Síntesis de series temporales de atenuación por lluvia en radioenlaces horizontales

En la actualidad, el uso de los sistemas de telecomunicaciones está aumentado a una gran velocidad. Debido a este aumento se hace imprescindible tener que crear sistemas de telecomunicaciones que trabajen en altas frecuencias. Al trabajar en estas frecuencias aparecen fenómenos atmosféricos que hacen que la señal se vea afectada, como la atenuación por lluvia o la atenuación por gases. Estos efectos podrían incluso hacer inviable la comunicación. Este Proyecto Fin de Grado tiene como objetivo la síntesis de series temporales de atenuación por lluvia en trayecto horizontal que afecta a un radioenlace de 38 GHz situado en Madrid. Para la realización del proyecto se utiliza como punto de partida los efectos de atenuación por lluvia en radioenlace horizontal descritos en la Recomendación UIT-R P.530-15. Los resultados obtenidos permiten examinar la validez del método propuesto en la Recomendación UIT-R P.1853-1 referente a la síntesis de las series temporales de atenuación troposférica. Este método propone un sintetizador para combinar los distintos efectos provocados por atenuación troposférica, que será utilizado para la realización del estudio. Este sintetizador ha sido implementado usado el programa Matlab. La primera parte del Proyecto expone los fundamentos teóricos sobre los que sustenta el presente trabajo. Comienza con la descripción de radioenlace, explicando sus distintos tipos y características. Después se muestran los distintos efectos de la propagación que afectan a un radioenlace. Por último, se presentan los fundamentos teóricos de series temporales, así como su implementación en un enlace de comunicaciones. En la segunda parte del Proyecto se explica el análisis de datos que se ha llevado a cabo. Primero se hace un análisis de la intensidad de lluvia según la Recomendación UIT-R P.837-6. Posteriormente se presentan los resultados de atenuación por lluvia obtenidos, haciendo una comparativa entre el modelo de la Recomendación UIT-R P.530-15, el modelo de la Recomendación UIT-R P.1853-1 y gráficas obtenidas a partir de datos experimentales. Como final del Proyecto se exponen las conclusiones que se han extraído durante su realización, así como posibles líneas futuras de investigación que continúen con el estudio realizado. ABSTRACT. The use of telecommunication systems is increasing rapidly. Due to this rising trend, it’s important to create telecommunication systems that work at high frequencies. However, some atmospheric impairments may well appear when working at these kinds of frequencies such as attenuation due to rain or gases which affect the signal. These effects could even render communication impossible. The aim of this Project is the synthesis of a time series, studying the effect of rain attenuation on a 38 GHz radio link located in Madrid. To carry out this study, the rain attenuation effects on horizontal radio links described in the ITU-R P.530-15 method are used as the starting point. The results obtained enable us to examine the validity of the practical application of the ITU R-P.1853-1 method for tropospheric attenuation time series synthesis. This method proposes a synthesizer which can combine the different effects caused by tropospheric attenuation that will be used for the study. This synthesizer has been implemented using the program Matlab. The first part of the Project starts by describing the theoretical foundations used during the Project performance. It begins with a description of radio links explaining their different types and characteristics. Following on from that, various propagation effects that affect the said radio link are shown. Finally, the theoretical foundations of the time series are presented, as well as its implementation in a communication link. In turn, the second part of the Project explains the data analysis that has been carried out. Firstly an analysis of the intensity of rain is made based on the ITU-R P.837-6 Recommendation. Subsequently, the results of the rain attenuation are presented, thus making a comparison between the ITU-R P.530-15 model, the ITU-R P.1853-1 model and graphs obtained from experimental data. Lastly, the Project shows the conclusions that have been drawn as well as propose future research lines that can be proposed so as to continue with the stud further.

Doubling the width of the platform of the San Pedro bridge in Spain

The San Pedro Bridge has six spans and is 750 m (2460 ft) long, 88 m (290 ft) high, 12 m (39 ft) wide, and curved with a radius of 700 m (2300 ft). It was built in 1993 using the cantilever method. Its super - structure is a prestressed concrete box girder with main spans of 150 m (490 ft). In 2008 and 2009, the width of the platform was enlarged to 23 m (75 ft) using five movable sets of scaffolding. The bridge remained open to traffic during construction. The original platform was widened 6 m (20 ft) on each side by connecting a new lightweight concrete cantilever to the original upper slab. These cantilevers were supported by steelstruts. The tie into the upper slab was made with new transverse post-tensioned tendons. The original superstructure was strengthened to resist the additional dead load of the expansion and live loads of the extra traffic. An additional new central web and a composite concrete-steel section were constructed and connected to the concrete box and central web using vertical high-strength post-tensioning bars. Also, external post-tensioning cables were implemented. It was also necessary to strengthen the connection of the original concrete box section to the piers. Detailed calculations were performed to evaluate the load distribution transmitted to the piers by the webs and by the original inclined concrete walls of the box girder. Finally, a detailed second-order-analysis of the complete structure was made to guarantee the resistance of the piers compared with actual loads

Proprioceptive force estimation and control for teleoperation in radioactive environments

A medida que se incrementa la energía de los aceleradores de partículas o iones pesados como el CERN o GSI, de los reactores de fusión como JET o ITER, u otros experimentos científicos, se va haciendo cada vez más imprescindible el uso de técnicas de manipulación remota para la interacción con el entorno sujeto a la radiación. Hasta ahora la tasa de dosis radioactiva en el CERN podía tomar valores cercanos a algunos mSv para tiempos de enfriamiento de horas, que permitían la intervención humana para tareas de mantenimiento. Durante los primeros ensayos con plasma en JET, se alcanzaban valores cercanos a los 200 μSv después de un tiempo de enfriamiento de 4 meses y ya se hacía extensivo el uso de técnicas de manipulación remota. Hay una clara tendencia al incremento de los niveles de radioactividad en el futuro en este tipo de instalaciones. Un claro ejemplo es ITER, donde se esperan valores de 450 Sv/h en el centro del toroide a los 11 días de enfriamiento o los nuevos niveles energéticos del CERN que harán necesario una apuesta por niveles de mantenimiento remotos. En estas circunstancias se enmarca esta tesis, que estudia un sistema de control bilateral basado en fuerza-posición, tratando de evitar el uso de sensores de fuerza/par, cuyo contenido electrónico los hace especialmente sensitivos en estos ambientes. El contenido de este trabajo se centra en la teleoperación de robots industriales, que debido a su reconocida solvencia y facilidad para ser adaptados a estos entornos, unido al bajo coste y alta disponibilidad, les convierte en una alternativa interesante para tareas de manipulación remota frente a costosas soluciones a medida. En primer lugar se considera el problema cinemático de teleoperación maestro-esclavo de cinemática disimilar y se desarrolla un método general para la solución del problema en el que se incluye el uso de fuerzas asistivas para guiar al operador. A continuación se explican con detalle los experimentos realizados con un robot ABB y que muestran las dificultades encontradas y recomendaciones para solventarlas. Se concluye el estudio cinemático con un método para el encaje de espacios de trabajo entre maestro y esclavo disimilares. Posteriormente se mira hacia la dinámica, estudiándose el modelado de robots con vistas a obtener un método que permita estimar las fuerzas externas que actúan sobre los mismos. Durante la caracterización del modelo dinámico, se realizan varios ensayos para tratar de encontrar un compromiso entre complejidad de cálculo y error de estimación. También se dan las claves para modelar y caracterizar robots con estructura en forma de paralelogramo y se presenta la arquitectura de control deseada. Una vez obtenido el modelo completo del esclavo, se investigan diferentes alternativas que permitan una estimación de fuerzas externas en tiempo real, minimizando las derivadas de la posición para minimizar el ruido. Se comienza utilizando observadores clásicos del estado para ir evolucionando hasta llegar al desarrollo de un observador de tipo Luenberger-Sliding cuya implementación es relativamente sencilla y sus resultados contundentes. También se analiza el uso del observador propuesto durante un control bilateral simulado en el que se compara la realimentación de fuerzas obtenida con las técnicas clásicas basadas en error de posición frente a un control basado en fuerza-posición donde la fuerza es estimada y no medida. Se comprueba como la solución propuesta da resultados comparables con las arquitecturas clásicas y sin embargo introduce una alternativa para la teleoperación de robots industriales cuya teleoperación en entornos radioactivos sería imposible de otra manera. Finalmente se analizan los problemas derivados de la aplicación práctica de la teleoperación en los escenarios mencionados anteriormente. Debido a las condiciones prohibitivas para todo equipo electrónico, los sistemas de control se deben colocar a gran distancia de los manipuladores, dando lugar a longitudes de cable de centenares de metros. En estas condiciones se crean sobretensiones en controladores basados en PWM que pueden ser destructivas para el sistema formado por control, cableado y actuador, y por tanto, han de ser eliminadas. En este trabajo se propone una solución basada en un filtro LC comercial y se prueba de forma extensiva que su inclusión no produce efectos negativos sobre el control del actuador. ABSTRACT As the energy on the particle accelerators or heavy ion accelerators such as CERN or GSI, fusion reactors such as JET or ITER, or other scientific experiments is increased, it is becoming increasingly necessary to use remote handling techniques to interact with the remote and radioactive environment. So far, the dose rate at CERN could present values near several mSv for cooling times on the range of hours, which allowed human intervention for maintenance tasks. At JET, they measured values close to 200 μSv after a cooling time of 4 months and since then, the remote handling techniques became usual. There is a clear tendency to increase the radiation levels in the future. A clear example is ITER, where values of 450 Sv/h are expected in the centre of the torus after 11 days of cooling. Also, the new energetic levels of CERN are expected to lead to a more advanced remote handling means. In these circumstances this thesis is framed, studying a bilateral control system based on force-position, trying to avoid the use of force/torque sensors, whose electronic content makes them very sensitive in these environments. The contents of this work are focused on teleoperating industrial robots, which due its well-known reliability, easiness to be adapted to these environments, cost-effectiveness and high availability, are considered as an interesting alternative to expensive custom-made solutions for remote handling tasks. Firstly, the kinematic problem of teloperating master and slave with dissimilar kinematics is analysed and a new general approach for solving this issue is presented. The solution includes using assistive forces in order to guide the human operator. Coming up next, I explain with detail the experiments accomplished with an ABB robot that show the difficulties encountered and the proposed solutions. This section is concluded with a method to match the master’s and slave’s workspaces when they present dissimilar kinematics. Later on, the research studies the dynamics, with special focus on robot modelling with the purpose of obtaining a method that allows to estimate external forces acting on them. During the characterisation of the model’s parameters, a set of tests are performed in order to get to a compromise between computational complexity and estimation error. Key points for modelling and characterising robots with a parallelogram structure are also given, and the desired control architecture is presented. Once a complete model of the slave is obtained, different alternatives for external force estimation are review to be able to predict forces in real time, minimizing the position differentiation to minimize the estimation noise. The research starts by implementing classic state observers and then it evolves towards the use of Luenberger- Sliding observers whose implementation is relatively easy and the results are convincing. I also analyse the use of proposed observer during a simulated bilateral control on which the force feedback obtained with the classic techniques based on the position error is compared versus a control architecture based on force-position, where the force is estimated instead of measured. I t is checked how the proposed solution gives results comparable with the classical techniques and however introduces an alternative method for teleoperating industrial robots whose teleoperation in radioactive environments would have been impossible in a different way. Finally, the problems originated by the practical application of teleoperation in the before mentioned scenarios are analysed. Due the prohibitive conditions for every electronic equipment, the control systems should be placed far from the manipulators. This provokes that the power cables that fed the slaves devices can present lengths of hundreds of meters. In these circumstances, overvoltage waves are developed when implementing drives based on PWM technique. The occurrence of overvoltage is very dangerous for the system composed by drive, wiring and actuator, and has to be eliminated. During this work, a solution based on commercial LC filters is proposed and it is extensively proved that its inclusion does not introduce adverse effects into the actuator’s control.

Laboratorio remoto eLab3D: un mundo virtual inmersivo para el aprendizaje de la electrónica

En este artículo se presenta un recurso educativo novedoso, basado en un laboratorio remoto, que permite a los estudiantes, a través de Internet, la realización de experimentos reales en el área de la electrónica. Se ha creado un mundo virtual 3D donde los usuarios, a través de sus avatares, pueden interaccionar con réplicas virtuales de instrumentos, placas de circuitos, componentes o cables y con compañeros y profesores, de forma similar a como lo harían en un laboratorio presencial. Este recurso ofrece múltiples posibilidades que pueden ser muy útiles en los diferentes niveles educativos. Se han llevado a cabo algunas experiencias educativas de utilización de la plataforma con el fin de valorar sus posibilidades docentes y los resultados obtenidos han sido muy positivos. ABSTRACT. This paper presents an innovative educational resource, based on a remote laboratory, which allows users to conduct real experiments through Internet in the area of electronics. A 3D virtual world has been created in which users, by means of their avatars, can interact with virtual replicas of instruments, circuit boards, components or cables and with peers and teachers, as they would in a traditional laboratory. This resource offers multiple possibilities that can be very useful in the different education levels. Some educative experiences have been carried out using the platform to evaluate its educational possibilities and the obtained results have been very positive.

Algoritmos de decisión para priorizar las acciones de restauración en el mantenimiento de redes

En la actualidad, cualquier compañía de telecomunicaciones que posea su propia red debe afrontar el problema del mantenimiento de la misma. Ofrecer un mínimo de calidad de servicio a sus clientes debe ser uno de sus objetivos principales. Esta calidad debe mantenerse aunque ocurran incidencias en la red. El presente trabajo pretende resolver el problema de priorizar el orden en que se restauran los cables, caminos y circuitos, dañados por una incidencia, dentro de una red troncal de transporte perteneciente a una operadora de telecomunicaciones. Tras un planteamiento detallado del problema y de todos los factores que influirán en la toma de decisión, en primer lugar se acomete una solución basada en Métodos Multicriterio Discretos, concretamente con el uso de ELECTRE I y AHP. A continuación se realiza una propuesta de solución basada en Redes Neuronales (con dos aproximaciones diferentes al problema). Por último se utiliza un método basado en la Optimización por Enjambre de Partículas (PSO), adaptado a un problema de permutación de enteros (ordenación), y con una forma particular de evaluar la mejor posición global del enjambre. Complementariamente se realiza una exposición de lo que es una empresa Operadora de telecomunicaciones, de sus departamentos y procesos internos, de los servicios que ofrece, de las redes sobre las que se soportan, y de los puntos clave a tener en cuenta en la implementación de sus sistemas informáticos de gestión integral. ABSTRACT: Nowadays, any telecommunications company that owns its own network must face the problem of maintaining it (service assurance). Provide a minimum quality of service to its customers must be one of its main objectives. This quality should be maintained although any incidents happen to occur in the network. This thesis aims to solve the problem of prioritizing the order in which the damaged cables, trails, path and circuits, within a backbone transport network, should be restored. This work begins with a detailed explanation about network maintenance issues and all the factors that influence decision-making problem. First of all, one solution based on Discrete Multicriteria methods is tried (ELECTRE I and AHP algorithms are used). Also, a solution based on neural networks (with two different approaches to the problem) is analyzed. Finally, this thesis proposes an algorithm based on Particle Swarm Optimization (PSO), adapted to a problem of integers permutation, and a particular way of evaluating the best overall position of the swarm method. In addition, there is included an exhibition about telecommunications companies, its departments, internal processes, services offered to clients, physical networks, and key points to consider when implementing its integrated management systems.

Criterios de dimensionamiento en estado límite de servicio de tableros de puentes con sistema de atirantamiento extradosado

Los criterios de dimensionamiento en tableros de hormigón pretensado están claramente establecidos para sistemas de pretensado convencional adherente o no, interior o exterior a la losa del tablero así como para sistemas de atirantamiento convencionales. Pero no para sistemas de atirantamiento no convencional como los puentes de atirantamiento extradosado. Con la realización de este trabajo, se pretende llegar a unos criterios de dimensionamiento en Estado Límite de Servicio (ELS), analizando los criterios de dimensionamiento ya existentes en sistemas de atirantamiento convencional, estableciendo las bases para su generalización al sistema de atirantamiento extradosado. El desarrollo del presente proyecto comenzará con una breve introducción sobre los sistemas estructurales con cables y tirantes, explicando sus características y comportamientos estructurales y dará paso al desarrollo del proyecto, utilizando para dicho estudio la modelización de un puente atirantado y otro dos extradosados, uno cuyos tirantes soportarán el 100% de la carga permanente mientras que en el otro soportarán el 60%. Se ha utilizado el programa de elementos finitos SAP 2000 v.17 para su modelación. Para llegar al dimensionamiento óptimo del puente extradosado, se han llevado a cabo diferentes cálculos para el predimensionamiento del pretensado, el efecto de la fluencia y el dimensionamiento final del pretensado. Llegando finalmente al coste que supondría el acero de cada una de las tipologías estudiadas.

Nuevas tendencias en el proyecto de buques cableros

Esta tesis tiene como objetivo actualizar los criterios de proyecto de los buques cableros, analizando las nuevas tendencias en el uso de este tipo de barcos. Se estudian las operaciones realizadas para la instalación y reparación de los cables submarinos. Del estudio de estas técnicas se extraen unos requisitos para el proyecto. Se comprueba cómo han evolucionado a lo largo de la historia las características de los buques cableros. Comparando esta evolución con las características necesarias extraídas del estudio de las operaciones de cables, se concluyen los aspectos fundamentales a tener en cuenta por los ingenieros navales en el proyecto de estos buques cableros. Se presentan: datos concretos para el dimensionamiento, equipamiento, distribución y disposición general; soluciones utilizadas y desarrolladas por el autor para la configuración del manejo del cable de tiro del arado submarino; y se propone una disposición general más acorde con las necesidades modernas de uso de estos barcos. Todo está basado en la experiencia del autor de más de 20 años operando buques cableros de la empresa Transoceanic Cableship Company, Ltd.1 y TE Conectivity SubCom SL (antes TEMASA2), ambas filiales de la empresa norteamericana de telecomunicaciones submarinas TE SubCom, por ello su utilidad práctica queda garantizada. ABSTRACT The goal of this thesis is to update the design criteria applied to cableships by analyzing the new trends in the use of this type of ships. The methods of procedure for the installation and repair of fiber optic submarine cables are also explored. The study of these techniques draws conclusions regarding the design requirements. The evolution of the characteristics of the cableships throughout the history is also approached. The primary aspect that has to be taken into account by the naval architect is also stated, when comparing the evolution and the necessary requirements extracted from the study of cable operations. Specific data for dimensioning, outfitting, distribution and arrangement are presented, as well as solutions found and developed by the author for the tow wire handling configuration. Furthermore, a new general arrangement more in line with the modern needs for the purpose of these ships is suggested. Everything is based in the author´s experience of over twenty years in the operation of cableships owned by Transoceanic Cableship Company Ltd. and TE Connectivity SubCom SL (formerly called TEMASA), both subsidiaries of the US based company TE SubCom. For this reason, the practical value of the study is guaranteed.

Influencia de la altura de lluvia en la atenuación por precipitación en comunicaciones por satélite

El objetivo principal de este Proyecto Fin de Grado es el estudio experimental de la atenuación, producida por las precipitaciones, en los enlaces Tierra – satélite en banda Ka. En particular, se ha realizado el estudio para una frecuencia de 19,701 GHz, que corresponde con la frecuencia de una de las balizas del satélite Hot Bird 13 de Eutelsat. Para la realización del estudio se dispone de datos experimentales recogidos por sondeos realizados en la estación meteorológica de Madrid - Barajas y de datos sinópticos. La primera parte del proyecto comienza con una descripción de los fundamentos teóricos de los distintos fenómenos que afectan a la propagación en un enlace por satélite, y se enumeran los distintos modelos de predicción más importantes. Posteriormente se describen en más detalle algunos modelos de predicción propuestos por la Unión Internacional de Telecomunicaciones (UIT). En la segunda parte del mismo se explica en detalle el proceso necesario para el procesado de los datos experimentales, con el fin de poder manejarlos de forma más sencilla a la hora de presentar los resultados pertinentes. En una tercera parte se recogen los resultados experimentales obtenidos para el caso de la altura de la isoterma a 0˚C y de la atenuación por lluvia. El capítulo dedicado al estudio de la altura de la isoterma a 0˚C se centra en obtener dicho dato a partir de los datos experimentales de los sondeos, que fueron procesados con anterioridad. Así mismo, se realizará una comparativa de estos datos experimentales con los proporcionados por la UIT en la Recomendación 839. En el capítulo dedicado al estudio de la atenuación producida por la lluvia se compararán los resultados obtenidos de forma experimental con los datos proporcionados por la UIT en la Recomendación 618. Por último se recoge en un capítulo las conclusiones obtenidas con la realización de este PFG y las líneas futuras de investigación. ABSTRACT. The main aim of this Final Degree Project is the experimental study of attenuation by precipitation, in links Earth – Ka band satellite. In particular, the study was performed at a frequency of 19.701 GHz, which corresponds to the frequency of one of the beacons of the satellite Hot Bird 13 (Eutelsat). There is experimental data and synoptic data provided, which was collected from surveys in the Madrid- Barajas weather station. The first part of the project outlines some theoretical foundations regarding different phenomena which affect propagation in a satellite link and includes the most important prediction models. Additionally, some prediction models proposed by the International Telecommunication Union (ITU) are outlined in detail. In the second part, the process for the processing of the experimental data is explained in detail. This process is necessary in order to be able to utilize the data more easily when presenting the results of this project. In the third part, the experimental results obtained in the study are presented for both cases: isotherm height at 0°C and rain attenuation. The chapter dedicated to the study of the isotherm height at 0°C focuses on obtaining the real isotherm height at 0°C from the experimental data processed previously. Furthermore, a comparison will be made between the experimental results and data proposed by ITU in Recommendation 839. The chapter dedicated to the study of rain attenuation focuses on making a comparison between the results from the experimental data and data proposed by ITU in Recommendation 618. Finally, there is a chapter which revises all the conclusions obtained throughout this project and provides recommendations for future research.

A Transportable Reflectarray Antenna for Satellite Ku-band Emergency Communications

The design of a Ku-band reconfigurable reflectarray antenna for emergency satellite communications is presented. Bidirectional high data rate satellite links are needed in emergency conditions where other telecommunication infrastructures are not available. In order to operate in this type of scenario, an antenna should be deployable, transportable, and easily repointable. The need of an automatic and fast satellite location and pointing system leads to a completely electronic reconfigurable antenna. The operative bandwidth is from 10.7 to 12.5 GHz for reception and from 14 up to 14.5 GHz for transmission (30% of relative bandwidth). The selected antenna architecture is based on a dual reflectarray system comprising a passive subreflectarray and an active main reflectarray made of reconfigurable 1-bit elementary cells based on PIN diodes.

RÁDIO, TECNOLOGIA E SOCIEDADE: O desenvolvimento da Radiodifusão Digital no Brasil

O trabalho trata da tecnologia de radiodifusão em processo de digitalização. Estabelece os elementos históricos fundamentais que contribuíram para a consolidação de uma indústria da comunicação, características demonstradas nos debates e argumentações em torno da digitalização do setor. Através da descrição das tecnologias digitais a serem empregadas na transição foi possível identificar os potenciais tecnológicos que são formatados socialmente nos atos pela padronização técnica, expondo as limitações e o teor conservador que uma nova tecnologia pode representar.(AU)

RÁDIO, TECNOLOGIA E SOCIEDADE: O desenvolvimento da Radiodifusão Digital no Brasil

O trabalho trata da tecnologia de radiodifusão em processo de digitalização. Estabelece os elementos históricos fundamentais que contribuíram para a consolidação de uma indústria da comunicação, características demonstradas nos debates e argumentações em torno da digitalização do setor. Através da descrição das tecnologias digitais a serem empregadas na transição foi possível identificar os potenciais tecnológicos que são formatados socialmente nos atos pela padronização técnica, expondo as limitações e o teor conservador que uma nova tecnologia pode representar.(AU)

Rádio, tecnologia e sociedade: o desenvolvimento da radiodifusão digital no Brasil

O trabalho trata da tecnologia de radiodifusão em processo de digitalização. Estabelece os elementos históricos fundamentais que contribuíram para a consolidação de uma indústria da comunicação, características demonstradas nos debates e argumentações em torno da digitalização do setor. Através da descrição das tecnologias digitais a serem empregadas na transição foi possível identificar os potenciais tecnológicos que são formatados socialmente nos atos pela padronização técnica, expondo as limitações e o teor conservador que uma nova tecnologia pode representar.

Regulation of the Actin Cytoskeleton by Thrombin in Human Endothelial Cells: Role of Rho Proteins in Endothelial Barrier Function

Endothelial barrier function is regulated at the cellular level by cytoskeletal-dependent anchoring and retracting forces. In the present study we have examined the signal transduction pathways underlying agonist-stimulated reorganization of the actin cytoskeleton in human umbilical vein endothelial cells. Receptor activation by thrombin, or the thrombin receptor (proteinase-activated receptor 1) agonist peptide, leads to an early increase in stress fiber formation followed by cortical actin accumulation and cell rounding. Selective inhibition of thrombin-stimulated signaling systems, including Gi/o (pertussis toxin sensitive), p42/p44, and p38 MAP kinase cascades, Src family kinases, PI-3 kinase, or S6 kinase pathways had no effect on the thrombin response. In contrast, staurosporine and KT5926, an inhibitor of myosin light chain kinase, effectively blocked thrombin-induced cell rounding and retraction. The contribution of Rho to these effects was analyzed by using bacterial toxins that either activate or inhibit the GTPase. Escherichia coli cytotoxic necrotizing factor 1, an activator of Rho, induced the appearance of dense actin cables across cells without perturbing monolayer integrity. Accordingly, lysophosphatidic acid, an activator of Rho-dependent stress fiber formation in fibroblasts, led to reorganization of polymerized actin into stress fibers but failed to induce cell rounding. Inhibition of Rho with Clostridium botulinum exoenzyme C3 fused to the B fragment of diphtheria toxin caused loss of stress fibers with only partial attenuation of thrombin-induced cell rounding. The implication of Rac and Cdc42 was analyzed in transient transfection experiments using either constitutively active (V12) or dominant-interfering (N17) mutants. Expression of RacV12 mimicked the effect of thrombin on cell rounding, and RacN17 blocked the response to thrombin, whereas Cdc42 mutants were without effect. These observations suggest that Rho is involved in the maintenance of endothelial barrier function and Rac participates in cytoskeletal remodeling by thrombin in human umbilical vein endothelial cells.

The metabolic implications of intracellular circulation

Two views currently dominate research into cell function and regulation. Model I assumes that cell behavior is quite similar to that expected for a watery bag of enzymes and ligands. Model II assumes that three-dimensional order and structure constrain and determine metabolite behavior. A major problem in cell metabolism is determining why essentially all metabolite concentrations are remarkably stable (are homeostatic) over large changes in pathway fluxes—for convenience, this is termed the [s] stability paradox. For muscle cells, ATP and O2 are the most perfectly homeostatic, even though O2 delivery and metabolic rate correlate in a 1:1 fashion. In total, more than 60 metabolites are known to be remarkably homeostatic in differing metabolic states. Several explanations of [s] stability are usually given by traditional model I studies—none of which apply to all enzymes in a pathway, and all of which require diffusion as the means for changing enzyme–substrate encounter rates. In contrast, recent developments in our understanding of intracellular myosin, kinesin, and dyenin motors running on actin and tubulin tracks or cables supply a mechanistic basis for regulated intracellular circulation systems with cytoplasmic streaming rates varying over an approximately 80-fold range (from 1 to >80 μm × sec−1). These new studies raise a model II hypothesis of intracellular perfusion or convection as a primary means for bringing enzymes and substrates together under variable metabolic conditions. In this view, change in intracellular perfusion rates cause change in enzyme–substrate encounter rates and thus change in pathway fluxes with no requirement for large simultaneous changes in substrate concentrations. The ease with which this hypothesis explains the [s] stability paradox is one of its most compelling features.

Identification of Two Type V Myosins in Fission Yeast, One of Which Functions in Polarized Cell Growth and Moves Rapidly in the Cell

We characterized the novel Schizosaccharomyces pombe genes myo4+ and myo5+, both of which encode myosin-V heavy chains. Disruption of myo4 caused a defect in cell growth and led to an abnormal accumulation of secretory vesicles throughout the cytoplasm. The mutant cells were rounder than normal, although the sites for cell polarization were still established. Elongation of the cell ends and completion of septation required more time than in wild-type cells, indicating that Myo4 functions in polarized growth both at the cell ends and during septation. Consistent with this conclusion, Myo4 was localized around the growing cell ends, the medial F-actin ring, and the septum as a cluster of dot structures. In living cells, the dots of green fluorescent protein-tagged Myo4 moved rapidly around these regions. The localization and movement of Myo4 were dependent on both F-actin cables and its motor activity but seemed to be independent of microtubules. Moreover, the motor activity of Myo4 was essential for its function. These results suggest that Myo4 is involved in polarized cell growth by moving with a secretory vesicle along the F-actin cables around the sites for polarization. In contrast, the phenotype of myo5 null cells was indistinguishable from that of wild-type cells. This and other data suggest that Myo5 has a role distinct from that of Myo4.