Applications for wireless sensor networks : tracking with binary proximity sensors

El interés cada vez mayor por las redes de sensores inalámbricos pueden ser entendido simplemente pensando en lo que esencialmente son: un gran número de pequeños nodos sensores autoalimentados que recogen información o detectan eventos especiales y se comunican de manera inalámbrica, con el objetivo final de entregar sus datos procesados a una estación base. Los nodos sensores están densamente desplegados dentro del área de interés, se pueden desplegar al azar y tienen capacidad de cooperación. Por lo general, estos dispositivos son pequeños y de bajo costo, de modo que pueden ser producidos y desplegados en gran numero aunque sus recursos en términos de energía, memoria, velocidad de cálculo y ancho de banda están enormemente limitados. Detección, tratamiento y comunicación son tres elementos clave cuya combinación en un pequeño dispositivo permite lograr un gran número de aplicaciones. Las redes de sensores proporcionan oportunidades sin fin, pero al mismo tiempo plantean retos formidables, tales como lograr el máximo rendimiento de una energía que es escasa y por lo general un recurso no renovable. Sin embargo, los recientes avances en la integración a gran escala, integrado de hardware de computación, comunicaciones, y en general, la convergencia de la informática y las comunicaciones, están haciendo de esta tecnología emergente una realidad. Del mismo modo, los avances en la nanotecnología están empezando a hacer que todo gire entorno a las redes de pequeños sensores y actuadores distribuidos. Hay diferentes tipos de sensores tales como sensores de presión, acelerómetros, cámaras, sensores térmicos o un simple micrófono. Supervisan las condiciones presentes en diferentes lugares tales como la temperatura, humedad, el movimiento, la luminosidad, presión, composición del suelo, los niveles de ruido, la presencia o ausencia de ciertos tipos de objetos, los niveles de tensión mecánica sobre objetos adheridos y las características momentáneas tales como la velocidad , la dirección y el tamaño de un objeto, etc. Se comprobara el estado de las Redes Inalámbricas de Sensores y se revisaran los protocolos más famosos. Así mismo, se examinara la identificación por radiofrecuencia (RFID) ya que se está convirtiendo en algo actual y su presencia importante. La RFID tiene un papel crucial que desempeñar en el futuro en el mundo de los negocios y los individuos por igual. El impacto mundial que ha tenido la identificación sin cables está ejerciendo fuertes presiones en la tecnología RFID, los servicios de investigación y desarrollo, desarrollo de normas, el cumplimiento de la seguridad y la privacidad y muchos más. Su potencial económico se ha demostrado en algunos países mientras que otros están simplemente en etapas de planificación o en etapas piloto, pero aun tiene que afianzarse o desarrollarse a través de la modernización de los modelos de negocio y aplicaciones para poder tener un mayor impacto en la sociedad. Las posibles aplicaciones de redes de sensores son de interés para la mayoría de campos. La monitorización ambiental, la guerra, la educación infantil, la vigilancia, la micro-cirugía y la agricultura son solo unos pocos ejemplos de los muchísimos campos en los que tienen cabida las redes mencionadas anteriormente. Estados Unidos de América es probablemente el país que más ha investigado en esta área por lo que veremos muchas soluciones propuestas provenientes de ese país. Universidades como Berkeley, UCLA (Universidad de California, Los Ángeles) Harvard y empresas como Intel lideran dichas investigaciones. Pero no solo EE.UU. usa e investiga las redes de sensores inalámbricos. La Universidad de Southampton, por ejemplo, está desarrollando una tecnología para monitorear el comportamiento de los glaciares mediante redes de sensores que contribuyen a la investigación fundamental en glaciología y de las redes de sensores inalámbricos. Así mismo, Coalesenses GmbH (Alemania) y Zurich ETH están trabajando en diversas aplicaciones para redes de sensores inalámbricos en numerosas áreas. Una solución española será la elegida para ser examinada más a fondo por ser innovadora, adaptable y polivalente. Este estudio del sensor se ha centrado principalmente en aplicaciones de tráfico, pero no se puede olvidar la lista de más de 50 aplicaciones diferentes que ha sido publicada por la firma creadora de este sensor específico. En la actualidad hay muchas tecnologías de vigilancia de vehículos, incluidos los sensores de bucle, cámaras de video, sensores de imagen, sensores infrarrojos, radares de microondas, GPS, etc. El rendimiento es aceptable, pero no suficiente, debido a su limitada cobertura y caros costos de implementación y mantenimiento, especialmente este ultimo. Tienen defectos tales como: línea de visión, baja exactitud, dependen mucho del ambiente y del clima, no se puede realizar trabajos de mantenimiento sin interrumpir las mediciones, la noche puede condicionar muchos de ellos, tienen altos costos de instalación y mantenimiento, etc. Por consiguiente, en las aplicaciones reales de circulación, los datos recibidos son insuficientes o malos en términos de tiempo real debido al escaso número de detectores y su costo. Con el aumento de vehículos en las redes viales urbanas las tecnologías de detección de vehículos se enfrentan a nuevas exigencias. Las redes de sensores inalámbricos son actualmente una de las tecnologías más avanzadas y una revolución en la detección de información remota y en las aplicaciones de recogida. Las perspectivas de aplicación en el sistema inteligente de transporte son muy amplias. Con este fin se ha desarrollado un programa de localización de objetivos y recuento utilizando una red de sensores binarios. Esto permite que el sensor necesite mucha menos energía durante la transmisión de información y que los dispositivos sean más independientes con el fin de tener un mejor control de tráfico. La aplicación se centra en la eficacia de la colaboración de los sensores en el seguimiento más que en los protocolos de comunicación utilizados por los nodos sensores. Las operaciones de salida y retorno en las vacaciones son un buen ejemplo de por qué es necesario llevar la cuenta de los coches en las carreteras. Para ello se ha desarrollado una simulación en Matlab con el objetivo localizar objetivos y contarlos con una red de sensores binarios. Dicho programa se podría implementar en el sensor que Libelium, la empresa creadora del sensor que se examinara concienzudamente, ha desarrollado. Esto permitiría que el aparato necesitase mucha menos energía durante la transmisión de información y los dispositivos sean más independientes. Los prometedores resultados obtenidos indican que los sensores de proximidad binarios pueden formar la base de una arquitectura robusta para la vigilancia de áreas amplias y para el seguimiento de objetivos. Cuando el movimiento de dichos objetivos es suficientemente suave, no tiene cambios bruscos de trayectoria, el algoritmo ClusterTrack proporciona un rendimiento excelente en términos de identificación y seguimiento de trayectorias los objetos designados como blancos. Este algoritmo podría, por supuesto, ser utilizado para numerosas aplicaciones y se podría seguir esta línea de trabajo para futuras investigaciones. No es sorprendente que las redes de sensores de binarios de proximidad hayan atraído mucha atención últimamente ya que, a pesar de la información mínima de un sensor de proximidad binario proporciona, las redes de este tipo pueden realizar un seguimiento de todo tipo de objetivos con la precisión suficiente. Abstract The increasing interest in wireless sensor networks can be promptly understood simply by thinking about what they essentially are: a large number of small sensing self-powered nodes which gather information or detect special events and communicate in a wireless fashion, with the end goal of handing their processed data to a base station. The sensor nodes are densely deployed inside the phenomenon, they deploy random and have cooperative capabilities. Usually these devices are small and inexpensive, so that they can be produced and deployed in large numbers, and so their resources in terms of energy, memory, computational speed and bandwidth are severely constrained. Sensing, processing and communication are three key elements whose combination in one tiny device gives rise to a vast number of applications. Sensor networks provide endless opportunities, but at the same time pose formidable challenges, such as the fact that energy is a scarce and usually non-renewable resource. However, recent advances in low power Very Large Scale Integration, embedded computing, communication hardware, and in general, the convergence of computing and communications, are making this emerging technology a reality. Likewise, advances in nanotechnology and Micro Electro-Mechanical Systems are pushing toward networks of tiny distributed sensors and actuators. There are different sensors such as pressure, accelerometer, camera, thermal, and microphone. They monitor conditions at different locations, such as temperature, humidity, vehicular movement, lightning condition, pressure, soil makeup, noise levels, the presence or absence of certain kinds of objects, mechanical stress levels on attached objects, the current characteristics such as speed, direction and size of an object, etc. The state of Wireless Sensor Networks will be checked and the most famous protocols reviewed. As Radio Frequency Identification (RFID) is becoming extremely present and important nowadays, it will be examined as well. RFID has a crucial role to play in business and for individuals alike going forward. The impact of ‘wireless’ identification is exerting strong pressures in RFID technology and services research and development, standards development, security compliance and privacy, and many more. The economic value is proven in some countries while others are just on the verge of planning or in pilot stages, but the wider spread of usage has yet to take hold or unfold through the modernisation of business models and applications. Possible applications of sensor networks are of interest to the most diverse fields. Environmental monitoring, warfare, child education, surveillance, micro-surgery, and agriculture are only a few examples. Some real hardware applications in the United States of America will be checked as it is probably the country that has investigated most in this area. Universities like Berkeley, UCLA (University of California, Los Angeles) Harvard and enterprises such as Intel are leading those investigations. But not just USA has been using and investigating wireless sensor networks. University of Southampton e.g. is to develop technology to monitor glacier behaviour using sensor networks contributing to fundamental research in glaciology and wireless sensor networks. Coalesenses GmbH (Germany) and ETH Zurich are working in applying wireless sensor networks in many different areas too. A Spanish solution will be the one examined more thoroughly for being innovative, adaptable and multipurpose. This study of the sensor has been focused mainly to traffic applications but it cannot be forgotten the more than 50 different application compilation that has been published by this specific sensor’s firm. Currently there are many vehicle surveillance technologies including loop sensors, video cameras, image sensors, infrared sensors, microwave radar, GPS, etc. The performance is acceptable but not sufficient because of their limited coverage and expensive costs of implementation and maintenance, specially the last one. They have defects such as: line-ofsight, low exactness, depending on environment and weather, cannot perform no-stop work whether daytime or night, high costs for installation and maintenance, etc. Consequently, in actual traffic applications the received data is insufficient or bad in terms of real-time owed to detector quantity and cost. With the increase of vehicle in urban road networks, the vehicle detection technologies are confronted with new requirements. Wireless sensor network is the state of the art technology and a revolution in remote information sensing and collection applications. It has broad prospect of application in intelligent transportation system. An application for target tracking and counting using a network of binary sensors has been developed. This would allow the appliance to spend much less energy when transmitting information and to make more independent devices in order to have a better traffic control. The application is focused on the efficacy of collaborative tracking rather than on the communication protocols used by the sensor nodes. Holiday crowds are a good case in which it is necessary to keep count of the cars on the roads. To this end a Matlab simulation has been produced for target tracking and counting using a network of binary sensors that e.g. could be implemented in Libelium’s solution. Libelium is the enterprise that has developed the sensor that will be deeply examined. This would allow the appliance to spend much less energy when transmitting information and to make more independent devices. The promising results obtained indicate that binary proximity sensors can form the basis for a robust architecture for wide area surveillance and tracking. When the target paths are smooth enough ClusterTrack particle filter algorithm gives excellent performance in terms of identifying and tracking different target trajectories. This algorithm could, of course, be used for different applications and that could be done in future researches. It is not surprising that binary proximity sensor networks have attracted a lot of attention lately. Despite the minimal information a binary proximity sensor provides, networks of these sensing modalities can track all kinds of different targets classes accurate enough.

Overall and local mechanical behaviour of dissimilar laser welded joints

The present investigation addresses the overall and local mechanical performance of dissimilar joints of low carbon steel (CS) and stainless steel (SS) thin sheets achieved by laser welding in case of heat source displacement from the weld gap centreline towards CS. Microstructure characterization and residua! strain scanning, carried out by neutron diffraction, were used to assess the joints features. It was found that the heat source position influences the base metals dilution and the residua! stress field associated to the welding process; the transverse residual stress is smaller than for the longitudinal component, of magnitudes close to the parent CS yield strength. Furthermore, compressive transverse residual stresses were encountered at the SS-weld interface. The tensile behavior of the joint different zones assessed by using a video-image based system (VIC-2D) reveals that the residual stress field, together with the positive difference in yield between the weld metal and the base materials protects the joint from being piastically deformed. The tensile loadings of flat transverse specimens generate the strain localization and failure in CS, far away from the weld.En este trabajo se exponen los resultados de una investigacion sobre el comportamiento mecanico de soldaduras disimiles acero inoxidable-acero al carbono, realizadas para unir chapas delgadas, desplazando la fuente de calor del eje longitudinal de la union soldada por laser sobre el acero al carbono. Se han determinado las caracteristicas microestructurales de la union soldada, las tensiones residuales generadas (mediante difraccion de neutrones) y las curvas tension-deformacion locales y globales, mediante medidas locales de deformacion empleando el sistema VIC-2D "video image correlation". El desplazamiento de la fuente de calor infiuye en la dilution de los metales base y el campo de tensiones residuales asociado al proceso de soldeo; las tensiones residuales medidas en direction longitudinal se aproximan al limite elastico del acero al carbono, mientras que las tensiones residuales transversales son menores, e incluso de compresion. El ensayo a traccion de la union soldada revela que las tensiones residuales y la diferencia de limite elastico entre los metales base y la soldadura propician que la rotura se produzca por inestabilidad plastica del acero al carbono, lejos de la soldadura, sin que la union plastifique.

Cycling Dof Factors: Molecular and functional characterization of Arabidopsis thaliana AtCDF3 and tomato (Solanum lycopersicum L.) SlCDF3 in response to abiotic stress

El tomate (Solanum lycopersicum L.) es considerado uno de los cultivos hortícolas de mayor importancia económica en el territorio Español. Sin embargo, su producción está seriamente afectada por condiciones ambientales adversas como, salinidad, sequía y temperaturas extremas. Para resolver los problemas que se presentan en condiciones de estrés, se han empleado una serie de técnicas culturales que disminuyen sus efectos negativos, siendo de gran interés el desarrollo de variedades tolerantes. En este sentido la obtención y análisis de plantas transgénicas, ha supuesto un avance tecnológico, que ha facilitado el estudio y la evaluación de genes seleccionados en relación con la tolerancia al estrés. Estudios recientes han mostrado que el uso de genes reguladores como factores de transcripción (FTs) es una gran herramienta para obtener nuevas variedades de tomate con mayor tolerancia a estreses abióticos. Las proteínas DOF (DNA binding with One Finger) son una familia de FTs específica de plantas (Yangisawa, 2002), que están involucrados en procesos fisiológicos exclusivos de plantas como: asimilación del nitrógeno y fijación del carbono fotosintético, germinación de semilla, metabolismo secundario y respuesta al fotoperiodo pero su preciso rol en la tolerancia a estrés abiótico se desconoce en gran parte. El trabajo descrito en esta tesis tiene como objetivo estudiar genes reguladores tipo DOF para incrementar la tolerancia a estrés abiotico tanto en especies modelo como en tomate. En el primer capítulo de esta tesis se muestra la caracterización funcional del gen CDF3 de Arabidopsis, así como su papel en la respuesta a estrés abiótico y otros procesos del desarrollo. La expresión del gen AtCDF3 es altamente inducido por sequía, temperaturas extremas, salinidad y tratamientos con ácido abscísico (ABA). La línea de inserción T-DNA cdf3-1 es más sensible al estrés por sequía y bajas temperaturas, mientras que líneas transgénicas de Arabidopsis 35S::AtCDF3 aumentan la tolerancia al estrés por sequía, osmótico y bajas temperaturas en comparación con plantas wild-type (WT). Además, estas plantas presentan un incremento en la tasa fotosintética y apertura estomática. El gen AtCDF3 se localiza en el núcleo y que muestran una unión específica al ADN con diferente afinidad a secuencias diana y presentan diversas capacidades de activación transcripcional en ensayos de protoplastos de Arabidopsis. El dominio C-terminal de AtCDF3 es esencial para esta localización y su capacidad activación, la delección de este dominio reduce la tolerancia a sequía en plantas transgénicas 35S::AtCDF3. Análisis por microarray revelan que el AtCDF3 regula un set de genes involucrados en el metabolismo del carbono y nitrógeno. Nuestros resultados demuestran que el gen AtCDF3 juega un doble papel en la regulación de la respuesta a estrés por sequía y bajas temperaturas y en el control del tiempo de floración. En el segundo capítulo de este trabajo se lleva a cabo la identificación de 34 genes Dof en tomate que se pueden clasificar en base a homología de secuencia en cuatro grupos A-D, similares a los descritos en Arabidopsis. Dentro del grupo D se han identificado cinco genes DOF que presentan características similares a los Cycling Dof Factors (CDFs) de Arabidopsis. Estos genes son considerados ortólogos de Arabidopsis CDF1-5, y han sido nombrados como Solanum lycopersicum CDFs o SlCDFs. Los SlCDF1-5 son proteínas nucleares que muestran una unión específica al ADN con diferente afinidad a secuencias diana y presentan diversas capacidades de activación transcripcional in vivo. Análisis de expresión de los genes SlCDF1-5 muestran diferentes patrones de expresión durante el día y son inducidos de forma diferente en respuesta a estrés osmótico, salino, y de altas y bajas temperaturas. Plantas de Arabidopsis que sobre-expresan SlCDF1 y SlCDF3 muestran un incremento de la tolerancia a la sequía y salinidad. Además, de la expresión de varios genes de respuesta estrés como AtCOR15, AtRD29A y AtERD10, son expresados de forma diferente en estas líneas. La sobre-expresión de SlCDF3 en Arabidopsis promueve un retardo en el tiempo de floración a través de la modulación de la expresión de genes que controlan la floración como CONSTANS (CO) y FLOWERING LOCUS T (FT). En general, nuestros datos demuestran que los SlCDFs están asociados a funciones aun no descritas, relacionadas con la tolerancia a estrés abiótico y el control del tiempo de floración a través de la regulación de genes específicos y a un aumento de metabolitos particulares. ABSTRACT Tomato (Solanum lycopersicum L.) is one of the horticultural crops of major economic importance in the Spanish territory. However, its production is being affected by adverse environmental conditions such as salinity, drought and extreme temperatures. To resolve the problems triggered by stress conditions, a number of agricultural techniques that reduce the negative effects of stress are being frequently applied. However, the development of stress tolerant varieties is of a great interest. In this direction, the technological progress in obtaining and analysis of transgenic plants facilitated the study and evaluation of selected genes in relation to stress tolerance. Recent studies have shown that a use of regulatory genes such as transcription factors (TFs) is a great tool to obtain new tomato varieties with greater tolerance to abiotic stresses. The DOF (DNA binding with One Finger) proteins form a family of plant-specific TFs (Yangisawa, 2002) that are involved in the regulation of particular plant processes such as nitrogen assimilation, photosynthetic carbon fixation, seed germination, secondary metabolism and flowering time bur their precise roles in abiotic stress tolerance are largely unknown. The work described in this thesis aims at the study of the DOF type regulatory genes to increase tolerance to abiotic stress in both model species and the tomato. In the first chapter of this thesis, we present molecular characterization of the Arabidopsis CDF3 gene as well as its role in the response to abiotic stress and in other developmental processes. AtCDF3 is highly induced by drought, extreme temperatures, salt and abscisic acid (ABA) treatments. The cdf3-1 T-DNA insertion mutant was more sensitive to drought and low temperature stresses, whereas the AtCDF3 overexpression enhanced the tolerance of transgenic plants to drought, cold and osmotic stress comparing to the wild-type (WT) plants. In addition, these plants exhibit increased photosynthesis rates and stomatal aperture. AtCDF3 is localized in the nuclear region, displays specific binding to the canonical DNA target sequences and has a transcriptional activation activity in Arabidopsis protoplast assays. In addition, the C-terminal domain of AtCDF3 is essential for its localization and activation capabilities and the deletion of this domain significantly reduces the tolerance to drought in transgenic 35S::AtCDF3 overexpressing plants. Microarray analysis revealed that AtCDF3 regulated a set of genes involved in nitrogen and carbon metabolism. Our results demonstrate that AtCDF3 plays dual roles in regulating plant responses to drought and low temperature stress and in control of flowering time in vegetative tissues. In the second chapter this work, we carried out to identification of 34 tomato DOF genes that were classified by sequence similarity into four groups A-D, similar to the situation in Arabidopsis. In the D group we have identified five DOF genes that show similar characteristics to the Cycling Dof Factors (CDFs) of Arabidopsis. These genes were considered orthologous to the Arabidopsis CDF1 - 5 and were named Solanum lycopersicum CDFs or SlCDFs. SlCDF1-5 are nuclear proteins that display specific binding to canonical DNA target sequences and have transcriptional activation capacities in vivo. Expression analysis of SlCDF1-5 genes showed distinct diurnal expression patterns and were differentially induced in response to osmotic, salt and low and high temperature stresses. Arabidopsis plants overexpressing SlCDF1 and SlCDF3 showed increased drought and salt tolerance. In addition, various stress-responsive genes, such as AtCOR15, AtRD29A and AtERD10, were expressed differently in these lines. The overexpression of SlCDF3 in Arabidopsis also results in the late flowering phenotype through the modulation of the expression of flowering control genes such CONSTANS (CO) and FLOWERING LOCUS T (FT). Overall, our data connet SlCDFs to undescribed functions related to abiotic stress tolerance and flowering time through the regulation of specific target genes and an increase in particular metabolites.

Robust Real-time Vision-based Aircraft Tracking From Unmanned Aerial Vehicles.

Aircraft tracking plays a key and important role in the Sense-and-Avoid system of Unmanned Aerial Vehicles (UAVs). This paper presents a novel robust visual tracking algorithm for UAVs in the midair to track an arbitrary aircraft at real-time frame rates, together with a unique evaluation system. This visual algorithm mainly consists of adaptive discriminative visual tracking method, Multiple-Instance (MI) learning approach, Multiple-Classifier (MC) voting mechanism and Multiple-Resolution (MR) representation strategy, that is called Adaptive M3 tracker, i.e. AM3. In this tracker, the importance of test sample has been integrated to improve the tracking stability, accuracy and real-time performances. The experimental results show that this algorithm is more robust, efficient and accurate against the existing state-of-art trackers, overcoming the problems generated by the challenging situations such as obvious appearance change, variant surrounding illumination, partial aircraft occlusion, blur motion, rapid pose variation and onboard mechanical vibration, low computation capacity and delayed information communication between UAVs and Ground Station (GS). To our best knowledge, this is the first work to present this tracker for solving online learning and tracking freewill aircraft/intruder in the UAVs.

A Ground-Truth Video Dataset for the Development and Evaluation of Vision-based Sense-and-Avoid systems

The importance of vision-based systems for Sense-and-Avoid is increasing nowadays as remotely piloted and autonomous UAVs become part of the non-segregated airspace. The development and evaluation of these systems demand flight scenario images which are expensive and risky to obtain. Currently Augmented Reality techniques allow the compositing of real flight scenario images with 3D aircraft models to produce useful realistic images for system development and benchmarking purposes at a much lower cost and risk. With the techniques presented in this paper, 3D aircraft models are positioned firstly in a simulated 3D scene with controlled illumination and rendering parameters. Realistic simulated images are then obtained using an image processing algorithm which fuses the images obtained from the 3D scene with images from real UAV flights taking into account on board camera vibrations. Since the intruder and camera poses are user-defined, ground truth data is available. These ground truth annotations allow to develop and quantitatively evaluate aircraft detection and tracking algorithms. This paper presents the software developed to create a public dataset of 24 videos together with their annotations and some tracking application results.

Real-Time Adaptive Multi-Classifier Multi-Resolution Visual Tracking Framework for Unmanned Aerial Vehicles

This paper presents a novel robust visual tracking framework, based on discriminative method, for Unmanned Aerial Vehicles (UAVs) to track an arbitrary 2D/3D target at real-time frame rates, that is called the Adaptive Multi-Classifier Multi-Resolution (AMCMR) framework. In this framework, adaptive Multiple Classifiers (MC) are updated in the (k-1)th frame-based Multiple Resolutions (MR) structure with compressed positive and negative samples, and then applied them in the kth frame-based Multiple Resolutions (MR) structure to detect the current target. The sample importance has been integrated into this framework to improve the tracking stability and accuracy. The performance of this framework was evaluated with the Ground Truth (GT) in different types of public image databases and real flight-based aerial image datasets firstly, then the framework has been applied in the UAV to inspect the Offshore Floating Platform (OFP). The evaluation and application results show that this framework is more robust, efficient and accurate against the existing state-of-art trackers, overcoming the problems generated by the challenging situations such as obvious appearance change, variant illumination, partial/full target occlusion, blur motion, rapid pose variation and onboard mechanical vibration, among others. To our best knowledge, this is the first work to present this framework for solving the online learning and tracking freewill 2D/3D target problems, and applied it in the UAVs.

Detecting, Localizing and Following Dynamic Objects with a Mini-UAV

This paper presents an approach for the detection, localization and following of dynamic terrestrial objects using a mini-UAV. The development is intended to be used for surveillance of large infrastructures. The detection algorithm is based on finding several pre-defined characteristics of the target, such as color, shape and size. The process used to localize the target, once it is detected, is based on an inversion of the Pinhole camera model. The task of following the Summit XL was designed to keep the target inside the field of view of the camera, and it was implemented in the form of a PID controller. The system has been tested both in simulation and with real robots, showing promising results.

Mammalian capping enzyme binds RNA and uses protein tyrosine phosphatase mechanism

Mammalian capping enzymes are bifunctional proteins with both RNA 5′-triphosphatase and guanylyltransferase activities. The N-terminal 237-aa triphosphatase domain contains (I/V)HCXXGXXR(S/T)G, a sequence corresponding to the conserved active-site motif in protein tyrosine phosphatases (PTPs). Analysis of point mutants of mouse RNA 5′-triphosphatase identified the motif Cys and Arg residues and an upstream Asp as required for activity. Like PTPs, this enzyme was inhibited by iodoacetate and VO43− and independent of Mg2+, providing additional evidence for phosphate removal from RNA 5′ ends by a PTP-like mechanism. The full-length, 597-aa mouse capping enzyme and the C-terminal guanylyltransferase fragment (residues 211–597), unlike the triphosphatase domain, bound poly (U) and were nuclear in transfected cells. RNA binding was increased by GTP, and a guanylylation-defective, active-site mutant was not affected. Ala substitution at positions required for the formation of the enzyme-GMP capping intermediate (R315, R530, K533, or N537) also eliminated poly (U) binding, while proteins with conservative substitutions at these sites retained binding but not guanylyltransferase activity. These results demonstrate that the guanylyltransferase domain of mammalian capping enzyme specifies nuclear localization and RNA binding. Association of capping enzyme with nascent transcripts may act in synergy with RNA polymerase II binding to ensure 5′ cap formation.

mRNA binding protein mrnp 41 localizes to both nucleus and cytoplasm

We have identified and molecularly characterized a human protein with a Mr of 40,880 Da. After UV irradiation of HeLa cells, this protein was cross-linked to poly(A)-containing mRNA and was therefore designated mrnp 41 (for mRNA binding protein of 41 kDa). Cell fractionation and immunoblotting showed mrnp 41 in both the cytoplasm and the nucleus and particularly in the nuclear envelope. Immunofluorescence microscopy localized mrnp 41 to distinct foci in the nucleoplasm, to the nuclear rim, and to meshwork-like structures throughout the cytoplasm. The cytoplasmic meshwork staining was disrupted by prior treatment of cells with the actin filament- or microtubule-disrupting drugs cytochalasin or nocodazole, respectively, suggesting association of mrnp 41 with the cytoskeleton. Double immunofluorescence with antibodies against mrnp 41 and the cytoplasmic poly(A) binding protein showed colocalization to the cytoplasmic meshwork. Immunogold electronmicroscopy confirmed mrnp 41’s cytoplasmic and nucleoplasmic localization and revealed a striking labeling of nuclear pore complexes. Together these data suggest that mrnp 41 may function in nuclear export of mRNPs and/or in cytoplasmic transport on, or attachment to, the cytoskeleton. Consistent with a role of mrnp 41 in nuclear export are previous reports that mutations in homologs of mrnp 41 in Schizosaccharomyces pombe, designated Rae1p, or in Saccharomyces cerevisiae, designated Gle2p, result in mRNA accumulation in the nucleus although it is presently not known whether these homologs are mRNA binding proteins as well.

Role of lipid polymorphism in G protein-membrane interactions: Nonlamellar-prone phospholipids and peripheral protein binding to membranes

Heterotrimeric G proteins (peripheral proteins) conduct signals from membrane receptors (integral proteins) to regulatory proteins localized to various cellular compartments. They are in excess over any G protein-coupled receptor type on the cell membrane, which is necessary for signal amplification. These facts account for the large number of G protein molecules bound to membrane lipids. Thus, the protein-lipid interactions are crucial for their cellular localization, and consequently for signal transduction. In this work, the binding of G protein subunits to model membranes (liposomes), formed with defined membrane lipids, has been studied. It is shown that although G protein α-subunits were able to bind to lipid bilayers, the presence of nonlamellar-prone phospholipids (phosphatidylethanolamines) enhanced their binding to model membranes. This mechanism also appears to be used by other (structurally and functionally unrelated) peripheral proteins, such as protein kinase C and the insect protein apolipophorin III, indicating that it could constitute a general mode of protein-lipid interactions, relevant in the activity and translocation of some peripheral (amphitropic) proteins from soluble to particulate compartments. Other factors, such as the presence of cholesterol or the vesicle surface charge, also modulated the binding of the G protein subunits to lipid bilayers. Conversely, the binding of G protein-coupled receptor kinase 2 and the G protein β-subunit to liposomes was not increased by hexagonally prone lipids. Their distinct interactions with membrane lipids may, in part, explain the different cellular localizations of all of these proteins during the signaling process.

Excitonic couplings and electronic coherence in bridged naphthalene dimers

The electronic excitations of naphthalene and a family of bridged naphthalene dimers are calculated and analyzed by using the Collective Electronic Oscillator method combined with the oblique Lanczos algorithm. All experimentally observed trends in absorption profiles and radiative lifetimes are reproduced. Each electronic excitation is linked to the corresponding real-space transition density matrix, which represents the motions of electrons and holes created in the molecule by photon absorption. Two-dimensional plots of these matrices help visualize the degree of exciton localization and explain the dependence of the electronic interaction between chromophores on their separation.

Arp2/3 Complex from Acanthamoeba Binds Profilin and Cross-links Actin Filaments

The Arp2/3 complex was first purified from Acanthamoeba castellanii by profilin affinity chromatography. The mechanism of interaction with profilin was unknown but was hypothesized to be mediated by either Arp2 or Arp3. Here we show that the Arp2 subunit of the complex can be chemically cross-linked to the actin-binding site of profilin. By analytical ultracentrifugation, rhodamine-labeled profilin binds Arp2/3 complex with a Kd of 7 μM, an affinity intermediate between the low affinity of profilin for barbed ends of actin filaments and its high affinity for actin monomers. These data suggest the barbed end of Arp2 is exposed, but Arp2 and Arp3 are not packed together in the complex exactly like two actin monomers in a filament. Arp2/3 complex also cross-links actin filaments into small bundles and isotropic networks, which are mechanically stiffer than solutions of actin filaments alone. Arp2/3 complex is concentrated at the leading edge of motile Acanthamoeba, and its localization is distinct from that of α-actinin, another filament cross-linking protein. Based on localization and actin filament nucleation and cross-linking activities, we propose a role for Arp2/3 in determining the structure of the actin filament network at the leading edge of motile cells.

A Role for the Lumenal Domain in Golgi Localization of the Saccharomyces cerevisiae Guanosine Diphosphatase

Integral membrane proteins (IMPs) contain localization signals necessary for targeting to their resident subcellular compartments. To define signals that mediate localization to the Golgi complex, we have analyzed a resident IMP of the Saccharomyces cerevisiae Golgi complex, guanosine diphosphatase (GDPase). GDPase, which is necessary for Golgi-specific glycosylation reactions, is a type II IMP with a short amino-terminal cytoplasmic domain, a single transmembrane domain (TMD), and a large catalytic lumenal domain. Regions specifying Golgi localization were identified by analyzing recombinant proteins either lacking GDPase domains or containing corresponding domains from type II vacuolar IMPs. Neither deletion nor substitution of the GDPase cytoplasmic domain perturbed Golgi localization. Exchanging the GDPase TMD with vacuolar protein TMDs only marginally affected Golgi localization. Replacement of the lumenal domain resulted in mislocalization of the chimeric protein from the Golgi to the vacuole, but a similar substitution leaving 34 amino acids of the GDPase lumenal domain intact was properly localized. These results identify a major Golgi localization determinant in the membrane-adjacent lumenal region (stem) of GDPase. Although necessary, the stem domain is not sufficient to mediate localization; in addition, a membrane-anchoring domain and either the cytoplasmic or full-length lumenal domain must be present to maintain Golgi residence. The importance of lumenal domain sequences in GDPase Golgi localization and the requirement for multiple hydrophilic protein domains support a model for Golgi localization invoking protein–protein interactions rather than interactions between the TMD and the lipid bilayer.

Differential Expression and Functions of Cortical Myosin IIA and IIB Isotypes during Meiotic Maturation, Fertilization, and Mitosis in Mouse Oocytes and Embryos

To explore the role of nonmuscle myosin II isoforms during mouse gametogenesis, fertilization, and early development, localization and microinjection studies were performed using monospecific antibodies to myosin IIA and IIB isotypes. Each myosin II antibody recognizes a 205-kDa protein in oocytes, but not mature sperm. Myosin IIA and IIB demonstrate differential expression during meiotic maturation and following fertilization: only the IIA isoform detects metaphase spindles or accumulates in the mitotic cleavage furrow. In the unfertilized oocyte, both myosin isoforms are polarized in the cortex directly overlying the metaphase-arrested second meiotic spindle. Cortical polarization is altered after spindle disassembly with Colcemid: the scattered meiotic chromosomes initiate myosin IIA and microfilament assemble in the vicinity of each chromosome mass. During sperm incorporation, both myosin II isotypes concentrate in the second polar body cleavage furrow and the sperm incorporation cone. In functional experiments, the microinjection of myosin IIA antibody disrupts meiotic maturation to metaphase II arrest, probably through depletion of spindle-associated myosin IIA protein and antibody binding to chromosome surfaces. Conversely, the microinjection of myosin IIB antibody blocks microfilament-directed chromosome scattering in Colcemid-treated mature oocytes, suggesting a role in mediating chromosome–cortical actomyosin interactions. Neither myosin II antibody, alone or coinjected, blocks second polar body formation, in vitro fertilization, or cytokinesis. Finally, microinjection of a nonphosphorylatable 20-kDa regulatory myosin light chain specifically blocks sperm incorporation cone disassembly and impedes cell cycle progression, suggesting that interference with myosin II phosphorylation influences fertilization. Thus, conventional myosins break cortical symmetry in oocytes by participating in eccentric meiotic spindle positioning, sperm incorporation cone dynamics, and cytokinesis. Although murine sperm do not express myosin II, different myosin II isotypes may have distinct roles during early embryonic development.

Differential Roles of Syntaxin 7 and Syntaxin 8 in Endosomal Trafficking

To understand molecular mechanisms that regulate the intricate and dynamic organization of the endosomal compartment, it is important to establish the morphology, molecular composition, and functions of the different organelles involved in endosomal trafficking. Syntaxins and vesicle-associated membrane protein (VAMP) families, also known as soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptors (SNAREs), have been implicated in mediating membrane fusion and may play a role in determining the specificity of vesicular trafficking. Although several SNAREs, including VAMP3/cellubrevin, VAMP8/endobrevin, syntaxin 13, and syntaxin 7, have been localized to the endosomal membranes, their precise localization, biochemical interactions, and function remain unclear. Furthermore, little is known about SNAREs involved in lysosomal trafficking. So far, only one SNARE, VAMP7, has been localized to late endosomes (LEs), where it is proposed to mediate trafficking of epidermal growth factor receptor to LEs and lysosomes. Here we characterize the localization and function of two additional endosomal syntaxins, syntaxins 7 and 8, and propose that they mediate distinct steps of endosomal protein trafficking. Both syntaxins are found in SNARE complexes that are dissociated by α-soluble NSF attachment protein and NSF. Syntaxin 7 is mainly localized to vacuolar early endosomes (EEs) and may be involved in protein trafficking from the plasma membrane to the EE as well as in homotypic fusion of endocytic organelles. In contrast, syntaxin 8 is likely to function in clathrin-independent vesicular transport and membrane fusion events necessary for protein transport from EEs to LEs.