Hsc70-induced changes in clathrin-auxilin cage structure suggest a role for clathrin light chains in cage disassembly

The molecular chaperone, Hsc70, together with its co-factor, auxilin, facilitates the ATP-dependent removal of clathrin during clathrin-mediated endocytosis in cells. We have used cryo-electron microscopy to determine the 3D structure of a complex of clathrin, auxilin401-910 and Hsc70 at pH 6 in the presence of ATP, frozen within 20 seconds of adding Hsc70 in order to visualize events that follow the binding of Hsc70 to clathrin and auxilin before clathrin disassembly. In this map, we observe density beneath the vertex of the cage that we attribute to bound Hsc70. This density emerges asymmetrically from the clathrin vertex, suggesting preferential binding by Hsc70 for one of the three possible sites at the vertex. Statistical comparison with a map of whole auxilin and clathrin previously published by us reveals the location of statistically significant differences which implicate involvement of clathrin light chains in structural rearrangements which occur after Hsc70 is recruited. Clathrin disassembly assays using light scattering suggest that loss of clathrin light chains reduces the efficiency with which auxilin facilitates this reaction. These data support a regulatory role for clathrin light chains in clathrin disassembly in addition to their established role in regulating clathrin assembly

Passive Hydro-actuated Unfolding of Ice Plant Seed Capsules as a Concept Generator for Autonomously Deforming Devices

In der Natur und ihren biologischen Systemen existieren zahlreiche Beispiele für gerichtete Bewegung durch spezifische Reaktion auf externe Stimuli. Diese potentiellen Quellen der Inspiration dienen oft als Vorbilder für energieeffiziente "Smart" Technologien. Vom Wasser getriebenen schnellen Zuschnappen der Venusfliegenfalle bis zum einfacheren ebenso hydroresponsiven Biegen der Weizengrannen, viele Pflanzen haben im Laufe der Evolution verschiedene Mechanismen entwickelt, um Wasser als Triebkraft ihrer Aktoren-Gewebe zu nutzen, die für spezifische und gerichtete Bewegung sowie die gewünschte Verformung sorgen. Das ist diesen Pflanzen möglich durch die Organisation ihrer Gewebe in ausgereiften, komplexen und hierarchisch organisierten Architekturen auf verschiedensten Skalen. Einige Arten der Familie Aizoaceae, auch bekannt als Mittagsblumen oder Ice plant, zeigen ein geniales Beispiel für solche passiven Betätigungssysteme, da sie einen "intelligenten" Mechanismus entwickelt haben, um ihre Schutzsamenkapseln öffnen zu lassen und die Samen nur in Anwesenheit von flüssigem Wasser (Regen) freizugeben. Schwerpunkt der ersten Phase dieser Arbeit war die Untersuchung der zu Grunde liegenden Mechanismen und der strukturellen und kompositorischen Basis von Wasser-getriebenen Bewegungen der Samenkapseln von Ice plant (Delosperma nakurense) auf ihren verschiedenen hierarchischen Ebenen. Fünf hygroskopische Kiele erwiesen sich als aktive "Muskeln", die zu einer reversiblen origamiartigen Entfaltung der Samenkapsel führen, wenn diese mit Wasser benetzt wird. Jeder Kiel besteht aus zwei wabenartigen Geweben, die aus hochgradig schwellfähigen und elliptisch-sechseckig geformten Zellen zusammengesetzt sind, die entlang einem inerten Träger organisiert sind. Als Hauptmotor der Aktuation wurde die signifikante Schwellung von hochgradig schwellfähigen zellulosereichen Innenschichten (CIL) im Lumen der Zellen identifiziert. Die Morphologie der CIL und deren physikochemische Reaktion auf Wasser wurde unter Verwendung einer Vielzahl von Techniken untersucht und damit gezeigt, dass der Entropiegewinn während der Wasserabsorption die Hauptantriebskraft für die Schwellung der Zellen ist. Die Umsetzung dieser relativ kleinen Energiebeiträge in eine konzertierte und komplexe makroskopische Bewegung, wurde durch ein optimiertes Design auf den verschiedenen Ebenen der hierarchischen Organisation des Systems erläutert. Das kooperative anisotropische Anschwellen der Zellen des hygroskopischen Gewebes führt durch das Timoschenko Doppelschicht-Biegeprinzip zu einer Umsetzung in eine Biegebewegung der Strukturen und letztlich zur Entfaltung der Samenkapseln. Inspiriert von den zugrunde liegenden Mechanismen in Ice plants, wurden zwei unterschiedliche Strategien entwickelt, um durch kleine Dehnungen im mikroskopischen Bereich eine vorprogrammierte Makro-Bewegung einer Wabenstruktur zu ermöglichen. Durch eine geschickte Anwendung dieses einfachen Prinzips, kann eine Mimik des biologischen Vorbilds im weiteren technischen Sinne zu zahlreichen Anwendungsbeispielen führen, wie als passive Schalter und Aktoren in der Biomedizin, Landschaftsgestaltung oder der Architektur.Numerous examples of actuated-movements with specific responses of the structure to external stimuli can be found in biological systems, which can be a potential source of inspiration for the design of energy-efficient "smart" devices. From the hydro-driven rapid snapping of the Venus fly trap leaves to simple hydro-responsive bending of wheat awns, various plants have evolved different mechanisms to utilize water as an actuator to undergo a desired deformation via sophisticated architecture at different hierarchical levels of their systems. Some species of the family Aizoaceae, also known as ice plants, show an ingenious example of such passive actuation systems, as they evolved a smart mechanism to open their protective seed capsules and release their seeds only in the presence of liquid water (rain). The scope of the first phase of the thesis was to investigate the underlying mechanism and the structural and compositional basis of the hydro-actuated movement of the ice plant seed capsules (Delosperma nakurense) at several hierarchical levels. Five hygroscopic keels were found to be the active muscles responsible for the reversible origami-like unfolding of the seed capsule upon wetting. Each keel consists of two honeycomb-like tissues made up of highly swellable hexagonal/elliptical shape cells running along an inert backing tissue. The significant swelling of a highly swellable cellulosic inner layer (CIL) inside the lumen of these cells was found to be the main engine of the actuation. The morphology and physicochemical response of the CIL to water was studied using a variety of techniques and it was shown that the entropic changes during water absorption were the main driving force for swelling of the cells. The translation of such relatively small available energy to the complex movement at a macro scale was explained by an optimized design at different hierarchical levels of the system. The cooperative anisotropic swelling of the cells in the hygroscopic tissue is translated into a flexing movement of the structure via simple Timoshenko’s bilayer bending principle, which then results in an unfolding of the seed capsules. Inspired by the underlying mechanism in ice plant, two different strategies were developed to translate small strains at micro scale into a pre-programmed macro movement of a honeycomb structure. Through a clever application of the same simple concepts, one can "mimic" the biological model system in a broader engineering sense, with potential applications of such passive switches in biomedicine, agricultural engineering or architectural design

Deep Space Network information system architecture study

The purpose of this article is to describe an architecture for the Deep Space Network (DSN) information system in the years 2000-2010 and to provide guidelines for its evolution during the 1990s. The study scope is defined to be from the front-end areas at the antennas to the end users (spacecraft teams, principal investigators, archival storage systems, and non-NASA partners). The architectural vision provides guidance for major DSN implementation efforts during the next decade. A strong motivation for the study is an expected dramatic improvement in information-systems technologies, such as the following: computer processing, automation technology (including knowledge-based systems), networking and data transport, software and hardware engineering, and human-interface technology. The proposed Ground Information System has the following major features: unified architecture from the front-end area to the end user; open-systems standards to achieve interoperability; DSN production of level 0 data; delivery of level 0 data from the Deep Space Communications Complex, if desired; dedicated telemetry processors for each receiver; security against unauthorized access and errors; and highly automated monitor and control

Functional requirements document for the Earth Observing System Data and Information System (EOSDIS) Scientific Computing Facilities (SCF) of the NASA/MSFC Earth Science and Applications Division, 1992

Five scientists at MSFC/ESAD have EOS SCF investigator status. Each SCF has unique tasks which require the establishment of a computing facility dedicated to accomplishing those tasks. A SCF Working Group was established at ESAD with the charter of defining the computing requirements of the individual SCFs and recommending options for meeting these requirements. The primary goal of the working group was to determine which computing needs can be satisfied using either shared resources or separate but compatible resources, and which needs require unique individual resources. The requirements investigated included CPU-intensive vector and scalar processing, visualization, data storage, connectivity, and I/O peripherals. A review of computer industry directions and a market survey of computing hardware provided information regarding important industry standards and candidate computing platforms. It was determined that the total SCF computing requirements might be most effectively met using a hierarchy consisting of shared and individual resources. This hierarchy is composed of five major system types: (1) a supercomputer class vector processor; (2) a high-end scalar multiprocessor workstation; (3) a file server; (4) a few medium- to high-end visualization workstations; and (5) several low- to medium-range personal graphics workstations. Specific recommendations for meeting the needs of each of these types are presented

Image-Guided Robot-Assisted Techniques with Applications in Minimally Invasive Therapy and Cell Biology

There are several situations where tasks can be performed better robotically rather than manually. Among these are situations (a) where high accuracy and robustness are required, (b) where difficult or hazardous working conditions exist, and (c) where very large or very small motions or forces are involved. Recent advances in technology have resulted in smaller size robots with higher accuracy and reliability. As a result, robotics is fi nding more and more applications in Biomedical Engineering. Medical Robotics and Cell Micro-Manipulation are two of these applications involving interaction with delicate living organs at very di fferent scales.Availability of a wide range of imaging modalities from ultrasound and X-ray fluoroscopy to high magni cation optical microscopes, makes it possible to use imaging as a powerful means to guide and control robot manipulators. This thesis includes three parts focusing on three applications of Image-Guided Robotics in biomedical engineering, including: Vascular Catheterization: a robotic system was developed to insert a catheter through the vasculature and guide it to a desired point via visual servoing. The system provides shared control with the operator to perform a task semi-automatically or through master-slave control. The system provides control of a catheter tip with high accuracy while reducing X-ray exposure to the clinicians and providing a more ergonomic situation for the cardiologists. Cardiac Catheterization: a master-slave robotic system was developed to perform accurate control of a steerable catheter to touch and ablate faulty regions on the inner walls of a beating heart in order to treat arrhythmia. The system facilitates touching and making contact with a target point in a beating heart chamber through master-slave control with coordinated visual feedback. Live Neuron Micro-Manipulation: a microscope image-guided robotic system was developed to provide shared control over multiple micro-manipulators to touch cell membranes in order to perform patch clamp electrophysiology. Image-guided robot-assisted techniques with master-slave control were implemented for each case to provide shared control between a human operator and a robot. The results show increased accuracy and reduced operation time in all three cases

Fatias de rede fim-a-fim : da extração de perfis de funções de rede a SLAs granulares

Orientador: Christian Rodolfo Esteve RothenbergTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de ComputaçãoResumo: Nos últimos dez anos, processos de softwarização de redes vêm sendo continuamente diversi- ficados e gradativamente incorporados em produção, principalmente através dos paradigmas de Redes Definidas por Software (ex.: regras de fluxos de rede programáveis) e Virtualização de Funções de Rede (ex.: orquestração de funções virtualizadas de rede). Embasado neste processo o conceito de network slice surge como forma de definição de caminhos de rede fim- a-fim programáveis, possivelmente sobre infrastruturas compartilhadas, contendo requisitos estritos de desempenho e dedicado a um modelo particular de negócios. Esta tese investiga a hipótese de que a desagregação de métricas de desempenho de funções virtualizadas de rede impactam e compõe critérios de alocação de network slices (i.e., diversas opções de utiliza- ção de recursos), os quais quando realizados devem ter seu gerenciamento de ciclo de vida implementado de forma transparente em correspondência ao seu caso de negócios de comu- nicação fim-a-fim. A verificação de tal assertiva se dá em três aspectos: entender os graus de liberdade nos quais métricas de desempenho de funções virtualizadas de rede podem ser expressas; métodos de racionalização da alocação de recursos por network slices e seus re- spectivos critérios; e formas transparentes de rastrear e gerenciar recursos de rede fim-a-fim entre múltiplos domínios administrativos. Para atingir estes objetivos, diversas contribuições são realizadas por esta tese, dentre elas: a construção de uma plataforma para automatização de metodologias de testes de desempenho de funções virtualizadas de redes; a elaboração de uma metodologia para análises de alocações de recursos de network slices baseada em um algoritmo classificador de aprendizado de máquinas e outro algoritmo de análise multi- critério; e a construção de um protótipo utilizando blockchain para a realização de contratos inteligentes envolvendo acordos de serviços entre domínios administrativos de rede. Por meio de experimentos e análises sugerimos que: métricas de desempenho de funções virtualizadas de rede dependem da alocação de recursos, configurações internas e estímulo de tráfego de testes; network slices podem ter suas alocações de recursos coerentemente classificadas por diferentes critérios; e acordos entre domínios administrativos podem ser realizados de forma transparente e em variadas formas de granularidade por meio de contratos inteligentes uti- lizando blockchain. Ao final deste trabalho, com base em uma ampla discussão as perguntas de pesquisa associadas à hipótese são respondidas, de forma que a avaliação da hipótese proposta seja realizada perante uma ampla visão das contribuições e trabalhos futuros desta teseAbstract: In the last ten years, network softwarisation processes have been continuously diversified and gradually incorporated into production, mainly through the paradigms of Software Defined Networks (e.g., programmable network flow rules) and Network Functions Virtualization (e.g., orchestration of virtualized network functions). Based on this process, the concept of network slice emerges as a way of defining end-to-end network programmable paths, possibly over shared network infrastructures, requiring strict performance metrics associated to a par- ticular business case. This thesis investigate the hypothesis that the disaggregation of network function performance metrics impacts and composes a network slice footprint incurring in di- verse slicing feature options, which when realized should have their Service Level Agreement (SLA) life cycle management transparently implemented in correspondence to their fulfilling end-to-end communication business case. The validation of such assertive takes place in three aspects: the degrees of freedom by which performance of virtualized network functions can be expressed; the methods of rationalizing the footprint of network slices; and transparent ways to track and manage network assets among multiple administrative domains. In order to achieve such goals, a series of contributions were achieved by this thesis, among them: the construction of a platform for automating methodologies for performance testing of virtual- ized network functions; an elaboration of a methodology for the analysis of footprint features of network slices based on a machine learning classifier algorithm and a multi-criteria analysis algorithm; and the construction of a prototype using blockchain to carry out smart contracts involving service level agreements between administrative systems. Through experiments and analysis we suggest that: performance metrics of virtualized network functions depend on the allocation of resources, internal configurations and test traffic stimulus; network slices can have their resource allocations consistently analyzed/classified by different criteria; and agree- ments between administrative domains can be performed transparently and in various forms of granularity through blockchain smart contracts. At the end of his thesis, through a wide discussion we answer all the research questions associated to the investigated hypothesis in such way its evaluation is performed in face of wide view of the contributions and future work of this thesisDoutoradoEngenharia de ComputaçãoDoutor em Engenharia ElétricaFUNCAM

COBE's search for structure in the Big Bang

The launch of Cosmic Background Explorer (COBE) and the definition of Earth Observing System (EOS) are two of the major events at NASA-Goddard. The three experiments contained in COBE (Differential Microwave Radiometer (DMR), Far Infrared Absolute Spectrophotometer (FIRAS), and Diffuse Infrared Background Experiment (DIRBE)) are very important in measuring the big bang. DMR measures the isotropy of the cosmic background (direction of the radiation). FIRAS looks at the spectrum over the whole sky, searching for deviations, and DIRBE operates in the infrared part of the spectrum gathering evidence of the earliest galaxy formation. By special techniques, the radiation coming from the solar system will be distinguished from that of extragalactic origin. Unique graphics will be used to represent the temperature of the emitting material. A cosmic event will be modeled of such importance that it will affect cosmological theory for generations to come. EOS will monitor changes in the Earth's geophysics during a whole solar color cycle

Augmented reality device for first response scenarios

A prototype of a wearable computer system is proposed and implemented using commercial off-shelf components. The system is designed to allow the user to access location-specific information about an environment, and to provide capability for user tracking. Areas of applicability include primarily first response scenarios, with possible applications in maintenance or construction of buildings and other structures. Necessary preparation of the target environment prior to system\u27s deployment is limited to noninvasive labeling using optical fiducial markers. The system relies on computational vision methods for registration of labels and user position. With the system the user has access to on-demand information relevant to a particular real-world location. Team collaboration is assisted by user tracking and real-time visualizations of team member positions within the environment. The user interface and display methods are inspired by Augmented Reality1 (AR) techniques, incorporating a video-see-through Head Mounted Display (HMD) and fingerbending sensor glove.*. 1Augmented reality (AR) is a field of computer research which deals with the combination of real world and computer generated data. At present, most AR research is concerned with the use of live video imagery which is digitally processed and augmented by the addition of computer generated graphics. Advanced research includes the use of motion tracking data, fiducial marker recognition using machine vision, and the construction of controlled environments containing any number of sensors and actuators. (Source: Wikipedia) *This dissertation is a compound document (contains both a paper copy and a CD as part of the dissertation). The CD requires the following system requirements: Adobe Acrobat; Microsoft Office; Windows MediaPlayer or RealPlayer