Ricerca di nuove strategie differenziative: Analisi degli effetti di stimoli fisici e molecolari su cellule mesenchimali umane isolate da tessuto adiposo

Introduzione. Le cellule mesenchimali derivate dal tessuto adiposo (hASC) rappresentano un importante strumento per la terapia cellulare, in quanto derivano da un tessuto adulto abbondante e facilmente reperibile. Con il dispositivo medico Lipogems l’isolamento di tali cellule è eseguito esclusivamente mediante sollecitazioni meccaniche. Il prodotto ottenuto è quindi minimamente manipolato e subito utilizzabile. Ad oggi, il condizionamento pro-differenziativo delle staminali è per lo più attuato mediante molecole di sintesi. Tuttavia, altri fattori possono modulare la fisiologia cellulare, come gli stimoli fisici e molecole naturali. Onde elettromagnetiche hanno indotto in modelli cellulari staminali l’espressione di alcuni marcatori di differenziamento e, in cellule adulte, una riprogrammazione, mentre estratti embrionali di Zebrafish sono risultati antiproliferativi sia in vitro che in vivo. Metodi. La ricerca di nuove strategie differenziative sia di natura fisica che molecolare, nel particolare onde acustiche ed estratti embrionali di Zebrafish, è stata condotta utilizzando come modello cellulare le hASC isolate con Lipogems. Onde acustiche sono state somministrate mediante l’utilizzo di due apparati di trasduzione, un generatore di onde meccaniche e il Cell Exciter . I trattamenti con gli estratti embrionali sono stati effettuati utilizzando diverse concentrazioni e diversi tempi sperimentali. Gli effetti sull’espressione dei marcatori di staminalità e differenziamento relativi ai trattamenti sono stati saggiati in RT-PCR quantitativa relativa e/o in qPCR. Per i trattamenti di tipo molecolare è stata valutata anche la proliferazione. Risultati e conclusioni. La meta-analisi dei dati delle colture di controllo mostra la stabilità d’espressione genica del modello. I trattamenti con i suoni inducono variazioni dell’espressione genica, suggerendo un ruolo regolatorio di tali stimoli, in particolare del processo di commitment cardiovascolare. Due degli estratti embrionali di Zebrafish testati inibiscono la proliferazione alle 72 ore dalla somministrazione. L’analisi d’espressione associata ai trattamenti antiproliferativi suggerisce che tale effetto abbia basi molecolari simili ai processi di differenziamento.

Fluctuation spectroscopy of single plasmonic nanostructures

Plasmonic nanoparticles are great candidates for sensing applications with optical read-out. Plasmon sensing is based on the interaction of the nanoparticle with electromagnetic waves where the particle scatters light at its resonance wavelength. This wavelength depends on several intrinsic factors like material, shape and size of the nanoparticle as well as extrinsic factors like the refractive index of the surrounding medium. The latter allows the nanoparticle to be used as a sensor; changes in the proximate environment can be directly monitored by the wavelength of the emitted light. Due to their minuscule size and high sensitivity this allows individual nanoparticles to report on changes in particle coverage.rnrnTo use this single particle plasmon sensor for future sensing applications it has to meet the demand for detection of incidents on the single molecule level, such as single molecule sensing or even the detection of conformational changes of a single molecule. Therefore, time resolution and sensitivity have to be enhanced as today’s measurement methods for signal read-out are too slow and not sensitive enough to resolve these processes. This thesis presents a new experimental setup, the 'Plasmon Fluctuation Setup', that leads to tremendous improvements in time resolution and sensitivity. This is achieved by implementation of a stronger light source and a more sensitive detector. The new setup has a time resolution in the microsecond regime, an advancement of 4-6 orders of magnitude to previous setups. Its resonance wavelength stability of 0.03 nm, measured with an exposure time of 10 ms, is an improvement of a factor of 20 even though the exposure time is 3000 times shorter than in previous reports. Thus, previously unresolvable wavelength changes of the plasmon sensor induced by minor local environmental alteration can be monitored with extremely high temporal resolution.rnrnUsing the 'Plasmon Fluctuation Setup', I can resolve adsorption events of single unlabeled proteins on an individual nanorod. Additionally, I monitored the dynamic evolution of a single protein binding event on a millisecond time scale. This feasibility is of high interest as the role of certain domains in the protein can be probed by a study of modified analytes without the need for labels possibly introducing conformational or characteristic changes to the target. The technique also resolves equilibrium fluctuations in the coverage, opening a window into observing Brownian dynamics of unlabeled macromolecules. rnrnA further topic addressed in this thesis is the usability of the nanoruler, two nanospheres connected with a spacer molecule, as a stiffness sensor for the interparticle linker under strong illumination. Here, I discover a light induced collapse of the nanoruler. Furthermore, I exploit the sensing volume of a fixed nanorod to study unlabeled analytes diffusing around the nanorod at concentrations that are too high for fluorescence correlation spectroscopy but realistic for biological systems. Additionally, local pH sensing with nanoparticles is achieved.

Analysis of retrieved shoulder prostheses

In the last years the number of shoulder arthroplasties has been increasing. Simultaneously the study of their shape, size and strength and the reasons that bring to a possible early explantation have not yet been examined in detail. The research carried out directly on explants is practically nonexistent, this means a poor understanding of the mechanisms leading the patient and so the surgeon, to their removal. The analysis of the mechanisms which are the cause of instability, dislocation, broken, fracture, etc, may lead to a change in the structure or design of the shoulder prostheses and lengthen the life of the implant in situ. The idea was to analyze 22 explants through three methods in order to find roughness, corrosion and surface wear. In the first method, the humeral heads and/or the glenospheres were examined with the interferometer, a machine that through electromagnetic waves gives information about the roughness of the surfaces under examination. The output of the device was a total profile containing both roughness and information on the waves (representing the spatial waves most characteristic on the surface). The most important value is called "roughness average" and brings the average value of the peaks found in the local defects of the surfaces. It was found that 42% of the prostheses had considerable peak values in the area where the damage was caused by the implant and not only by external events, such as possibly the surgeon's hand. One of the problems of interest in the use of metallic biomaterials is their resistance to corrosion. The clinical significance of the degradation of metal implants has been the purpose of the second method; the interaction between human body and metal components is critical to understand how and why they arrive to corrosion. The percentage of damage in the joints of the prosthetic components has been calculated via high resolution photos and the software ImageJ. The 40% and 50% of the area appeared to have scratches or multiple lines due to mechanical artifacts. The third method of analysis has been made through the use of electron microscopy to quantify the wear surface in polyethylene components. Different joint movements correspond to different mechanisms of damage, which were imprinted in the parts of polyethylene examined. The most affected area was located mainly in the side edges. The results could help the manufacturers to modify the design of the prostheses and thus reduce the number of explants. It could also help surgeons in choosing the model of the prosthesis to be implanted in the patient.

X-ray phase contrast imaging of metallized and non-metallized biological samples

Imaging of biological samples has been performed with a variety of techniques for example electromagnetic waves, electrons, neutrons, ultrasound and X-rays. Also conventional X-ray imaging represents the basis of medical diagnostic imaging, it remains of limited use in this application because it is based solely on the differential absorption of X-rays by tissues. Coherent and bright photon beams, such as those produced by third-generation synchrotron X-ray sources, provide further information on subtle X-ray phase changes at matter interfaces. This complements conventional X-ray absorption by edge enhancement phenomena. Thus, phase contrast imaging has the potential to improve the detection of structures on images by detecting those structures that are invisible with X-ray absorption imaging. Images of a weakly absorbing nylon fibre were recorded in in-line holography geometry using a high resolution low-noise CCD camera at the ESRF in Grenoble. The method was also applied to improve image contrast for images of biological tissues. This paper presents phase contrast microradiographs of vascular tree casts and images of a housefly. These reveal very fine structures, that remain invisible with conventional absorption contrast only.

Contribution to the Analysis, Design and Assessment of Compact Antenna Test Ranges At Millimeter Wavelengths - Contribución al Análisis, Diseño y Caracterización de Sistemas de Medida de Antenas en Rango Compacto para Frecuencias Milimétricas

This Doctoral Thesis entitled Contribution to the analysis, design and assessment of compact antenna test ranges at millimeter wavelengths aims to deepen the knowledge of a particular antenna measurement system: the compact range, operating in the frequency bands of millimeter wavelengths. The thesis has been developed at Radiation Group (GR), an antenna laboratory which belongs to the Signals, Systems and Radiocommunications department (SSR), from Technical University of Madrid (UPM). The Radiation Group owns an extensive experience on antenna measurements, running at present four facilities which operate in different configurations: Gregorian compact antenna test range, spherical near field, planar near field and semianechoic arch system. The research work performed in line with this thesis contributes the knowledge of the first measurement configuration at higher frequencies, beyond the microwaves region where Radiation Group features customer-level performance. To reach this high level purpose, a set of scientific tasks were sequentially carried out. Those are succinctly described in the subsequent paragraphs. A first step dealed with the State of Art review. The study of scientific literature dealed with the analysis of measurement practices in compact antenna test ranges in addition with the particularities of millimeter wavelength technologies. Joint study of both fields of knowledge converged, when this measurement facilities are of interest, in a series of technological challenges which become serious bottlenecks at different stages: analysis, design and assessment. Thirdly after the overview study, focus was set on Electromagnetic analysis algorithms. These formulations allow to approach certain electromagnetic features of interest, such as field distribution phase or stray signal analysis of particular structures when they interact with electromagnetic waves sources. Properly operated, a CATR facility features electromagnetic waves collimation optics which are large, in terms of wavelengths. Accordingly, the electromagnetic analysis tasks introduce an extense number of mathematic unknowns which grow with frequency, following different polynomic order laws depending on the used algorithmia. In particular, the optics configuration which was of our interest consisted on the reflection type serrated edge collimator. The analysis of these devices requires a flexible handling of almost arbitrary scattering geometries, becoming this flexibility the nucleus of the algorithmia’s ability to perform the subsequent design tasks. This thesis’ contribution to this field of knowledge consisted on reaching a formulation which was powerful at the same time when dealing with various analysis geometries and computationally speaking. Two algorithmia were developed. While based on the same principle of hybridization, they reached different order Physics performance at the cost of the computational efficiency. Inter-comparison of their CATR design capabilities was performed, reaching both qualitative as well as quantitative conclusions on their scope. In third place, interest was shifted from analysis - design tasks towards range assessment. Millimetre wavelengths imply strict mechanical tolerances and fine setup adjustment. In addition, the large number of unknowns issue already faced in the analysis stage appears as well in the on chamber field probing stage. Natural decrease of dynamic range available by semiconductor millimeter waves sources requires in addition larger integration times at each probing point. These peculiarities increase exponentially the difficulty of performing assessment processes in CATR facilities beyond microwaves. The bottleneck becomes so tight that it compromises the range characterization beyond a certain limit frequency which typically lies on the lowest segment of millimeter wavelength frequencies. However the value of range assessment moves, on the contrary, towards the highest segment. This thesis contributes this technological scenario developing quiet zone probing techniques which achieves substantial data reduction ratii. Collaterally, it increases the robustness of the results to noise, which is a virtual rise of the setup’s available dynamic range. In fourth place, the environmental sensitivity of millimeter wavelengths issue was approached. It is well known the drifts of electromagnetic experiments due to the dependance of the re sults with respect to the surrounding environment. This feature relegates many industrial practices of microwave frequencies to the experimental stage, at millimeter wavelengths. In particular, evolution of the atmosphere within acceptable conditioning bounds redounds in drift phenomena which completely mask the experimental results. The contribution of this thesis on this aspect consists on modeling electrically the indoor atmosphere existing in a CATR, as a function of environmental variables which affect the range’s performance. A simple model was developed, being able to handle high level phenomena, such as feed - probe phase drift as a function of low level magnitudes easy to be sampled: relative humidity and temperature. With this model, environmental compensation can be performed and chamber conditioning is automatically extended towards higher frequencies. Therefore, the purpose of this thesis is to go further into the knowledge of millimetre wavelengths involving compact antenna test ranges. This knowledge is dosified through the sequential stages of a CATR conception, form early low level electromagnetic analysis towards the assessment of an operative facility, stages for each one of which nowadays bottleneck phenomena exist and seriously compromise the antenna measurement practices at millimeter wavelengths.

Estudio de la variación del contenido de humedad en el Pinus pinaster Ait. por medio de la técnica no destructiva del georradar / Assessment of the variation of the moisture content in the Pinus pinaster Ait using the non destructive gpr technique

La variación en el contenido de humedad (CH) tiene una influencia significativa tanto en las propiedades físico- químicas de la madera, como en sus propiedades electromagnéticas y por tanto afecta a las características de la propagación de las ondas. En concreto, en este trabajo se estudia la capacidad del georradar (GR) empleando una antena de 1.6GHz de frecuencia central para registrar las variaciones que se producen en la velocidad y en la amplitud de las ondas electromagnéticas cuando se propagan en unas viguetas de madera de Pinus pinaster Ait de uso estructural cuyo CH va disminuyendo. Se ha comprobado como cuando el CH descendía la velocidad de propagación y las amplitudes, tanto de la onda directa como de la reflejada aumentaba. Los altos factores de correlación encontrados demuestran que el GR es una técnica capaz de evaluar, de forma no destructiva, el CH de la madera de uso estructural. The moisture content variations in wood have a significant influence in wood?s physicochemical properties, as well as in its electromagnetic properties and to specific effects upon waves? characteristics. In particular, this paper focuses on the analysis of the Ground-penetrating Radar?s (GPR) using an antenna of 1.6 GHz central frequency capacity to register the velocity and the amplitude of the electromagnetic waves? variation during the drying process of Pinus pinaster Ait timber joists. The results showed that when timber MC descended, the propagation velocity and amplitude of both the direct and the reflected wave increased. The high correlation found between the variables studied demonstrates GPR efficiency and the innovative application of this technique as a non-destructive evaluation tool for timber structures, particularly when studying its moisture content.

Evaluación de la madera estructural para la rehabilitación de las Naves Cros (Valencia, España) por medio de diferentes técnicas no destructivas

El proyecto de rehabilitación de una de las naves del complejo fabril de la industria química ?CROS? en Valencia se llevó a cabo con el criterio de mantener, en la medida de lo posible, los elementos estructurales presentes en la nave. Con este objetivo se realizaron una serie de ensayos no destructivos (END) in situ. Estos ensayos permitieron evaluar la calidad de la madera, determinar qué elementos estructurales debían ser sustituidos y comprobar la aptitud de los que iban a ser reutilizados. Los END empleados en este estudio fueron los siguientes: (1) Identificación de la especie por técnicas anatómicas, (2) Clasificación resistente por método visual, (3) Estimación de humedad por la técnica de resistencia eléctrica; (4) Obtención de velocidades de propagación ultrasónicas (5) Resistógrafía y (6) Alteración de la propagación de ondas electromagnéticas por medio de Georradar. Para la calibración de estos END se tomó una muestra de piezas y se hicieron ensayos destructivos bajo condiciones controladas en laboratorio. En el trabajo que aquí se presenta se muestra la metodología empleada durante el proceso de toma de datos, de análisis de resultados y de cruce de la información obtenida a partir de cada uno de los ensayos hasta llegar a un diagnóstico para los elementos analizados. The assessment of structural timber was requested in the rehabilitation project of the Naves of the chemical industry "CROS". The criterion was to maintain as much as possible timber of the structure and to make only partial replacements. In order not to damage the existing structure and to assess the quality of the existing timber, a series of non-destructive testing (NDT) in the entire structure were performed: (1) Identification of the species by anatomical techniques, (2) Strength grading by visual method, (3) Estimation of moisture content by the technique of electrical resistance, (4) Acquisition of ultrasonic propagation velocities (5) Resistography and (6) Record of the propagation of electromagnetic waves by means of Ground-penetrating radar. Following, a sample group was chose to carry out destructive testing in the lab and compare the NDT results with those obtained with the standard UNE-EN408 (modules of strength, stiffness and density). In the present work, the results provided by each of the NDT techniques are detailed and above all, what is more important, the validity of these after they have been contrasted with the destructive standard tests.

Influence of Atmospheric Conditions on mmWave CATR Facilities: Formulation and Compensation Scheme

The performance of a CATR relies on the planarity of the synthesized test wave, which is generated within a bounded volume for which specifications are drawn. Millimetre-wave facilities deal with the classical limitations of this frequency band, among which two become critical in our analysis: time-extensive acquisition campaigns and impact of environmental variables. Both features become more evident when increasing the frequency of operation. The variation in atmospheric variables, such as humidity, temperature and pressure has an influence over the performance of all the elements of the facility. The instrumentation behavior is influenced both by the warming up process, and the ambience conditions that surround the equipment. On the changes of the atmosphere itself, they affect the electromagnetic wave propagation, given the physical link between the conditions of the atmosphere and its electric properties as an electromagnetic waves propagation medium

Characterization of Cavities Using the GPR, LIDAR and GNSS Techniques

The study of the many types of natural and manmade cavities in different parts of the world is important to the fields of geology, geophysics, engineering, architectures, agriculture, heritages and landscape. Ground-penetrating radar (GPR) is a noninvasive geodetection and geolocation technique suitable for accurately determining buried structures. This technique requires knowing the propagation velocity of electromagnetic waves (EM velocity) in the medium. We propose a method for calibrating the EM velocity using the integration of laser imaging detection and ranging (LIDAR) and GPR techniques using the Global Navigation Satellite System (GNSS) as support for geolocation. Once the EM velocity is known and the GPR profiles have been properly processed and migrated, they will also show the hidden cavities and the old hidden structures from the cellar. In this article, we present a complete study of the joint use of the GPR, LIDAR and GNSS techniques in the characterization of cavities. We apply this methodology to study underground cavities in a group of wine cellars located in Atauta (Soria, Spain). The results serve to identify construction elements that form the cavity and group of cavities or cellars. The described methodology could be applied to other shallow underground structures with surface connection, where LIDAR and GPR profiles could be joined, as, for example, in archaeological cavities, sewerage systems, drainpipes, etc.

Sobre la solución del problema de la radiación electromagnética

En la presente Tesis Doctoral se propone una posible solución al problema de la radiación electromagnética. Este es un problema científico estudiado desde hace mucho tiempo y que hasta el presente no se ha podido resolver de manera exacta. Aquí se redefine nuevamente el problema y se plantea el mismo partiendo un modelo de radiador formado por dos placas planas circulares paralelas, que por sus reducidas dimensiones eléctricas, se considerará para el estudio como un dipolo simétrico corto. Se obtienen las soluciones de las ecuaciones de Maxwell para la radiación electromagnética emitida por dicho sistema y se propone lo que sería el principio de formación de las ondas electromagnéticas. Con ayuda de este se ofrece una posible interpretación de las soluciones obtenidas. Luego a dicho sistema se le realizan correcciones en sus dimensiones a fin de ajustar las soluciones obtenidas al caso real de un dipolo simétrico de media onda. Además se analiza de una manera sencilla la naturaleza material del campo electromagnético y su estructura, lo que hace posible proponer una manera original de comprender las ecuaciones de Maxwell. Finalmente se propone un modelo sencillo mediante el cual se puede demostrar la Ley de Planck para la distribución espectral de la energía electromagnética radiada por los cuerpos calientes, pero desde las soluciones de las ecuaciones de Maxwell para la radiación de las antenas. Con esto podría ser aceptado que la solución del problema de la radiación electromagnética es única tanto para las antenas como para los cuerpos calientes en la naturaleza. ABSTRACT In this PhD thesis a possible solution to the problem of electromagnetic radiation is proposed. This is a scientific problem which has been studied for a long time and to this day has still not exactly been solved. The problem is redefined here again and it is stated using a model formed by two flat parallel circular plates as the radiator system, which due to its reduced electrical dimensions will be considered for the study as a short symmetrical dipole. The solutions of the Maxwell equations for emitted electromagnetic radiation are obtained by said system, and that which could be the principle formation of electromagnetic waves is proposed. Using this principle, a possible interpretation of the solutions obtained is presented. Then the system will perform corrections in size to adjust the solutions obtained to the real case of a symmetrical half-wave dipole. The material nature of the electromagnetic field and its structure is also analyzed in a simple way, making it possible to present an original way to understand the Maxwell equations. Finally, a simple model which can be used to prove Planck's law of spectral distribution of the electromagnetic energy radiated by hot bodies is proposed, but parting from the solutions of the Maxwell equations for the antennas radiation. With this it could be accepted that the solution to the problem of electromagnetic radiation is unique to both antennas and hot bodies on nature.

The effects of traffic radar guns on law enforcement officers : hearing before the Ad Hoc Subcommittee on Consumer and Environmental Issues of the Committee on Governmental Affairs, United States Senate, One Hundred Second Congress, second session, August 10, 1992.

Includes bibliographical references.

A Kinetic Vlasov Model for Plasma Simulation Using Discontinuous Galerkin Method on Many-Core Architectures

Thesis (Ph.D.)--University of Washington, 2016-03

Effective cell-centred time-domain Maxwell's equations numerical solvers

This research work analyses techniques for implementing a cell-centred finite-volume time-domain (ccFV-TD) computational methodology for the purpose of studying microwave heating. Various state-of-the-art spatial and temporal discretisation methods employed to solve Maxwell's equations on multidimensional structured grid networks are investigated, and the dispersive and dissipative errors inherent in those techniques examined. Both staggered and unstaggered grid approaches are considered. Upwind schemes using a Riemann solver and intensity vector splitting are studied and evaluated. Staggered and unstaggered Leapfrog and Runge-Kutta time integration methods are analysed in terms of phase and amplitude error to identify which method is the most accurate and efficient for simulating microwave heating processes. The implementation and migration of typical electromagnetic boundary conditions. from staggered in space to cell-centred approaches also is deliberated. In particular, an existing perfectly matched layer absorbing boundary methodology is adapted to formulate a new cell-centred boundary implementation for the ccFV-TD solvers. Finally for microwave heating purposes, a comparison of analytical and numerical results for standard case studies in rectangular waveguides allows the accuracy of the developed methods to be assessed. © 2004 Elsevier Inc. All rights reserved.

Explicit derivation of the relativistic mass-energy relation for internal kinetic and potential energies of a composite system

A straightforward derivation of relativistic expressions for the mechanical momentum, kinetic and total energies, and mass-energy equivalence (including potential energy) which does not require any knowledge of the energy-momentum relation for electromagnetic waves or consideration of elastic collisions, but is directly based on Newton's second law and Lorentz's transformations, is presented in this paper. The existence of an invariant force is shown to be important for the validity of the relativistic mechanics.

Optical Bloch equations and enhanced decay of Rabi oscillations in strong driving fields

Optical Bloch equations are widely used for describing dynamics in a system consisting molecules, electromagnetic waves, and a thermal bath. We analyze applicability of these equations to a single molecule imbedded in a solid matrix. Classical Bloch equations and the limits of their applicability are derived from more general master equations. Simple and intuitively appealing picture based on stochastic Bloch equations shows that at low temperatures, contrary to common believes, a strong driving field can not only suppress but can also increase decay rates of Rabi oscillations. A physical system where predicted effects can be observed experimentally is suggested. (c) 2005 Elsevier B.V. All rights reserved.