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At least 1,009 records · Page 56

Increasing Importance of Material Electrical Interaction with the Space Environment

The electrical properties of materials have always been important for spacecraft in charging environments. However, in recent years consideration of interactions of materials and systems with the plasma environment has become more and more important in spacecraft design. This has primarily been driven by independent factors including increase in power and high voltage power systems, operation of tethered satellites, and science requirements for electrostatic clean spacecraft. Increased need for power has led to increased operating voltages for spacecraft. The Upper Atmospheric Research Satellite (UARS) was one of the first to operate at near 100 V solar array potential and demonstrate that the spacecraft floated nearly the entire voltage negative of the ionospheric plasma. The high voltage, 160 V, of the solar arrays on the International Space Station (ISS) led to the requirement to have a plasma contactor to control structure potential relative to the local plasma. Issues such as sputtering, dielectric breakdown, capacitive energy storage in the structure, space debris impact induced arcs and other arcing mechanisms had to be addressed. Recently commercial satellites, driven to higher voltages for efficiency, have experienced arcing problems which led to severe, permanent power degradation. The first tethered satellite, Tethered Satellite System (TSS), was deployed from the Space Shuttle. A conductive coating was developed which provided a low resistivity and also the required solar absorptivity and emittance. Other tether systems are being designed which will have similar requirements but also long life and "bare tether" designs are also being built for flight experiments. The wire requires an electrically conductive coating with proper thermal control properties, which a bare wire doesn't possess. Increasing sophistication of scientific instruments and measurements which scientists want to make have led to increasing requirements for conducting thermal control coatings to provide electrostatic cleanliness. These issues, which are increasing the importance of electrical interaction of materials to the space environment, will be discussed in this paper as well as research into material development to meet thermal and electrical requirements and research to understand phenomena regarding such interactions.

Carruth, M. R., Jr.↗

Control of the boundary layer separation about an airfoil by active surface heating

Application of active control to separated flow on the RC(6)-08 airfoil at high angle of attack by localized surface heating is numerically simulated by integrating the compressible two-dimensional nonlinear Navier-Stokes equations solver. Active control is simulated by local modification of the temperature boundary condition over a narrow strip on the upper surface of the airfoil. Both mean and perturbed profiles are favorably altered when excited with the same natural frequency of the shear layer by moderate surface heating for both laminar and turbulent separation. The shear layer is found to be very sensitive to localized surface heating in the vicinity of the separation point. The excitation field at the surface sufficiently altered both the local as well as the global circulation to cause a significant increase in lift and reduction in drag.

Maestrello, Lucio↗

Experimental Tests of Lateral Bedload Transport Induced by a Yawed Submerged Vane Array in Open-Channel Flows

This work proposes the use of an array of yawed porous vanes to control the lateral bedload transport by locally steering bedform migration and maximize the amount of sediments redirected toward a potential sediment extraction system or bypass channel. A laboratory experiment was conducted in a quasifield-scale channel with an array of permeable vanes installed on one side, in live-bed conditions under bedload dominant regime, i.e., negligible suspended load. A baseline experiment without vanes was also performed for comparison. The evolution of migrating bedforms of different scales was tracked in space and time using a high-resolution, state-of-the-art laser scanning device. The bedload transport rate in the streamwise direction was first calculated using bedforms’ geometry and migration velocity, and then spatially distributed over the entire monitored area using a new Eulerian-averaged grid-mapping method. This allowed us to introduce a new methodology to estimate the lateral bedload transport using control volume theory and applying mass conservation. Quantitative assessments of lateral bedload transport along the channel yield consistent results, suggesting that the vanes effectively move sediments laterally as intended. Under the investigated setup, the maximum lateral sediment transport rate ranges from 9% to 18% of the whole domain-averaged streamwise transport rate. The developed methodology also allowed to identify the location where sediment capture could be maximized for the given vane spatial distribution.

42 ENGINEERING↗

A New Distributed Model-Free Control Strategy to Diminish Distribution System Voltage Violations

This paper proposes a new distributed model-free control (MFC) strategy for dynamic voltage control to diminish distribution systems' voltage violations. The objective is to maintain all critical load bus voltages within the acceptable ANSI Range A (+/- 5% of nominal). The distributed MFC strategy, which only requires local voltage measurements from designated load buses, controls online the reactive power generation of available synchronous generator (SG)-based and photovoltaic (PV)-based distributed generators (DGs). The distributed MFC strategy is computationally efficient and does not require modelling of the different system components and disturbances. Time-domain dynamic simulations are conducted for the 21-bus test distribution system fed by multiple DGs to verify the performance of the proposed MFC strategy, and the results are compared against the conventional model-based microgrid voltage stabilizer (MGVS) control strategy. The simulation results show that the distributed MFC strategy provides minimal voltage violations and achieves the dynamic voltage stability of the system under diverse disturbances.

Hatipoglu, Kenan↗

Autonomy Architectures for a Constellation of Spacecraft

Until the past few years, missions typically involved fairly large expensive spacecraft. Such missions have primarily favored using older proven technologies over more recently developed ones, and humans controlled spacecraft by manually generating detailed command sequences with low-level tools and then transmitting the sequences for subsequent execution on a spacecraft controller. This approach toward controlling a spacecraft has worked spectacularly on previous missions, but it has limitations deriving from communications restrictions - scheduling time to communicate with a particular spacecraft involves competing with other projects due to the limited number of deep space network antennae. This implies that a spacecraft can spend a long time just waiting whenever a command sequence fails. This is one reason why the New Millennium program has an objective to migrate parts of mission control tasks onboard a spacecraft to reduce wait time by making spacecraft more robust. The migrated software is called a "remote agent" and has 4 components: a mission manager to generate the high level goals, a planner/scheduler to turn goals into activities while reasoning about future expected situations, an executive/diagnostics engine to initiate and maintain activities while interpreting sensed events by reasoning about past and present situations, and a conventional real-time subsystem to interface with the spacecraft to implement an activity's primitive actions. In addition to needing remote planning and execution for isolated spacecraft, a trend toward multiple-spacecraft missions points to the need for remote distributed planning and execution. The past few years have seen missions with growing numbers of probes. Pathfinder has its rover (Sojourner), Cassini has its lander (Huygens), and the New Millenium Deep Space 3 (DS3) proposal involves a constellation of 3 spacecraft for interferometric mapping. This trend is expected to continue to progressively larger fleets. For example, one mission proposed to succeed DS3 would have 18 spacecraft flying in formation in order to detect earth-sized planets orbiting other stars. A proposed magnetospheric constellation would involve 5 to 500 spacecraft in Earth orbit to measure global phenomena within the magnetosphere. This work describes and compares three autonomy architectures for a system that continuously plans to control a fleet of spacecraft using collective mission goals instead of goals or command sequences for each spacecraft. A fleet of self-commanding spacecraft would autonomously coordinate itself to satisfy high level science and engineering goals in a changing partially-understood environment making feasible the operation of tens or even a hundred spacecraft (such as for interferometry or plasma physics missions). The easiest way to adapt autonomous spacecraft research to controlling constellations involves treating the constellation as a single spacecraft. Here one spacecraft directly controls the others as if they were connected. The controlling "master" spacecraft performs all autonomy reasoning, and the slaves only have real-time subsystems to execute the master's commands and transmit local telemetry/observations. The executive/diagnostics module starts actions and the master's real-time subsystem controls the action either locally or remotely through a slave. While the master/slave approach benefits from conceptual simplicity, it relies on an assumption that the master spacecraft's executive can continuously monitor the slaves' real-time subsystems, and this relies on high-bandwidth highly-reliable communications. Since unintended results occur fairly rarely, one way to relax the bandwidth requirements involves only monitoring unexpected events in spacecraft. Unfortunately, this disables the ability to monitor for unexpected events between spacecraft and leads to a host of coordination problems among the slaves. Also, failures in the communications system can result in losing slaves. The other two architectures improve robustness while reducing communications by progressively distributing more of the other three remote agent components across the constellation. In a teamwork architecture, all spacecraft have executives and real-time subsystems - only the leader has the planner/scheduler and mission manager. Finally, distributing all remote agent components leads to a peer-to-peer approach toward constellation control.

Barrett, Anthony↗

Local prediction of Laser Powder Bed Fusion porosity by short-wave infrared imaging thermal feature porosity probability maps

We report that local thermal history can significantly vary in parts during metal Additive Manufacturing (AM), leading to local defects. However, the sequential layer-by-layer nature of AM facilitates in-situ part voxelmetric observations that can be used to detect and correct these defects for part qualification and quality control. The challenge is to relate this local radiometric data with local defect information to estimate process error likelihood in future builds. This paper uses a Short-Wave Infrared (SWIR) camera to record the temperature history for parts manufactured with Laser Powder Bed Fusion (LPBF) processes. The porosity from a cylindrical specimen is measured by ex-situ micro-computed tomography (μCT). Specimen data from the SWIR camera, combined with the μCT data, are used to generate thermal feature-based porosity probability maps. The porosity predictions made by various SWIR thermal feature-porosity probability maps of a specimen with a complex geometry are scored against the true porosity obtained via μCT. The receiver operating characteristic curves constructed from the predictions for the complex sample demonstrate the porosity probability mapping methodology’s potential for in-situ based porosity detection.

36 MATERIALS SCIENCE↗

Biology Inspired Approach for Communal Behavior in Sensor Networks

Research in wireless sensor network technology has exploded in the last decade. Promises of complex and ubiquitous control of the physical environment by these networks open avenues for new kinds of science and business. Due to the small size and low cost of sensor devices, visionaries promise systems enabled by deployment of massive numbers of sensors working in concert. Although the reduction in size has been phenomenal it results in severe limitations on the computing, communicating, and power capabilities of these devices. Under these constraints, research efforts have concentrated on developing techniques for performing relatively simple tasks with minimal energy expense assuming some form of centralized control. Unfortunately, centralized control does not scale to massive size networks and execution of simple tasks in sparsely populated networks will not lead to the sophisticated applications predicted. These must be enabled by new techniques dependent on local and autonomous cooperation between sensors to effect global functions. As a step in that direction, in this work we detail a technique whereby a large population of sensors can attain a global goal using only local information and by making only local decisions without any form of centralized control.

Jones, Kennie H.↗

Automation and AI in Space Drilling

Future planetary surface sampling missions, such as delving past the near-surface ice layers on Mars in search of organics and possibly signs of past/extant life, will require lightweight, low-mass planetary drilling and sample handling. Unlike terrestrial drills, these exploration drills must work dry (without drilling muds or gas), blind (no prior local or regional seismic or other surveys), and light (very low downward force or weight on bit, and perhaps 100 W available from solar power or batteries). Given the lightspeed transmission delays to Mars and outward, an exploratory planetary drill cannot be controlled directly from Earth. Drills that penetrate deeper than a few centimeters are likely to get stuck if operated open-loop (the MSL drill only penetrates 5 cm, and the MER Rock Abrasion Tools 5 mm by comparison), so some form of local drill control is required. In the relatively near-term, human crews cannot be presumed to be available for surface instrument teleoperation. Therefore highly automated drill and sample-transfer operations will be required, to explore the subsurface with the ability to safe robotic drilling systems and recover and continue on from the most probable fault conditions. Current automation, scheduling and diagnostic approaches will be discussed that roughly track the actions and roles of humans in terrestrial manual drilling operations.

drill automation↗

Tuning counterion chemistry to reduce carrier localization in doped semiconducting carbon nanotube networks

Understanding and controlling the impact that electrostatic interactions have on the transport of injected charge carriers is important for the utilization of pi-conjugated semiconductors in opto-electronic applications. Here, we explore the impact of dopant chemical and electronic structure on the doping efficacy and charge carrier transport in semiconducting single-walled carbon nanotube (s-SWCNT) networks using molecular charge-transfer dopants based on functionalized icosahedral dodecaborane (DDB) clusters. Calculations indicate that localization of electron density on the DDB core reduces the coulombic interactions that contribute to hole localization in the s-SWCNTs, thereby improving charge carrier transport. The enhanced delocalization produces an increase in the electrical conductivity and thermopower at lower charge carrier densities, yielding enhanced thermoelectric transport and a thermoelectric power factor that surpasses the previous best in class for enriched s-SWCNT thin-film networks. This strategy can be applied broadly across pi-conjugated semiconductors to tune and enhance performance in a variety of energy harvesting devices.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Limitations in Representation of Physical Processes Prevents Successful Simulation of PM2.5 During KORUS-AQ

High levels of fine particulate matter (PM2.5) pollution in East Asia often exceed local air quality standards. Observations from the Korea United States-Air Quality (KORUS-AQ) field campaign in May and June 2016 showed that development of extreme pollution (haze) occurred through a combination of long-range transport and favorable meteorological conditions that enhanced local production of PM2.5. Atmospheric models often have difficulty simulating PM2.5 chemical composition during haze, which is of concern for the development of successful control measures. We use observations from KORUS-AQ to examine the ability of the GEOS-Chem chemical transport model to simulate PM2.5 composition throughout the campaign and identify the mechanisms driving the pollution event. At the surface, the model underestimates sulfate by -64% but overestimates nitrate by +36%. The largest underestimate in sulfate occurs during the pollution event, where models typically struggle to generate elevated sulfate concentrations due to missing heterogeneous chemistry in aerosol liquid water in the polluted boundary layer. Hourly surface observations show that the model nitrate bias is driven by an overestimation of the nighttime peak. In the model, nitrate formation is limited by the supply of nitric acid, which is biased by +100% against aircraft observations. We hypothesize that this is due to a large missing sink, which we implement here as a factor of five increase in dry deposition. We show that the resulting increased deposition velocity is consistent with observations of total nitrate as a function of photochemical age. The model does not account for factors such as the urban heat island effect or the heterogeneity of the built-up urban landscape resulting in insufficient model turbulence and surface area over the study area that likely results in insufficient dry deposition. Other species such as NH3 could be similarly affected but were not measured during the campaign. Nighttime production of nitrate is driven by NO2 hydrolysis in the model, while observations show that unexpectedly elevated nighttime ozone (not present in the model) should result in N2O5 hydrolysis as the primary pathway. The model is unable to represent nighttime ozone due to an overly rapid collapse of the afternoon mixed layer and excessive titration by NO. We attribute this to missing nighttime heating driving deeper nocturnal mixing that would be expected to occur in a city like Seoul. This urban heating is not considered in air quality models run at large enough scales to treat both local chemistry and long-range transport. Key model failures in simulating nitrate, mainly overestimated daytime nitric acid, incorrect representation of nighttime chemistry, and an overly shallow and insufficiently turbulent nighttime mixed layer, exacerbate the model’s inability to simulate the buildup of PM2.5 during haze pollution. To address the underestimate in sulfate most evident during the haze event, heterogeneous aerosol uptake of SO2 is added to the model which previously only considered aqueous production of sulfate from SO2 in cloud water. Implementing a simple parameterization of this chemistry improves the model abundance of sulfate but degrades the SO2 simulation implying that emissions are underestimated. We find that improving model simulations of sulfate has direct relevance to determining local vs. transboundary contributions to PM2.5. During the haze pollution event, the inclusion of heterogeneous aerosol uptake of SO2 decreases the fraction of PM2.5 attributable to long-range transport from 66% to 54%. Locally-produced sulfate increased from 1% to 25% of locally-produced PM2.5, implying that local emissions controls would have a larger effect than previously thought. However, this additional uptake of SO2 is coupled to the model nitrate prediction which affects the aerosol liquid water abundance and chemistry driving sulfate-nitrate-ammonium partitioning. An additional simulation of the haze pollution with heterogeneous uptake of SO2 to aerosol and simple improvements to the model nitrate simulation results in 30% less sulfate due to 40% less nitrate and aerosol water, and results in an underestimate of sulfate during the haze event. Future studies need to better consider the impact of model physical processes such as dry deposition and nighttime boundary layer mixing on the simulation of nitrate and the effect of improved nitrate simulations on the overall simulation of secondary inorganic aerosol (sulfate+nitrate+ammonium) in East Asia. Foreign emissions are rapidly changing, increasing the need to understand the impact of local emissions on PM2.5 in South Korea to ensure continued air quality improvements.

Katherine R. Travis↗

Non-linear coherent mode interactions and the control of shear layers

A nonlinear integral formulation, based on local linear stability considerations, is used to study the collective interactions between discrete wave-modes associated with large-scale structures and the mean flow in a developing shear layer. Aspects of shear layer control are examined in light of the sensitivity of these interactions to the initial frequency parameter, modal energy contents and modal phases. Manipulation of the large-scale structure is argued to be an effective means of controlling the flow, including the small-scale turbulence dominated region far downstream. Cases of fundamental, 1st and 2nd subharmonic forcing are discussed in conjunction with relevant experiments.

Nikitopoulos, D. E.↗

Regional surrogates for predictive control of digital twins

Digital twins of complex systems must involve a model that is fast, generalizable, and usable for real-time control. For example, high-fidelity nonlinear multiphysics simulations can capture laser-material interactions, but are too slow for optimization or model predictive control (MPC). Reduced-order models, used to accelerate such computation, frequently fail to generalize to unseen inputs or control states. We show theoretically that this failure is intrinsic, i.e., that a learned model is non-unique outside the sampled subspace when its low-rank structure arises from limited excitation and clustered eigenvalues, rather than from a user-imposed truncation alone. Motivated by this result, we propose a control-ready regional surrogate-construction framework for both autonomous and nonautonomous dynamics; it employs Koopman lifting to represent nonlinearities, while preserving spatial locality. We illustrate our approach by constructing a control-ready surrogate for the digital twin of a thermal component of additive-manufacturing process. Our surrogate, localized in space through a von Neumann stencil, is learned from noisy high-fidelity simulations that emulate thermal-camera images collected during the manufacturing. It is linear in thermo-physically augmented states so that MPC reduces to a convex quadratic program. The surrogate requires no online correction, generalizes to unseen scan paths and power profiles of the laser, and is more than three orders of magnitude faster than a finite-difference solver. Furthermore, when the MPC sequence computed on the digital twin is applied to this solver, closed-loop temperature regulation is recovered, showing that the surrogate preserves control-relevant input-output behavior.

Data-driven model↗

Organizational Principles of Hyporheic Exchange Flow and Biogeochemical Cycling in River Networks Across Scales

Hyporheic zones increase freshwater ecosystem resilience to hydrological extremes and global environmental change. However, current conceptualizations of hyporheic exchange, residence time distributions, and the associated biogeochemical cycling in streambed sediments do not always accurately explain the hydrological and biogeochemical complexity observed in streams and rivers. Specifically, existing conceptual models insufficiently represent the coupled transport and reactivity along groundwater and surface water flow paths, the role of autochthonous organic matter in streambed biogeochemical functioning, and the feedbacks between surface-subsurface ecological processes, both within and across spatial and temporal scales. While simplified approaches to these issues are justifiable and necessary for transferability, the exclusion of important hyporheic processes from our conceptualizations can lead to erroneous conclusions and inadequate understanding and management of interconnected surface water and groundwater environments. This is particularly true at the landscape scale, where the organizational principles of spatio-temporal dynamics of hyporheic exchange flow (HEF) and biogeochemical processes remain largely uncharacterized. This article seeks to identify the most important drivers and controls of HEF and biogeochemical cycling based on a comprehensive synthesis of findings from a wide range of river systems. We use these observations to test current paradigms and conceptual models, discussing the interactions of local-to-regional hydrological, geomorphological, and ecological controls of hyporheic zone functioning. This improved conceptualization of the landscape organizational principles of drivers of HEF and biogeochemical processes from reach to catchment scales will inform future river research directions and watershed management strategies.

54 ENVIRONMENTAL SCIENCES↗

Aerodynamic control using forebody blowing and suction

Aerodynamic control using pneumatic forebody flow control was studied. Three methods of control were investigated: (1) blowing from a localized jet, (2) blowing from a slot, and (3) surface suction. Flow visualization and yawing moment measurements were performed on F/A-18 models in a water tunnel. The results show that all the methods were effective in controlling the forebody flow over a wide range of angles of attack and sideslip. The advantages and limitations of each of the control methods were discussed. The experiments suggested that all the control methods work basically on the principle of separation control. Based on the results of the water tunnel tests, the blowing or suction mass flow requirements appear to be within the limits of typical engine-bleed available from a modern fighter engine.

Ng, T. Terry↗

Resilient Autonomous Wind Farms: Preprint

With the advent of an increasing number of control strategies that seek to optimize wind turbine performance on a farm-level, taking account of individual wind turbine information to achieve wind farm-level objectives has become an increasingly important goal. Methods for controlling wind turbines on an individual and farm level have seen significant development, and an abundance of new implementations for gathering and using data from turbines have created potential for novel control mechanisms which can further optimize the performance and delivery characteristics of a wind farm. A key element of making these wind farms more efficient is to develop reliable algorithms that use local sensor information that is already being collected, such as supervisory control and data acquisition (SCADA) data, local meteorological stations, and nearby radars/sodars/lidars. Making use of information from all wind turbines in a wind farm can enable such approaches as determining the atmospheric conditions across the farm, improving fault-finding, and enabling more efficient overall control of farm-wide optimizations through mechanisms such as wake-steering. However, these approaches typically involve a centralized communications and control center. In order to ensure the resilient operation of the farm, it is necessary to develop an approach which distributes the calculation and communication amongst multiple nodes throughout the farm. In this fashion, a redundant, robust, and secure network can be created, which can tolerate faults in calculation, communication, and even external attacks which seek to disrupt the operation of the wind farm. This paper introduces the use of the Raft Byzantine Fault Tolerance algorithm in the implementation of autonomous control of a wind farm. This implementation will allow for fault tolerance for malfunctioning nodes, sensors, transmitters, and connectors. This approach is equally extensible to account for malicious actors. It will be shown to achieve overall consensus, provided the number of faults/malicious nodes is less than 3$n$+1, where $n$ is the number of turbine cluster faults which may occur, and to be robust in the face of multiple arbitrary faults.

autonomous↗

Lunar rovers and local positioning system

Telerobotic rovers equipped with adequate actuators and sensors are clearly necessary for extraterrestrial construction. They will be employed as substitutes for humans, to perform jobs like surveying, sensing, signaling, manipulating, and the handling of small materials. Important design criteria for these rovers include versatility and robustness. They must be easily programmed and reprogrammed to perform a wide variety of different functions, and they must be robust so that construction work will not be jeopardized by parts failures. The key qualities and functions necessary for these rovers to achieve the required versatility and robustness are modularity, redundancy, and coordination. Three robotic rovers are being built by CSC as a test bed to implement the concepts of modularity and coordination. The specific goal of the design and construction of these robots is to demonstrate the software modularity and multirobot control algorithms required for the physical manipulation of constructible elements. Each rover consists of a transporter platform, bus manager, simple manipulator, and positioning receivers. These robots will be controlled from a central control console via a radio-frequency local area network (LAN). To date, one prototype transporter platform frame was built with batteries, motors, a prototype single-motor controller, and two prototype internal LAN boards. Software modules were developed in C language for monitor functions, i/o, and parallel port usage in each computer board. Also completed are the fabrication of half of the required number of computer boards, the procurement of 19.2 Kbaud RF modems for inter-robot communications, and the simulation of processing requirements for positioning receivers. In addition to the robotic platform, the fabrication of a local positioning system based on infrared signals is nearly completed. This positioning system will make the rovers into a moving reference system capable of performing site surveys. In addition, a four degree mechanical manipulator especially suited for coordinated teleoperation was conceptually designed and is currently being analyzed. This manipulator will be integrated into the rovers as their end effector. Twenty internal LAN cards fabricated by a commercial firm are being used, a prototype manipulator and a range finder for a positioning system were built, a prototype two-motor controller was designed, and one of the robots is performing its first telerobotic motion. In addition, the robots' internal LAN's were coordinated and tested, hardware design upgrades based on fabrication and fit experience were completed, and the positioning system is running. The rover system is able to perform simple tasks such as sensing and signaling; coordination systems which allow construction tasks to begin were established, and soon coordinated teams of robots in the laboratory will be able to manipulate common objects.

Avery, James↗

Cascading a systolic array and a feedforward neural network for navigation and obstacle avoidance using potential fields

A technique is developed for vehicle navigation and control in the presence of obstacles. A potential function was devised that peaks at the surface of obstacles and has its minimum at the proper vehicle destination. This function is computed using a systolic array and is guaranteed not to have local minima. A feedfoward neural network is then used to control the steering of the vehicle using local potential field information. In this case, the vehicle is a trailer truck backing up. Previous work has demonstrated the capability of a neural network to control steering of such a trailer truck backing to a loading platform, but without obstacles. Now, the neural network was able to learn to navigate a trailer truck around obstacles while backing toward its destination. The network is trained in an obstacle free space to follow the negative gradient of the field, after which the network is able to control and navigate the truck to its target destination in a space of obstacles which may be stationary or movable.

Plumer, Edward S.↗

Streptococcus pneumoniae PstS production is phosphate responsive and enhanced during growth in the murine peritoneal cavity

Differential display-PCR (DDPCR) was used to identify a Streptococcus pneumoniae gene with enhanced transcription during growth in the murine peritoneal cavity. Northern dot blot analysis and comparative densitometry confirmed a 1.8-fold increase in expression of the encoded sequence following murine peritoneal culture (MPC) versus laboratory culture or control culture (CC). Sequencing and basic local alignment search tool analysis identified the DDPCR fragment as pstS, the phosphate-binding protein of a high-affinity phosphate uptake system. PCR amplification of the complete pstS gene followed by restriction analysis and sequencing suggests a high level of conservation between strains and serotypes. Quantitative immunodot blotting using antiserum to recombinant PstS (rPstS) demonstrated an approximately twofold increase in PstS production during MPC from that during CCs, a finding consistent with the low levels of phosphate observed in the peritoneum. Moreover, immunodot blot and Northern analysis demonstrated phosphate-dependent production of PstS in six of seven strains examined. These results identify pstS expression as responsive to the MPC environment and extracellular phosphate concentrations. Presently, it remains unclear if phosphate concentrations in vivo contribute to the regulation of pstS. Finally, polyclonal antiserum to rPstS did not inhibit growth of the pneumococcus in vitro, suggesting that antibodies do not block phosphate uptake; moreover, vaccination of mice with rPstS did not protect against intraperitoneal challenge as assessed by the 50% lethal dose.

Non-NASA Center↗