This thesis purposes and develops inductively coupled LC (inductive-capacitive) pH sensors based on pH-sensitive electrode pair. The LC resonator circuit is based on a varactor and measures the low frequency potential difference. For wireless pH monitoring, the resonator circuit is integrated with a pH-sensitive electrode pair. This sensor demonstrates a linear response over 2 to 12 pH dynamic range, 0.1 pH accuracy and long-term stability. Accurate measurement of pH using electrode-based sensors is affected by temperature variation. A technique of simultaneously measuring two parameters, pH and temperature, with a single RLC resonator based sensor is presented. An algorithm is developed, which applies both pH and temperature measurement to incorporate temperature compensation in pH measurement. For in-fluid applications, an encapsulation method is applied to the LC resonator based sensor to reduce the influence of medium permittivity and conductivity on the sensor measurement. Non-invasive way to obtain reliable pH information from bacterial culture bioprocesses is demonstrated with the fluid embeddable sensor. The pH sensor is remodeled to an acidic and basic volatile sensor by embedding the electrodes in a hydrogel host electrolyte. Tests demonstrate that the volatile sensor has a detection limit of 1.5 ppm and 2 ppm for ammonia and acetic acid vapor, respectively. Application of the volatile sensor to fish spoilage monitoring shows that the sensor is capable of detecting the product rejection level with good sensitivity in real-time. It is important to develop low cost wireless passive pH sensor technologies for embedded applications such as bioprocess and food spoilage monitoring. The electrode-based passive LC sensor approach employed in this thesis overcomes drawbacks of some of the early developed passive pH sensors and can lead to an inexpensive implementation using printed electronics technology.

The thesis describes development and application of coupled resonant coil sensors, which is of growing interest for remote monitoring applications. An interrogation technique, which improves the accuracy and interrogation range of coupled resonant coil sensors, is introduced. The method uses time-domain gating to produce measurements that are dominated by the response of the sensor coil and are immune to surrounding object interference. For application in structural health monitoring a low cost embeddable coupled coil sensor, which is able to monitor the corrosion potential of reinforcement steel is presented. Results of an accelerated corrosion test using the sensor indicate that corrosion potential can be monitored with a resolution less than 10 mV and a sensitivity of 0.76 kHz/mV. The last part describes a coupled-coil pH sensor based on pH electrode potential measurement. A linear response over a 4 to 10 pH dynamic range and 50 kHz/pH sensitivity are achieved with a 0.1 pH resolution and 30 s response time.

After nearly a decade of development, human-implantable sensors for detection of muscle activity have recently been demonstrated in the literature. The implantable sensors are powered and communicate wirelessly through the skin using coupled inductor coils. The focus of the present work has been the development of a new approach to modeling the inductively coupled link by using the finite element method (FEM) to simulate a three-dimensional representation of the coils and surrounding magnetic field. The validity of the simulation is tested by comparison to analytically-developed formulas for self-inductance, ac resistance and mutual inductance of the coils. Determination of these parameters is necessary for calculation of the coupling coefficient between the coils, and to fully define the lumped circuit model of the link. This 3D FEM approach is novel and attractive because it is able to encompass physical geometric parameters and material properties that have been traditionally been a challenge to determine. In particular the contribution of a ferrite-core, and the case of non-symmetrical relative coil positioning can be evaluated. / Graduate

Detta examensarbete har utförts hos Vattenfall Eldistribution på avdelningen Kontroll och Skydd med målsättningen att utforma och utvärdera en nätmodell avsedd för jordslutningssimuleringar i PSCAD. Ombyggnationen från luftledningar till markkablar har medfört att kapacitansen i distributionsnätet ökat, vilket ställer högre krav på jordfelsutrustning och på nätanalyser för att kunna försäkra att en säkerställd frånkoppling kan ske enligt de myndighetskrav som föreligger. Syftet med arbetet har varit att undersöka hur en nätmodell bör utformas för analys av stationära in-svängningsförlopp och utvärdera hur stor noggrannhet som kan förväntas gentemot verkliga jordfelsprov vid felresistanser på främst 3 kΩ och 5 kΩ. Nätmodellen har ut-formats efter ett verkligt impedansjordat mellanspänningsnät med π-länkar i PSCAD och utifrån de nätuppgifter som förekommer i Vattenfalls näthanteringsprogram Netbas. Simuleringsresultaten har jämförts mot resultat från det verkliga nätets jord-felsprover vid olika inställningar på den centrala kompenseringsutrustningen som är placerad mellan den matande transformatorns nollpunkt och jord. Jordslutningssimuleringarna visar, trots antaganden och en viss osäkerhet omkring de verkliga nollföljdskomponenterna, godtyckliga simuleringsresultat vid avstämt och snedavstämt nät motsvarande ± 30 A gentemot ledningarnas kapacitiva strömmar vid en felresistans på 3 kΩ. Jordslutningssimuleringarnas händelseförlopp överensstäm-mer överlag väl mot det verkliga nätet samtidigt som jordslutningar vid 5 kΩ medför en större procentuell avvikelse. Särskilt framträdande avvikelser kunde urskiljas vid analys av fasvinkeln mellan nollföljdsspänning och nollföljdsström. En analys av nätmodellens resultat och troliga orsaker till uppkomna simulerings-avvikelser gentemot det verkliga nätet indikerar på att nätmodellens tillförlitlighet bör kunna optimeras ytterligare om noggrannare hänsyn tas gällande nollföljdsimpedanser, dc-komponenter och de toleranser som förekommer i det verkliga nätets avstäm-ningsutrustning och mätkretsar. / This thesis has been carried out at Vattenfall Eldistribution at the department Control and Protection with the objective to design and evaluate a network model for ground fault simulations in PSCAD. The reconversion from overhead lines to underground cables has led to increased capacitances in the distribution network and this places greater demands on the feeder protection unit and network analyzes in order to assure that faulted feeders are disconnected according to regulatory requirements. The aim of this work has been to determine how a network model could be designed for analysis of stationary signal characteristics and evaluate how great accuracy the power system model have compared to real earth fault test results. Earth fault simulations are performed with fault resistances of 3 kΩ and 5 kΩ. The power system model have been created to emulate a real impedance grounded network according to the π-model in PSCAD based on system information from Vattenfalls network management program Netbas. Results from the simulations have been compared against results obtained from real earth faults from the physical network with different settings on the central compensation equipment placed between the transformers neutral and ground. Simulations show, despite assumptions and some uncertainty about the actual zero sequence components similar results when fault resistance was 3 kΩ, both when compensation coil are fully tuned and out of tune ± 30 A corresponding to the feeder capacitance. The overall signal sequence conform quite well to the real network but at the same time simulations with 5 kΩ obtains greater deviations when results are represented in percentage. Particularly prominent abnormalities could be identified in the phase angle between zero sequence voltage and zero sequence current. An analysis of deviations from the simulations in the digital network model against the real system indicates that the model probably could be further optimized if zero sequence impedances, dc components, and tolerances that occur in the real systems reactive compensation equipment and measuring circuits are taken into account.

Esta dissertação apresenta modelagem matemática e um método de solução numérica para problemas de conformação eletromagnética de chapas metálicas finas circulares utilizando uma bobina em espiral plana. O método foca especificamente o cálculo do campo eletromagnético gerado pela bobina e análise do circuito que modela o sistema de estampagem eletromagnética. A bobina plana é aproximada por círculos concêntricos carregando a corrente de descarga dos capacitores. Os cálculos das correntes induzidas e perfil da força eletromagnética na chapa e acoplamentos magnéticos entre a bobina e chapa metálica são realizados para o instante inicial, antes da deformação plástica da chapa. O método utiliza a lei de Biot-Savart, sendo que a solução das integrais obtidas para indução magnética é realizada através de métodos numéricos, considerando as simetrias do problema. Para verificação da modelagem matemática, da solução numérica e comprovação da viabilidade técnica deste processo, um dispositivo de estampagem eletromagnética foi desenvolvido e diversos experimentos foram realizados com chapas de alumínio. Os parâmetros investigados foram a espessura da chapa a ser deformada e a influência da presença de vácuo na cavidade da ferramenta. Os resultados experimentais demonstram concordância com os resultados da modelagem proposta. A presença de vácuo também demonstrou um incremento na deformação da chapa. A rotina desenvolvida no software Matlab fornece informações importantes para o processo e permite que se faça ajustes no dispositivo. / This dissertation presents a mathematical model and numerical method to solve the problems of electromagnetic forming of thin circular metal sheets by using a flat spiral coil. The method focuses specifically on the calculation of the electromagnetic field generated by the coil flat and circuit analysis modeling system electromagnetic forming. The flat coil is approximated by concentric circles carrying a discharge current of capacitors. The calculations of induced currents and profile of the electromagnetic force on the plate and magnetic coupling between the coil and sheet metal are made to the initial time, before the plastic deformation of the plate. The method uses the law of Biot-Savart, and the solution of the integral obtained for magnetic induction is performed by numerical methods, considering the symmetries of the problem. To verify the mathematical model, the numerical solution and proving the technical feasibility of this process, a electromagnetic forming device was developed and several experiments were made with aluminum plates. The parameters investigated were the thickness of the plate to be deformed and the influence of the vacuum in the cavity of the tool. The experimental results show agreement with the results of the proposed model. The presence of vacuum also showed an increase in the deformation of the plate. A routine developed in software Matlab provide important information for the process and allow to make adjustments on the device.

A percussion performance by a skilled musician will often extend beyond a written score in terms of expressiveness. This assertion is clearly evident when comparing a human performance with one that has been rendered by some form of automaton that expressly follows a transcription. Although music notation enforces a significant set of constraints, it is the responsibility of the performer to interpret the piece and “bring it to life” in the context of the composition, style, and perhaps with a historical perspective. In this sense, the sheet music serves as a general guideline upon which to build a credible performance that can carry with it a myriad of subtle nuances. Variations in such attributes as timing, dynamics, and timbre all contribute to the quality of the performance that will make it unique within a population of musicians. The ultimate goal of this research is to gain a greater understanding of these subtle nuances, while simultaneously developing a set of stochastic motion models that can similarly approximate minute variations in multiple dimensions on a purpose-built robot. Live or recorded motion data, and algorithmic models will drive an articulated robust multi-axis mechatronic system that can render a unique and audibly pleasing performance that is comparable to its human counterpart using the same percussion instruments. By utilizing a non-invasive and flexible design, the robot can use any type of drum along with different types of striking implements to achieve an acoustic richness that would be hard if not impossible to capture by sampling or sound synthesis. The flow of this thesis will follow the course of this research by introducing high-level topics and providing an overview of related work. Next, a systematic method for gesture acquisition of a set of well-defined percussion scores will be introduced, followed by an analysis that will be used to derive a set of requirements for motion control and its associated electromechanical subsystems. A detailed multidiscipline engineering effort will be described that culminates in a robotic platform design within which the stochastic motion models can be utilized. An analysis will be performed to evaluate the characteristics of the robotic renderings when compared to human reference performances. Finally, this thesis will conclude by highlighting a set of contributions as well as topics that can be pursued in the future to advance percussion robotics. / Graduate / 2019-12-10

Idag befinner sig produktionsföretag under konkurrenskraftiga och snabba förändringar i en global marknad. Det utmanar företag som jobbar med materialtillverkning och distributionsnät på en global nivå till att ständigt förbättra deras verksamhet. Aktörerna som verkar inom en och samma supply chain bör enas för att kunna bli konkurrenskraftiga och flexibla, som respons på globaliseringen. Genom att företag skapar gemensamma mål, delar information, risker och vinster kan alla aktörer inom kedjan gynnas. Denna fallstudie har genomförts på en av Oxelösunds Hamns verksamheter, nämligen SSAB:s Stålhamn, som de driver åt SSAB i Oxelösund kommun. Stålhamnen har en viktig roll i SSAB:s supply chain eftersom den agerar som SSAB:s skeppningshamn där material som kommer från SSAB i Borlänge och Oxelösund lagras tills de ska skeppas vidare till andra hamnar i världen. Idag upplever Oxelösunds Hamn ineffektiv information- och materialflöde inom Stålhamnens verksamhet. Syftet med studien är att undersöka materialhantering i form av vilka transport- och lagringsmetoder som sker inne i en av Stålhamnens verksamheter, Coilsterminalen. Därefter klargörs vilka slöserier som finns samt hitta förbättringar för att effektivisera information- och materialflödet vid hanteringar av plåtrullar i Coilsterminalen. Studien har primärt genomförts genom observationer, ostrukturerade och några strukturerade intervjuer samt strukturerade mejlkonversationer och insamling av data. Fokuset låg på att hitta de möjliga effektiviseringsområden i praktiken för att sedan hitta teori och litteratur som belyser de upphittade bristerna. Några teorier som kommer att betonas på grund av deras relevans är Lean, lager- och supply chain teori. Studiens resultat och analys visar att utmaningarna som hindrar effektivisering i Coilsterminalen är baserad på interna och externa faktorer. Interna faktorer består av onödig förflyttning av material, omarbetning i kontor och outnyttjat arbetsskift. Externa faktorer orsakas på grund av leverantörer i form av osäkerhet av informationsinput samt material input/output. Studien resulterar i att den optimala lösningen är att samarbetet mellan aktörerna i kedjan förbättras så att flöde av information blir bättre och förminska osäkerheten i leverans och utleverans. Vidare uppmuntrar studien Oxelösund Hamn att samla in data, i form av statistik, om dagliga händelser i Stålhamnen så att de belyser problem och kostnader som är dolda. Oxelösund Hamn uppmuntras också införa det outnyttjade arbetsskiftet under natten och förminskning av lagret. / Today, manufacturing companies are under competitive and rapid changes in the global market. It challenges companies working with material manufacturing and distribution networks at a global level to continually improve their business. The actors operating within a single supply chain should unite to increase their ability to compete and remain flexible, as a response to the effects of globalization. By creating business goals, sharing information, risks, and profits, all operators in the chain can benefit. This case study has been carried out in one of Oxelösund Harbor's operations, namely SSAB's Steel Port, which they operate for SSAB in Oxelösund municipality. The steel harbor has an important role in SSAB's supply chain because it acts as SSAB's port of shipment where materials coming from SSAB in Borlänge and Oxelösund are stored until they are shipped to other ports in the world. Today, Oxelösund Harbor is experiencing inefficient information and material flow within the Steel Harbor's operations. The purpose of the study is to investigate material handling in terms of which transport and storage methods are carried out in one of the steel ports operations, the coil terminal. Thereafter, the study will clarify what kind of waste there is and find improvements to streamline information and material flow when handling coils in the coil terminal. The study was primarily conducted through observations, unstructured and to a lesser extent structured interviews, structured email conversations and data collection. The focus was on finding the possible areas that could become more efficient in practice and then finding the theory and literature that highlight the shortcomings found. Some theories that will be emphasized, because of their relevance, are Lean, inventory and supply chain theory. The study's results and analysis show that the challenges that hinder the efficiency of the coil terminal are based on internal and external factors. Internal factors consist of unnecessary movement of materials, reworking in office and unused work shifts. External factors are caused by vendors in the form of information insufficiency and material input/output. The results of the study conclude that the optimal solution is that the cooperation between the operators in the chain is improved so that flow of information becomes better and reduce uncertainty in delivery and shipping. Furthermore, the study encourages Oxelösund Harbor to collect data, in the form of statistics, concerning daily events in Stålhamnen, highlighting problems and costs that are hidden and start utilizing the unused work shift during the night, as well as reducing the stock.

Most of the chemical reactions of living cells are catalyzed by protein enzymes. These enzymes are very efficient and display a high degree of specificity with respect to the reaction catalyzed. Cellular activities depend critically on the precise three-dimensional structure and function of thousands of enzymes. Many enzymes require binding of metal ions or small organic molecules for their function. The organic molecules that are indispensible components of catalysis by proteins are called coenzymes. Pyridoxal 5ʹ-phosphate (PLP) is a versatile coenzyme found in all living cells. PLP-dependent enzymes play a key role in the function of most of the enzymes catalyzing reactions in the metabolic pathways of amino acid synthesis and degradation. The enzyme pyridoxal kinase serves to make available the co-enzyme PLP to apo-PLP dependent enzymes. Because of their key role in cellular function and their medical importance, the structure and function of PLP-dependent enzymes have been extensively investigated. In the past decade, detailed investigations on the structure and function of several PLP-dependent enzymes have been carried out in our laboratory. The enzymes studied are B. subtilis serinehydroxymethyl transferase (SHMT), S. typhimurium acetylornithine aminotransferase (AcOAT), S. typhimurium and E. coli diaminopropionate ammonia lyase (DAPAL), S. typhimurium D-serine dehydratase (DSD), S. typhimurium D-cysteine desulfhydrase (DCyD) and S. typhimurium arginine decarboxylase (ArgD). The extensive studies conducted on PLP-dependent enzymes in our laboratory during the past decade has not only resulted in deeper understanding of their structure and function but also raised several new questions regarding substrate recognition, reaction specificity, role of active site residues in the catalytic reaction, mechanism of catalysis and potential applications of these enzymes. This thesis is an attempt to answer some of these questions. The thesis also presents the structure and function of a new protein, Salmonella typhimurium pyridoxal kinase, the enzyme that provides PLP for PLP-dependent enzymes. Single crystal X-ray diffraction technique is the most powerful tool currently available for the elucidation of the three-dimensional structures of proteins and other biological macromolecules and for revealing the relationship between their structure and function. X-ray diffraction studies have provided in depth understanding of the topology of secondary structural elements in the three-dimensional structures of proteins, the hierarchical organization of protein domains, structural basis for the substrate specificity of enzymes, intricate details of mechanisms of enzyme catalyzed reactions, allosteric regulation of enzyme activity, mechanisms of feed-back inhibition, structural basis of protein stability, symmetry of oligomeric proteins and their possible biological implications and a myriad of other biochemical and biophysical properties of proteins. The work reported in this thesis is primarily based on X-ray diffraction studies. X-ray crystal structure investigations are complemented by spectral and biochemical studies on the catalyzed reactions. The thesis begins with an introduction to PLP-dependent enzymes and presentation of a brief summary of the earlier work carried out in our laboratory on PLP-dependent enzymes (Chapter 1). A brief description of earlier functional classification of PLP-dependent enzymes and the more recent classification of these enzymes into the four groups based on their three-dimensional structure is provided. Although enzymes belonging to these four structural classes have evolved from independent evolutionary lineages, they share some common features near their active sites and in the mode of PLP binding. Earlier work carried out elsewhere on pyridoxal kinase and its key role in maintaining PLP at a low concentration in the cytosol is presented. Different mechanisms that have been proposed for the transfer of PLP from pyridoxal kinase to other apo PLP-dependent enzymes are briefly described. The experimental procedures and computational methods used during the course of these investigations to obtain the results reported in chapters 3-6 are presented in Chapter 2. Most of these methods are applicable to the isolation of plasmids, cloning, over expression, protein purification, mutant construction, crystallization, X-ray diffraction data collection and processing, structure elucidation and refinement, validation and structural analysis presented in the next three chapters. Various programs and protocols used for data processing, structure determination, refinement, model building, structure validation and analysis are also briefly described. In chapter 3, the role of a number of active site residues in the reaction catalyzed by EcDAPAL, a fold type II PLP-dependent enzyme, the structure of which was determined earlier in the laboratory is explored by mutational, biochemical and structural analyses. Earlier studies had established the probable role of Asp120 and Lys77 in the reaction leading to the breakdown of D-DAP and L-DAP, respectively (Bisht et al., 2012). To further validate the earlier observations, a number of active site mutants were generated for Asp 120 (D120N, D120C, D120S and D120T), Asp 189 (D189N, D189C, D189S and D189T), Lys77 (K77T, K77H, K77R and K77A), His 123 (H123L) and Tyr 168 (Y168F). The structure of D120N mutant crystal obtained after soaking in crystallization cocktail containing D-DAP revealed the presence of an intact external aldimine complex at the active site supporting the earlier proposal that Asp120 is the base abstracting the Cα proton from the D-isomer of DAP. Biochemical and structural observations suggested that none of the Asp189 mutants may bind PLP and were catalytically inactive suggesting an essential role for Asp189 in catalysis. In contrast to type I PLP-dependent enzymes, none of the Lys 77 mutants of EcDAPAL could bind PLP either covalently or non-covalently and were inactive with both the isomers of DAP. Thus, Lys77 appears to be important for both PLP binding and catalysis. H123L mutant formed an external aldimine with D-DAP and a gem-diamine complex with L-DAP indicating that this residue is also crucial for catalysis. These studies have provided additional support to the catalytic mechanism of EcDAPAL proposed earlier. The next Chapter 4 explores the structure, function and catalytic mechanism of Salmonella typhimurium DAPAL (StDAPAL). The protein was purified from a construct carrying a hexa-histidine tag at the C-terminus by Ni-NTA chromatography. The purified protein was demonstrated to be homogeneous by SDS-PAGE and MALDI-TOF. Crystals of StDAPAL belonging to the C-centred monoclinic space group (C121) with four molecules in the asymmetric unit were obtained by the micro batch method and used for collecting X-ray diffracting data. The crystal structure was determined by molecular replacement using the homologous enzyme from E. coli (PDB code 4D9M, Bisht et al., 2012), which shares a sequence identity of 50% with the S. typhimurium enzyme as the phasing model in the program Phaser (McCoy et al., 2007) of the CCP4 suite. The model was refined with Refmac5 of CCP4 suite to R and Rfree values of 25.5% and 30.9%, respectively. A superposition of the structure so obtained over EcDAPAL revealed that the two structures are very similar. A sulfate molecule bound to the active site of StDAPAL could be located. The position of the sulfate corresponds to that of the carboxyl group of aminoacrylate intermediate of EcDAPAL (4D9M). The PLP was bound to Lys78 as an internal aldimine. Since the active sites of the two protomers in fold type II PLP-dependent enzymes are independent, it might be possible to obtain functional monomers of EcDAPAL. With this view, mutation of a conserved Trp (Trp399) present in the dimeric interface resulted in the destabilization of the dimeric interface and partial conversion of the dimeric protein to a monomeric protein. However, the monomeric species of EcDAPALW399R was unable to bind PLP and hence did not possess any catalytic activity. This highlights the importance of dimeric organization for efficient binding of PLP as well as for the activity of the enzyme. A remarkable difference between EcDAPAL and StDAPAL is the absence of a disulfide bond between residues Cys271 and Cys299 in StDAPAL equivalent to the bond formed between Cys265 and Cys291 in EcDAPAL. Mutation of Cys265 and Cys291 of EcDAPAL to Ser did not affect the activity of the enzyme towards either of the isomers of the substrate indicating that the disulfide bond is not crucial for enzyme activity. The stability of the loop corresponding residues 261-295 of EcDAPAL was believed to be promoted by the disulfide bond. However, the equivalent loop was found to be ordered in StDAPAL even though the disulfide bond is absent. In contrast to StDAPAL, EcDAPAL did not show any metal dependent activity. The previous two chapters dealt with fold type II PLP-dependent enzymes. In contrast, Chapter 5 deals with revisiting the structure and function of a fold type I PLP-dependent enzyme, Salmonella typhimurium arginine decarboxylase (StADC). ADC is a very large polypeptide in comparison with other fold type I enzymes. It is induced when the bacterium is subjected to low pH and plays a major role in protecting the cells from acid stress. The structure of StADC was determined but not satisfactorily refined by Dr. S. R. Bharat earlier. The X-ray diffraction data collected by Bharat needed to be improved and the structure needed to be further refined and compared with the homologous E. coli enzyme. Therefore, the entire process of data processing, structure solution and refinement was repeated. The refined structure of StADC was found to correspond to the apo form of the enzyme with only a phosphate molecule occupying the position equivalent to that of 5’ phosphate of PLP observed in EcADC holo enzyme structure. This allowed examination of structural changes that accompany PLP binding and formation of an internal aldimine. The apo to holo transition in StADC involves the movement and ordering of two loops consisting of residues 151-164 and 191-196 which are in the linker and PLP binding domains of the protein, respectively. Phosphate binding by itself appears to be insufficient for these structural changes. These two loops are close to the PLP binding site of the other protomer of the dimer. Hence, these movements are probably important for the catalytic function of the enzyme. Holo ADC has been found as a decamer in other studies. The decameric form of the apo-StADC suggests that PLP binding may not be essential for the oligomeric state of the protein. ADC appears to reduce proton concentration inside the cell in two ways; (i) by surface charge neutralization and (ii) by arginine decarboxylation by extracting a proton from the cytoplasm. The resulting product agmatine is exchanged for extra cellular arginine by arginine-agmatine antiporter. The low sequence identity and lack of structural similarity of the inducible and constitutive forms of ADC from S. typhimurium shows that these are unlikely to be products of divergent evolution. The final chapter 6 of the thesis presents the work carried out on S. typhimurium pyridoxal kinase (PLK). In the salvage pathway of pyridoxal 5’phosphate (PLP), PLP is produced as the product of the reaction catalyzed by PLK using PL, PN and PM as substrates. Thus, PLK plays the critical role of ensuring availability of PLP to the large number of PLP-dependent enzymes. S. typhimurium PLK was purified to homogeneity, crystallized in its native as well as ligand bound forms. It was necessary to circumvent an unusual problem caused by spots arising from a contaminant crystal to obtain the structure of the native crystals of PLK that belonged to the P212121 space group with two protomers in the crystal asymmetric unit. It was then straight forward to determine the ligand bound structures of StPLK (space group P43212) obtained by co-crystallization with ATP, PL and Mg2+ by molecular replacement using the wild type structure as the phasing model. The structures obtained by co-crystallization revealed the presence of ADP, Mg2+ and a PL bound to the active site Lys233 via a Schiff base (internal aldimine). This is the first structure in which the presence of an internal aldimine in the active site of PLK has been observed. Formation of the internal aldimine might be one way to prevent the release of excess PLP and protecting the cell from PLP induced toxicity. The enzyme was shown to be inhibited by the product which will also help in maintaining PLP concentration at low levels. It was also demonstrated that PLK interacts with apo-PLP-dependent enzymes. This observation supports possible direct transfer of PLP from PLK to PLP-dependent enzymes. The thesis ends with an appendix where the work carried out during the course of the thesis work but not as part of the thesis is briefly described.

Esta dissertação apresenta modelagem matemática e um método de solução numérica para problemas de conformação eletromagnética de chapas metálicas finas circulares utilizando uma bobina em espiral plana. O método foca especificamente o cálculo do campo eletromagnético gerado pela bobina e análise do circuito que modela o sistema de estampagem eletromagnética. A bobina plana é aproximada por círculos concêntricos carregando a corrente de descarga dos capacitores. Os cálculos das correntes induzidas e perfil da força eletromagnética na chapa e acoplamentos magnéticos entre a bobina e chapa metálica são realizados para o instante inicial, antes da deformação plástica da chapa. O método utiliza a lei de Biot-Savart, sendo que a solução das integrais obtidas para indução magnética é realizada através de métodos numéricos, considerando as simetrias do problema. Para verificação da modelagem matemática, da solução numérica e comprovação da viabilidade técnica deste processo, um dispositivo de estampagem eletromagnética foi desenvolvido e diversos experimentos foram realizados com chapas de alumínio. Os parâmetros investigados foram a espessura da chapa a ser deformada e a influência da presença de vácuo na cavidade da ferramenta. Os resultados experimentais demonstram concordância com os resultados da modelagem proposta. A presença de vácuo também demonstrou um incremento na deformação da chapa. A rotina desenvolvida no software Matlab fornece informações importantes para o processo e permite que se faça ajustes no dispositivo. / This dissertation presents a mathematical model and numerical method to solve the problems of electromagnetic forming of thin circular metal sheets by using a flat spiral coil. The method focuses specifically on the calculation of the electromagnetic field generated by the coil flat and circuit analysis modeling system electromagnetic forming. The flat coil is approximated by concentric circles carrying a discharge current of capacitors. The calculations of induced currents and profile of the electromagnetic force on the plate and magnetic coupling between the coil and sheet metal are made to the initial time, before the plastic deformation of the plate. The method uses the law of Biot-Savart, and the solution of the integral obtained for magnetic induction is performed by numerical methods, considering the symmetries of the problem. To verify the mathematical model, the numerical solution and proving the technical feasibility of this process, a electromagnetic forming device was developed and several experiments were made with aluminum plates. The parameters investigated were the thickness of the plate to be deformed and the influence of the vacuum in the cavity of the tool. The experimental results show agreement with the results of the proposed model. The presence of vacuum also showed an increase in the deformation of the plate. A routine developed in software Matlab provide important information for the process and allow to make adjustments on the device.

Submitted by Daniel Araujo Gonçalves null (daniel.araujogoncalves@gmail.com) on 2016-08-01T13:43:54Z No. of bitstreams: 1 Goncalves_Tese_versão_final.pdf: 3648542 bytes, checksum: 7acd793a874f48c8a008a28e1e550948 (MD5) / Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-08-02T19:00:04Z (GMT) No. of bitstreams: 1 goncalves_da_dr_ilha.pdf: 3648542 bytes, checksum: 7acd793a874f48c8a008a28e1e550948 (MD5) / Made available in DSpace on 2016-08-02T19:00:04Z (GMT). No. of bitstreams: 1 goncalves_da_dr_ilha.pdf: 3648542 bytes, checksum: 7acd793a874f48c8a008a28e1e550948 (MD5) Previous issue date: 2016-07-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Neste trabalho foram desenvolvidos quatro métodos analíticos visando a determinação de elementos químicos em diversas amostras. O primeiro estudo consistiu na determinação de Cr e Mn em placentônios provenientes de bovinos por espectrometria de emissão atômica em filamento de tungstênio (WC AES). O tempo de integração de sinal para 30 ms, proporcionou LOD 45 vezes menor para Mn e 150 vezes menor para Cr com relação a 500 ms. Testes de adição e recuperação apresentaram desvio padrão (SD) de ± 0,02 mg L-1 e recuperações de 91 a 102%. No segundo foram realizadas quantificações de Cr em cápsulas medicinais produzidas na China, a WC AES. A faixa linear de trabalho para determinação de Cr por WC AES foi de 10-100 μg L-1. Os limites de detecção foram de 3 μg L-1, o que possibilitou determinações em um nível 28 vezes menor que o valor máximo de Cr recomendado em medicamentos pela Farmacopeia Americana (USP). A precisão (% RSD) foi calculada na faixa de 5,2 – 7,4% e não foram significativamente diferentes através da aplicação de um teste ANOVA fator único a nível de confiança de 95% (p > 0,05) quando comparados com a técnica de Espectrometria de emissão óptica com plasma acoplado indutivamente (ICP OES). No terceiro estudo, um Espectrometro de Emissão Óptica com plasma induzido por micro-ondas – MIP OES (MP-AES, Agilent 4200), foi usado em combinação com o método de análise por diluição de padrão (SDA) para determinar Al, Cr, Co, Cu, Fe, Mn, Ni e Zn em amostras de café, chá verde, energético, cerveja, whiskey e cachaça. Não foi necessária preparação da amostra, a não ser uma simples diluição em HNO3 1% v v-1. A eficiência do SDA foi avaliada por experimentos de adição e recuperação e os resultados foram comparados com os métodos tradicionais de calibração externa (EC), o padrão interno (IS) e adição de padrão (AS). As recuperações médias por SDA para vários elementos avaliadas neste trabalho foram entre 90 e 99%. Para o método tradicional de adições padrão, por exemplo, as recuperações médias entre 101 e 122% foram obtidos para os mesmos analitos e amostras. No quarto estudo um MIP OES (MP-AES, Agilent 4200) foi utilizado para a determinação de Cd, Cr, Cu, Ni e Pb na matriz de óleo fusel onde emprega-se a calibração externa preparada em solução de 1% HNO3 e 1-propanol. Soluções contendo 5 e 50% da matriz v v-1 foram diluidas em 1-propanol e solução de óleo fúsel puro foram analisados. Não foram observados efeitos de matriz quando empregou-se a calibração com soluções de 1-propanol mesmo para determinações na amostra não diluída, com recuperações entre 90-109%. Os LODs para Cd, Cr, Cu, Ni e Pb em 1-propanol foram de 0,03; 0,009; 0,01; 0,007 e 0,04 mg L-1, respectivamente. O procedimento descrito para o MIP OES é uma abordagem simples, exata e precisa para rastrear análise de elementos de óleo fúsel. / In this work were developed four analytical methods for the determination of chemical elements in several samples. The first study consisted in determination of Cr and Mn in placenton from bovine animals by Atomic emission spectrometry in tungsten filament (WC AES). Signal integration time for 30 ms, provided LOD 45 times lower for Mn and 150 times lower for Cr with respect to 500 ms. Addition and recovery tests presented standard deviation (SD) of ± 0,02 mg L-1 and recoveries of 91 to 102%. In the second were carried out baseline measurements of Cr in medicinal capsule produced in China by WC AES. The linear range for determination of Cr by WC AES was 10 - 100 μg L-1 . The limits of detection were 3 μg L-1 , which allowed determinations in a level 28 times smaller than the maximum value of Cr recommended medicines for the U.S. Pharmacopeial Convention (USP). The precision (% RSD) was estimated in the range of 5,2 – 7,4% and were not significantly different by applying a test ANOVA single factor at the level of confidence 95% (p > 0,05) when compared with Optical emission spectrometry with inductively coupled plasma (ICP OES). In the third study, a Spectrometer of Optical Emission plasma induced by microwave – MIP OES (MP-AES, Agilent 4200), was used in combination with the method of analysis for standard dilution (SDA) to determine Al, Cr, Co, Cu, Fe, Mn, Ni e Zn in samples of coffee, green tea, energetic, beer, whiskey and cachaça brazilian. It didn't require sample preparation except a simple dilution in HNO3 1% v v-1 . The efficiency of the SDA was evaluated by experiments of addition and recovery, and the results were compared with the traditional methods of external calibration (EC), the internal standard (IS) and addition of default (AS). Average recoveries for SDA to various elements evaluated in this study were between 90 and 99%. For the traditional method of standard additions, for example, the average recoveries between 101 and 122% were obtained for the same analytes and samples. In the fourth study a MIP OES (MP-AES, Agilent 4200), was used for the determination of Cd, Cr, Cu, Ni e Pb in the array of fusel oil where is used the external calibration solution prepared 1% HNO3 and1-propanol. Solutions containing 5 and 50% of the matrix v v-1 were dissolved in 1-propanol and fusel oil pure solution were analyzed. Matrix effects were not observed when it was calibration with 1-propanol solutions even for undiluted sample determinations, with recoveries between 90 - 109%. The LODs for Cd, Cr, Cu, Ni and Pb in 1-propanol were of 0,03; 0,009; 0,01; 0,007 and 0,04 mg L-1 , respectively. The procedure described for MIP OES is a simple, accurate and precise approach to trace elements analysis of fusel oil. / CAPES PDSE: 99999.002566/ 2014-01