Este trabalho apresenta um estudo sobre sistemas operacionais de tempo real (RTOS) utilizados na implementação da infraestrutura de controle digital para sistemas mecatrônicos, mas serve também como referência para outros sistemas que possuam restrições de tempo. Além de ter um caráter experimental, onde foram medidos e analisados dados como o pior tempo de resposta dos sistemas e a latência para tratamento de interrupções, este trabalho de pesquisa ainda contempla a implementação e uso de RTOS em situações práticas, bem como contempla a construção de uma plataforma geral de pesquisa que servirá de base para futuros trabalhos no laboratório de mecatrônica. Os sistemas analisados neste trabalho foram o VxWorks, QNX, Linux, RTAI, Windows XP, Windows CE e \'mü\'C/OS-II. Outro produto gerado durante este trabalho foi um Live CD para auxiliar na implementação e ensino de conceitos e sistemas de tempo real. / This work presents a study about real time operating systems (RTOS) that are utilized as infrastructure to create digital control systems for mechatronics systems, and also for systems that have critical time constraints. Parameters like worst case response time and interrupt latency were measured for each operating system. This research project also covers the implementation and use of RTOS in practical situations. A general research platform for robotics and real time research was also developed and will be used for future works in the Mechatronics Laboratory. The tested systems were VxWorks, QNX, Linux, RTAI, Windows XP, Windows CE and \'mü\'C/OS-II. Another product released during this work was a Live CD to aid the implementation and teaching of real time systems and concepts.

Este estudo teve como objetivo detectar a presença do vírus da raiva em diferentes órgãos de morcegos do gênero Artibeus empregando as técnicas moleculares como RT-PCR, hnRT-PCR e Real Time RT-PCR. De aproximadamente 4000 espécimes de morcegos recebidas no Instituto Pasteur para o diagnóstico da raiva, foram selecionados 30 morcegos do gênero Artibeus, com resultados positivos para raiva pelas técnicas tradicionais de IFD e inoculação em células N2A utilizando suspensões feitas a partir do SNC. Para as técnicas moleculares, foram retirados glândulas salivares, bexigas urinárias, rins, pulmões e conteúdos fecais e ainda foram lavadas as calotas cranianas dos espécimes. Os órgãos e conteúdos fecais foram diluídos a 1:10 (P/V) e as bexigas urinárias a 1:20 (P/V). As suspensões foram inoculadas em células N2A para o isolamento viral. Foi realizada a extração do RNA total usando o TRIzol®, foram realizadas a transcrição reversa seguida da PCR e hnRT-PCR com utilização de primers específicos para o gene codificante da proteína N. A partir do produto da transcrição reversa foi realizada a técnica de Real Time RT-PCR, utilizando primers e sonda específicos para variante antigênica 3. Das 30 suspensões de lavado cerebral, 28 (93,33%) resultaram positivos, na inoculação em cultura de células, seguido de glândulas salivares (36,67%), bexigas (16,67%) e conteúdos fecais (3,33%). Os resultados encontrados da sensibilidade nas técnicas de RT-PCR, hnRT-PCR e Real Time RT-PCR foram 56,25%, 82,57% e 82,19% quando avaliadas as 180 amostras analisadas. A comparação das técnicas de hnRT-PCR e Real Time RT-PCR feita pelo teste exato de Fisher quanto a proporção de positivos detectados mostrou que para o lavado cerebral, órgãos e conteúdos fecais a proporção foi igual (P>0,05). Em relação à positividade os resultados encontrados nas técnicas de hnRT-PCR e Real Time RT-PCR foram 100% em lavado cerebral; 90% e 93,33% em glândulas salivares; 83,33% e 90% em bexigas; 80% e 93,33% em rins; 76,67% e 50% em pulmões e 43,33% em ambas as técnicas em conteúdos fecais. Esses resultados sugerem que tanto as técnicas de hnRT-PCR como Real Time RT-PCR podem ser utilizadas como métodos complementares para o diagnóstico da raiva e são sensíveis o bastante para o uso em estudos de patogênese. A técnica de Real Time RT-PCR realizada neste estudo se mostrou eficiente em detectar o RABV em diferentes órgãos e tecidos extraneurais com a vantagem de ser uma técnica mais rápida e sensível. / This study was aimed to detect the presence of rabies virus in different organs of the genus Artibeus bats using molecular techniques such as RT-PCR, hnRT-PCR, and the Real Time RT-PCR. From about 4,000 specimens of bats received for rabies diagnosis at the Pasteur Institute, 30 bats of the genus Artibeus were then selected. The selected bats presented positive results by the traditional DFA and N2A-cells inoculation test using brain tissue suspensions. Samples of salivary glands, urinary bladders, kidneys, lungs, and fecal contents and washings of the skulls were collected for the molecular techniques testing. The organs and the fecal contents were diluted at 1:10 (w/v) and the urinary bladder, at 1:20 (w/v) and these suspensions were inoculated into N2A cells for viral isolation. The extraction of the total RNA was performed by using TRIzol® and followed by the reverse transcription and the PCR and the hnRT-PCR were performed by using specific primers for the gene encoding the protein N. The product obtained by the reverse transcription technique was submitted to the Real Time RT-PCR technique, using primers and probe specific for antigenic variant 3 of the rabies virus. Of the 30 suspensions of the brain washings, 28 (93.33%) were positive in N2A cell culture inoculation, followed by the suspensions of the salivary glands (36.67%), bladders (16.67%) and fecal contents (3.33%). For the 180 samples evaluated, the results of sensitivity found for the RT-PCR, hnRT-PCR and Real Time RT-PCR techniques were 56.25%, 82.57%, and 82.19%, respectively. A comparison of hnRT-PCR and Real Time RT-PCR techniques performed by Fisher\'s exact test showed that the proportion of positives detected by the brain washings, organs and of the fecal content was non-significant (P> 0.05). Regarding the results found in hnRT-PCR and Real Time RT-PCR techniques, 100% positives were in brain washing, 90% and 93.33% in salivary glands, 83.33% and 90% in bladders, 80% and 93.33% in kidneys, 76.67% and 50% in lungs and 43.33% for both techniques on fecal contents. These results suggest that both hnRT-PCR and Real-Time PCR techniques can be used as complementary methods for the diagnosis of rabies and are sensitive enough for use in pathogenesis studies. The Real Time RT-PCR technique performed in this study proved to be faster and more sensitive and effective in detecting RABV in different organs and extra neural tissues of bats.

Artificial intelligence (AI) is already a fundamental component of computer games. In this context is emotions a growing part in simulating real life. The proposed emotional state module, provides a way for the game agents to select an action in real-time virtual environments. The modules function has been tested with the open-source strategy game ORTS. This thesis proposes a new approach for the design of an interacting network, similar to a spreading activation system, of emotional states that keeps track of emotion intensities changing and interacting over time. The network of emotions can represent any number of persisting states, such as moods, emotions and drives. Any emotional signal can affect every state positively or negatively. The states' response to emotional signals are influenced by the other states represented in the network. The network is contained within an emotional state module. This interactions between emotions are not the focus of much research, neither is the representation model. The focus tend to be on the mechanisms eliciting emotions and on how to express the emotions.

In the last few years, the number of real-time applications has increased. These applications are sensitive and require the methods to utilize existing network capacity efficiently to meet performance requirements and achieve the maximum throughput to overcome delay, jitter and packet loss. In such cases, when the network needs to support highly interactive traffic like packet-switched voice, the network congestion is an issue that can lead to various problems. If the level of congestion is high enough, the users may not be able to complete their calls and have existing calls dropped or may experience a variety of delays that make it difficult to participate smooth conversation. In this paper, we investigate the effect of priority levels on soft real-time performance. We use the priority queues to help us manage the congestion, handle the interactive traffic and improve the over all performance of the system. We consider switched multi-hop network with priority queues. All the switches and end-nodes control the real-time traffic with “Earlier Deadline First” scheduling. The performance of the network is characterized in terms of the average delay, the deadline missing ratio and the throughput. We will analyze these parameters with both the bursty traffic and evenly distributed traffic. We will analyze different priority levels and will see how the increase in priority level increases the performance of the soft real-time system.

The manufacturing industries play a significant role in contributing to the economy of a country. Among various manufacturing industries, the semiconductor manufacturing (SM) industries is one of the fastest growing industries in the world having worldwide sales of $31 billion in the month of December 2016. Semiconductors are required by large number of industries, including Telecommunications, Medical Electronics, Automobile, Defence and Aerospace, Consumer Electronics, etc.,. Today, without semiconductors, the technology that we count on every day would not be possible. Because of these, the demand for SM industry is increased rapidly. In addition, most of the semiconductor based products‘ life is very short. Due to these, SM industry is highly competitive industry. Thus, to utilize the resources effectively, to handle the huge demand, and to deliver the product on-time, efficient scheduling is important in SM industry. SM process can be broadly classified into Wafer Fabrication (called as wafer fab), Wafer Probing, Assembly, and Final Testing. Scheduling is more important in wafer fab due to complex operations involving with multiple types of machines and re-entrant, expensive machines, and time-consuming process involved. Thus, this study concerns about scheduling in wafer fab, particularly diffusion operation. The diffusion operation, carried out on batch processing machine, heavily impacts the production rate of wafer fab and in turn the SM industry. This is due to the fact that, diffusion operation requires relatively longest processing time among all the operations in the wafer fab. Due to these, diffusion operation is the bottleneck operations in the wafer fab. Based on the detailed literature review, this study addresses a new research problem on dynamic scheduling (DS) of diffusion furnace(s) by considering together the various real-life problem characteristics: Non-identical parallel diffusion furnaces, Machine eligibility restriction, Incompatible-job families, Job and/or resource related real time events, and Non-agreeable release time and due-date. In addition, due to the importance of on-time delivery this study deals with five due-date based scheduling objectives: Total weighted tardiness (TWT), Number of tardy jobs (NT), On time delivery (OTD) rate, Total earliness and lateness (TE/L) and Maximum lateness (Lmax) as a single objective as well as multi objectives. Here, the multi objectives are developed, considering all the five due-date based scheduling objectives in a linear form by randomly assigning equal and unequal weights to each of the due-date based single objectives considered in this study. With these, the main objective of this thesis is to study the impact of job and/or resource related real time events (JR-RTE) on the relative efficiency of the proposed dynamic scheduling algorithms for diffusion furnace(s) while optimizing each of the due-date based scheduling objectives considered in this study. The research problem considered in this study is decomposed into five phases. From the analysis of the literature, it is observed that, there is no earlier study has the mathematical models for dynamic scheduling (DS) of diffusion furnaces to optimize all the due-date based scheduling objectives, considered in this study. Due to this, in the first phase, fourteen (0-1) mixed integer linear programming (MILP) models are proposed for DS of diffusion furnaces (seven models for DS of single diffusion furnace and seven models for DS of non-identical parallel diffusion furnaces) to optimize the due-date based single objectives: TWT, NT, OTD rate, TE/L, and Lmax and multi objectives: MO1 and MO2. All the proposed (0-1) MILP models are demonstrated for its workability by developing a suitable numerical example, LINGO set code (which generates each of the proposed (0-1) MILP model for any given data), and solving using LINGO solver. Further, based on the analysis of the literature a suitable experimental design is proposed and generated 15 small-scale test data. The computational complexity of each of the proposed (0-1) MILP models is discussed empirically by solving 15 small-scale test data. Due to the computational intractability of the proposed (0-1) MILP models for DS of diffusion furnaces, the second phase of the research focuses on a simple alternative approach based on dispatching rules, as the analysis of the literature reveals that dispatching rules are heavily used in the SM industry. However, there is no study in the literature presenting a comparative analysis of various dispatching rules particularly due-date based dispatching rules (DDR) for DS of diffusion furnace(s) to optimize various due-date based scheduling objectives. Accordingly, in the second phase, this study proposes a simple Greedy Algorithm (GA) based on DDR (called as GA-DDR) for Dynamic Scheduling of Single Diffusion Furnace (DS-SDF). Further, this study proposes twenty variants of GA-DDR considering various due-date based dispatching rules such as Earliest Due-Date, Flow Due-Date, Operational Due-Date, Modified Operational Due-Date, Critical Ratio, Minimum Slack First, Cost OVER Time, ten versions of Apparent Tardiness Cost (ATC) [including a new ATC rule proposed in this study] & five versions of Batch Apparent Tardiness Cost (BATC) [including a new BATC rule proposed in this study] for DS-SDF. All these twenty variants of GA-DDR are implemented in Turbo C. An experimental design is proposed in this phase for generating large-scale test data. Accordingly, 270 large-scale problem instances (representing 27 problem configurations and 10 instances per configurations) are generated. With these, a series of computational experiments are carried out to understand the relative efficiency of the twenty proposed variants of GA-DDR as follows: The efficiency of each of the twenty proposed variants of GA-DDR for DS-SDF with respect to each of the scheduling objectives considered in this study is analysed in comparison with optimal objective function value obtained from the corresponding (0-1) MILP models for 15 small-scale problem instances using the standard performance measures: Average Relative Percentage Deviation (ARPD) and Maximum Relative Percentage Deviation (MRPD). Further, for each of the 270 problem instances the efficiency of each of the twenty proposed variants of GA-DDR for DS-SDF with respect to each of the scheduling objectives is analysed in comparison with estimated objective function value, which is computed by giving the twenty feasible solutions obtained for each instances as input to Weibull distribution, (i) empirically using the performance measures: ARPD, MRPD, Integrated Rank (IRANK), & Global comparison based on Worst Solution (GCWS), and (ii) statistically by using the performance measures: Mean, Median, and 95% confidence interval. From the overall analysis, at the end of the second phase of the study, six efficient variants of GA-DDR among the twenty proposed variants of GA-DDR are identified for DS-SDF and discussed the insights for their better performance. In these six efficient variants of GA-DDR, two variants of GA-DDR uses the new ATC rule and/or BATC rule proposed by the author of this thesis. The second phase of the research considers only dynamic arrival of jobs in all the twenty variants of GA-DDR. But, in the real-life various unexpected job related real time events: rush job, due-date change, early/late arrival of job, change in job priority, and job cancellation and/or resource related real time events: machine breakdown, operator illness, tool failure, shortage of material, and defective material will occur in addition to the dynamic arrival of jobs. From the literature, it is observed that, all the studies in the dynamic scheduling of diffusion furnaces consider only future arrival of jobs and no study considering real time events. Further, to the best of our knowledge, the research studies on discrete processing machines develop various rescheduling algorithm or modify the existing algorithm whenever real time events occur while taking the scheduling decision. However, due to the longest operation time requirements at diffusion furnace and the computerized tracking system in the shop floor of wafer fab, we strongly propose a research hypothesis that modifying appropriately the work-in-process (WIP) data and/or the availability time of the corresponding diffusion furnace(s) for next scheduling depending upon the occurrence of job and/or resource related real time events respectively by utilizing the existing computerized tracking system in the shop floor is sufficient, and changing any proposed efficient algorithms for DS-SDF is not required. This hypothesis is proved both empirically and statistically in the third phase of this research, considering the twenty proposed variants of GA-DDR for DS-SDF and the proposed experimental design. Accordingly, this study propose a formal researchable hypothesis that there is no impact of JR-RTE on the relative efficiency of the twenty proposed variants of GA-DDR for DS-SDF while optimizing each of the due-date based scheduling objectives considered in this study. For testing the proposed hypothesis, this study proposed adjusted GA-DDR (with JR-RTE) for each of the proposed GA-DDR, in which there is step to update the WIP data if job related event occurs, and/or the next available time of corresponding diffusion furnace(s) for scheduling the same if resource related event occurs, before finalizing the scheduling decision. Each of the 270 large-scale problem instances generated using the proposed experimental design is solved by each of the 20 adjusted variants of GA-DDR (with JR-RTE). The comparison on the relative efficiency of each of the 20 proposed variants of GA-DDR and adjusted GA-DDR (with JR-RTE) is carried out using the performance measures: ARPD and MRPD [that is, ARPD(GA-DDR) vs. ARPD(adjusted GA-DDR with JR-RTE), and MRPD(GA-DDR) vs. MRPD(adjusted GA-DDR with JR-RTE)] while optimizing each of the seven scheduling objectives considered in this study. The empirical analysis of the comparisons reveals that there is no change in the relative efficiency of each of the 20 proposed variants of GA-DDR and the corresponding 20 adjusted variants of GA-DDR (with JR-RTE) while optimizing each of the scheduling objectives considered in this study. Further, this study proved the proposed hypothesis statistically by conducting the Spearman‘s rank order correlation between each of the 20 variants of GA-DDR and adjusted GA-DDR (with JR-RTE) for DS-SDF while optimizing each of the seven due-date based scheduling objectives considered in this study. From the empirical and statistical analyses carried out in the third phase of the study indicated that, no need to adjust the proposed variants of GA-DDR for any occurrences of real time events for obtaining efficient schedule. The SM industry normally would have more than one non-identical diffusion furnaces and that too in parallel. Due to some technical reasons, some jobs are processed only in specific diffusion furnace(s) available in the shop floor (this is called as machine eligibility restriction in scheduling theory). Hence, the impact of JR-RTE on the dynamic scheduling (DS) of non-identical parallel diffusion furnaces (NPDF) with machine eligibility restriction (MER) is addressed in the fourth phase of this study. In the fourth phase of the research study, the twenty proposed variants of GA-DDR for DS-SDF extended appropriately for DS-NPDF with MER [called as Extended GA-DDR (EGA-DDR)]. Further, a few new problem parameters required for NPDF with MER are identified from the literature and extended the proposed experimental design and generated 270 problem instances for representing NPDF with MER. For testing the proposed hypothesis on the impact of JR-RTE on DS-NPDF with MER, exactly the similar research processes carried out for comparing GA-DDR vs. adjusted GA-DDR (with JR-RTE) is followed for comparing EGA-DDR vs. adjusted EGA-DDR (with JR-RTE). Both empirical and statistical analyses clearly proved that there is no impact of JR-RTE on the relative efficiency of the twenty variants of EGA-DDR for DS-NPDF with MER while optimizing each of the due-date based scheduling objectives considered in this study and no need to adjust the variants of EGA-DDR for any occurrences of real time events for obtaining efficient schedule. So far, the study addressed the development of efficient GA-DDR and EGA-DDR for DS-SDF and DS-NPDF with MER respectively and studied the impact of JR-RTE on the relative efficiency of these proposed GA-DDR and EGA-DDR. Now, in the final phase of the research study, the impact of JR-RTE on the meta heuristics: Simulated Annealing (SA) and Tabu Search (TS), one at a time, for DS-SDF while minimizing TWT are studied. Accordingly, the required parameters for these two meta heuristics are identified from the literature and the meta heuristics: SA and TS, considering each of the six solutions obtained from the six efficient variants of GA-DDR respectively as initial solution are implemented. From the analysis of the solutions obtained, for each of the 270 problem instances, from each of the six efficient variants of GA-DDR and from each of the meta heuristics: SA and TS, it appears that the six efficient proposed variants of GA-DDR seems to be robust in terms of both quality and computational time requirements in obtaining efficient solution. Further, to study the impact of JR-RTE on meta heuristics: SA and TS, this study considers (a) six solutions obtained from each of the six efficient variants of GA-DDR for DS-SDF as the initial solution and obtained six final solutions respectively from each of the meta heuristics, and (b) six solutions obtained from each of the six adjusted variants of GA-DDR (with JR-RTE) for DS-SDF as the initial solution and obtained six final solutions respectively from each of the meta heuristics. For each of the meta heuristics, these two sets of final solutions, obtained for each of the 270 problem instances, are compared empirically and statistically, based on various performance measures considered in this study, and proved the research hypothesis defined in this study. The major research contributions of this study are as follows - By analyzing the literature on scheduling diffusion furnaces and the real-life situation in scheduling diffusion furnaces, a new research problem on dynamic scheduling (DS) of diffusion furnaces with incompatible-job families, machine eligibility restriction, non-agreeable release time and due-date, considering job and/or resource related real time events (JR-RTE) along with dynamic job arrival to optimize due-date based scheduling objectives: TWT, NT, OTD rate, TE/L, and Lmax as a single objective as well as multi objective was defined. - Seven (0-1) MILP models for each of DS-SDF and DS-NPDF were proposed for optimizing each of the seven due-date based scheduling objectives considered in this study and the computational complexity was observed. - Due to the computational complexity of the proposed (0-1) MILP models and the popularity of the dispatching rules in the semiconductor manufacturing industry, this study proposed and compared the twenty variants of (i) greedy algorithm based on due-date based dispatching rules (GA-DDR) for DS-SDF, and (ii) Extended GA-DDR for DS-NPDF with machine eligibility restriction (MER). - The impact of JR-RTE on the twenty proposed variants of (a) GA-DDR for DS-SDF, and (b) EGA-DDR for DS-NPDF with MER was studied and observed that modifying the data appropriately by utilizing the existing computerized tracking system available in the shop floor is sufficient and rescheduling or modifying the existing algorithms are not required when the occurrences of JR-RTE happens. - Finally, single solution based meta heuristics: Simulated Annealing (SA) and Tabu Search (TS), considering each of the six solution obtained from each of the six efficient variants of GA-DDR proposed in this study as initial solution respectively, were proposed for DS-SDF to minimize TWT. Performance analysis of the solution obtained from each of the six efficient variants of GA-DDR and from each of the meta heuristics were carried out and observed that efficient variants of GA-DDR seems to be robust in terms of both quality and computational time requirements in obtaining efficient solution. In addition, the impact of JR-RTE on the meta heuristics: SA and TS were studied and proved the research hypothesis proposed in this study. Although, this study considers many real-life problem characteristics, there are certain limitations in this study. Though this study proposed mathematical model for DS-NPDF, the required additional constraint on Machine Eligibility is not considered in this study. Further, the impact of JR-RTE on the meta heuristics: SA and TS were studied considering only DS-SDF and not for DS-NPDF with MER. In addition to overcoming the limitations mentioned here, there are many immediate future research directions for the problem studied in this thesis such as proposing the greedy algorithms for scheduling diffusion operation along with upstream or downstream operation, and proposing population based meta heuristics for the research problem defined in this study.

This thesis addresses a number of problems in computer vision, image processing, and geometry processing, and presents novel solutions to these problems. The overarching theme of the techniques presented here is a multi-scale approach, leveraging mathematical tools to represent images and surfaces at different scales, and methods that can be adapted from one type of domain (eg., the plane) to another (eg., the sphere). The main problem addressed in this thesis is known as stereo reconstruction: reconstructing the geometry of a scene or object from two or more images of that scene. We develop novel algorithms to do this, which work for both planar and spherical images. By developing a novel way to formulate the notion of disparity for spherical images, we are able effectively adapt our algorithms from planar to spherical images. Our stereo reconstruction algorithm is based on a novel application of distance transforms to multi-scale matching. We use matching information aggregated over multiple scales, and enforce consistency between these scales using distance transforms. We then show how multiple spherical disparity maps can be efficiently and robustly fused using visibility and other geometric constraints. We then show how the reconstructed point clouds can be used to synthesize a realistic sequence of novel views, images from points of view not captured in the input images, in real-time. Along the way to this result, we address some related problems. For example, multi-scale features can be detected in spherical images by convolving those images with a filterbank, generating an overcomplete spherical wavelet representation of the image from which the multiscale features can be extracted. Convolution of spherical images is much more efficient in the spherical harmonic domain than in the spatial domain. Thus, we develop a GPU implementation for fast spherical harmonic transforms and frequency domain convolutions of spherical images. This tool can also be used to detect multi-scale features on geometric surfaces. When we have a point cloud of a surface of a particular class of object, whether generated by stereo reconstruction or by some other modality, we can use statistics and machine learning to more robustly estimate the surface. If we have at our disposal a database of surfaces of a particular type of object, such as the human face, we can compute statistics over this database to constrain the possible shape a new surface of this type can take. We show how a statistical spherical wavelet shape prior can be used to efficiently and robustly reconstruct a face shape from noisy point cloud data, including stereo data.

Células tronco mesenquimais de tecido adiposo, são uma promissora ferramenta para aplicações clínicas em terapias celular e regenerativa, em vista da facilidade de sua extração e da maior quantidade de células por unidade de massa de tecido quando comparado a outras fontes clássicas de células mesenquimais como medula óssea. O protocolo clássico de extração e purificação dessas células, depende de sua adesão em plástico e xeno-materiais demandando muito tempo para ser utilizado por médicos para auxiliar pacientes em procedimentos de emergência. Estas células são capazes se diferenciar em diversos tipos celulares, o que as torna boas candidatas para terapia celular, embora sua capacidade de transdiferenciação para fenótipos neuronais seja ainda discutida. Neste trabalho demonstramos um novo processo para isolar essas células na base de epitopos específicos expressos (assinatura molecular de superfície) utilizando aptâmeros como ligantes de alta afinidade para estes sitios. Aptâmeros, moléculas de DNA simples fita identificadas a partir de uma biblioteca combinatória de sequencias de DNA simples-fita foram identificados por ciclos reiterativos de seleção in vitro (SELEX) utilizando células tronco do lipoaspirado como alvo. Dois aptâmeros isolados, denominados APT9 e APT11, foram capazes de identificar subpopulações (15,8 e 23,7% respectivamente) dentre as células tronco mesenquimais (classicamente CD29+/CD90+/CD45-) e separá-las usando nano-partículas magnéticas acopladas aos aptâmeros. Além disso, seguindo uma indução para diferenciação neuronal, as células tronco mesenquimais passam a apresentar morfologia neuronal e apresentam expressão e atividade de diversos receptores de neurotransmissores, avaliados por PCR real-time e imageamento de variações da concentração de cálcio intracelular ápos stimulação com vários agonistas de receptores metatrópicos e ionotrópicos. Ao longo da diferenciação, os níveis transcricionais de mRNA de receptores de cininas (B1 e B2), nicotínicos (alfa 7), muscarínicos (M1, M3 e M4), glutamatérgicos (AMPA2 e mGluR2), purinérgicos (P2Y1 e P2Y4) e GABAergicos (GABA-A, subunidade 3) e da óxido nítrico sintase neural aumentaram quando comparados aos níveis das células não diferenciadas, enquanto que os níveis de expressão de outros receptores incluindo purinérgicos P2X1, P3X4, P2X7 e P2Y6 e muscarínico M5 diminuíram. Os níveis de atividade das classes dos receptores estudados, por imageamento de variações da concentração de cálcio intrac, aumentaram para a maioria dos agonistas analisados durante a diferenciação neuronal com exceção para respostas induzidas por glutamato e NMDA. Células diferenciadas expressavam altos níveis de antígenos específicos de neurônios como β3-tubulina, NF-H, NeuN e MAP-2 indicando uma diferenciação em fenótipo neuronal bem sucedida. Desta maneira, esta tese, ao identificar aptâmeros, prove uma inovadora solução para médicos usarem as células tronco mesenquimais dentro de uma sala de cirurgia, através de um método que é capaz de purificar essas células em um tempo clínico viável, com pureza e sem contato com contaminantes. Além disso, nós mostramos aqui que com um protocolo como o proposto para diferenciação neuronal, nós poderíamos induzir essas células para se diferenciar em neurônios, através da ativação de fatores de transcrição específicos, levando às células tronco mesenquimais a serem possivelmente utilizadas em terapias celulares de reparo neuronal. / Adipose mesenchymal stem cells are promising tools for clinical applications in cellular and regeneration therapies, in view of easiness of extraction and higher amount of isolated stem cells per mass of tissue when compared to other classical mesenchymal stem cell sources including bone marrow. The classical protocol to extract and purify these cells, depending on plastic adherence and xeno-materials, is too time consuming to be used by physicians to help patients at emergency procedures. These cells are able to differentiate into various cell types, making them good candidates for cell therapy, however their capability for transdifferentiation into neural phenotypes is yet discussed. Here we show a novel process to isolate these cells using their surface molecular signature and aptamers, ssDNA molecules identified through the SELEX technique, denominated APT9 and APT11 that are able to identify subpopulations (15,8 and 23,7% respectively) within the mesenchymal stem cells (classically CD29+/CD90+/CD45-) and separate them using magnetic nano-particles attached to the aptamers. Moreover, following induction to neural differentiation, mesenchymal cells presents neuronal morphology and present expression and activity of several neurotransmitter receptors, as evaluated by real-time PCR and calcium imaging. During this process, mRNA transcription levels of bradykinin (B1 and B2), cholinergic (alpha 7), muscarinic (M1, M3 and M4), glutamatergic (AMPA2 and mGlu2), purinergic (P2Y1 and P2Y4) and GABAergic (GABA-A, subunit 3) receptors and neuronal nitric oxide synthase were augmented when compared to levels of undifferentiated cells, while the expression levels of other receptors including purinergic P2X1, P2X4, P2X7 and P2Y6 and muscarinic M5 receptors were down-regulated. Activity levels of the studied receptor classes, as studied by calcium imaging, increased for most of the agonists analyzed during the neuronal differentiation with the exception for glutamate- and NMDA-induced receptor responses. Differentiated cells expressed high levels of neuron-specific antigens such as β3-tubulin, NF-H, NeuN and MAP-2, indicating a successful differentiation into neuronal phenotypes. This thesis, by identifying aptamers, provides a novel solution for physicians to use mesenchymal stem cells inside a surgery room, by using a method that are able to purify the cells in a clinical viable time, with purity and no contact with contaminats. Furthermore, we show here that with a protocol as provided for neuronal differentiation, we could induce these cells to differentiate into neurons, by activating specific transcription factors,making mesenchymal stem cells to possibly be used in neuronal repair cell therapies.

The thesis deals with a development and application of laser interferometer system for the online error calibration of CNC machine tool. The term „online“ means, that laser interferometer is permanently built in the machine tool. Therefore particular error measurements can be done in very short time period.

The aim of the diploma thesis is to make functional application for design of DAQ system for productive machine by using LabVIEW and platforms cRIO. The first part is focused on productive machine. After the thesis continues with specific examples. These examples have impact on machining accuracy and their measurements. The second part of diploma thesis is focused on making measurement program. LabVIEW FPGA Module, LabVIEW Real-Time Module are used in the program.

The diploma thesis deals with file sharing among users through peer-to-peer transfer in a web environment. The aim was to create a simple application which would provide a group of users with an opportunity to share files among them without any presence of third party or limitation. I have met the target using modern web technologies, of which the most important was WebRTC library through which peer-to-peer transfer has been implemented. These technologies were supported by optimased algorithm and the most recent tools for processing and storage of shared files. The result is a real-time application enabling users to create rooms, in which they can share their files. This product was subjected to speed tests, thanks to which the application turned out to be the fastest in the tested environment contrary to solutions with similar nature or any other commonly used tools for file sharing.