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NASA NTRS2003

Self-Supervised Dynamical Systems

Some progress has been made in a continuing effort to develop mathematical models of the behaviors of multi-agent systems known in biology, economics, and sociology (e.g., systems ranging from single or a few biomolecules to many interacting higher organisms). Living systems can be characterized by nonlinear evolution of probability distributions over different possible choices of the next steps in their motions. One of the main challenges in mathematical modeling of living systems is to distinguish between random walks of purely physical origin (for instance, Brownian motions) and those of biological origin. Following a line of reasoning from prior research, it has been assumed, in the present development, that a biological random walk can be represented by a nonlinear mathematical model that represents coupled mental and motor dynamics incorporating the psychological concept of reflection or self-image. The nonlinear dynamics impart the lifelike ability to behave in ways and to exhibit patterns that depart from thermodynamic equilibrium. Reflection or self-image has traditionally been recognized as a basic element of intelligence. The nonlinear mathematical models of the present development are denoted self-supervised dynamical systems. They include (1) equations of classical dynamics, including random components caused by uncertainties in initial conditions and by Langevin forces, coupled with (2) the corresponding Liouville or Fokker-Planck equations that describe the evolutions of probability densities that represent the uncertainties. The coupling is effected by fictitious information-based forces, denoted supervising forces, composed of probability densities and functionals thereof. The equations of classical mechanics represent motor dynamics that is, dynamics in the traditional sense, signifying Newton s equations of motion. The evolution of the probability densities represents mental dynamics or self-image. Then the interaction between the physical and metal aspects of a monad is implemented by feedback from mental to motor dynamics, as represented by the aforementioned fictitious forces. This feedback is what makes the evolution of probability densities nonlinear. The deviation from linear evolution can be characterized, in a sense, as an expression of free will. It has been demonstrated that probability densities can approach prescribed attractors while exhibiting such patterns as shock waves, solitons, and chaos in probability space. The concept of self-supervised dynamical systems has been considered for application to diverse phenomena, including information-based neural networks, cooperation, competition, deception, games, and control of chaos. In addition, a formal similarity between the mathematical structures of self-supervised dynamical systems and of quantum-mechanical systems has been investigated.

Zak, Michail↗

NASA NTRS1998

Autonomous Satellite Command and Control through the World Wide Web: Phase 3

NASA's New Millenium Program (NMP) has identified a variety of revolutionary technologies that will support orders of magnitude improvements in the capabilities of spacecraft missions. This program's Autonomy team has focused on science and engineering automation technologies. In doing so, it has established a clear development roadmap specifying the experiments and demonstrations required to mature these technologies. The primary developmental thrusts of this roadmap are in the areas of remote agents, PI/operator interface, planning/scheduling fault management, and smart execution architectures. Phases 1 and 2 of the ASSET Project (previously known as the WebSat project) have focused on establishing World Wide Web-based commanding and telemetry services as an advanced means of interfacing a spacecraft system with the PI and operators. Current automated capabilities include Web-based command submission, limited contact scheduling, command list generation and transfer to the ground station, spacecraft support for demonstrations experiments, data transfer from the ground station back to the ASSET system, data archiving, and Web-based telemetry distribution. Phase 2 was finished in December 1996. During January-December 1997 work was commenced on Phase 3 of the ASSET Project. Phase 3 is the subject of this report. This phase permitted SSDL and its project partners to expand the ASSET system in a variety of ways. These added capabilities included the advancement of ground station capabilities, the adaptation of spacecraft on-board software, and the expansion of capabilities of the ASSET management algorithms. Specific goals of Phase 3 were: (1) Extend Web-based goal-level commanding for both the payload PI and the spacecraft engineer; (2) Support prioritized handling of multiple PIs as well as associated payload experimenters; (3) Expand the number and types of experiments supported by the ASSET system and its associated spacecraft; (4) Implement more advanced resource management, modeling and fault management capabilities that integrate the space and ground segments of the space system hardware; (5) Implement a beacon monitoring test; (6) Implement an experimental blackboard controller for space system management; (7) Further define typical ground station developments required for Internet-based remote control and for full system automation of the PI-to-spacecraft link. Each of those goals is examined in the next section. Significant sections of this report were also published as a conference paper.

Cantwell, Brian↗

NASA NTRS2002

Cancer Risk Assessment for Space Radiation

Predicting the occurrence of human cancer following exposure to any agent causing genetic damage is a difficult task. This is because the uncertainty of uniform exposure to the damaging agent, and the uncertainty of uniform processing of that damage within a complex set of biological variables, degrade the confidence of predicting the delayed expression of cancer as a relatively rare event within any given clinically normal individual. The radiation health research priorities for enabling long-duration human exploration of space were established in the 1996 NRC Report entitled "Radiation Hazards to Crews of Interplanetary Missions: Biological Issues and Research Strategies". This report emphasized that a 15-fold uncertainty in predicting radiation-induced cancer incidence must be reduced before NASA can commit humans to extended interplanetary missions. That report concluded that the great majority of this uncertainty is biologically based, while a minority is physically based due to uncertainties in radiation dosimetry and radiation transport codes. Since that report, the biologically based uncertainty has remained large, and the relatively small uncertainty associated with radiation dosimetry has increased due to the considerations raised by concepts of microdosimetry. In a practical sense, however, the additional uncertainties introduced by microdosimetry are encouraging since they are in a direction of lowered effective dose absorbed through infrequent interactions of any given cell with the high energy particle component of space radiation. The biological uncertainty in predicting cancer risk for space radiation derives from two primary facts. 1) One animal tumor study has been reported that includes a relevant spectrum of particle radiation energies, and that is the Harderian gland model in mice. Fact #1: Extension of cancer risk from animal models, and especially from a single study in an animal model, to humans is inherently uncertain. 2) One human database is predominantly used for assessing cancer risk caused by space radiation, and that is the Japanese atomic bomb survivors. Fact #2: The atomic-bomb-survivor database, itself a remarkable achievement, contains uncertainties. These include the actual exposure to each individual, the radiation quality of that exposure, and the fact that the exposure was to acute doses of predominantly low-LET radiation, not to chronic exposures of high-LET radiation expected on long-duration interplanetary manned missions.

Richmond, Robert C.↗

NASA NTRSDate not supplied

Sea Surface Air Pressure Retrieval with MBARS

Global meteorological observations are essential in initialization and assimilation of numerical weather prediction (NWP) models. Many key weather variables such as temperature, humidity, and wind can be relatively well observed by in-situ and remote techniques from surface, suborbital and orbital platforms. However, sea surface air pressure cannot be well-sampled. Over open oceans, the pressure can only be observed by very limited buoys, ships, and oceanic platforms. There is no operational remote sensing method available for the crucial dynamic variable of the Earth’s climate and weather systems. Advanced techniques for global sea level pressure (SLP) observations are urgently needed for improving NWP models, particularly for improvement of severe weather forecasts. Recent studies on the space and airborne Microwave BArometric Radar and Sounder (MBARS) systems provide a great potential to fill the SLP observational gap. The systems studied have three channels with center frequencies at 65.5, 67.76 and 70 GHz. These channels are designed to operate at atmospheric oxygen (O2) absorption bands for column O2 mass measurements. When sea surface returns of spectrally closely-spaced paired channels are measured, surface reflection and atmospheric attenuation other than O2 are very similar, but the difference in O2 absorption is substantial. Thus, the differential absorption loss of this frequency pair leads to measures of atmospheric O2 amount. For the residual absorption loss caused by other agents, a third frequency having the same spectral space to the weak absorption frequency of the first pair can be used, which forms the second pair and produces the same residual as the first pair. Removal of the residual of this second pair from the first pair can further mitigate environmental impacts. Thus, this 3-channel approach will provide precise O2 amount retrievals. Since oxygen is well mixed in the atmosphere, the column air mass or SLP can be obtained. Currently, MBARS is rapidly maturing its advanced technology. An upcoming airborne field flight campaign is planned in late 2024. The 3-channel approach will be use in SLP retrievals. This MBARS team will discuss the physics-based SLP retrieval technique and report latest progresses in this upcoming AGU annual meeting.

O2 differential absorption↗

NASA NTRS2014

Virtualization - A Key Cost Saver in NASA Multi-Mission Ground System Architecture

With science team budgets being slashed, and a lack of adequate facilities for science payload teams to operate their instruments, there is a strong need for innovative new ground systems that are able to provide necessary levels of capability processing power, system availability and redundancy while maintaining a small footprint in terms of physical space, power utilization and cooling.The ground system architecture being presented is based off of heritage from several other projects currently in development or operations at Goddard, but was designed and built specifically to meet the needs of the Science and Planetary Operations Control Center (SPOCC) as a low-cost payload command, control, planning and analysis operations center. However, this SPOCC architecture was designed to be generic enough to be re-used partially or in whole by other labs and missions (since its inception that has already happened in several cases!)The SPOCC architecture leverages a highly available VMware-based virtualization cluster with shared SAS Direct-Attached Storage (DAS) to provide an extremely high-performing, low-power-utilization and small-footprint compute environment that provides Virtual Machine resources shared among the various tenant missions in the SPOCC. The storage is also expandable, allowing future missions to chain up to 7 additional 2U chassis of storage at an extremely competitive cost if they require additional archive or virtual machine storage space.The software architecture provides a fully-redundant GMSEC-based message bus architecture based on the ActiveMQ middleware to track all health and safety status within the SPOCC ground system. All virtual machines utilize the GMSEC system agents to report system host health over the GMSEC bus, and spacecraft payload health is monitored using the Hammers Integrated Test and Operations System (ITOS) Galaxy Telemetry and Command (TC) system, which performs near-real-time limit checking and data processing on the downlinked data stream and injects messages into the GMSEC bus that are monitored to automatically page the on-call operator or Systems Administrator (SA) when an off-nominal condition is detected. This architecture, like the LTSP thin clients, are shared across all tenant missions.Other required IT security controls are implemented at the ground system level, including physical access controls, logical system-level authentication authorization management, auditing and reporting, network management and a NIST 800-53 FISMA-Moderate IT Security plan Risk Assessment Contingency Plan, helping multiple missions share the cost of compliance with agency-mandated directives.The SPOCC architecture provides science payload control centers and backup mission operations centers with a cost-effective, standardized approach to virtualizing and monitoring resources that were traditionally multiple racks full of physical machines. The increased agility in deploying new virtual systems and thin client workstations can provide significant savings in personnel costs for maintaining the ground system. The cost savings in procurement, power, rack footprint and cooling as well as the shared multi-mission design greatly reduces upfront cost for missions moving into the facility. Overall, the authors hope that this architecture will become a model for how future NASA operations centers are constructed!

Ground System Architecture↗

NASA NTRS2015

Forest Disturbance Across the Conterminous United States from 1985-2012: The Emerging Dominance of Forest Decline

Evidence of shifting dominance among major forest disturbance agent classes regionally to globally has been emerging in the literature. For example, climate-related stress and secondary stressors on forests (e.g., insect and disease, fire) have dramatically increased since the turn of the century globally, while harvest rates in the western US and elsewhere have declined. For shifts to be quantified, accurate historical forest disturbance estimates are required as a baseline for examining current trends. We report annual disturbance rates (with uncertainties) in the aggregate and by major change causal agent class for the conterminous US and five geographic subregions between 1985 and 2012. Results are based on human interpretations of Landsat time series from a probability sample of 7200 plots (30 m) distributed throughout the study area. Forest disturbance information was recorded with a Landsat time series visualization and data collection tool that incorporates ancillary high-resolution data. National rates of disturbance varied between 1.5% and 4.5% of forest area per year, with trends being strongly affected by shifting dominance among specific disturbance agent influences at the regional scale. Throughout the time series, national harvest disturbance rates varied between one and two percent, and were largely a function of harvest in the more heavily forested regions of the US (Mountain West, Northeast, and Southeast). During the first part of the time series, national disturbance rates largely reflected trends in harvest disturbance. Beginning in the mid-90s, forest decline-related disturbances associated with diminishing forest health (e.g., physiological stress leading to tree canopy cover loss, increases in tree mortality above background levels), especially in the Mountain West and Lowland West regions of the US, increased dramatically. Consequently, national disturbance rates greatly increased by 2000, and remained high for much of the decade. Decline-related disturbance rates reached as high as 8% per year in the western regions during the early-2000s. Although low compared to harvest and decline, fire disturbance rates also increased in the early- to mid-2000s. We segmented annual decline-related disturbance rates to distinguish between newly impacted areas and areas undergoing gradual but consistent decline over multiple years. We also translated Landsat reflectance change into tree canopy cover change information for greater relevance to ecosystem modelers and forest managers, who can derive better understanding of forest-climate interactions and better adapt management strategies to changing climate regimes. Similar studies could be carried out for other countries where there are sufficient Landsat data and historic temporal snapshots of high-resolution imagery.

Cohen, Warren B.↗

NASA NTRSDate not supplied

Accuracy Assessment of Two Gps Fidelity Prediction Services in Urban Terrain

Low altitude flight in urban areas is susceptible to degraded GNSS-based navigation system performance due to terrain interference with radio signals from orbital positioning satellites. Predictive navigation performance fidelity tools are needed a) in preflight planning to assist in the creation of safe flight paths and b) in-flight to provide contingency management agents with the navigation risk of proximal flight corridors. Two navigation fidelity prediction services are validated by comparison with over 6000 readings from GNSS sensors collected along a five-mile path through urban areas of Corpus Christi, Texas, on three dates in 2022. Predictions are based on satellite line of sight through 3D terrain data collected in 2018. Each service predicts a set of navigation fidelity metrics over a user-specified time period. One metric estimated by both is the number of visible satellites. A direct comparison of the number of predicted visible satellites with the number sensed by the receiver is used to validate the prediction services. Results show an exact match in the number of predicted satellites for 60% of the measurements, and a match within +/- 4 satellites for 95% of the measurements. As expected, agreement improves away from vertical blocking terrain. Most cases of mismatch are due a lower predicted count than measured (false negatives), and can be accounted for by receiver pickup of stray signals caused by multipath propagation. About 10% of mismatches are false positives and are mostly accounted for by foliage effects. The two services predict visibility of the same set of satellites 80% of the time, differ by two or less satellites 95% of the time, and can compute predictions for one hour of observations in one minute or less. Validation is analyzed statistically and in detailed case studies of selected observation times. The prediction services validated in this study run fast enough for preflight safety planning. The more stringent challenge of inflight navigation fidelity prediction for contingency management requires both a speedup of the current level of modeling and equally fast stray signal modeling.

Andrew Moore↗

NASA NTRSDate not supplied

Accuracy Assessment of Two GPS Fidelity Prediction Services in Urban Terrain

Andrew J. Moore↗

NASA NTRS2012

Impact of Aerosols on Convective Clouds and Precipitation

Aerosols are a critical.factor in the atmospheric hydrological cycle and radiation budget. As a major agent for clouds to form and a significant attenuator of solar radiation, aerosols affect climate in several ways. Current research suggests that aerosols have a major impact on the dynamics, microphysics, and electrification properties of continental mixed-phase convective clouds. In addition, high aerosol concentrations in urban environments could affect precipitation variability by providing a significant source of cloud condensation nuclei (CCN). Such pollution . effects on precipitation potentially have enormous climatic consequences both in terms of feedbacks involving the land surface via rainfall as well as the surface energy budget and changes in latent heat input to the atmosphere. Basically, aerosol concentrations can influence cloud droplet size distributions, the warm-rain process, the cold-rain process, cloud-top heights, the depth of the mixed-phase region, and the occurrence of lightning. Recently, many cloud resolution models (CRMs) have been used to examine the role of aerosols on mixed-phase convective clouds. These modeling studies have many differences in terms of model configuration (two- or three-dimensional), domain size, grid spacing (150-3000 m), microphysics (two-moment bulk, simple or sophisticated spectral-bin), turbulence (1st or 1.5 order turbulent kinetic energy (TKE)), radiation, lateral boundary conditions (i.e., closed, radiative open or cyclic), cases (isolated convection, tropical or midlatitude squall lines) and model integration time (e.g., 2.5 to 48 hours). Among these modeling studies, the most striking difference is that cumulative precipitation can either increase or decrease in response to higher concentrations of CCN. In this presentation, we review past efforts and summarize our current understanding of the effect of aerosols on convective precipitation processes. Specifically, this paper addresses the following topics: observational evidence of the effect of aerosols on precipitation processes, and results from (CRM) simulations. Note that this presentation is mainly based on a recent paper published in Geophy. Rev. (Tao et al. 2012).

Tao, Wei-Kuo↗

NASA NTRS2004

A Survey of Formal Methods for Intelligent Swarms

Swarms of intelligent autonomous spacecraft, involving complex behaviors and interactions, are being proposed for future space exploration missions. Such missions provide greater flexibility and offer the possibility of gathering more science data than traditional single spacecraft missions. The emergent properties of swarms make these missions powerful, but simultaneously far more difficult to design, and to assure that the proper behaviors will emerge. These missions are also considerably more complex than previous types of missions, and NASA, like other organizations, has little experience in developing or in verifying and validating these types of missions. A significant challenge when verifying and validating swarms of intelligent interacting agents is how to determine that the possible exponential interactions and emergent behaviors are producing the desired results. Assuring correct behavior and interactions of swarms will be critical to mission success. The Autonomous Nano Technology Swarm (ANTS) mission is an example of one of the swarm types of missions NASA is considering. The ANTS mission will use a swarm of picospacecraft that will fly from Earth orbit to the Asteroid Belt. Using an insect colony analogy, ANTS will be composed of specialized workers for asteroid exploration. Exploration would consist of cataloguing the mass, density, morphology, and chemical composition of the asteroids, including any anomalous concentrations of specific minerals. To perform this task, ANTS would carry miniaturized instruments, such as imagers, spectrometers, and detectors. Since ANTS and other similar missions are going to consist of autonomous spacecraft that may be out of contact with the earth for extended periods of time, and have low bandwidths due to weight constraints, it will be difficult to observe improper behavior and to correct any errors after launch. Providing V&V (verification and validation) for this type of mission is new to NASA, and represents the cutting edge in system correctness, and requires higher levels of assurance than other (traditional) missions that use a single or small number of spacecraft that are deterministic in nature and have near continuous communication access. One of the highest possible levels of assurance comes from the application of formal methods. Formal methods are mathematics-based tools and techniques for specifying and verifying (software and hardware) systems. They are particularly useful for specifying complex parallel systems, such as exemplified by the ANTS mission, where the entire system is difficult for a single person to fully understand, a problem that is multiplied with multiple developers. Once written, a formal specification can be used to prove properties of a system (e.g., the underlying system will go from one state to another or not into a specific state) and check for particular types of errors (e.g., race or livelock conditions). A formal specification can also be used as input to a model checker for further validation. This report gives the results of a survey of formal methods techniques for verification and validation of space missions that use swarm technology. Multiple formal methods were evaluated to determine their effectiveness in modeling and assuring the behavior of swarms of spacecraft using the ANTS mission as an example system. This report is the first result of the project to determine formal approaches that are promising for formally specifying swarm-based systems. From this survey, the most promising approaches were selected and are discussed relative to their possible application to the ANTS mission. Future work will include the application of an integrated approach, based on the selected approaches identified in this report, to the formal specification of the ANTS mission.

Truszkowski, Walt↗

NASA NTRS1999

Three-Dimensional, Transgenic Cell Models to Quantify Space Genotoxic Effects

The space environment contains radiation and chemical agents known to be mutagenic and carcinogenic to humans. Additionally, microgravity is a complicating factor that may modify or synergize induced genotoxic effects. Most in vitro models fail to use human cells (making risk extrapolation to humans more difficult), overlook the dynamic effect of tissue intercellular interactions on genotoxic damage, and lack the sensitivity required to measure low-dose effects. Currently a need exists for a model test system that simulates cellular interactions present in tissue, and can be used to quantify genotoxic damage induced by low levels of radiation and chemicals, and extrapolate assessed risk to humans. A state-of-the-art, three-dimensional, multicellular tissue equivalent cell culture model will be presented. It consists of mammalian cells genetically engineered to contain multiple copies of defined target genes for genotoxic assessment,. NASA-designed bioreactors were used to coculture mammalian cells into spheroids, The cells used were human mammary epithelial cells (H184135) and Stratagene's (Austin, Texas) Big Blue(TM) Rat 2 lambda fibroblasts. The fibroblasts were genetically engineered to contain -a high-density target gene for mutagenesis (60 copies of lacl/LacZ per cell). Tissue equivalent spheroids were routinely produced by inoculation of 2 to 7 X 10(exp 5) fibroblasts with Cytodex 3 beads (150 micrometers in diameter). at a 20:1 cell:bead ratio, into 50-ml HARV bioreactors (Synthecon, Inc.). Fibroblasts were cultured for 5 days, an equivalent number of epithelial cells added, and the fibroblast/epithelial cell coculture continued for 21 days. Three-dimensional spheroids with diameters ranging from 400 to 600 micrometers were obtained. Histological and immunohistochemical Characterization revealed i) both cell types present in the spheroids, with fibroblasts located primarily in the center, surrounded by epithelial cells; ii) synthesis of extracellular matrix; and iii,, mitotic cells located throughout the spheroids. Spheroidal integrity and cell viability were retained for the 30-day test period after removal of spheroids from the bioreactor. Potential utility of this three-dimensional, transgenic model for genotoxicity was initially assessed by exposure of spheroids to 0-2 Gy neon at dose rates of 0.3 to 1.5 Gy/min (National Institute of Radiological Sciences, Chiba, Japan). Quantification of mutation at the lacl gene revealed a linear dose response for mutation induction. Limited sequencing analysis of mutant clones revealed higher frequencies of deletions and multiple base sequence changes with increasing dose. These results suggest that our three-dimensional, transgenic model is applicable to a wide variety of studies involving the quantification, identification, and characterization of genotoxicity incurred in space and on Earth. This model uniquely allows investigation of the interaction of relevant factors, namely cell-to-cell interactions and the mechanistic interaction of microgravity with radiation insults and DNA repair. Using this three-dimensional model will allow us to obtain dual genotoxic information (i.e., mutation rate plus chromosome aberration data) from the same system so that one endpoint can be used to reference the other, thereby increasing the fidelity of the data set. Moreover, the tissue-equivalent nature of the three-dimensional model provides high confidence for relevance of risk assessment, i.e., the establishment of quality factors directly applicable to the microgravity environment.

Gonda, S. R.↗

NASA NTRS2009

Microbial Risk Assessment

Historically, microbiological spaceflight requirements have been established in a subjective manner based upon expert opinion of both environmental and clinical monitoring results and the incidence of disease. The limited amount of data, especially from long-duration missions, has created very conservative requirements based primarily on the concentration of microorganisms. Periodic reevaluations of new data from later missions have allowed some relaxation of these stringent requirements. However, the requirements remain very conservative and subjective in nature, and the risk of crew illness due to infectious microorganisms is not well defined. The use of modeling techniques for microbial risk has been applied in the food and potable water industries and has exceptional potential for spaceflight applications. From a productivity standpoint, this type of modeling can (1) decrease unnecessary costs and resource usage and (2) prevent inadequate or inappropriate data for health assessment. In addition, a quantitative model has several advantages for risk management and communication. By identifying the variable components of the model and the knowledge associated with each component, this type of modeling can: (1) Systematically identify and close knowledge gaps, (2) Systematically identify acceptable and unacceptable risks, (3) Improve communication with stakeholders as to the reasons for resource use, and (4) Facilitate external scientific approval of the NASA requirements. The modeling of microbial risk involves the evaluation of several key factors including hazard identification, crew exposure assessment, dose-response assessment, and risk characterization. Many of these factors are similar to conditions found on Earth; however, the spaceflight environment is very specialized as the inhabitants live in a small, semi-closed environment that is often dependent on regenerative life support systems. To further complicate modeling efforts, microbial dose-response characteristics may be affected by a potentially dysfunctional crew immune system during a mission. In addition, microbial virulence has been shown to change under certain conditions during spaceflight, further complicating dose-response characterization. An initial study of the applicability of microbial risk assessment techniques was performed using Crew Health Care System (CHeCS) operational data from the International Space Station potable water systems. The risk of infection from potable water was selected as the flight systems and microbial ecology are well defined. This initial study confirmed the feasibility of using microbial risk assessment modeling for spaceflight systems. While no immediate threat was detected, the study identified several medically significant microorganisms that could pose a health risk if uncontrolled. The study also identified several specific knowledge gaps in making a risk assessment and noted that filling these knowledge gaps is essential as the risk estimates may change by orders of magnitude depending on the answers. The current phase of the microbial risk assessment studies focuses on the dose-response relationship of specific infectious agents, focusing on Salmonella enterica Typhimurium, Pseudomonas spp., and Escherichia coli, as their evaluation will provide a better baseline for determining the overall hazard characterization. The organisms were chosen as they either have been isolated on spacecraft or have an identified route of infection during a mission. The characterization will utilize dose-response models selected either from the peer-reviewed literature and/or by using statistical approaches. Development of these modeling and risk assessment techniques will help to optimize flight requirements and to protect the safety, health, and performance of the crew.

Ott, C. M.↗

NASA NTRS2000

Enhanced expression of extracellular calcium sensing receptor in monocyte-differentiated versus undifferentiated HL-60 cells: potential role in regulation of a nonselective cation channel

Human promyelocytic leukemia cells (HL-60) have been used widely as a model for studying the differentiation of hematopoietic progenitor cells in vitro. After treatment with phorbol-12-myristate-13-acetate (PMA) or 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], HL-60 cells differentiate into cells with the phenotype of monocytes/macrophages. We previously showed that peripheral blood monocytes and the murine J774 monocytic cell line express the CaR, and myeloid progenitors in the bone marrow and myeloid cells in peripheral blood other than monocytes express lower levels of the CaR. Therefore, we investigated whether undifferentiated HL-60 cells express a functional G protein-coupled, extracellular calcium (Ca(2+)(o))-sensing receptor (CaR) and if the expression of the CaR increases as these cells differentiate along the monocytic lineage. The use of reverse transcription-polymerase chain reaction (RT-PCR) with CaR-specific primers, followed by sequencing of the amplified products, identified an authentic CaR transcript in undifferentiated HL-60 cells. Both immunocytochemistry and Western blot analysis using a CaR-specific antiserum detected low levels of CaR protein expression in undifferentiated HL-60 cells. The levels of CaR protein increased considerably following treatment of the cells with PMA (50 nM) or 1,25(OH)(2)D(3) (100 nM) for 5 days. Northern analysis using a CaR-specific riboprobe identified CaR transcripts in undifferentiated HL-60 cells, but CaR mRNA levels did not change appreciably after treatment with either agent, suggesting that upregulation of CaR protein occurs at a translational level. PMA-treated HL-60 cells expressed a nonselective cation channel (NCC), and the calcimimetic CaR activator, NPS R-467, but not its less active stereoisomer, NPS S-467, as well as the polycationic CaR agonist, neomycin, activated this NCC, demonstrating that the CaR expressed in these cells is functionally active. Therefore, HL-60 cells exhibit an increase in CaR protein expression, occurring at a translational level during their differentiation into cells with a monocyte/macrophage phenotype in response to treatment with PMA or 1, 25(OH)(2)D(3), which is functionally linked to activation of a nonselective cation channel.

NASA Discipline Musculoskeletal↗