288 research outputs found
Microvision: Static analysis-based approach to visualizing microservices in augmented reality
Microservices are supporting digital transformation; however, fundamental
tools and system perspectives are missing to better observe, understand, and
manage these systems, their properties, and their dependencies. Microservices
architecture leans toward decentralization, which yields many advantages to
system operation; it, however, brings challenges to their development.
Microservices lack a system-centric perspective to better cope with system
evolution and quality assessment. In this work, we explore
microservice-specific architecture reconstruction based on static analysis.
Such reconstruction typically results in system models to visualize selected
system-centric perspectives. Conventional models are limited in utility when
the service cardinality is high. We consider an alternative data visualization
using 3D space using augmented reality. To begin testing the feasibility of
deriving such perspectives from microservice systems, we developed and
implemented prototype tools for software architecture reconstruction and
visualization of compared perspectives
Microservice Architecture Reconstruction and Visualization Techniques: A Review
Microservice system solutions are driving digital transformation; however,
fundamental tools and system perspectives are missing to better observe,
understand, and manage these systems, their properties, and their dependencies.
Microservices architecture leads towards decentralization, which implies many
advantages to system operation; it, however, brings challenges to their
development. Microservice systems often lack a system-centric perspective that
would help engineers better cope with system evolution and quality assessment.
In this work, we explored microservice-specific architecture reconstruction
based on static analysis. Such reconstruction typically results in system
models to visualize selected system-centric perspectives. Conventional models
involve 2D methods; however, these methods are limited in utility when services
proliferate. We considered various architectural perspectives relevant to
microservices and assessed the relevancy of the traditional method, comparing
it to alternative data visualization using 3D space. As a representative of the
3D method, we considered a 3D graph model presented in augmented reality. To
begin testing the feasibility of deriving such perspectives from microservice
systems, we developed and implemented prototype tools for software architecture
reconstruction and visualization of compared perspectives. Using these
prototypes, we performed a small user study with software practitioners to
highlight the potentials and limitations of these innovative visualizations
used for common practitioner reasoning and tasks
Isoform-specific subcellular localization and function of protein kinase A identified by mosaic imaging of mouse brain.
Protein kinase A (PKA) plays critical roles in neuronal function that are mediated by different regulatory (R) subunits. Deficiency in either the RIβ or the RIIβ subunit results in distinct neuronal phenotypes. Although RIβ contributes to synaptic plasticity, it is the least studied isoform. Using isoform-specific antibodies, we generated high-resolution large-scale immunohistochemical mosaic images of mouse brain that provided global views of several brain regions, including the hippocampus and cerebellum. The isoforms concentrate in discrete brain regions, and we were able to zoom-in to show distinct patterns of subcellular localization. RIβ is enriched in dendrites and co-localizes with MAP2, whereas RIIβ is concentrated in axons. Using correlated light and electron microscopy, we confirmed the mitochondrial and nuclear localization of RIβ in cultured neurons. To show the functional significance of nuclear localization, we demonstrated that downregulation of RIβ, but not of RIIβ, decreased CREB phosphorylation. Our study reveals how PKA isoform specificity is defined by precise localization
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Imaging calcium microdomains within entire astrocyte territories and endfeet with GCaMPs expressed using adeno-associated viruses.
Intracellular Ca(2+) transients are considered a primary signal by which astrocytes interact with neurons and blood vessels. With existing commonly used methods, Ca(2+) has been studied only within astrocyte somata and thick branches, leaving the distal fine branchlets and endfeet that are most proximate to neuronal synapses and blood vessels largely unexplored. Here, using cytosolic and membrane-tethered forms of genetically encoded Ca(2+) indicators (GECIs; cyto-GCaMP3 and Lck-GCaMP3), we report well-characterized approaches that overcome these limitations. We used in vivo microinjections of adeno-associated viruses to express GECIs in astrocytes and studied Ca(2+) signals in acute hippocampal slices in vitro from adult mice (aged ∼P80) two weeks after infection. Our data reveal a sparkling panorama of unexpectedly numerous, frequent, equivalently scaled, and highly localized Ca(2+) microdomains within entire astrocyte territories in situ within acute hippocampal slices, consistent with the distribution of perisynaptic branchlets described using electron microscopy. Signals from endfeet were revealed with particular clarity. The tools and experimental approaches we describe in detail allow for the systematic study of Ca(2+) signals within entire astrocytes, including within fine perisynaptic branchlets and vessel-associated endfeet, permitting rigorous evaluation of how astrocytes contribute to brain function
Advances in molecular probe-based labeling tools and their application to multiscale multimodal correlated microscopies
The need to determine the precise subcellular distribution of specific proteins and macromolecular complexes in cells and tissues has been the major driving force behind the development of new molecular-genetic and chemical-labeling approaches applicable to high-resolution, correlated, multidimensional microscopy. This short review is intended to provide an overview of recently developed and widely used electron microscopy (EM)-compatible probes, including tetracysteine tags, mini singlet oxygen generator (MiniSOG), time-specific tag for the age measurement of proteins (TimeSTAMP) with MiniSOG, and enhanced ascorbate peroxidase (APEX). We describe how these highly specific and genetically introduced EM probes are now used, in conjunction with lower resolution light microscopic methods, to obtain wide field or dynamic records of live preparation or of large maps in 3D using recently developed laboratory-scale X-ray microscopes. The article is intended to enable researchers through a high-level view of the toolbox of labels available today for studies aiming to analyze dynamic subcellular and molecular processes in cell culture systems as well as in animal tissues—and ultimately allow investigators to determine the precise location of macromolecular complexes by EM
Hydrometeorological Analysis and Support Function at the Southeast River Forecast Center
Proceedings of the 2001 Georgia Water Resources Conference, April 26 and 27, 2001, Athens, Georgia.In the late 1980s, the National Weather Service (NWS) launched its ten year, 4.5 billion dollar Modernization and Restructuring (MAR) program to take advantage of rapidly advancing scientific and computer technologies. The implementation of MAR is complete and has succeeded in modernizing the hydrometeorological operations of the NWS. As a result of MAR, all 13 River Forecast Centers (RFC) across the U.S. restructured their operations and upgraded computer technology. The RFCs have extended their hours into the evening and nearly doubled their staffs, which included the hiring of three meteorologists at each RFC. These meteorologists have become part of a new function at RFCs, known as the Hydrometeorological Analysis and Support (HAS)function, to manage the greatly increased flow of hydrometeorological data for input into the hydrologic models. The HAS forecaster is primarily responsible for the production of the Quantitative Precipitation Forecast (QPF), the verification activities associated with the QPF, the comparison and quality control of radar and rain gage data, and the production of hydrometeorological discussions.Sponsored and Organized by: U.S. Geological Survey, Georgia Department of Natural Resources, Natural Resources Conservation Service, The University of Georgia, Georgia State University, Georgia Institute of TechnologyThis book was published by the Institute of Ecology, The University of Georgia, Athens, Georgia 30602-2202. The views and statements advanced in this publication are solely those of the authors and do not represent official views or policies of The University of Georgia, the U.S. Geological Survey, the Georgia Water Research Institute as authorized by the Water Resources Research Act of 1990 (P.L. 101-397) or the other conference sponsors
Spatial constraints dictate glial territories at murine neuromuscular junctions
Spatial competition between glial cells causes them to partition neuromuscular junctions into discrete domains within a synapse
Cellular and subcellular localization of the neuron-specific plasma membrane calcium ATPase PMCA1a in the rat brain
Regulation of intracellular calcium is crucial both for proper neuronal function and survival. By coupling ATP hydrolysis with Ca2+ extrusion from the cell, the plasma membrane calcium-dependent ATPases (PMCAs) play an essential role in controlling intracellular calcium levels in neurons. In contrast to PMCA2 and PMCA3, which are expressed in significant levels only in the brain and a few other tissues, PMCA1 is ubiquitously distributed, and is thus widely believed to play a “housekeeping” function in mammalian cells. Whereas the PMCA1b splice variant is predominant in most tissues, an alternative variant, PMCA1a, is the major form of PMCA1 in the adult brain. Here, we use immunohistochemistry to analyze the cellular and subcellular distribution of PMCA1a in the brain. We show that PMCA1a is not ubiquitously expressed, but rather is confined to neurons, where it concentrates in the plasma membrane of somata, dendrites and spines. Thus, rather than serving a general “housekeeping” function, our data suggest that PMCA1a is a calcium pump specialized for neurons, where it may contribute to the modulation of somatic and dendritic Ca2+ transients
Disentangling astroglial physiology with a realistic cell model in silico
Electrically non-excitable astroglia take up neurotransmitters, buffer extracellular K+ and generate Ca2+ signals that release molecular regulators of neural circuitry. The underlying machinery remains enigmatic, mainly because the sponge-like astrocyte morphology has been difficult to access experimentally or explore theoretically. Here, we systematically incorporate multi-scale, tri-dimensional astroglial architecture into a realistic multi-compartmental cell model, which we constrain by empirical tests and integrate into the NEURON computational biophysical environment. This approach is implemented as a flexible astrocyte-model builder ASTRO. As a proof-of-concept, we explore an in silico astrocyte to evaluate basic cell physiology features inaccessible experimentally. Our simulations suggest that currents generated by glutamate transporters or K+ channels have negligible distant effects on membrane voltage and that individual astrocytes can successfully handle extracellular K+ hotspots. We show how intracellular Ca2+ buffers affect Ca2+ waves and why the classical Ca2+ sparks-and-puffs mechanism is theoretically compatible with common readouts of astroglial Ca2+ imaging
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