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Orientadores: Jacobus Willibrordus Swart, Pierre Kaufmann / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de Computação / Made available in DSpace on 2018-08-16T16:19:31Z (GMT). No. of bitstreams: 1 Marins_CarlosNazarethMotta_D.pdf: 3218841 bytes, checksum: 93bb7dd5ba9269c6004bfb826f4c53fc (MD5) Previous issue date: 2010 / Resumo: Nesta tese foram propostas, analisadas e implementadas soluções completas de sistemas para disseminação de referência temporal com alta resolução através de enlaces de rádio tendo como foco o atendimento das necessidades do novo sistema de geo-referenciamento denominado GEOLOCAL. As soluções encontradas atendem outras aplicações que requerem sincronismo remoto de tempo. O trabalho foi desenvolvido em diferentes áreas que englobam contadores de alta precisão; relógios com resolução de nano segundo; enlaces de rádio para disseminação de tempo e determinação de distância; geradores de seqüências codificadas para detecção e identificação de campos de contagem; supressores digitais de jitter com uso de média de amostras; e sincronizadores de marcha para osciladores baseados em quartzo. Os contadores de alta precisão foram pesquisados para construção do relógio de alta resolução. Como resultado foi obtida uma solução inédita empregando linhas de retardo com contadores independentes. Esta configuração possibilita aliar alta resolução de contagem com uso de osciladores de referência operando em baixa freqüência sem que a incerteza do processo seja aumentada em função dos erros de não linearidade de interpolação. Os enlaces de rádio foram construídos com interface síncrona entre as bases de transmissão e recepção empregando modulação FSK (Frequency Shift Keying) e os erros nas medidas de tempo de propagação nos circuitos causados pela influência de jitter foram minimizados através do cálculo de média das amostras tomadas em cada campo de contagem. No intuito de viabilizar a implementação de um sistema de baixo custo, foi desenvolvida uma técnica inovadora de sincronização de marcha entre elementos osciladores de quartzo do tipo OCXO (Oven Controlled Crystal Oscillator). Esta técnica de sincronização permite que o sistema opere com estabilidades equivalentes a referências baseadas em rubídio. Os sistemas eletrônicos digitais foram construídos com solução FPGA (Field Programmable Gate Array). / Abstract: This thesis present new proposals for time dissemination systems, which were analyzed and implemented as complete solutions providing high resolution in radio links focusing in the requirements of the new geolocation system known as GEOLOCAL. The solutions found are useful for other applications equiring time synchronization. The work has been developed in different areas that include high precision counters; nanosecond resolutions clocks; radio links for time dissemination and measurements of distance; coded sequence generators for detection and identification of counting fields, digital jitter suppressors using samples averaging and clock synchronizers for quartz based oscillators. High precision counters have been investigated for the purpose to construct a high resolution clock. As a result it has been obtained a unique solution using delay lines with independent counters. This configuration allows the obtaining high count resolution with the use of low frequency reference oscillators, without increasing the system uncertainties due to interpolation non-linearity errors. The radio links were constructed with synchronous interface between the transmission and reception bases using FSK (Frequency Shift Keying) modulation. The measurements errors due to jitter in circuit propagation were minimized by calculating the average of samples taken every count set. In order to implement a low cost system, it has been developed an innovative clock synchronization technique between quartz oscillator elements to the type OCXO (Oven Controlled Crystal Oscillator). This synchronization technique produced stabilities equivalent to those obtained with rubidium-referred oscillators. The digital systems have been constructed using FPGA (Field Programmable Gate Array). / Doutorado / Telecomunicações e Telemática / Doutor em Engenharia Elétrica

The function of biomacromolecules is controlled by their structure and conformational flexibility. Investigating the structure of biologically important macromolecules can, therefore, yield information that could explain their complex biological function. In addition to X ray crystallography and nuclear magnetic resonance (NMR) methods, pulsed electron paramagnetic resonance (EPR) methods, in particular the pulsed electron electron double resonance (PELDOR) technique has, during the last decade, become a valuable tool for structural determination of macromolecules. Long range distance constraints obtained from pulsed EPR measurements, make it possible to carry out structural refinements on structures from NMR and X ray methods. In addition, EPR yields distance distributions that give information about structural flexibility. The use of EPR for structural studies of biomacromolecules requires in most cases site specific incorporation of paramagnetic centres known as spin labelling. To date, spin labelling nucleic acids has required complex spin labelling chemistry. The first application of a site directed and noncovalent spin labelling method for distance measurements on DNA is described. It is demonstrated that noncovalent spin labelling with a rigid spin label can afford detailed information on internal DNA dynamics using PELDOR. Furthermore, it is shown that noncovalent spin labelling can be used to study DNA protein complexes. PELDOR can also yield information about spin label orientation. Therefore, spin labels with limited flexibility can be used to measure the relative orientation of the spin labelled sites. Although information on orientation can be obtained from 9.7 GHz PELDOR measurements in selected applications, measurements at 97 GHz or higher, increases orientation selection. It is shown that PELDOR measurements on semi rigid and rigid nitroxide biradicals using a home built high power 97 GHz EPR spectrometer (Hiper) and model based simulations yield quantitative information on spin label orientations and dynamics. The most widely used spin labels for EPR studies on biomacromolecules are the aminoxyl (nitroxide) radicals. The major drawbacks of nitroxide spin labels include low sensitivity for distance measurements, fast spin spin relaxation in solution and limited stability in reducing environments. Carbon centered triarylmethyl (trityl) radicals have properties that could eliminate some of the limitations of nitroxide spin labels. To evaluate the use of trityl spin labels for nanometer distance measurements, models systems with trityl and nitroxide spin labels were measured using PELDOR and Double Quantum Coherence (DQC). This study shows that trityl spin labels yield reliable information on interlabel distances and dynamics, establishing the trityl radical as a viable spin label for structural studies on biomacromolecules.

Os Medidores de Distância por Triangulação a Laser são sistemas optoeletrônicos destinados à medição de pequenas distâncias ou deslocamentos, sem contato físico direto. Estes equipamentos possuem um grande campo de aplicações industriais. Este trabalho descreve as diversas configurações que este equipamento pode se apresentar, discutindo suas vantagens relativas. São estudadas também as suas partes e os componentes mais indicados. O projeto detalhado de um medidor usando sensor tipo CCD, e apresentado. Algumas aplicações industriais deste modelo são relatadas e diversos métodos e sugestões de uso são apresentadas, com base nas experiências realizadas pelo autor. / Laser Triangulation Rangefinders are non-contact, optoelectronic devices for measurement of small displacements or distances. These devices offer a broad range of industrial applications. This work explains common configurations, parts and main components, comparing their advantages. A detailed project on a CCD based Laser Rangefinder is presented. Some industrial applications for that project are related, discussing practical aspects based on authors experience. Some potential applications are also suggested.

Os Medidores de Distância por Triangulação a Laser são sistemas optoeletrônicos destinados à medição de pequenas distâncias ou deslocamentos, sem contato físico direto. Estes equipamentos possuem um grande campo de aplicações industriais. Este trabalho descreve as diversas configurações que este equipamento pode se apresentar, discutindo suas vantagens relativas. São estudadas também as suas partes e os componentes mais indicados. O projeto detalhado de um medidor usando sensor tipo CCD, e apresentado. Algumas aplicações industriais deste modelo são relatadas e diversos métodos e sugestões de uso são apresentadas, com base nas experiências realizadas pelo autor. / Laser Triangulation Rangefinders are non-contact, optoelectronic devices for measurement of small displacements or distances. These devices offer a broad range of industrial applications. This work explains common configurations, parts and main components, comparing their advantages. A detailed project on a CCD based Laser Rangefinder is presented. Some industrial applications for that project are related, discussing practical aspects based on authors experience. Some potential applications are also suggested.

EPR spectroscopy of site-directed spin labeled proteins is extremely informative in the studies of protein dynamics; however, it is difficult to interpret the spectra in terms of the conformational dynamics in atomic detail.In the present work we aimed to investigate the site-specific structural dynamics of proteins by using MD simulations upon analyzing and interpreting the EPR data. The major goal of this work is to know how far the computer simulations can meet the experiments. As a first step, MD simulations are performed to identify the location and orientation of the tyrosine radical in the R2 subunit of ribonucleotide reductase. The MD results show that the tyrosine is moving away from the diiron center in its radical state. This data is in agreement with EPR results and suggests reorientation of the tyrosine radical when compared to its neutral state. In further studies, the behavior of a methanethiosulfonate spin label, R1, in various environments of the protein is characterized by using MD simulations. RMSD analysis and angle ß distributions of the nitroxide show that R1 in buried sites in a protein helix is significantly immobile and in surface exposed sites it is highly mobile. Analyses of MD data suggest that internal rotations of x4 and x5 dihedrals of R1 are dominant in the R1 dynamics.Our studies also show that interaction with the surrounding residues show significant influence on the dynamics of R1. MD simulations data of the vinculin tail protein, both in water and in vacuo, are compared to the experimental results for further analysis of 12 different R1 sites in various environments.In a study on the photosynthetic reaction center(RC),MD is used to identify the location of the R1 binding site (H156)and thereby exploring the conformational dynamics in the RC protein upon light activation. The distance between the primary quinone, QA, and H156R1 determined from MD is in reasonable agreement with that measured by EPR.

This project was carried out for a Company in the mining and construction business, and focused on the development of a radar based material monitoring solution for conveyor belts. A minor prestudy showed that there are possible customer values related to material monitoring solutions for equipment relevant for the Company, and that such solutions using radar possibly can fulfil the application requirements. The main focus of the project was chosen based on this prestudy. The methods used were mainly experiments with the developed prototype, and a study of literature. The prototype was developed with an aim to measure mass flow and to monitor the material surface for deviances. Millimeter-wave pulsed coherent radar sensors were used, and data processing approaches included Kalman filtering. Results showed that mass flow can be calculated using the tested senors and data processing models, as the calculated mass flow was well in line with a belt scale reference, but that additional data is needed to prove consistency in the results. Results also showed that trends in the material surface can be observed in the radar data, indicating the possibilities to detect deviances. From a technical perspective, results and the possibilities for further development and improvement show that radar based solutions can be used for applications relevant for the Company. / Detta examensarbete genomfördes för ett företag inomgruv- och anläggningsbranschen och handlade om utvecklingen av en radarbaserad prototyp för mätningar av materialet på transportband. En mindre förstudie visade på möjliga kundvärden relaterade till sådana mätningar, samt att radar har potentialen att uppfylla kravspecifikationen för flera applikationer. Huvudfokuset för projektet valdes sedan utifrån denna förstudie. Metoderna som användes var främst experiment med den utvecklade prototypen samt litteraturstudier. Prototypen utvecklades med syfte att mäta massflöde samt att upptäcka avikelser i materialets yta. Radarsensorer med en frekvens på 60GHz användes, och för databehandling användes bland annat ett Kalman-filter. Resultaten visade att massflöde kan beräknas med de testade sensorerna och databehandlingsmodellerna, eftersom det beräknade massflödet låg i linje med referensdata från en bandvåg. Ytterligare data behövs emellertid för att visa hur resultaten påverkas av förändrade omständigheter, såsom storleken på fragmenten som mäts. Studien visade också att mönster i materialytan kan observeras i radardatan, vilket indikerar att det också är möjligt upptäcka avvikelser. Ur ett teknisktperspektiv visar resultaten och möjligheterna till vidareutveckling tillsammans att radarbaserade lösningar kan användas för applikationer relevanta för företaget.

A kinetic model of electronic energy migration within pairs of photophysically non-identical fluorophores has been developed. The model applies to fluorescent groups that exhibit different photophysical and spectral properties when attached to different positions in a macromolecule. The energy migration within such asymmetric pairs is partially reversible, which leads to the case of partial donor-donor energy migration (PDDEM). The model of PDDEM is an extension of the recently developed donor-donor energy migration model (DDEM, F. Bergström et al, PNAS 96 (1999) 12477), and applies to quantitative measurements of energy migration rates and distances within macromolecules. One important distinction from the DDEM model is that the distances can be obtained from fluorescence lifetime measurements. A model of fluorescence depolarisation in the presence of PDDEM is also presented. To experimentally test the PDDEM approach, different model systems were studied. The model was applied to measure distances between rhodamine and fluorescein groups within on-purpose synthesised molecules that were solubilised in lipid bilayers. Moreover, distances were measured between BODIPY groups in mutant forms of the plasminogen activator inhibitor of type 2 (PAI-2). Measurements of both the fluorescence intensity decays and the time-resolved depolarisation were performed. The obtained distances were in good agreement with independent determinations. Finally, the PDDEM within pairs of donors is considered, for which both donors exhibit a nonexponential fluorescence decay. In this case it turns out that the fluorescence relaxation of a coupled system contains distance information even if the photophysics of the donors is identical. It is also demonstrated that the choice of relaxation model has a negligible effect on the obtained distances. The latter conclusion holds also for the case of donor-acceptor energy transfer.