When a moving dot, which is tracked by the eyes and enclosed in a moving framework, suddenly stops while the enclosing framework continues its motion, the dot is seen to describe a curved path. This illusion can be explained only by assuming that extra-retinal signals are taken into account in interpreting retinal information. The form of the illusion, and the fact that the phenomenal path cannot be explained on the basis of positional information alone, suggests that the perceived path is computed by integrating (instantaneous) velocity information over time. A vector addition model embodying a number of simplifying assumptions is found to qualitatively fit the experimental data. A number of follow-up studies are suggested.

The hearing process involves a series of physical events in which acoustic waves in the outer ear are transduced into acousto-mechanical motions of the middle ear, and then into chemo-electro-mechanical reactions of the inner ear sensors that are interpreted by the brain. Air in the ear canal has low mechanical impedance, whereas the mechanical impedance at the center of the eardrum, the umbo, is high. The eardrum or Tympanic Membrane (TM) must act as a transformer between these two impedances; otherwise, most of the energy will be reflected rather than transmitted. The acousto-mechanical transformer behavior of the TM is determined by its geometry, internal fibrous structure, and mechanical properties. Therefore, full-field-of-view techniques are required to quantify shape, sound-induced displacements, and mechanical properties of the TM. Shapes of the mammalian TMs are in millimeter ranges, whereas their acoustically-induced motions are in nanometer ranges, therefore, a clinically-applicable system with a measuring range spanning six orders of magnitude needs to be realized. In this Dissertation, several full-field measuring modalities are developed, to incrementally address the questions regarding the geometry, kinematics, and dynamics of the sound-induced energy transfer through the mammalian TMs. First, a digital holographic system with a measuring range spanning several orders of magnitude is developed and shape and 1D sound-induced motions of the TM are measured with dual-wavelength holographic contouring and single sensitivity vector holographic interferometry, respectively. The sound-induced motions of the TMs are hypothesized to be similar to those of thin-shells (with negligible tangential motions) and therefore, 3D sound-induced motions of the TM are estimated by combining measurements of shape and 1D motions. In order to test the applicability of the thin-shell hypothesis, and to obtain further details of complex spatio-temporal response of the TMs, holographic systems with multiple illumination directions are developed and shape and acoustically-induced vibrational patterns of the TMs are quantified in full 3D. Furthermore, to move toward clinical applications and in-vivo measurements, high-speed single-shot multiplexing holographic system are developed and 3D sound-induced motions of the TM are measured simultaneously in one single frame of the camera. Finally, MEMS-based high-resolution force sensing capabilities are integrated with holographic measurements to relate the kinematics and dynamics of the acousto-mechanical energy transfer in the hearing processes. The accuracy and repeatability of the measuring systems are tested and verified using artificial samples with geometries similar to those of human TMs. The systems are then used to measure shape, 3D sound-induced motions, and forces of chinchilla and human cadaveric TM samples at different tonal frequencies (ranging from 400 Hz to 15 kHz) simultaneously at more than 1 million points on its surface. A general conclusion is that the tangential motions are significantly (8-20 dB) smaller than the motions perpendicular to the TM plane, which is consistent with the thin-shell hypothesis of the TM. Force measurements reveal that frequency-dependent forces of the TM, are also spatially dependent so that the maximum magnitudes of the force transfer function of the umbo occurs at frequencies between 1.6 to 2.3 kHz, whereas the maximum values for other points on the TM surface occurs at higher frequency ranges (4.8 to 6.5 kHz). The Dissertation is divided into two Parts, each contains several Chapters. In the first Part, general overviews of the physiology of the human middle ear, along with brief summaries of previous studies are given, and basics of holographic interferometry are described. In the second Part, developments and implementations achieved in completion of this work are described in the form of a series of manuscripts. Finally, conclusions and recommendations for future work are provided.

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Made available in DSpace on 2016-06-02T19:56:57Z (GMT). No. of bitstreams: 1 6615.pdf: 4694610 bytes, checksum: 288df2441cd04d5e5e4c36da49c66cb9 (MD5) Previous issue date: 2015-03-05 / Financiadora de Estudos e Projetos / Mesenchymal stromal cells (MSCs) are required by the scientific community in the development and enhancement of therapeutic techniques in different fields of medicine. The MSCs are present in small concentrations in tissues, which makes necessary the expansion in vitro for enable studies and therapeutic applicability. These are cells with high sensitivity to environmental conditions of cultivation. So, for increase productivity in vitro is used the technology of bioreactors in the development of processes in order to produce high cell densities in less time, with reduce use of resources and maintaining a safe operation. The new concepts of "disposable bioreactors", as the wave-induced motion bioreactor or Wave bioreactor, with possibility operating in a closed system, controlled and automated, reduced investment cost and operation, less risk of contamination, higher level biosecurity, added to the fact of being a underexplored technology and already approved by the FDA (Food and Drugs Administration) becomes a highly attractive alternative bioprocessing for cultivation of animal cells in large scale. In this context, the present work aims to develop a protocol for cultivation of MSCs in the Wave Bioreactor System 2/10. Experiments were performed to characterize the CEMs's culture behavior in the Wave bioreactor to obtain high cell productivity while maintaining the therapeutic potential of the CEMs. The experiments were carried out with 2 L Cellbag and Cultispher-S microcarrier with 300 ml of α-MEM medium culture supplemented with glucose, glutamine, and arginine and 15% v/v fetal bovine serum at 37 ° C and pH between 6,9-7,4. In the preliminary experiments it was verified that most of the inoculated cells did not adhere to the microcarriers. It was shown that such behavior is due to low relation between adhesion area (AMC = total projected area of the microcarriers) and wet surface area of Cellbag (ASMCellbag), which in the normal condition of operation results an adhesion between 25,7 and 61,7% of the inoculated cells. To solve the problem were performed experiments reducing ACellbag which enabled improvements in cell adhesion by up to 100%. It was also found low performance of the cell expansion phase, presumably linked to operational problems like: microcarriers segregation in certain regions of the bioreactor causing depletion of nutrients, formation of aggregates of MCs colonized with cells and adhesion of MCs to Cellbag. In addition, it was observed that reducing CEM/MC ratio at the start of the culture, the cell expansion factor could be increased to values equal to or greater than 10. These results show that the Wave bioreactor has good potential for expansion of MSCs and that the same can be improved. / As células estromais mesenquimais (CEMs) estão sendo visadas pela comunidade científica no desenvolvimento e aprimoramento de técnicas terapêuticas em diferentes ramos da medicina. As CEMs estão presentes em pequenas concentrações nos tecidos, o que torna necessário a sua expansão in vitro para viabilizar pesquisas e a aplicabilidade terapêutica. Tratam-se de células com elevada sensibilidade em relação às condições do ambiente de cult ivo. Assim, para o aumento da produtividade in vitro utiliza-se a tecnologia de biorreatores no desenvolvimento de processos com objetivo de produzir altas densidades celulares em curto tempo, de forma econômica e respeitando as normas impostas pelos órgãos reguladores. O novo conceito de biorreator descartável, como o do biorreator com movimento induzido em forma de ondas, ou biorreator Wave, apresenta possibilidade de operação em sistema fechado segundo as boas práticas de fabricação (BPF), controlado e automatizado. O custo de investimento e operação reduzido, com menor risco de contaminação, maior nível de biossegurança, somado ao fato de utilizar uma tecnologia pouco explorada e já aprovada pelo FDA (Food and Drugs Administration) se transforma numa alternativa de bioprocessamento altamente atrativa para cultivo de células animais em larga escala. Nesse contexto, o presente trabalho tem por meta avaliar o desempenho do biorreator Wave 2/10 na expansão das CEMs. Para tal, foram realizados experimentos visando caracterizar o comportamento do cultivo nesse biorreator a fim de obter alta produtividade celular mantendo a potencialidade terapêutica das CEMs. Os experimentos foram realizados com saco plástico (doravante Cellbag) de 2 L e microcarregador (MC) Cultispher-S com 300 mL me io de cultivo α-MEM suplementado com glicose, glutamina e arginina e 15% v/v de soro fetal bovino a 37°C e pH entre 6,9-7,4. Nos experimentos preliminares constatou-se que grande parte das células inoculadas não aderiam aos microcarregadores. Comprovou-se que tal comportamento se devia à baixa relação entre área de adesão (AMC = área total projetada dos microcarregadores) e área de superfície molhada da Cellbag (ASMCellbag) que na condição normal de operação resultava numa adesão entre 25,7 e 61,7% das células inoculadas. Para melhorar a adesão foram realizados experimentos reduzindo a ASMCellbag, o que possibilitou melhoria na adesão celular em até 100%. Na etapa de expansão celular verificou-se baixo desempenho, presumivelmente vinculado a problemas de operação como: segregação de microcarregadores em determinadas regiões do biorreator provocando o esgotamento de nutrientes, formação de agregados de MCs colonizados com células e adesão dos MCs à Cellbag. Em adição, notou-se que diminuindo a relação CEM/MC no início do cultivo a expansão celular podia ser aumentada para valores iguais ou maiores que 10. Ao todo, os resultados mostraram que o biorreator Wave possui bom potencial para a expansão de CEMs e que o mesmo ainda pode ser melhorado.

Tall slender buildings are easily set in motion by wind and earthquakes but by estimating the buildings horizontal natural frequencies in the design phase, these motions can be kept within acceptable boundaries. There are many parameters that decides the natural frequency of a building and it can therefore be difficult to calculate it. There are a few ways though to estimate horizontal natural frequencies of tall buildings and two methods have been tested in this report. Both methods give the frequency of a clamped-free cantilever but one of them requires a single degree of freedom system whilst the other handles a multi degree of freedom system. The methods are called SDOF method and MDOF method in this report. A fictional building was created for this project to be the reference object in the comparison between the two methods SDOF and MDOF. The walls and floors of the building was designed with the support of both an acoustic engineer and a structural engineer to create a realistic building. A building’s natural frequency is dependent of the self-weight, stiffness and height of the building and it was therefore important to design these components with care. The fictional building is called House 1 and is a 10 story, almost square building about 20 m wide and broad and 30 m high. This report does not only compare the natural frequencies obtained from the two different calculation methods, but it also shows the difference in frequency in timber and concrete structures. Shear walls constitutes the horizontal stabilization system of the fictional building and both a CLT core and a concrete core is designed and compared. It is only the walls that comes in two different versions, the floorings consist of CLT boards for both structures tested. The horizontal natural frequencies of House 1 were about 2 Hz and 3 Hz for the CLT version and concrete version respectively. It was expected to get frequencies within that range considering the height of House 1. The CLT core having a lower frequency than the concrete core was also expected since concrete is a stiffer material than wood. To be able to make a fair comparison between the SDOF method and the MDOF method, House 1 was designed with the same dimensions and stiffness on all floors because the SDOF method requires that. The results from the two methods are almost identical with only 0.3 Hz and 0.4 Hz difference for the concrete and CLT respectively. For a shear wall structure with a consistent stiffness, weight and dimension, any of the two methods can be used to estimate the horizontal natural frequency. However, it is not realistic for a building of 30 m or higher, to have the same dimensions on the load bearing structure on all floors which makes the MDOF method more accurate in more cases than the SDOF method. / Höga slanka byggnader kan sättas i svajande rörelser av vind och jordbävningar, men genom att uppskatta byggnadernas horisontella egenfrekvenser i den tidiga konstruktionsfasen kan dessa rörelser hållas inom acceptabla gränser. Det är många parametrar som bestämmer byggnadens egenfrekvens och det kan därför vara svårt att beräkna den. Det finns dock några sätt att uppskatta horisontella egenfrekvenser hos höga byggnader och två metoder har testats i denna rapport. Båda metoderna ger frekvensen av en fast inspänd konsolbalk men en av dem kräver ett enfrihetsgradsystem medan den andra kan hantera ett system med flera frihetsgrader. Metoderna kallas SDOF-metoden och MDOF-metoden i denna rapport. En fiktiv byggnad skapades i detta projekt för att vara referensobjekt i jämförelsen mellan de två metoderna SDOF och MDOF. Byggnadens väggar och golv konstruerades med stöd av både en akustiker och en konstruktör för att skapa en realistisk byggnad. Byggnadens egenfrekvens är beroende av byggnadens egenvikt, styvhet och höjd och det var därför viktigt att utforma dessa komponenter med omsorg. Den fiktiva byggnaden kallas House 1 och är en 10 vånings-, nästan fyrkantig byggnad ca 20 m lång och bred och 30 m hög. Denna rapport jämför inte bara egenfrekvenserna erhållna från de två olika beräkningsmetoderna, den visar även skillnaden i frekvens i trä- och betongkonstruktioner. Skjuvväggar utgör det horisontella stabiliseringssystemet för den fiktiva byggnaden och både en KL-kärna och en betongkärna har utformats och jämförts. Det är bara väggarna som skiljer de två olika versionerna åt, bjälklagen består av KL-skivor i båda fallen. De horisontella egenfrekvenserna hos House 1 var ca 2 Hz och 3 Hz för KL-version respektive betongversion. Frekvenser inom detta område var väntade med tanke på höjden av House 1. Att KL-kärnan skulle ha en lägre frekvens än betongkärnan förväntades också eftersom betong är ett styvare material än trä. För att kunna göra en rättvis jämförelse mellan SDOF-metoden och MDOF-metoden, var House 1 utformad med samma dimension och styvhet på alla våningsplan eftersom SDOF-metoden kräver det. Resultaten från de två metoderna är nästan identiska med endast 0,3 Hz och 0,4 Hz skillnad för betong respektive KL. För en skjuvväggskonstruktion med en kontinuerlig styvhet, vikt och dimension kan båda de två metoderna användas för att uppskatta den horisontella egenfrekvensen. Det är dock inte realistiskt för en byggnad på 30 m eller högre att ha samma dimensioner på den lastbärande konstruktionen på alla våningar vilket gör MDOF-metoden mer korrekt i fler fall än SDOF-metoden.