In this work, we used atomic force spectroscopy to obtain the elastic modulus of melanoma and keratinocytes fixed cells, with the purpose to determine the initial conditions for studies of confluente cultures of these cells in the future. The cell lines used were HaCaT cells and WM1366 melanoma cell, the last one is derived from a radial growth melanoma and both were analyzed, parental WM1366 cells (WM1366 shSCR cells) and galectin-3 silenced WM1366 cells (WM1366 shGal3). Cells were located and images of them were obtained by AFM contact mode under liquid conditions. Single force curves acquired in the central region of cells were used to determine the elastic modulus by the Hertzian contact model for the pyramidal tip, allowing to establish a comparison patter between cancer and normal cells. It was found that the melanoma cell (21.8 ± 0.5 kPa) exhibit smaller elastic modulus than keratinocytes cells (31.9 ± 0.4 kPa). For WM1366 shGal3 was found a elastic modulus of 16.1 ± 0.6 kPa, therefore, we found that for large indentation depth it is possible to distinguish between the same melanoma cell line, which represents general alterations in the organization of the cytoskeleton induced by the presence or absence of the galectin-3 protein. On the other hand, to detect local elastic modulus variations along the cell and to identify subcellular regions characterized by specific stiffness associated with local structures, we took elasticity maps in which a single force curve is acquired in each probe position. In order to interpret these maps, the cell was sliced into several different heights, curves of each height section were analyzed and represented in histograms, adjusted by the binomial distribution function. It was observed that the gradient of elastic modulus in cells from the nuclear region towards the cell periphery is more pronounced in cells devoid of galectin-3 than parental cells. The increased elastic modulus in the pericellular region of cells devoid of galectin-3 suggests that the organization of the extracellular matrix in these areas is different than those observed around HaCaT and shSCR WM1366 cells. / Neste trabalho, utilizamos espectroscopia de força atômica para obtenção do módulo elástico de células fixadas de melanoma e queratinócitos, com o objetivo de determinar as condições iniciais de estudos a serem realizados futuramente de culturas de células confluentes do mesmo tipo. As linhagem celulares utilizadas foram as células HaCaT e as células de melanoma WM1366, sendo a última derivada de um melanoma de crescimento radial sendo analisadas tanto as células parentais (células WM1366 shSCR) e as células WM1366 silenciadas com galectina-3 (WM1366 shGal3). As células foram localizadas e imageadas no modo AFM contato em meio líquido. Curvas de força adquiridas na região central das células foram utilizadas para determinar o módulo elástico, a partir do modelo de contato hertziano por uma ponta piramidal, permitindo estabelecer um padrão para comparação entre células normais e cancerígenas. Verificou-se que a célula de melanoma exibe menor módulo de elasticidade (21.8 ± 0.5 kPa) do que as células de queratinócitos (31.9 ± 0.4 kPa). Para as células WM1366 shGal3 foi encontrado um módulo elástico de 16.1 ± 0.6 kPa. Portanto, verificou-se que, para grandes profundidades de indentação, é possível distinguir entre a mesma linhagem de melanoma, células que apresentam alterações gerais na organização do citoesqueleto induzidas pela presença ou ausência da proteína galectina-3. Por outro lado, para detectar variações locais do módulo elástico ao longo da célula e identificar regiões subcelulares, caracterizadas por rigidez específica associada a estruturas locais, foram obtidos mapas de elasticidade nos quais uma única curva de força é adquirida em cada posição da sonda. Para interpretar estes mapas, a célula foi dividida em regiões de diferentes alturas e curvas de cada seção de altura foram analisadas e representadas em histogramas, ajustadas pela função de distribuição binomial. Observou-se que o gradiente de módulo de elasticidade em células da região nuclear em direção à periferia celular é mais acentuado em células desprovidas de galectina-3 do que em células parentais. O aumento do módulo de elasticidade na região pericelular das células desprovidas de galectina-3 sugere que a organização da matriz extracelular nestas áreas é diferente das observadas em torno das células HaCaT e shSCR WM1366.

Cada vez mais, a competividade fortalece a filosofia de administração empresarial baseada na orientação para o cliente e para o lucro. A busca de novas formas de relacionamento com os clientes requer instrumentos que possam personalizar inclusive os preços que compõem a equação de valor de cada um. O que temos visto, na prática, são ações que não consideram a dinâmica das forças do mercado, representada pelas curvas de oferta e demanda, e por isso expõem a organização ao risco de propor um processo de troca que não atenda ao preceito de que deve beneficiar tanto o vendedor quanto o comprador, como fator mais importante para a manutenção de um relacinamento de longo prazo. Os instrumentos apresentados como equações matemamáticas, pretendem viabilizar a adoção de forma monitorada com os objetos da gestão econômica. Como benefício complementar, os intrumentos por requererem regras claras, objetivas e bem delimitadas para os processos de negociação, permitem compartilhar de maneira justa, a responsabilidade sobre o atingimento dos resultados através de transações comerciais. / Competitiveness fortifies the entrepreneurial management philosophy that prioritizes more and more, customers and profit. The search for new relationship between companies and customers requires appropriate tools to even, embody the prices that composes the equation of value. We have noticed according to the procedures, actions disrespecting the dynamic of market forces, represented by supply and demand curves and, as a consequence, the companies can be involved in a trading process that can not serve neither the seller nor the buyer, damaging the relationship in the long run. The algorithms presented as mathematical equation aim at enabling the approval of the dynamic prices in trading processes to support their own flexibility in a controlled way that is connected to the goals of the economic management. As a complementary benefit, the algorithms require clear, delimitative and objective rules for the dealing processes and make possible sharing in a fair way, the responsability for the achievement of the results through the trading negotiations.

L’objectif principal est d’étudier l’influence de la microstructure sur le comportement macroscopique des roches hétérogènes, particulièrement, la roche de schiste de Vaca Muerta. Des modèles micromécaniques sont développés par des techniques d’homogénéisation bien adaptées. La microstructure de cette roche a été étudiée afin de définir le volume élémentaire représentatif indispensable pour la mise en œuvre d’une approche micromécanique. Le matériau est modélisé comme un milieu hétérogène, constitué à l’échelle mésoscopique d’une matrice composite dans laquelle sont plongés différents types d’inclusions. A une échelle plus petite, des fins grains de calcite et kérogéne sont immergés dans une matrice argileuse poreuse. L’homogénéisation du comportement instantané est établie dans le contexte de l’élastoplasticité de la matrice argileuse et de la décohésion progressive des inclusions: la transition d’échelle nano-micro aboutit au domaine de résistance de la phase argileuse où la phase solide est considérée comme un matériau cohésif-frottant obéissant au critère de Drucker-Prager. La méthode de Hill est utilisée comme moyen d’homogénéisation pour les transitions micro-méso et méso-macro. Ensuite, le comportement à long terme est défini en terme de la dégradation de la microstructure et en particulier, de la matrice argileuse. Finalement, un modèle micromécanique simplifié est développé pour étudier le comportement à long terme, des roches argileuses et de schistes, dans le contexte de la viscoplasticité de la matrice argileuse. En se basant sur ce modèle, pour l’application industrielle, nous étudions l’interaction entre la roche Vaca Muerta et les grains de proppants. / The main objective of this thesis is to study the influence of microstructure on the macroscopic mechanical behavior of heterogeneous rocks, particularly Vaca Muerta shale rock. In this context, micromechanical models are developed by well-adapted homogenization techniques. The microstructure of this rock has been extensively studied in order to define the representative elementary volume indispensable for the implementation of a micromechanical approach. The studied material is modeled as a heterogeneous medium, made up, at mesoscale, of a composite matrix in which are dipped different types of inclusions. On a smaller scale, fine grains of calcite and kerogen are immersed in a porous clay matrix. The homogenization of non-linear instantaneous behavior is established in the context of clay matrix elastoplasticity and the progressive debonding of mineral inclusions: the nano-micro transition leads to the strength domain of the porous clay phase where the solid phase is considered to be a cohesive-frictional material that obeys to the classic Drucker-Prager criterion. Hill’s incremental method is used as a homogenization means at micro-meso and meso-macro transitions. Next, the long-term behavior of the studied material is defined in terms of microstructure degradation. Finally, a simplified micomechanical model is developed to study long-term behavior of clayey and shale rocks in the context of clay matrix viscoplasticity. Based on this model, for the industrial application of the thesis, we study the interaction between Vaca Muerta shale rock and spherical grains of proppants.

Nowadays, inspection process planning (IPP) for large volume metrology (LVM) attracts increasing attention in manufacturing and assembly industries such as aerospace and automotive, where large and complex assemblies and fabrications with complex surfaces are employed. Inspection is conventionally considered as a quality control manner. But there is changing shift to processes that are more related to the early design stage aiming to increase product performance and reduce costs by automation and elimination of rework. This is especially evident in the standardisation and implementation of Geometric Dimensioning and Tolerancing (GD&T) of new products and systems at the design stage. This study proposes a GD&T based systematic framework for the IPP of LVM systems within a digital environment. Orientating to solve the “what to measure” and “how to measure” problems in IPP, the prototype system has seven functional core modules including: tolerance feature analysis, instrument selection, inspection point selection, accessibility and visibility analysis, instrument setup and configuration, clustering analysis and measurement sequencing. An optimized inspection plan is output for the designer to evaluate the product design as well as for guiding the metrologist and process planner to conduct the inspection process. Heuristic rules, evolutionary algorithms and modern computational graphic techniques have been adopted to facilitate the supported functions. Coupled with state of art metrology systems, metrology and CAD software, the framework is able to work effectively and efficiently by means of incorporating international standards and industrial best practice. It is the first attempt to successfully minimise manual activities in the planning process for LVM systems, which results in improved efficiency, enhanced decision making and a better inspection plan overall. Two case studies have been conducted to validate the functionalities of the prototype system.

Introdução: A tomografia de impedância elétrica (TIE) é uma técnica de imagem não invasiva que mensura o potencial elétrico na superfície torácica através de eletrodos colocados ao redor do tórax. Essas medidas são transformadas em uma imagem bidimensional da distribuição da impedância elétrica no tórax. Esse instrumento detecta modificações da densidade pulmonar e distribuição do volume corrente. Entretanto, é necessário melhor avaliação da sua eficiência em estimar o volume corrente (VT) a cada ciclo ventilatório. O objetivo do estudo é avaliar a correlação do volume estimado pela TIE com o VT aplicado durante o suporte ventilatório mecânico. Métodos: Inicialmente, foram estudados cinco suínos da raça Landrace. Os animais foram sedados, intubados e monitorados com a TIE. Foram aplicados volumes incrementais (100, 250, 500, 750 e 1000 mL) com a seringa de precisão em ZEEP e com PEEP de 5, 10 e 20 cmH2O, antes e depois da promoção de lesão pulmonar com lavagem com solução salina. Posteriormente, outros cinco animais foram monitorados com a tomografia computadorizada de tórax por raios X (TC) e a TIE. Foram aplicados volumes incrementais (250, 500 e 1000 mL) com a seringa de precisão em ZEEP e com PEEP de 10 e 20 cmH2O, também antes e depois da promoção de lesão pulmonar. A amplitude da variação de impedância mensurada pela TIE foi convertida em volume e foi calculado o volume de ar na TC. Análises de correlação e concordância foram realizadas com o programa \"R\" (© R Foundation for Statistical Computing). Resultados: Em ZEEP, o coeficiente de correlação entre os volumes estimado pela TIE e calculado pela TC foram, respectivamente, 0,90 e 0,96 na comparação com a seringa de precisão. O coeficiente de determinação entre a TIE e a seringa de precisão foi progressivamente reduzindo (0,90; 0,89 e 0,81 com PEEP de 5; 10 e 20, respectivamente) com o aumento do volume pulmonar promovido pela elevação da PEEP. A TC mostrou um deslocamento progressivo do conteúdo aéreo no sentido caudal com o aumento do volume pulmonar. Conclusões: Há uma boa correlação entre o volume estimado pela TIE e o VT aplicado durante o suporte ventilatório mecânico quando utilizados volumes e pressões usuais na prática clínica / Rationale: Electrical impedance tomography (EIT) is a noninvasive imaging tool that reconstructs a cross-sectional image of the lung\'s regional conductivity using electrodes placed circumferentially around the thorax. It is able to detect changes of lung air content and tidal volume (VT) distribution. However, better evaluation of its capacity to quantify VT variations is necessary. The aim of our study was to assess the correlation between volume estimated by EIT and tidal volume applied at different positive end-expiratory pressures (PEEP). Methods: Initially, in an experimental study five mechanically ventilated pigs monitored by EIT were studied. VT increments (100, 250, 500, 750 and 1000 mL) were applied with a calibrated syringe at zero end-expiratory pressure (ZEEP) and PEEP levels of 5, 10 and 20 cmH2O before and after lung-injury (induced by saline lavage). Another five pigs was monitored by EIT and x-ray computed tomography (CT). VT increments (250, 500 and 1000 mL) were applied with a calibrated syringe at zero end-expiratory pressure (ZEEP) and PEEP levels of 10 and 20 cmH2O before and after lung-injury. Lung air volume was calculated at CT scan and the amplitude of impedance change measured by EIT was converted to volume (mL). Correlation and agreement analysis was performed at \"R\" program (© R Foundation for Statistical Computing). Results: At ZEEP, volume estimated by EIT and volume calculated at CT obtained r2 = 0.90 and 0.96 respectively, when compared to calibrated syringe. The coefficient of correlation between EIT and calibrated syringe impaired (0.90; 0.89 and 0.81 with PEEP of 5; 10 and 20, respectively) with increase of the lung volume due to increased PEEP. CT showed a progressive displacement of the air content to the caudal thoracic levels with the increase of the lung volume. Conclusion: EIT is able to estimate tidal volume during mechanical ventilatory support when used volumes and pressures usually applied at bedside

A esquizofrenia (SZ) é uma doença mental crônica e grave, que compromete o funcionamento psicossocial do indivíduo nos mais variados graus. Atinge 1% da população mundial, considerando todo seu espectro de sintomas (DSM-IV). O pobre funcionamento cognitivo é um dos principais fatores que explicam as elevadas taxas de prejuízos e encargos associados à esquizofrenia. A etiologia da SZ é desconhecida, tendo muitas hipóteses etiológicas como fatores genéticos, epidemias virais durante a gestação, época de nascimento, traumatismos de parto, infecções perinatais, condições neurológicas ou neuropsiquiátricas que geram sintomas tipo esquizofrênicos ou desenvolvimento anormal (avaliados por testes psicológicos, estudos de neuroimagem e neuropatológicos que sugerem alterações no desenvolvimento cerebral). A fisiopatologia da SZ pode ser resultante de uma desregulação na plasticidade sináptica por alterações de neurotofinas, radicais livres e processos inflamatórios. Existe uma larga evidência que os radicais livres podem ter um papel importante na fisiopatologia da SZ, podendo induzir danos na membrana celular, em proteínas e DNA. Problemas com estresse oxidativo, como o aumento da peroxidação lipídica foram relatados previamente em pacientes com SZ em primeiro episódio, virgens de tratamento e naqueles cronicamente medicados. As citocinas inflamatórias têm sido estudadas como importantes participantes na etiologia e desenvolvimento das doenças psiquiatricas. Seu papel ainda não é bem estabelecido, porém diversas alterações têm sido vistas nas doenças psiquiátricas. Dentre as citocinas, destacam-se as interleucinas (IL) e o fator de necrose tumoral alfa (TNF-α), que podem ter ação inflamatória e anti-inflamatória. Dentre as próinflamatórias, podemos destacar a IL-6 e o TNF-α. Alteração de IL na SZ tem sido relatada nesses últimos anos, relacionada à etiologia e à atividade da doença. Pacientes em episódio agudo da doença apresentaram aumento dos níveis séricos de IL pró-inflamatórias sugerindo atividade inflamatória sistêmica. Identificar, além dos sintomas clínicos, possíveis alterações bioquímicas e de neuroimagem em pacientes com SZ pode ajudar em futuras intervenções tanto para identificar, como para prevenir ou atenuar o curso da SZ. Estudos que permitam avançar no entendimento da psicopatologia deste grupo de pacientes são de grande importância, na medida em que proporcionarão futuras abordagens terapêuticas. Está bem estabelecido que a matéria cinzenta cortical e o volume de córtex préfrontal estão diminuídos em pacientes com SZ. Entretanto, os fatores que levam à perda de tecido não estão claras. Uma hipótese para esse fato é que o estado próinflamatório aumentado em SZ está relacionado com a diminuição volumétrica da massa cinzenta. O objetivo deste estudo piloto foi correlacionar os níveis séricos de IL-6 com o volume cortical total de pacientes com SZ e controles. Foram selecionados 36 pacientes com SZ (28 do sexo masculino, com idade média de 37,17 ± 12,05; anos de doença 15,56 ± 11,75), 35 controles pareados idade (21 do sexo masculino, idade média= 36,97 ± 13,04). As imagens foram adquiridas por um equipamento de ressonância magnética Philips Achieva 1.5T no Hospital de Clínicas de Porto Alegre, Brasil. Todas as imagens foram processadas usando o pipeline automatizado de FreeSurfer v5.1. Concluímos que a IL-6 está negativamente correlacionada com o volume cortical total (p= 0,027; rho= -0,370) nos pacientes com esquizofrenia, tal correlação não foi vista nos controles (p= 0,235, rho= -0,206). Nosso resultado sugere que a ativação inflamatória crônica em pacientes com SZ pode estar relacionada com a diminuição volumétrica total do córtex. / Schizophrenia (SZ) is a chronic and severe mental illness, which affects the psychosocial functioning of the individual in many degrees. It reaches 1% of the population, considering all its spectrum of symptoms (DSM-IV). Poor cognitive functioning is one of the main factors responsible for the high rates of disability and costs associated with schizophrenia. The etiology of SZ is unknown, and many etiological assumptions are taken, as genetic factors, viral epidemics during pregnancy, time of birth, birth trauma, perinatal infections, neuropsychiatric or neurological conditions that produce symptoms like schizophrenia or unnatural development (assessed by psychological tests, neuroimaging and neuropathological studies that suggest changes in brain development). The pathophysiology of SZ may be due to a deregulation in synaptic plasticity caused by changes in neurotrophins, free radicals and inflammatory processes. There is a wide evidence that free radicals may have a main role in the pathophysiology of SZ, and can induce damage into the membrane cell, in proteins and DNA. Problems with oxidative stress, such as increased lipid peroxidation have been previously reported in treatment virgem patients with SZ in first episodes and in those chronically treated. And inflammatory cytokines have been studied as important parts in the etiology of psychiatric diseases’ development. Its role is not well established, however a number of changes have been noticed in psychiatric illnesses. Among the cytokines, the Interleukins (IL) and the tumor necrosis factor alpha (TNF-α) stand out, these two may have inflammatory and anti-inflammatory action. Among the pro-inflammatory, we can highlight IL-6 and TNF-α. IL change in the SZ has been reported in these last few years, related to the etiology and disease activity. Patients with acute episode of the disease showed increased serum levels of IL proinflammatory suggesting systemic inflammatory activity. Identify not only the clinical symptoms, possible biochemical and neuroimaging abnormalities in patients with SZ can help in future interventions both to identify and prevent or slow down the course of SZ. Studies to enable progress in the understanding of psychopathology this group of patients are of great importance to the extent that provide future therapeutic approaches. It is well established that cortical gray matter and the prefrontal cortex volume are reduced in patients with SZ. However, the factors that lead to tissue loss are unclear. One possible explanation is that the increased proinflammatory state in SZ is related to the volumetric reduction of the gray matter. The objective of this pilot study was to correlate serum levels of IL-6 in the hole cortex volume of schizophrenic patients and controls. We selected 36 patients with SZ (28 male, average age 37.17±12.05; years of illness 15.56±11.75), 35 matched controls (21 male, average age= 36.97±13.04). Images were obtained by an MRI equipment, brand Philips Achieva 1.5T at Hospital de Clinicas de Porto Alegre, Brazil. All images were processed using automated pipeline FreeSurfer v5.1. We concluded thatIL-6 is negatively correlated with the total cortical volume in patients (p= 0.027, rho= -0.370), this correlation was not seen in controls (p= 0.235; rho= -0.206). Our results suggest that chronic inflammatory activation in patients with SZ can be related to the total volumetric reduction of the cortex.

Nearly 50 million people are afflicted with epilepsy, worldwide. These patients suffer from unprovoked seizures, where neurons in the cerebral cortex under go uncontrolled, hypersynchronous firing of neurons. 30\% of patients with epilepsy do not respond to drug treatments. For these patients, surgical treatment involving the removal or disconnection of brain matter is one of the only alternatives. Such surgical treatments often rely on long-term monitoring of neuronal activity in the brain using subdurally implanted surface electrodes to locate the epileptic focus, but these clinical methods for mapping neuronal activity suffer from low spatial resolutions and poor noise, which can limit the success of surgical treatments where an error of even 1 mm can be critical. The work described here involves the development of an implantable system for performing optical recordings of intrinsic signal (ORIS) on the surface of the brain. By taking advantage of the unique absorption spectrum of hemoglobin, cerebral blood volume (CBV) can be measured via reflectivity changes in the brain at at specific wavelengths of light. Due to the metabolic demands of the brain, the exaggerated neuronal activity and spiking associated with epileptic seizures can be detected indirectly through changes in CBV. While high resolution ORIS measurements have been recorded using externally mounted CCD sensors, this work presents some of the first developments in producing a fully implantable ORIS sensor. Progress in the development of an implantable ORIS sensor described here includes: an implantable organic light emitting diode (OLED) and organic photodetector (OPD) integrated on a highly flexible parylene-c substrate, an implantable sensor using a microLED array embedded on a flexible polyimide substrate, and the application of quantum dots to microLEDs for optical down-conversion. Successful in vivo detection of seizures is achieved with high signal-to-noise using these methods. Additionally, spatial localization of seizure activity is performed using the microLED array. These developments represent crucial first steps in the development of a full 2D neuronal mapping system using implantable ORIS devices.

Walkability indices are currently used for a wide range of research and commercial applications. Few studies have examined the relationship between walkability indices and measured pedestrian volume or walking rates, nor explored moderators of pedestrian volume such as weather. With 14 years of traffic study data from the City of Ottawa, a spatial auto-regressive (SAR) multi-level model (MLM) was used to understand the proportion of variance in walking explained by the commercial Walk Score® index and selected weather variables. Modeling revealed that a significant proportion of pedestrian volume at a given location in Ottawa, including its spatial lag, was explained by the corresponding Walk Score® value and its spatial lag (51.45%). Furthermore, weather expressed as a combination of ‘felt’ temperature, presence or absence of precipitation, and percent cloud cover, accounted for 2.79% of the variance in walking. These findings indicate that walkability indices may provide value as cost-effective engineering and urban planning tools.

The main aim of this research was to investigate the mechanism of cavitation in conforming and partially conforming tribological conjunctions. The effect of cavitation on load carrying capacity and frictional performance of is also investigated. This is important with regards to fuel efficiency in internal combustion (IC) engines. Friction accounts for 15–20% of IC engine losses. The piston–cylinder system contributes to 40–50% of these, with the compression ring(s) being responsible for most of this. This is because the primary function of the ring is to seal the combustion chamber, thus small emerging gaps lead to increased friction. In fact, compression ring(s) expend 3–5% of engine input fuel energy. The share of frictional losses of engine bearings is approximately 20–25%. Traditionally, prediction of performance of tribological conjunctions has been studied using Reynolds equation. When the effect of cavitation is considered, various cavitation algorithms with associated boundary conditions for lubricant rupture and reformation are proposed. These include Elrod, and Elrod and Coyne algorithms, as well as boundary conditions such as Swift-Stieber, JFO and Prandtl-Hopkins. There are a number of assumptions embodied in these approaches, as well as the use of Reynolds equation itself. These approaches do not uphold the continuity of mass and momentum in multi-phase flow, in cavitation beyond the lubricant film rupture. A detailed methodology for multi-phase flow, comprising simultaneous solution of Navier-Stokes, energy and lubricant rheological state equations is developed.

Estágio realizado na Sonae MC, e orientado pela Dr. Ana Cristina Leandro / Tese de Mestrado Integrado. Engenharia Industrial e Gestão. Faculdade de Engenharia. Universidade do Porto. 2012