of 61 of transducers passed the test and 12 did not pass it.  Finally it was observed, that the reason for not passing the evaluation test was mostly due to dead elements in a row in the centre of the transducers. Thanks to the software program of FirstCall aPerio, the results of the evaluation test are immediately visible, through the report. It is useful in early discovering the source of error in transducer and minimizing of wrong diagnosis. The testing system gives a possibility to test the transducer without US equipment and to collect the database information. Keywords: ultrasound, FirstCall aPerio Testing Accreditation Program, ultrasound transducers, evaluation test, wrong diagnosis.

This report evaluates possibilities to combine volumetric ultrasound (us) data together with the recent work published on advanced skin rendering techniques. We focus mainly on how to filter us data and localize surfaces within us data. We also evaluate recent skin rendering techniques in order to have a good understanding of what is needed from the us for rendering realistic skin. us data is acquired using sonography and have a low signal-to-noise ratio by nature, this makes it harder to extract surfaces compared to other medical data acquisition methods such as ct and mr. This report present an algorithm which implements a variational classification technique to emphasize surfaces within us and using a rbf network to fit an implicit function to these surfaces. Using this approach presented we have successfully extract smooth meshes from the noisy us data.

There is increasing interest in right ventricular function as an important determinant of cardiac output. However, the chamber is difficult to study, because of its shape and relationship to the left ventricle. Invasive studies, radionuclide studies and two-dimensional echocardiography are all useful approaches, but all have serious limitations. Systolic time intervals, best measured by pulsed Doppler ultrasound in the proximal pulmonary artery, offer one method of assessing right ventricular systolic function. Previous "normal" ranges, however, could be criticised on many counts. I present data from carefully checked normal controls and compare to previous publications, and explore variability and relationships between the various systolic time intervals. Most variables have skewed frequency distributions; the ranges are somewhat wider than previously described; most heart rate corrections are found to have serious limitations; and the effect of age is explored. Complete heart block offers a model to study the the effects of varying atrioventricular intervals whilst the ventricular rate is held unphysiologically steady by an artificial pacemaker. Given the current controversy about the merits of single- versus dual-chamber pacing, the issue is of topical interest also. The effect of varying the "P-R" interval within the physiological range is explored, and "optimal" ranges identified. A curious "nadir" effect, previously unknown, was discovered. When P waves followed paced QRS complexes at about 50-100ms, forward flow into the pulmonary artery (as judged from systolic time intervals) fell in most patients, and in some subjects virtually ceased. As a small included invasive part of the study showed, this was accompanied by falls in RV systolic pressure and rises in right atrial pressure. This study demonstrates that right ventricular systolic time intervals can be used to study right ventricular function in pacing situations, and is further evidence of the unsatisfactory nature of single-chamber ventricular pacing.

Le système visuel cérébral inclut le corps genouillé latéral, le colliculus supérieur et le cortex visuel. Ces structures ont été étudiées depuis les années 50 et cette étude a été possible par l’apparition de l’imagerie par résonnance magnétique fonctionnelle ainsi que par différentes techniques optiques. Pour évaluer la résolution spatiotemporelle de la technique d’imagerie ultrasonore, nous avons mesuré les réponses visuelles des structures cérébrales sur le rat anesthésié en étudiant leur dynamique respective soit sur un seul plan, soit par reconstruction tridimensionnelle. Après avoir validé cette approche technologique sur le petit animal, nous avons appliqué cette technique sur le cortex visuel du primate non-humain éveillé. Ainsi, nous avons pu reconstruire les cartes rétinotopiques du cortex visuel en surface mais aussi en profondeur. Nous avons mis en évidence, les bandes de dominance oculaire dans V1. En accord avec certaines données anatomiques, cette mesure fonctionnelle suggère la présence d’une dominance oculaire au-delà de la couche 4, dans les couches 3 et 5 de V1. En conclusion, ces travaux de thèse ont permis de montrer l’intérêt spécifique de l’imagerie fonctionnelle par ultrasons sur le rongeur et le primate non-humain, anesthésiés et vigiles. Cette nouvelle technique d'imagerie permet la visualisation fonctionnelle à haute résolution spatiotemporelle en profondeur des aires visuelles. Cette illustration sur le système visuel met en lumière l’intérêt prometteur du déploiement de cette technique pour l’exploration fonctionnelle de structures inaccessibles en imagerie optique de surface et en dessous de la résolution de l’IRMf
In the brain, the visual system includes the Lateral Geniculate Nucleus, the Superior Colliculus and the visual cortex. These structures have been studied since the 50’s and these studies have relied on functional MRI but also on the development of functional surface microscopic imaging techniques. To examine the spatiotemporal resolution of the ultrafast ultrasound imaging technique, we measured visual responses in in the cerebral structures on anesthetized rats in one imaging plan or with a 3D reconstruction. After having validated our technology on rodents, we have applied the technique to awake non-human primate visual cortex. We reconstructed the retinotopic maps of visual cortex, at the surface and in depth. In addition, we could resolve the ocular dominance columns within V1, which represent a major functional structuration in the primary visual cortex. This functional observation suggests the presence of ocular dominance beyond layer 4 in layers 3 and 5 of V1. In conclusion, this thesis work has demonstrated that the functional ultrafast ultrasound imaging can measure activity in the visual system for both rats and non-human primates, anesthetized and awake. This new imaging technique can provide functional measures with a high spatiotemporal resolution in the deep tissue. This illustration on the visual system highlights the potential of the technique to measure functional units not accessible to optical surface analysis and below the resolution of fMRI. Future studies will have to define if this technique can still provide a better resolution than fMRI in deeper structures than 1cm

Foi realizado um estudo experimental da influência do ultra-som terapêutico na regeneração do nervo ciático submetido a esmagamento controlado, em ratos. Foram empregados no estudo 20 ratos da linhagem Wistar, de peso médio em torno de 250 g, divididos em dois grupos, conforme o tipo de procedimento realizado: 1) somente esmagamento, dez ratos; e 2) esmagamento e irradiação com ultra-som, 10 ratos. Sob anestesia, o nervo ciático era exposto na coxa direita do animal e esmagado com um dispositivo especialmente idealizado e confeccionado para essa finalidade, com uma carga fixa de 15 Kg, por 10 minutos, num segmento de 5 mm proximal à sua bifurcação. A irradiação com o ultra-som pulsado (1:5, freqüência de 1 MHz, intensidade de 0.4 W/'CM POT.2', duração de 2 minutos) era iniciada já no primeiro dia pós-operatório e realizada por dez dias consecutivos. Os resultados foram avaliados pela análise funcional da impressão plantar dos animais obtidas numa pista de marcha e determinação do índice funcional do ciático (De Medinaceli e Bain, Mackinnon e Hunter), e pela morfometria, através do cálculo da densidade de fibras nervosas, após sacrifício dos animais no vigésimo primeiro dia pós-operatório. Os resultados foram submetidos a análise estatística e mostraram que, nas condições do trabalho, o ultra-som acelerou a regeneração do nervo, demonstrável com significância no vigésimo primeiro dia pós-operatório
An experimental study of the influence of therapeutic ultrasound on the regeneration of the sciatic nerve submitted to controlled crush injury was carried out in rats. Twenty female Wistar rats weighing 250 g on average were used and distributed in two groups according to the procedure: 1) crush injury only, ten rats; 2) crush injury followed by ultrasound irradiation, ten rats. Under general anesthesia the sciatic nerve was exposed on the right thigh and crushed with a device especially devised and built for that purpose, with a 15 kg constant load for 10 minutes, affecting a 5 mm-long segment of the nerve proximal to its bifurcation. Pulsed ultrasound irradiation (1:5, 1 MHz frequency, 0.4 W/'CM POT.2' intensity, 2 minutes duration) was started the day after the operation and repeated for ten consecutive days. The results were evaluated by functional footprint analysis and determination of the sciatic functional index (DeMedinaceli's and Bain, Mackinnon and Hunter's methods) at weekly intervals, and by morphometry (nerve fiber density) of the resected sciatic nerve after killing the animals on the 21st postoperative day. Results were submitted to statistical evaluation and showed that, in the conditions of the study, ultrasound accelerated nerve regeneration, as demonstrated with significance on the 21st postoperative day

Mes travaux de thèse portent sur l’application de l’imagerie fUS (functional ultrasound imaging) à l’imagerie cérébrale préclinique chez le petit animal. Le but était de transformer cette technique d’imagerie cérébrale récente en un véritable outil de quantification de l’état cérébral. Les objectifs principaux ont été de démontrer la faisabilité de l’imagerie fUS chez le petit animal non anesthésié ainsi que de passer du modèle rat au modèle souris - modèle de choix en imagerie préclinique en neurosciences - de surcroît de façon non invasive. J’ai tout d’abord mis au point une nouvelle séquence d’imagerie ultrasonore ultrarapide (Multiplane Wave imaging), permettant d’améliorer le rapport signal-à-bruit des images grâce à l’augmentation virtuelle de l’amplitude du signal émis, sans diminuer la cadence ultrarapide d’acquisition. Dans un deuxième temps j’ai démontré la possibilité d’imager le cerveau de la souris et du jeune rat anesthésiés par échographie Doppler ultrarapide, de manière transcrânienne et complètement non invasive, sans chirurgie ni injection d’agents de contraste. J’ai ensuite mis au point un montage expérimental, une séquence ultrasonore et un protocole expérimental permettant de réaliser de l’imagerie fUS de manière minimalement invasive chez des souris éveillées et libres de leurs mouvements. Enfin, j’ai démontré la possibilité d’utiliser le fUS pour étudier la connectivité fonctionnelle du cerveau au repos (sans stimulus) chez des souris éveillées ou sédatées. L’imagerie fUS et la combinaison « modèle souris » + « minimalement invasif » + « animal éveillé » + « connectivité fonctionnelle » constituent un outil précieux pour la communauté des neuroscientifiques travaillant sur des modèles animaux pathologiques ou de nouvelles molécules pharmacologiques
My work focuses on the application of fUS (functional ultrasound) imaging to preclinical brain imaging in small animals. The goal of my thesis was to turn this recent vascular brain imaging technique into a quantifying tool for cerebral state. The main objectives were to demonstrate the feasibility of fUS imaging in the non-anaesthetized small rodents and to move from rat model imaging to mouse model imaging –most used model for preclinical studies in neuroscience-, while developing the least invasive imaging protocols. First, I have developed a new ultrafast ultrasonic imaging sequence (Multiplane Wave imaging), improving the image signal-to-noise ratio by virtually increasing emitted signal amplitude, without reducing the ultrafast framerate. Then, I have demonstrated the possibility to use ultrafast Doppler ultrasound imaging to image both the mouse brain and the young rat brain, non-invasively and through the intact skull, without surgery or contrast agents injection. Next, I have developed an experimental setup, an ultrasound sequence and an experimental protocol to perform minimally invasive fUS imaging in awake and freely-moving mice. Finally, I have demonstrated the possibility to use fUS imaging to study the functional connectivity of the brain in a resting state in awake or sedated mice, still in a transcranial and minimally invasive way. fUS imaging and the combination of "mouse model" + "minimally invasive" + "awake animal" + "functional connectivity" represent a very promising tool for the neuroscientist community working on pathological animal models or new pharmacological molecules

The aim of this study was to define new quantitative parameters of dynamic Pelvic Floor Muscle (PFM) function using 2D transperineal (TP) ultrasound imaging combined with image processing methods (IPM). Ultrasound and digital vaginal data were obtained from a volunteer convenience sample of 23 continent and 9 Stress Urinary Incontinent (SUI) women recruited from the San Francisco Bay Area community, California, U.S.A. Two methods of IPM were used; speckle tracking for measuring displacement of the ano-rectal angle (ARA) and symphysis pubis (SP); and segmentation IPM for measuring displacement of the urethra. Good to excellent inter and intra observer reliability was established for processing the ultrasound images on stored audio-visual imaging files (AVI). Intraclass correlation coefficient (ICC) values: 0.61 to 0.99; 95% confidence intervals (CI) 0.08→0.84 to 0.97→0.99; standard error of measurement (SEM) 0.01to 0.25 cm. There was good agreement between measures on Bland and Altman analysis: mean difference 0.06 to -0.21 cm; 95% CI 0.02→0.45cm to -0.01→0.20cm. Validity of the IPM was confirmed with digital palpation, and furthermore indicated good sensitivity and specificity. Technical and methodological limitations of the IPM, and suggestions for future studies were discussed. Several research questions were addressed using TP ultrasound combined with IPM that enabled the dynamic evaluation of pelvic floor (PF) displacement throughout an entire manoeuvre, rather than limiting the quantification from static images at rest to the end of the manoeuvre. In this way, IPM determined unique quantitative data regarding the automatic displacement, velocity and acceleration of the ARA and urethra during a cough, Valsalva, voluntary PFM contraction, The Knack and a Transversus Abdominis contraction. During automatic events that raised intra-abdominal pressure (IAP), normal PFM function produced timely compression of the PF and additional external support to the urethra, reducing the displacement, velocity, and acceleration. In women with SUI, who have weaker urethral attachments, this shortening contraction did not occur; consequently, the urethra of women with SUI moved further and faster for a longer duration. Caution regarding the generalisability of this study is warranted due to the small number of women in the SUI group and the significant difference in parity between groups. However this non-invasive physiological measurement tool demonstrated a new way of assessing the PFM. It is envisaged that this study will provide the foundation for future studies with larger parity matched populations and eventually improve the rehabilitation of women with SUI and other PF disorders.

Aim: To measure fetal cardiac output prior to labour and assess the risk of adverse obstetric and neonatal outcome in singleton pregnancies with appropriately grown for gestational age (AGA) fetuses at term. Methods: This was a prospective observational study conducted at Queen Charlotte’s and Chelsea Hospital, London UK. Fetal cardiac output and fetal cerebroplacental ratio (CPR) was measured within 72 hours before birth in 200 nulliparous women having singleton pregnancies with AGA fetuses. Scan details were not available to the clinicians and delivery was managed per the local protocol and guidelines. Obstetric and neonatal outcomes were obtained from case notes and correlated with the ultrasound findings. Results: Delivery was vaginal in 129 (64.5%) cases and by caesarean section in 71 (35.5%), including 34 (17.0%) for fetal distress and 37 (18.5%) for failure to progress. Fetuses delivered by caesarean section for fetal distress, compared to the remaining fetuses, had a lower median left cardiac output(LCO) (152.3 vs. 191.7 mL/min/kg; p=0.003), higher difference in the median ratio between the right to left cardiac output (RCO to LCO ratio) 1.925 vs. 1.340; p=0.002) and lower CPR (1.222 vs. 1.607; p < 0.0001). In screening for emergency caesarean section for fetal distress, for a 10% false positive rate, the detection rate with the RCO to LCO ratio was higher that with the LCO (41% vs. 29%) and with the CPR (41% vs. 27%). Similarly, the positive predictive value for the RCO to LCO ratio (45%) was higher than LCO (37%) and the CPR (35%). Conclusion: In AGA fetuses at term that develop intrapartum distress, there is evidence of prelabour redistribution of the cardiac output. The RCO to LCO ratio is superior to the LCO and CPR in predicting intrapartum fetal distress. Such assessments may be useful in stratifying patients for the intensity of monitoring during labour.

Pour répondre à des impératifs croissants de fiabilité et de sûreté, les procédés mis en œuvre dans le cadre du contrôle non destructif sont en constante évolution. Au vu de la complexité des techniques utilisées, la simulation prend une part importante dans leur développement. Nous présentons des travaux ayant abouti à un outil rapide de simulation du champ émis par un traducteur ultrasonore plan quelconque dans des configurations complexes de contrôle non destructif impliquant des géométries maillées sans arêtes saillantes, des matériaux isotropes ou anistropes, homogènes ou hétérogènes et des trajectoires d'ondes pouvant comporter des rebonds et des transmissions. Les fronts d'ondes ultrasonores sont approximés à l'aide d'interpolateurs polynomiaux locaux à des pinceaux de rayons ultrasonores. Ceux-ci sont obtenus au moyen d'un algorithme de recherche de surface par lancer de pinceaux et subdivisions successives. Ils permettent le calcul des grandeurs utiles à la constitution de la réponse impulsionnelle en chaque point d'un échantillonnage du traducteur respectant le critère de Shannon. De cette façon, nous pouvons calculer une réponse impulsionnelle qui, convoluée au signal d'excitation du traducteur, donne le champ ultrasonore. Les performances des simulations ont été accrues par l'exploitation du parallélisme de tâches et des instructions SIMD dans les parties les plus coûteuses du calcul. Enfin, un outil de calcul progressif continu a été développé pour permettre une visualisation interactive d'images de champ. Il exploite une méthode de reconstruction d'images et ordonnance les calculs de champ de manière à accélérer la convergence des images produites
In order to fulfill increasing reliability and safety requirements, non destructive testing techniques are constantly evolving and so does their complexity. Consequently, simulation is an essential part of their design. We developed a tool for the simulation of the ultrasonic field radiated by any planar probes into non destructive testing configurations involving meshed geometries without prominent edges, isotropic and anisotropic, homogeneous and heterogeneous materials, and wave trajectories that can include reflections and transmissions. We approximate the ultrasonic wavefronts by using polynomial interpolators that are local to ultrasonic ray pencils. They are obtained using a surface research algorithm based on pencil tracing and successive subdivisions. Their interpolators enable the computation of the necessary quantities for the impulse response computation on each point of a sampling of the transducer surface that fulfills the Shannon criterion. By doing so, we can compute a global impulse response which, when convoluted with the excitation signal of the transducer, results in the ultrasonic field. The usage of task parallelism and of SIMD instructions on the most computationally expensive steps yields an important performance boost. Finally, we developed a tool for progressive visualization of field images. It benefits from an image reconstruction technique and schedules field computations in order to accelerate convergence towards the final image

Elastography, which is based on applying pressure and estimating the resulting deformation, involves the forward problem to obtain the strain distributions and inverse problem to construct the elastic distributions consistent with the obtained strains on observation points. This thesis focuses on the former problem whose solution is used as an input to the latter problem. The aim is to provide the inverse problem community with accurate strain estimates of a coronary artery vessel wall. In doing so, a new ultrasonic image-based elastography approach is developed. Because the accuracy and quality of the estimated strain fields depend on the resolution level of the ultrasound image and to date best resolution levels obtained in the literature are not enough to clearly see all boundaries of the artery, one of the main goals is to acquire high-resolution coronary vessel wall ultrasound images at different pressures. For this purpose, first an experimental setup is designed to collect radio frequency (RF) signals, and then image formation algorithm is developed to obtain ultrasound images from the collected signals. To segment the noisy ultrasound images formed, a geodesic active contour-based segmentation algorithm with a novel stopping function that includes local phase of the image is developed. Then, region-based information is added to make the segmentation more robust to noise. Finally, elliptical deformable template is applied so that a priori information regarding the shape of the arteries could be taken into account, resulting in more stable and accurate results. The use of this template also implicitly provides boundary point correspondences from which high-resolution, size-independent, non-rigid and local strain fields of the coronary vessel wall are obtained.

Thesis (M.S.)--Worcester Polytechnic Institute.
Keywords: angular decorrelation function; impulse response; BUA; BMD; ultrasound; coral; trabecular bone; osteoporosis. Includes bibliographical references (p. 189-191).

Heart disease is one of the primary causes of morbidity and mortality for the adult population over the age of 65. Furthermore, ailments such as hypertension can affect as many as 50% of the adult population over the age of 45. If left untreated, these ailments eventually precipitate the onset of diastolic dysfunction and heart failure. Diastolic dysfunction is the alteration or impairment of performance in either the left or right ventricle of the heart. Although there has been a marked increase in study of this disease, there is still an apparent difficulty to diagnose patients. Flow visualization techniques have been commonly employed to study the development of these diseases as they relate to the filling process of the ventricles. One method, Echo Particle Image Velocimetry (Echo-PIV) is a relatively new method for cardiac flow chamber visualization, with the potential to provide physicians with a cost-effective and safe method for obtaining high temporal resolution recordings for extending knowledge on the filling processes in cardiac chamber flow. This work presents a new approach to extending the capabilities of Echo-PIV for more accurate measurement of cardiac flows for patients with poor quality recordings. Currently, much of the literature notes that temporal resolution and poor acoustic windows results in exclusion from study. These recordings are more representative of the contrast-enhancement studies used by physicians to better identify chamber walls. When applying standard PIV cross-correlation techniques, measurements tend to fail due to image noise and artifacts. By implementing a Moving Ensemble (MWE) with Product of Correlation (PoC) processing scheme, measurement accuracy, reliability, and robustness can be obtained for measurement in left ventricular filling assessment.
Master of Science