Potassium channel-interacting protein 1 (KChIP1), a Ca²⁺ sensor protein, regulates the function of A-type Kv4 potassium channels and increases their cell surface expression. Myristoylation at the N-terminus of KChIP1 has been suggested to facilitate membrane-binding, but was not sufficient for stable membrane assaociation. The aim of the present study is to investigate whether EF-hand motifs of KChIP1 are crucial for membranal targeting in addition to the N-terminal myristoyl group, and how the membrane association of KChIP1 is influenced by lipid compositions. According to hydropathy profile, EF-hands 3 and 4 of KChIP1 showed highly hydrophobicity. After deleting EF-hands 3 and 4, the altered microenvironment of Trp residue and decreased hydrophobicity were found in truncated KChIP1, but it still maintained £\-helix structure. Furthermore, truncated KChIP1 exhibited lower lipid-binding ability, affecting intracellular membrane localization and was almost diminished underlying increasing membrane permeability by digitonin in cells, suggesting that intact EF-hands 3 and 4 may be related to the anchorage of KChIP1 on cellular membrane. KChIP1, but not mutant, specifically bound with phosphatidylserine by lipid binding assay and the FTIR spectra showed the change of £\-helix structure by binding lipid large unilamellar vesicles was dependent on phosphatidylserine. Either phosphatidylserine or potassium channels enhanced KChIP1 to form tetramer for targeting to phospholipids by using chemical cross-linking assay. Taken together, our data highly suggest that intact of EF-hands 3 and 4 should structurally and functionally involve in fulfilling the physiological activity of KChIP1.

In this thesis closed-loop control strategies over a communication network with multiple clients are developed. To accomplish this objective, a steel-ball magnetic-levitation system, a DC motor speed-control system, and an autonomous wheelchair robot referred to as Clients 1, 2, and 3, respectively were used as Networked-Based-control (NCS) test beds to validate the proposed strategies. For real-time operation, Linux with Real-Time Application Interface (RTAI) and Control and Measurement Interface (Comedi) were used as the operating system for Clients 1 and 2. Client 3's software was written in Microsoft Visual Basic 6.0 on the Windows XP operating system (OS). User datagram protocol (UDP) was used as the communication network protocol in this research due to its better real-time performance instead of transmission control protocol (TCP). Although UDP has no guarantee for transferring data, it has smaller overheads and less time delay than TCP. Since the robotic wheelchair and the server are run on different OSs, Samba was used to put both systems into the same LAN with a fast data-transmission speed. Using Samba, the round-trip communication time between the robotic wheelchair and the server is only 11.2 ms whereas 30.8 ms is taken without using Samba. When the server receives the sensor data from multiple clients at the same time, the NCS stability may be deteriorated due to the limitation of the system bandwidth. The NCS stability is affected by the sampling period of the system, and the reduction of the sampling period improves the control loop's performance. However, a shorter sampling period requires more network bandwidth to transmit more sensor data or control data, which increases the network traffic load. Using the PING test, the transmission time for each control loop was measured. The processing time for each system was also measured by a time-stamp function, and the operation time for each control loop was obtained. In order to maintain the NCS stable, several combinations of the sampling periods for each client are suggested and verified. The bandwidth utilization of Client 1 is set to be 43.5% and the range of the bandwidth utilization of Client 2 with guaranteed stability was found to be between 9.1% and 45.3%. Thus, the bandwidth utilization of Client 3 is from 11.8% to 46.8%. The multiple-client NCS test bed could maintain its stability within these ranges of the bandwidth utilizations of all clients.

No description available.

Doctor of Philosophy / Department of Chemistry / Emily McLaurin / Quantum dots are nanocrystalline semiconductors in which the size is so small that optoelectronic properties are size dependent. QDs have a lot of applications in displays, solar cells, lasers, light emitting diodes, etc. The optoelectronic properties of QDs depend on their size, composition, the shape of the particles and also the surface chemistry of the QDs. Phosphine based precursors have been mostly used in the synthesis of QDs. Due to the lack of tunable reactivity, this class of precursors, QDs with different shape are obtained by under different reaction conditions. With that, branched QDs are less likely to be obtained in one step reaction using phosphine based precursors. To synthesis QDs with a branched structure, in a single step synthesis, mixtures of precursors with different reactivity were used. Using dichalcogenides mixture, CdSe₁-xSx hyperbranched supra-quantum dots (HSQDs) where synthesized in a one-step microwave-assisted synthesis and shape evolution mechanism of formation of NCs studied. It is shown that the NCs formed in three steps of nucleation, aggregation, and growth. By controlling the reaction conditions, simple branched tetrapod NCs are prepared, but the obtained NCs have no emission due to unpassivated surface and defects which work as trap. To obtain luminescent NCs obtained through doping. Hyperbranched Mn²+:ZnSe₁-xSx NCs also prepared using a mixture of Ph₂Se₂ and Me₂S₂. The shape evolution mechanism of the formation of NCs was studied and it is shown that the NCs are formed via oriented attachment of initially formed nanoparticles. The NCs used for thiol sensing, and it observed that they have a better sensitivity and detection limit than spherical QDs. Although hyperbranched NCs have higher sensitivities over nonbranched NCs but, the spherical NCs have better detection limit and can dispersed in aqueous medium by ZnS shell growth followed by silica shell formation. To study the effect of ZnS shell thickness on sensing property of NCs, a set of spherical Mn:ZnSe@ZnS with different ZnS shell thickness were prepared and used for thiol sensing. It observed that in organic medium, thinner ZnS layer gives the highest sensitivity and QDs with thick ZnS shell layer have less sensitivity. For measurement in aqueous medium, QDs transferred to PBS buffer after formation of silica shell over QDs. It observed that NCs with a thin ZnS shell layer lose their emission and sensing completely. Thick ZnS shell protects NCs in the silica shell formation step but they show very low sensitivity to thiol compounds as well. ZnS shell with medium thickness gives the best sensitivity in an aqueous medium. The emission of Mn:ZnSe@ZnS QDs originated from d-d electron transition of Mn(II) ions and is independent to the size of QDs. To extend our study to QDs with band edge emission, preparation of luminescent InP QDs by post-synthetic modification is studied. InP NCs were synthesized using heat up method and successive injection of precursors. Narrow size distribution NCs obtained after size selection precipitation. Emissive NCs obtained after etching using InCl3 and fluoride containing salts. The study showed that more InCl3 case more etching and presence of fluoride-containing salt is necessary for band edge emission of the NCs.

The importance of real-time communication at the device level in a factory automation setup is a widely researched area. This research is an effort to experimentally verify if Ethernet can be used as a real-time communication standard in a factory automation setup, by observing the effects of packet delays, packet loss, and network congestion on the performance of a networked control system (NCS). The NCS experimental setup used in this research involves real-time feedback control of multiple plants like DC motors and a magnetic-levitation system connected to one or more controllers. A multi-client-multi-server architecture on a local area network (LAN) was developed using user datagram protocol (UDP) as the communication protocol. Key observations are as follows. (1) The multi-client-single-server system showed the highest packet delays compared to single-client-single-server architecture. (2) In the singleclient- single-server system, as the Ethernet link utilization increased beyond 82 percent, the average packet delays and steady-state error of the DC motor speed-control system increased by 2231 percent and 304 percent, respectively. (3) Even under high link utilization, adding an additional server to the NCS reduced average packet delays considerably. (4) With large packet sizes, higher packet rates were automatically throttled by Ethernet’s flow control mechanism affecting the real-time communication negatively. (5) In the multiclient- multi-server architecture, average packet delays at higher packet rates, and at higher packet lengths were found to be 40 percent lesser than the those of the single-clientsingle- server system and 87.5 percent lesser than those of the multi-client-single-server system.

Sistemas de controle distribuídos cujas malhas são fechadas através de uma rede de comunicação são chamados de sistemas de controle distribuídos em redes de comunicação (NCS - Networked Control System). Este tipo de arquitetura permite a divisão do sistema de controle em módulos interconectados através da rede de comunicação, proporcionando a divisão do processamento, a redução de custo e de peso, além de facilitar o diagnóstico e manutenção do sistema e de aumentar a sua exibilidade e agilidade; e por isso seu emprego na indústria está se tornando comum (e.g., y-by-wire e drive-by-wire). Porém, a distribuição do processamento e a inserção de uma rede de comunicação aumenta a complexidade da análise e do projeto deste tipo de sistema. Um dos fatores que contribui para esse aumento da complexidade é a presença de atrasos aleatórios nos sinais de controle, causados pela dinâmica do sistema computacional (conjunto de hardware e software) que serve como plataforma para implementação do sistema de controle digital. Este trabalho faz um estudo sobre este tipo de sistema sob a perspectiva destes sinais com atrasos. Para isso, faz-se uso dos toolboxes para MATLAB: TrueTime e Jitterbug. Através destas ferramentas, mostra-se a existência de uma relação de compromisso entre o desempenho do controle e o desempenho do sistema computacional. Através deste estudo, é proposto uma solução de um sistema de controle do tipo NCS para um ROV (do inglês Remotely Operated Vehicle), modelado através de 6 equações diferenciais desacopladas não-lineares. Este tipo de veículo tem uma relevância econômica significativa para o Brasil, visto que é utilizado em operações de manutenção e instalação de plataformas de extração do petróleo que está depositado em profundidades que variam de mil a 2 mil metros. Para este NCS proposto, são utilizados controladores do tipo PI com estrutura feedback-feedfoward cujos parâmetros de projeto são obtidos em função dos atrasos inseridos pelo sistema computacional. / Distributed control systems wherein the control loops are closed through a communication network are called Networked Control Systems (NCSs). This type of architecture allows the control systems division into modules interconnected through the communication network, providing the processing division, reduction of cost and weight, and facilitates the systems diagnosis and maintenance, and increases their exibility and agility. Therefore its use in industry is becoming common (eg, y-by-wire and drive-by-wire). However, the processing distribution and the communication network insertion increase the system analysis and design complexity. One of the factors that contributes to this increased complexity is the presence of random time delays, caused by the dynamics of the computer system (set of hardware and software) used as a platform for digital control system implementation. This work deals with the networked control systems under these random time delays view. For this, it is used two MATLAB toolboxes: Jitterbug and TrueTime. With these tools, it is shown the existence of a relationship between the performance of control and performance of computer system. With this study, it proposed a solution of a NCS for a ROV (Vehicle Operated Remotely), modeled by 6 differential nonlinear decoupled equations. This type of vehicle has a significant economic relevance for Brazil, as it is used in maintenance and installation of platforms for oil extraction deposited at depths ranging from thousand to 2 thousand meters. For this proposed NCS are adopted PI controllers with feedfoward-feedback structure whose parameters design are given in terms of delay inserted by the computer system.

O foco do trabalho está direcionado para a construção da linhagem knock-out do gene ncsA de A. fumigatus, bem como a caracterização do fenótipo da linhagem DncsA e a investigação de interações deste gene com vias de sinalização importantes para o desenvolvimento do fungo. A linhagem mutante ncsA foi construída através da transformação do WT (Dku80) com o gene marcador pyrG de A. fumigatus. Foi confirmada a deleção por Southern blot e as alterações fenotípicas da linhagem foram analisadas. O gene ncsA em A. fumigatus não é essencial e a linhagem DncsA apresentou-se sensível EGTA e SDS, alé, de tolerante a altas concentrações de íons cálcio. Na linhagem NcsA:mRFP foi localizado a proteína NcsA no citoplasma das células e sua localização não é alterada pela em resposta ao aumento da concentração de cálcio. A linhagem DncsA foi sensível a drogas que afetam a biossíntese e a manutenção da membrana plasmática, tais como voriconazole, anfotericina e itraconazole. O crescimento polarizado em presença de lovastatina nessa linhagem foi consideravelmente mais afetado que no tipo selvagem. O Spitzenkörper não foi visualizado no mutante DncsA, e existe uma significante diminuição de estruturas vacuolares e endossomos. Os resultados obtidos em ensaios de TRPCR em tempo real sugerem que NcsA modula a expressão dos genes pmcA e pmcB, que codificam proteínas ATPases transportadoras de íons cálcio. Os ensaios de virulência no modelo animal revelaram que a mutação do gene ncsA não causa perda de virulência no A. fumigatus. / Here, we characterize the A. fumigatus Neuronal Calcium Sensor, ncsA homologue. We showed that ncsA is not an essential gene and ncsA growth was decreased in the presence of EGTA and SDS. Furthermore, the ncsA mutant is more resistant to calcium chloride. NcsA:mRFP localizes to the cytoplasm that its cellular localization is not affected by the cellular response to calcium chloride. The ncsA mutant strain is more sensitive to voriconazole, itraconazole, and the ergosterol intercalating agent, amphotericin. Polar growth in the DncsA mutant strain was also considerably more affected by lovastatin than in the wild type mutant strain. The Spitzenkörper cannot be visualized in the DncsA mutant, and there is a significant decrease of the endosome/vacuole structures. NcsA supports pmcA and pmcB expression therefore reduced expression of these ion pumps, and also of other genes involved in the response to calcium in A. fumigatus. The ncsA inactivation mutation is not causing loss of virulence in a low dose murine infection when compared to the corresponding wild type strain.

Sistemas de controle distribuídos cujas malhas são fechadas através de uma rede de comunicação são chamados de sistemas de controle distribuídos em redes de comunicação (NCS - Networked Control System). Este tipo de arquitetura permite a divisão do sistema de controle em módulos interconectados através da rede de comunicação, proporcionando a divisão do processamento, a redução de custo e de peso, além de facilitar o diagnóstico e manutenção do sistema e de aumentar a sua exibilidade e agilidade; e por isso seu emprego na indústria está se tornando comum (e.g., y-by-wire e drive-by-wire). Porém, a distribuição do processamento e a inserção de uma rede de comunicação aumenta a complexidade da análise e do projeto deste tipo de sistema. Um dos fatores que contribui para esse aumento da complexidade é a presença de atrasos aleatórios nos sinais de controle, causados pela dinâmica do sistema computacional (conjunto de hardware e software) que serve como plataforma para implementação do sistema de controle digital. Este trabalho faz um estudo sobre este tipo de sistema sob a perspectiva destes sinais com atrasos. Para isso, faz-se uso dos toolboxes para MATLAB: TrueTime e Jitterbug. Através destas ferramentas, mostra-se a existência de uma relação de compromisso entre o desempenho do controle e o desempenho do sistema computacional. Através deste estudo, é proposto uma solução de um sistema de controle do tipo NCS para um ROV (do inglês Remotely Operated Vehicle), modelado através de 6 equações diferenciais desacopladas não-lineares. Este tipo de veículo tem uma relevância econômica significativa para o Brasil, visto que é utilizado em operações de manutenção e instalação de plataformas de extração do petróleo que está depositado em profundidades que variam de mil a 2 mil metros. Para este NCS proposto, são utilizados controladores do tipo PI com estrutura feedback-feedfoward cujos parâmetros de projeto são obtidos em função dos atrasos inseridos pelo sistema computacional. / Distributed control systems wherein the control loops are closed through a communication network are called Networked Control Systems (NCSs). This type of architecture allows the control systems division into modules interconnected through the communication network, providing the processing division, reduction of cost and weight, and facilitates the systems diagnosis and maintenance, and increases their exibility and agility. Therefore its use in industry is becoming common (eg, y-by-wire and drive-by-wire). However, the processing distribution and the communication network insertion increase the system analysis and design complexity. One of the factors that contributes to this increased complexity is the presence of random time delays, caused by the dynamics of the computer system (set of hardware and software) used as a platform for digital control system implementation. This work deals with the networked control systems under these random time delays view. For this, it is used two MATLAB toolboxes: Jitterbug and TrueTime. With these tools, it is shown the existence of a relationship between the performance of control and performance of computer system. With this study, it proposed a solution of a NCS for a ROV (Vehicle Operated Remotely), modeled by 6 differential nonlinear decoupled equations. This type of vehicle has a significant economic relevance for Brazil, as it is used in maintenance and installation of platforms for oil extraction deposited at depths ranging from thousand to 2 thousand meters. For this proposed NCS are adopted PI controllers with feedfoward-feedback structure whose parameters design are given in terms of delay inserted by the computer system.

O foco do trabalho está direcionado para a construção da linhagem knock-out do gene ncsA de A. fumigatus, bem como a caracterização do fenótipo da linhagem DncsA e a investigação de interações deste gene com vias de sinalização importantes para o desenvolvimento do fungo. A linhagem mutante ncsA foi construída através da transformação do WT (Dku80) com o gene marcador pyrG de A. fumigatus. Foi confirmada a deleção por Southern blot e as alterações fenotípicas da linhagem foram analisadas. O gene ncsA em A. fumigatus não é essencial e a linhagem DncsA apresentou-se sensível EGTA e SDS, alé, de tolerante a altas concentrações de íons cálcio. Na linhagem NcsA:mRFP foi localizado a proteína NcsA no citoplasma das células e sua localização não é alterada pela em resposta ao aumento da concentração de cálcio. A linhagem DncsA foi sensível a drogas que afetam a biossíntese e a manutenção da membrana plasmática, tais como voriconazole, anfotericina e itraconazole. O crescimento polarizado em presença de lovastatina nessa linhagem foi consideravelmente mais afetado que no tipo selvagem. O Spitzenkörper não foi visualizado no mutante DncsA, e existe uma significante diminuição de estruturas vacuolares e endossomos. Os resultados obtidos em ensaios de TRPCR em tempo real sugerem que NcsA modula a expressão dos genes pmcA e pmcB, que codificam proteínas ATPases transportadoras de íons cálcio. Os ensaios de virulência no modelo animal revelaram que a mutação do gene ncsA não causa perda de virulência no A. fumigatus. / Here, we characterize the A. fumigatus Neuronal Calcium Sensor, ncsA homologue. We showed that ncsA is not an essential gene and ncsA growth was decreased in the presence of EGTA and SDS. Furthermore, the ncsA mutant is more resistant to calcium chloride. NcsA:mRFP localizes to the cytoplasm that its cellular localization is not affected by the cellular response to calcium chloride. The ncsA mutant strain is more sensitive to voriconazole, itraconazole, and the ergosterol intercalating agent, amphotericin. Polar growth in the DncsA mutant strain was also considerably more affected by lovastatin than in the wild type mutant strain. The Spitzenkörper cannot be visualized in the DncsA mutant, and there is a significant decrease of the endosome/vacuole structures. NcsA supports pmcA and pmcB expression therefore reduced expression of these ion pumps, and also of other genes involved in the response to calcium in A. fumigatus. The ncsA inactivation mutation is not causing loss of virulence in a low dose murine infection when compared to the corresponding wild type strain.

Modern heterogeneous networks present a great challenge for network operators and engineers from a management and configuration perspective. The Tail-f Systems’ Network Control System (NCS) is a network management framework that addresses these challenges. NCS offers centralized network configuration management functionality, along with providing options for extending the framework with additional features. The devices managed by NCS are stored in its Configuration Database (CDB). However, currently there is no mechanism for automatically adding network devices to the configuration of NCS, thus each device’s management parameters have to be entered manually. The goal of this master’s thesis project is to develop a software module for NCS that simplifies the process of initial NCS configuration by allowing NCS to automatically add network devices to the NCS CDB. Apart from developing the software module for discovery, this project aims to summarize existing methods and to develop new methods for automated discovery of network devices with the main focus on differentiating between different types of devices. A credential-based device discovery method was developed and utilized to make advantage of known credentials to access devices, which allows for more precise discovery compared to some other existing methods. The selected methods were implemented as a component of NCS to provide device discovery functionality. Another focus of this master’s thesis project was the development of an approach to network topology discovery and its representation. The aim is to provide both a logical Internet Protocol (IP) network topology and a physical topology of device interconnections. The result is that we are able to automatically discover and store the topology representation as a data structure, and subsequently generate a visualization of the network topology. / Moderna heterogena nätverk utgör en stor utmaning för operatörer och ingenjörer att hantera och konfigurera. Tail-f Systems NCS produkt är ett ramverk för nätverks konfiguration som addresserar dessa utmaningar. NCS är ett centraliserat nätverks konfigurations verktyg. NCS är användbart som det är, men kan också byggas ut av användaren med ytterligare funktioner. De enheter som hanteras med NCS lagras i konfigurationsdatabasen (CDB). För närvarande finns det ingen automatiserad mekanism för att addera nätverksenheter till NCS, och varje enhets parametrar måste fyllas i manuellt. Detta examensarbetes mål är att utveckla en mjukvarumodul för NCS som förenklar NCS konfiguration genom att automatiskt lägga nätverksenheter till NCS CDB. Förutom att utveckla programvara för enhetsidentifiering, syftar detta projekt till att sammanfatta befintliga metoder och utveckla nya metoder för automatiserad nätverksenhetsidentifiering med huvudfokus på att skilja mellan olika typer av enheter. En metod baserad på förkonfigurerade autenticeringsuppgifter har utvecklats och den används för att precist kunna identifiera olika typer av nätverkselement. De valda metoderna har implementerats som en optionell modul till NCS som erbjuder enhetsidentifieringsfunktionalitet. Ytterligare ett fokus för detta examensarbete har varit att utveckla metoder för identifieraing av nätverkstopologin, och modeller för hur topologin ska representeras.  Vi har syftat till att identifiera både den logiska IP nätverkstopologin (L3) och den fysiska topologin av sammankopplade enheter (L2). Den viktigaste uppgiften har varit att identifiera och lagra topologi representation som en datastruktur, och dessutom generera en visualisering av nätverkstopologin.