Quantitative 3·D Echocardiography of The Heart and The Coronary Vessels

The recognition of the existence of ultrasound is credited to L. Spallanzani (1729- 1799). In recent years, ultrasound has been used as an imaging modality in medicine. I. Edler and C.H. Hertz produced the first ultrasound images of the heart in 1953. In the 1960's great progress was made in the clinical application of ultrasound when real-time two-dimensional ultrasound scanners were developed. In 1968, J. Somer constructed the first electronic phased-array scanner and this technology is still the most widely used in ultrasound equipment. In 1974 F.E. Barber and colleagues produced a duplex scanner which integrated imaging with pulsed-wave Doppler measurements. C. Kasai and colleagues constmcted in 1982 the color-coded Doppler flow imaging system based on autocorrelation detection, providing a noninvasive "angiogram" simulation of normal and abnormal blood flow on a "beat-to-beat" basis. Transesophageal echocardiography became available to clinicians in 1985 due to the developments of 1. Soquet who invented the mono- and biplane electronic phased-array probel Echocardiography has become one of the most commonly used diagnostic imaging techniques in cardiology. The development of commercial 3-D echocardiographic equipment began in the early 1990's. In 1993 a technique allowing acquisition of tomographic parallel sliced data set of echocardiographic images of the heart with a lobster tail TEE probe, was 2 developed by the German based company "TomTec GmbH". The TEE probe had an imaging element which could be controlled by computer applying a stepping motor. They also developed an interface to the patient to record the respiration and R-R intervals. This allowed the acquisition of ultrasound images ECG-triggered and gated, which reduced motion artifacts caused by beat-to-beat and respiratory variations in cardiac dimensions and position. After the acquisition of a tomographic data set, the images were post-processed and with application of software interpolation algorithms, gaps in the data set could be filled. This post-processed data set could then be used to reconstruct 3-D volume rendered images of the heart. 3-D ultrasound provides cardiac images which more closely mimic actual anatomy'than 2-D cross-sectional linages, and may thus be easier to interpret

Glioma on Chips Analysis of glioma cell guidance and interaction in microfluidic-controlled microenvironment enabled by machine learning

In biosystems, chemical and physical fields established by gradients guide cell migration, which is a fundamental phenomenon underlying physiological and pathophysiological processes such as development, morphogenesis, wound healing, and cancer metastasis. Cells in the supportive tissue of the brain, glia, are electrically stimulated by the local field potentials from neuronal activities. How the electric field may influence glial cells is yet fully understood. Furthermore, the cancer of glia, glioma, is not only the most common type of brain cancer, but the high-grade form of it (glioblastoma) is particularly aggressive with cells migrating into the surrounding tissues (infiltration) and contribute to poor prognosis. In this thesis, I investigate how electric fields in the microenvironment can affect the migration of glioblastoma cells using a versatile microsystem I have developed. I employ a hybrid microfluidic design to combine poly(methylmethacrylate) (PMMA) and poly(dimethylsiloxane) (PDMS), two of the most common materials for microfluidic fabrication. The advantages of the two materials can be complemented while disadvantages can be mitigated. The hybrid microfluidics have advantages such as versatile 3D layouts in PMMA, high dimensional accuracy in PDMS, and rapid prototype turnaround by facile bonding between PMMA and PDMS using a dual-energy double sided tape. To accurately analyze label-free cell migration, a machine learning software, Usiigaci, is developed to automatically segment, track, and analyze single cell movement and morphological changes under phase contrast microscopy. The hybrid microfluidic chip is then used to study the migration of glioblastoma cell models, T98G and U-251MG, in electric field (electrotaxis). The influence of extracellular matrix and chemical ligands on glioblastoma electrotaxis are investigated. I further test if voltage-gated calcium channels are involved in glioblastoma electrotaxis. The electrotaxes of glioblastoma cells are found to require optimal laminin extracellular matrices and depend on different types of voltage-gated calcium channels, voltage-gated potassium channels, and sodium transporters. A reversiblysealed hybrid microfluidic chip is developed to study how electric field and laminar shear can condition confluent endothelial cells and if the biomimetic conditions affect glioma cell adhesion to them. It is found that glioma/endothelial adhesion is mediated by the Ang1/Tie2 signaling axis and adhesion of glioma is slightly increased to endothelial cells conditioned with shear flow and moderate electric field. In conclusion, robust and versatile hybrid microsystems are employed for studying glioma biology with emphasis on cell migration. The hybrid microfluidic tools can enable us to elucidate fundamental mechanisms in the field of the tumor biology and regenerative medicine.Okinawa Institute of Science and Technology Graduate Universit

Three-Dimensional Intravascular UItrasound Assessment of Coronary Lumen and Atherosclerotic Plaque Dimensions

Since the introduction of coronary balloon angioplasty in the clinical arena, percutaneous catheter- based interventions are perfornled with coronary angiographic guidance, depicting the lumen of an entire coronary artery in certain angiographicviews. Subsequently, quantitative coronary angiography was developed as an instnullent for off-line quantitative analysis of the acute and long-tenn effects of catheter-based and phanl1acological approaches on atherosclerotic lesions and on lesion recurrence following angioplasty. Despite some inherent limitations, tills analysis method became generally accepted for on-line guidance of balloon angioplasty and alternative catheterbased techniques. Thereafter, intravascular ultrasound (IVUS) was introduced as a new imaging method that provided deeper insights into the pathology of coronary artery disease by defining vessel wall geometry and the major components of the atherosclerotic plaque. Although invasive, IVUS is safe and allows in vivo a more comprehensive assessment of the plaque than the 'luminal silhouette' furnished by coro

OPTICAL GEOMETRY CALIBRATION METHOD FOR COMPUTED TOMOGRAPHY AND APPLICATIONS OF COMPACT MICROBEAM RADIATION THERAPY

Digital tomosynthesis is a type of limited angle tomography that allows for 3D information reconstructed from a set of X-ray projection images taken at various angles using an X-ray tube, a mechanical arm to rotate the tube, and a digital detector. Tomosynthesis reconstruction requires the knowledge of the precise location of the detector with respect to each X-ray source. Current clinical tomosynthesis methods use a physically coupled source and detector so the geometry is always known and is always the same. This makes it impractical for mobile or field operations. We demonstrated a free form tomosynthesis and free form computed tomography (CT) with a decoupled source and detector setup that uses a novel optical method for accurate and real-time geometry calibration. We accomplish this by using a camera to track the motion of the source relative to the detector. A checkerboard pattern is positioned on or next to the detector using an extension arm in such a way that the pattern will not move relative to the detector. A camera is mounted on the source in a way that the pattern is visible during imaging and will not move relative to the source. The image of the pattern captured by the camera is then used to determine the relative camera/pattern position and orientation by analyzing the pattern distortion. This allows for accurate, real time geometry calibration of the X-ray source relative to the detector. This method opens the doors for inexpensive upgrades to existing 2D imaging systems and an even more exciting application of a mobile, hand-held CT imaging system.Doctor of Philosoph

ALDH4A1 is an atherosclerosis auto-antigen targeted by protective antibodies

Cardiovascular disease (CVD) is the leading cause of mortality in the world, with most CVD-related deaths resulting from myocardial infarction or stroke. The main underlying cause of thrombosis and cardiovascular events is atherosclerosis, an inflammatory disease that can remain asymptomatic for long periods. There is an urgent need for therapeutic and diagnostic options in this area. Atherosclerotic plaques contain autoantibodies, and there is a connection between atherosclerosis and autoimmunity. However, the immunogenic trigger and the effects of the autoantibody response during atherosclerosis are not well understood. Here we performed high-throughput single-cell analysis of the atherosclerosis-associated antibody repertoire. Antibody gene sequencing of more than 1,700 B cells from atherogenic Ldlr and control mice identified 56 antibodies expressed by in-vivo-expanded clones of B lymphocytes in the context of atherosclerosis. One-third of the expanded antibodies were reactive against atherosclerotic plaques, indicating that various antigens in the lesion can trigger antibody responses. Deep proteomics analysis identified ALDH4A1, a mitochondrial dehydrogenase involved in proline metabolism, as a target antigen of one of these autoantibodies, A12. ALDH4A1 distribution is altered during atherosclerosis, and circulating ALDH4A1 is increased in mice and humans with atherosclerosis, supporting the potential use of ALDH4A1 as a disease biomarker. Infusion of A12 antibodies into Ldlr mice delayed plaque formation and reduced circulating free cholesterol and LDL, suggesting that anti-ALDH4A1 antibodies can protect against atherosclerosis progression and might have therapeutic potential in CVD.Ministerio de Economía y Competitividad (SVP-2014-068289); P.D. was supported by an AECC grant (AIO 2012, Ayudas a Investigadores en Oncología 2012); A.S.-B. is a Juan de la Cierva researcher (IJC2018-035279-I); I.M.-F. was a fellow of the research training program funded by Ministerio de Economía y Competitividad (SVP-2014-068216); and A.R.R. and J.V. are supported by Centro Nacional de Investigaciones Cardiovasculares (CNIC). The project leading to these results has received funding from la Caixa Banking Foundation under the project code HR17-00247 and from SAF2016-75511-R and PID2019-106773RB-I00 grants to A.R.R. (Plan Estatal de Investigación Científica y Técnica y de Innovación 2013–201

Terahertz Plasmonic Devices

Terahertz (THz) devices are designed to operate from 0.1-10 THz. The THz spectra have unique properties such as penetration through soft materials and reflecting from hard materials, which make THz technologies, a prime candidate for imaging. Plasmons are longitudinal charge oscillations in carrier rich materials. Plasmons can be generated over the channel of transistors inducing a voltage between the source-drain when conditions are satisfied. In this thesis, plasmonic devices operating in the THz region have been studied both theoretically and experimentally investigating GaN/AlGaN and Graphene based transistors. First, we report on a detailed study of dispersion properties of uniform grating gate THz plasmonic crystals, asymmetric dual grating gate plasmonic crystals and with symmetry-breaking defect-like cavities in order to understand the physics behind THz plasmons. For the first time, we defined the dispersion of plasmons in terms of effective plasmonic index. By adding an additional grating on top of the grating gate with a different periodicity, doubles the amount of absorption. Plasmons can be excited when polarization is perpendicular to the gate. We then showed focusing and exciting of THz plasmons polarization independent using circular grating lenses. Sub-micron THz ring resonators are presented showing THz guiding in plasmonic waveguides. So far, resonant sensing has been observed only at cryogenic temperatures since electron mobility is high enough at low temperatures to sustain resonant plasmonic excitation at the channel of the detector. Recently, graphene attracted the attention of the researchers because of its high mobility at room temperature. Room temperature detection has been attempted and achieved, however the detectors have very small responsivity with non-resonant behavior since the graphene is sandwiched and fabrication of such detectors in large scale is impossible with the methods used. Here, we present a resonant room temperature detection of THz with upside down free standing graphene FETs having more than a 400 quality factor, a record high number in the field which is up to 50 times higher than GaN detectors and hundreds of responsivity values with a maximum around 400 V/W which is record high for graphene (10,000 times higher than previously reported graphene detector)

A portable device for time-resolved fluorescence based on an array of CMOS SPADs with integrated microfluidics

[eng] Traditionally, molecular analysis is performed in laboratories equipped with desktop instruments operated by specialized technicians. This paradigm has been changing in recent decades, as biosensor technology has become as accurate as desktop instruments, providing results in much shorter periods and miniaturizing the instrumentation, moving the diagnostic tests gradually out of the central laboratory. However, despite the inherent advantages of time-resolved fluorescence spectroscopy applied to molecular diagnosis, it is only in the last decade that POC (Point Of Care) devices have begun to be developed based on the detection of fluorescence, due to the challenge of developing high-performance, portable and low-cost spectroscopic sensors. This thesis presents the development of a compact, robust and low-cost system for molecular diagnosis based on time-resolved fluorescence spectroscopy, which serves as a general-purpose platform for the optical detection of a variety of biomarkers, bridging the gap between the laboratory and the POC of the fluorescence lifetime based bioassays. In particular, two systems with different levels of integration have been developed that combine a one-dimensional array of SPAD (Single-Photon Avalanch Diode) pixels capable of detecting a single photon, with an interchangeable microfluidic cartridge used to insert the sample and a laser diode Pulsed low-cost UV as a source of excitation. The contact-oriented design of the binomial formed by the sensor and the microfluidic, together with the timed operation of the sensors, makes it possible to dispense with the use of lenses and filters. In turn, custom packaging of the sensor chip allows the microfluidic cartridge to be positioned directly on the sensor array without any alignment procedure. Both systems have been validated, determining the decomposition time of quantum dots in 20 nl of solution for different concentrations, emulating a molecular test in a POC device.[cat] Tradicionalment, l'anàlisi molecular es realitza en laboratoris equipats amb instruments de sobretaula operats per tècnics especialitzats. Aquest paradigma ha anat canviant en les últimes dècades, a mesura que la tecnologia de biosensor s'ha tornat tan precisa com els instruments de sobretaula, proporcionant resultats en períodes molt més curts de temps i miniaturitzant la instrumentació, permetent així, traslladar gradualment les proves de diagnòstic fora de laboratori central. No obstant això i malgrat els avantatges inherents de l'espectroscòpia de fluorescència resolta en el temps aplicada a la diagnosi molecular, no ha estat fins a l'última dècada que s'han començat a desenvolupar dispositius POC (Point Of Care) basats en la detecció de la fluorescència, degut al desafiament que suposa el desenvolupament de sensors espectroscòpics d'alt rendiment, portàtils i de baix cost. Aquesta tesi presenta el desenvolupament d'un sistema compacte, robust i de baix cost per al diagnòstic molecular basat en l'espectroscòpia de fluorescència resolta en el temps, que serveixi com a plataforma d'ús general per a la detecció òptica d'una varietat de biomarcadors, tancant la bretxa entre el laboratori i el POC dels bioassaigs basats en l'anàlisi de la pèrdua de la fluorescència. En particular, s'han desenvolupat dos sistemes amb diferents nivells d'integració que combinen una matriu unidimensional de píxels SPAD (Single-Photon Avalanch Diode) capaços de detectar un sol fotó, amb un cartutx microfluídic intercanviable emprat per inserir la mostra, així com un díode làser UV premut de baix cost com a font d'excitació. El disseny orientat a la detecció per contacte de l'binomi format pel sensor i la microfluídica, juntament amb l'operació temporitzada dels sensors, permet prescindir de l'ús de lents i filtres. Al seu torn, l'empaquetat a mida de l'xip sensor permet posicionar el cartutx microfluídic directament sobre la matriu de sensors sense cap procediment d'alineament. Tots dos sistemes han estat validats determinant el temps de descomposició de "quantum dots" en 20 nl de solució per a diferents concentracions, emulant així un assaig molecular en un dispositiu POC