Orientador: Willian Fernando Zambuzzi / Resumo: Modelos in vitro têm facilitado a análise da fisiologia celular sob condições experimentais diversas; trata-se de modelo alternativo ao uso de animais de experimentação, que vem sendo difundido e aceito amplamente em pesquisa científica. Além disso, estes modelos têm levado à avanços significativos na compreensão das interações mútuas e adaptativas entre células e substratos. Neste trabalho, nosso objetivo foi analisar eventos moleculares responsáveis pela adaptação de preosteoblastos em substrato orgânico composto por componentes da Matriz Extracelular (MEC), sob condição de diferenciação celular. Metodologicamente, pré-osteoblastos (MC3T3-E1 50x103 células/ml) foram semeados sobre uma fina camada de Matrigel® gelificada e mantidos por 10 dias (37oC, 5% de CO2 em ambiente úmido) sob condição de diferenciação (meio de cultivo contendo 50 µg de ácido ascórbico e 10 mM de ß- glicerofosfato), com renovação do meio de cultivo a cada 3 dias. Alterações morfológicas foram monitoradas em microscópio invertido e mecanismos moleculares acompanhados pela análise global da atividade de quinases, através de arranjo de peptídeos (Pepchip). Curiosamente, nossos resultados mostraram mudanças morfológicas significantes durante adaptação celular as quais foram acompanhadas pela atuação de vias de sinalização celular distintas, responsáveis pela sobrevivência (Eixo PI3K-Akt) e proliferação (Eixo Retinoblastoma-ciclinas) celulares, além de proteínas envolvidas com metabolismo energético e comun... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In vitro models have been proposed to analyze cellular physiology under various experimental conditions. It is an alternative model instead using experimental animals that have been widespread and widely accepted in scientific research. Moreover, it has led to significant advances in the understanding of mutual and adaptive interactions between cells and substrates. In this work, our aim was to analyze adaptive events of osteoblasts cultured on an organic substrate composed by components of the extracellular matrix (ECM) in the first 10 days of cultivation on differentiation condition. Methodologically, pre-osteoblasts (MC3T3-E1 pre-osteoblasts, 50x103 cells / ml) were seeded on a thin gelified Matrigel® layer and maintained for 10 days under standard cell culture conditions (37 ° C, 5% CO2 in a humid environment) under differentiation conditions (culture media containing 50 ug of ascorbic acid and 10 mM beta-glycerophosphate). The culture medium was changed every 3 days. Morphological changes were monitored using an inverted microscope and molecular mechanisms followed by comprehensive analysis of kinase activity by peptides arrangement (Pepchip). Interestingly, our results showed significant morphological changes during cell adaptation which were accompanied by distinct signaling pathways involving proteins responsible for survival (PI3K-Akt axis) and cell proliferation (Retinoblastoma-cyclins axis), in addition of proteins involved in energy metabolism and cellular communi... (Complete abstract click electronic access below) / Mestre

At present companies are facing to a problem of treating huge amount of company`s content. There is needed to provide correct metadata to the documents to enable its qualified classifying during a whole life cycle of the document. The target of this diploma thesis puts emphasis on a proposal of the classifying methodology of the customer`s documents, while intercepting a proposal of the process of classifying, testing and support or even enhancement. Topic of this thesis is unique while it is based on the combination of the IT and librarian`s technologies. Methodology also consists a proposal of a subject retrieval language and a proposal of IT services, which are going to ensure the quality of the provided classification. The thesis is devided into two parts. The theoretical one explains the basic terms, theory of classifying and standards, legislature, which all together have the important influence on the field. The practical part includes itself the methodology.

Enterprise content management (ECM) is an emerging research area that is beginning to find attention in academia. While the private sector has a growing industry and community for ECM, academia is starting to address this with direct links to the better-established areas of information systems and enterprise resource planning systems. ECM has been viewed as a higher-level concept of methods and strategies pertaining to content management in the context of the enterprise. Like many other organizational wide systems, ECM systems are complex, difficult to implement and risk failing to meet expected success measures. Definitions for what exactly constitutes an ECM system are still evolving. The major issues with ECM systems are that they are increasingly being implemented by organizations in an attempt to address the unmanageable amount of unstructured content over its lifecycle, compliance pressures, collaboration needs, content integrity and continuity, and controlling costs. However, the implementation problems are many and diverse, such as determining content and business processes to be included, determining technologies to fit the organizational needs, how to integrate with existing systems, and managing organizational culture and change for acceptance. There is currently little academic research in the area of ECM, and research determining the key factors that contribute to successful implementations of these systems is absent. This research addressed the existing gap in ECM research and investigated the key success factors for the implementations of ECM systems with the objectives of identifying a set of success factors. Guided by research in related areas and through developing a theoretical framework and the resulting research model, the study used a qualitative case study method to identify ECM implementation factors and their relationship to organizational culture and people, business processes, technology and organizational content. The results of this research were twofold, first by contributing needed research in the ECM area and second to aid organizations in the implementation of ECM systems by identifying key factors.

In Electrochemical Machining (ECM), where the material removal takes place based on the anodic dissolution of the workpiece material, the working distance is one of the most important parameters. Especially in Jet Electrochemical Machining (Jet-ECM), where a micro nozzle is moved over the initial surface of the workpiece in order to apply an electrolytic free jet to produce the desired shapes, the distance between the nozzle and the workpiece becomes even more important. On the one hand a small working distance is aspired to achieve high current densities resulting in a high efficiency of the process. On the other hand the working distance needs to be large enough to avoid damages on the micro nozzle caused by electrical discharges or mechanical contact. Hence, the adjustment of the working gap is essential to realize a precise, effective and secure Jet-ECM process. The control of the gap size is done based on the data gathered before machining by surface measurement. Until now, the initial surface has been detected by electrostatic probing through moving the nozzle stepwise to the work piece surface and detect the voltage drop between the nozzle and the work piece. With this strategy, only a limited number of points can be detected within adequate time. Hence, in most cases only three points of the initial surface are detected in order to adjust the working distance according to the planar inclination of the workpiece. The coordinates of the three detected points are used to calculate the normal vector of the initial surface. In recent studies, another strategy was analysed, which is realized by dividing the surface into smaller areas and respectively calculating the normal vector of each area in order to obtain more accurate data of the initial surface. A further strategy is to use probing along the machining path of the tool and to gather the coordinates of a number of points along the path. The above mentioned methods usually do not ensure the precise control of the gap size especially for the surfaces with complex geometry with locally confined convex and concave shapes and are highly affected by the size of the probe. In this study, the application of a laser scanner is investigated for the measurement of the workpiece surface before machining to gather the required data for the adjustment of the working distance during Jet-EC machining of complicated surfaces.

Jet Electrochemical Machining (Jet-ECM) is a manufacturing technique that applies a free electrolyte jet to generate the desired shapes [1]. Since the principle of the technique is the same as other techniques of Electrochemical Machining where the material removal takes place based on the anodic dissolution of workpiece, the working distance, which is the distance between nozzle’s front surface and the workpiece surface, is one important parameter of the process. The working distance affects the current density and consequently the geometry removal. The control of the working distance can be done based on the data gathered before and during machining by surface measurement [2]. This measurement usually is done by using electrostatic probing to detect a limited amount of points of the initial workpiece surface. Since electrostatic probing is comparatively slow, laser triangulation represents an alternative technique to detect a larger amount of points before machining within significantly shorter time [3]. In addition to electrostatic probing and laser triangulation, the actual working distance can be measured during the machining process to realize constant working distance. This can be done by detecting electrical signals like the actual total current. This method can be combined with pre-machining measurement by laser triangulation in order to ensure the prevention of any collision between the nozzle and the workpiece. In this study, on-machine metrology techniques for measuring the working gap as well as current measurements will be compared. Besides, the advantages and disadvantages of these techniques will be systematized. In further studies, the possibility of combining the techniques will be investigated to enhance Jet-ECM with more accurate measurement techniques.

The behavior of cancer cells is strongly influenced by the properties of extracellular microenvironments, including topology, mechanics and composition. As topological and mechanical properties of the extracellular matrix are hard to access and control for in-depth studies of underlying mechanisms in vivo, defined biomimetic in vitro models are needed. Herein we show, how pore size and fibril diameter of collagen I networks distinctively regulate cancer cell morphology and invasion. Three-dimensional collagen I matrices with a tight control of pore size, fibril diameter and stiffness were reconstituted by adjustment of concentration and pH value during matrix reconstitution. At first, a detailed analysis of topology and mechanics of matrices using confocal laser scanning microscopy, image analysis tools and force spectroscopy indicate pore size and not fibril diameter as the major determinant of matrix elasticity. Secondly, by using two different breast cancer cell lines (MDA-MB-231 and MCF-7), we demonstrate collagen fibril diameter - and not pore size - to primarily regulate cell morphology, cluster formation and invasion. Invasiveness increased and clustering decreased with increasing fibril diameter for both, the highly invasive MDA-MB-231 cells with mesenchymal migratory phenotype and the MCF-7 cells with amoeboid migratory phenotype. As this behavior was independent of overall pore size, matrix elasticity is shown to be not the major determinant of the cell characteristics. Our work emphasizes the complex relationship between structural-mechanical properties of the extracellular matrix and invasive behavior of cancer cells. It suggests a correlation of migratory and invasive phenotype of cancer cells in dependence on topological and mechanical features of the length scale of single fibrils and not on coarse-grained network properties.

Extracellular matrix stiffening of breast tissues has been clinically correlated with malignant transformation and poor prognosis. An increase of collagen fibril diameter and lysyl-oxidase mediated crosslinking has been observed in advanced tumor stages. Many current reports suggest that the local mechanical properties of single fibrillar components dominantly regulate cancer cell behavior. Here, we demonstrate by an independent control of fibril diameter and intrafibrillar crosslinking of threedimensional (3D) collagen matrices that fibril bending stiffness instructs cell behavior of invasive and non-invasive breast cancer cells. Two types of collagen matrices with fibril diameter of either 650 nm or 800 nm at a similar pore size of 10 µm were reconstituted and further modified with the zero-length crosslinker 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide at concentrations of 0, 20, 100 and 500 mM. This approach yields a set of collagen matrices with overlapping variation of matrix elasticity. Within this set of matrices we could prove the common assumption that matrix elasticity of collagen networks is bending dominated with a linear dependence on fibril bending stiffness. We derive that the measured variation of matrix elasticity is directly correlated to the variation of fibril bending stiffness, being independently controlled either by fibril diameter or by intrafibrillar crosslinking. We use these defined matrices to demonstrate that the adjustment of fibril bending stiffness allows to instruct the behavior of two different breast cancer cell lines, invasive MDA-MB-231 (human breast carcinoma) and non-invasive MCF-7 cells (human breast adenocarcinoma). Invasiveness and spreading of invasive MDA-MB-231 cells as well as clustering of non-invasive MCF-7 cells is thereby investigated over a broad parameter range. Our results demonstrate and quantify the direct dependence of cancer cell phenotypes on the matrix mechanical properties on the scale of single fibrils.

No description available.

No description available.

The Expectation confirmation model (ECM) is commonly employed to investigate technological advances and customer satisfaction with chatbots. However, customer satisfaction is a multifaceted concept intertwined with emotions and subjective perceptions, comprehending customer satisfaction fully can therefore be challenging. Consequently, the purpose of this exploratory study was to develop and refine the concept of customer satisfaction by answering the research question: How does the integration of underlying factors to the Expectation Confirmation Model (ECM) affect customer satisfaction with chatbot services?. The study incorporated underlying factors such as anthropomorphic behavior, brand perception, trust and word of mouth, which were integrated into the existing components of the ECM. Through a unique research approach involving 20 semi-structured interviews and observations, this study captured and reflected on the subjective impressions and emotions of customers while using chatbots. An array of findings are presented, such as the importance of distinguishing between usability and problem-solving efficiency of chatbots performance, the emphasis on accurate answers over human-like characteristics for chatbots, and the impact of experience, familiarity and age on customer satisfaction with chatbots. In conclusion, this study advocates for further qualitative research to explore  the potential impact of other underlying factors' on customer satisfaction for ECM.