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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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All sourcesNASA NTRSDOE OSTI
NASA NTRS1999

Soil Analysis Micro-Mission Concepts Derived from the MSP 2001 Mars Environmental Compatibility Assessment (MECA)

The Mars Environmental Compatibility Assessment (MECA) will evaluate the Martian environment for soil and dust-related hazards to human exploration as part of the Mars Surveyor Program 2001 Lander. The integrated MECA payload contains a wet-chemistry laboratory, a microscopy station, an electrometer to characterize the electrostatic environment, and arrays of material patches to study abrasion and adhesion. Heritage will be all-important for low cost micro-missions, and adaptations of instruments developed for the Pathfinder, '98 and '01 Landers should be strong contenders for '03 flights. This talk has three objectives: (1) Familiarize the audience with MECA instrument capabilities; (2) present concepts for stand-alone and/or mobile versions of MECA instruments; and (3) broaden the context of the MECA instruments from human exploration to a comprehensive scientific survey of Mars. Due to time limitations, emphasis will be on the chemistry and microscopy experiments. Ion-selective electrodes and related sensors in MECA's wet-chemistry laboratory will evaluate total dissolved solids, redox potential, pH, and the concentration of many soluble ions and gases in wet Martian soil. These electrodes can detect potentially dangerous heavy-metal ions, emitted pathogenic gases, and the soil's corrosive potential, and experiments will include cyclic voltammetry and anodic stripping. For experiments beyond 2001, enhancements could allow multiple use of the cells (for mobile experiments) and reagent addition (for quantitative mineralogical and exobiological analysis). MECA's microscopy station combines optical and atomic-force microscopy (AFM) in an actively focused, controlled illumination environment to image particles from millimeters to nanometers in size. Careful selection of substrates allows controlled experiments in adhesion, abrasion, hardness, aggregation, magnetic and other properties. Special tools allow primitive manipulation (brushing and scraping) of samples. Soil particle properties including size, shape, color, hardness, adhesive potential (electrostatic and magnetic), will be determined using an array of sample receptacles and collection substrates. The simple, rugged atomic-force microscope will image in the submicron size range and has the capability of performing a particle-by-particle analysis of the dust and soil. Future implementations might enhance the optical microscopy with spectroscopy, or incorporate advanced AFM techniques for thermogravimetric and chemical analysis.

Hecht, M. H.↗
NASA NTRS2021

A GTP-synthesizing ribozyme selected by metabolic coupling to an RNA polymerase ribozyme

Synthesis of RNA in early life forms required chemically activated nucleotides, perhaps in the same form of nucleoside 5′-triphosphates (NTPs) as in the contemporary biosphere. We show the development of a catalytic RNA (ribozyme) that generates the nucleoside triphosphate guanosine 5′-triphosphate (GTP) from the nucleoside guanosine and the prebiotically plausible cyclic trimetaphosphate. Ribozymes were selected from 1.6 × 1014 different randomized sequences by metabolically coupling 6-thio GTP synthesis to primer extension by an RNA polymerase ribozyme within 1016 emulsion droplets. Several functional RNAs were identified, one of which was characterized in more detail. Under optimized reaction conditions, this ribozyme produced GTP at a rate 18,000-fold higher than the uncatalyzed rate, with a turnover of 1.7-fold, and supported the incorporation of GTP into RNA oligomers in tandem with an RNA polymerase ribozyme. These results are discussed in the context of early life forms.

Arvin Akoopie↗
NASA NTRS2001

A Study of the Response of Deep Tropical Clouds to Mesoscale Processes: Modeling Strategies and Simulations of TOGA-COARE Convective Systems - Part 1

Interactions between deep tropical clouds over the western Pacific warm pool and the larger-scale environment are key to understanding climate change. Cloud models are an extremely useful tool in simulating and providing statistical information on heat and moisture transfer processes between cloud systems and the environment, and can therefore be utilized to substantially improve cloud parameterizations in climate models. In this paper, the Goddard Cumulus Ensemble (GCE) cloud-resolving model is used in multi-day simulations of deep tropical convective activity over the Tropical Ocean-Global Atmosphere Coupled Ocean-Atmosphere Response Experiment (TOGA COARE). Large-scale temperature and moisture advective tendencies, and horizontal momentum from the TOGA-COARE Intensive Flux Array (IFA) region, are applied to the GCE version which incorporates cyclical boundary conditions. Sensitivity experiments show that grid domain size produces the largest response to domain-mean temperature and moisture deviations, as well as cloudiness, when compared to grid horizontal or vertical resolution, and advection scheme. It is found that a minimum grid-domain size of 500 km is needed to adequately resolve the convective cloud features. The control experiment shows that the atmospheric heating and moistening is primarily a response to cloud latent processes of condensation/evaporation, and deposition/sublimation, and to a lesser extent, melting of ice particles. Air-sea exchange of heat and moisture is found to be significant, but of secondary importance, while the radiational response is small. The simulated rainfall and atmospheric heating and moistening, agrees well with observations, and performs favorably to other models simulating this case.

Johnson, Daniel E.↗
NASA NTRS2016

Mechanotransductive Regulation of Gap-Junction Activity Between MLO-Y4 Osteocyte-Like and MC3T3-E1 Osteoblast-Like Cells in Three-Dimensional Co-Culture.

Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesion/attachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by immunohistological observation. Increased CX43 expression is also observed with the three-dimensional co-cultures with MC3T3-E1 MCS stimulation but the increased gap-junction protein presence was limited to the osteoblast-osteocyte interface region. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggests an osteocyte-osteoblast gap-junction signaling feedback mechanism may regulate mechanotransduction of apoptosis initiation and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.

mechanical loading↗
NASA NTRS2016

Mechanotransductive Regulation of Gap-Junction Activity Between MLO-Y4 Osteocyte-Like and MC3T3-E1 Osteoblast-Like Cells in Three-Dimensional Co-Culture

Cell and animal studies conducted onboard the International Space Station and formerly on Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. However the intercellular communicative mechanisms associated with the regulation of bone synthesis and bone resorption cells are still largely unknown. Connexin-43 (CX43), a gap junction protein, is hypothesized to play a significant role in osteoblast and osteocyte signaling. The purpose of this investigation was to evaluate within a novel three-dimensional microenvironment how the osteocyte-osteoblast gap-junction expression changes when cultures are exposed to exaggerated mechanical load. MLO-Y4 osteocyte-like cells were cultured on a 3D-Biotek polystyrene insert and placed in direct contact with an MC3T3-E1 pre-osteoblast co-cultured monolayer and exposed to 48 h of mechanical stimulation (pulsatile fluid flow (PFF) or monolayer cyclic stretch (MCS)) then evaluated for viability, proliferation, metabolism, and CX43 expression. Mono-cultured MLO-Y4 and MC3T3-E1 control experiments were conducted under PFF and MCS stimulation to observe how strain application stimuli (PFF cell membrane shear or MCS cell focal adhesionattachment loading) initiates different signaling pathways or downstream regulatory controls. TotalLive cell count, viability and metabolic reduction (Trypan Blue, LIVEDead and Alamar Blue analysis respectively) indicate that mechanical activation of MC3T3-E1 cells inhibits proliferation while maintaining an average 1.04E4 reductioncell metabolic rate, *p0.05 n4. MLO-Y4s in monolayer culture increase in number when exposed to MCS loading but the percent of live cells within the population is low (46.3 total count, *p0.05 n4), these results may indicate an apoptotic signaling cascade. PFF stimulation of the three-dimensional co-cultures elicits a universal increase in CX43 in MLO-Y4 and MC3T3-E1 cells, illustrated by immunohistological observation. Increased CX43 expression is also observed with the three-dimensional co-cultures with MC3T3-E1 MCS stimulation but the increased gap-junction protein presence was limited to the osteoblast-osteocyte interface region. Previously reported PCR evaluation of osteogenic markers further corroborate that the co-cultured populations communicative networks play a role in translating mechanical signals to molecular messaging. These findings suggests an osteocyte-osteoblast gap-junction signaling feedback mechanism may regulate mechanotransduction of apoptosis initiation and transcription of cytokine signaling proteins responsible for stem cell niche recruitment much more directly than previously believed.

Juran, C. M.↗
NASA NTRS1975

Design and test status for life support applications of SPE oxygen generation systems

An advanced six-man rated oxygen generation system has been fabricated and tested as part of a NASA/JSC technology development program for a long lived, manned spacecraft life support system. Details of the design and tests results are presented. The system is based on the Solid Polymer Electrolyte (SPE) water electrolysis technology and its nominal operating conditions are 2760 kN/sq m (400 psia) and 355 K (180 F) with an electrolysis module current density capability up to 350 mA/sq cm (326 ASF). The system is centered on a 13-cell SPE water electrolysis module having a single cell active area of 214 sq cm (33 sq in) and it incorporates instrumentation and controls for single pushbutton automatic startup/shutdown, component fault detection and isolation, and self-contained sensors and controls for automatic safe emergency shutdown. The system has been tested in both the orbital cyclic and continuous mode of operation. Various parametric tests have been completed to define the system capability for potential application in spacecraft environmental systems.

Titterington, W. A.↗
NASA NTRS2012

Sensor-Only System Identification for Structural Health Monitoring of Advanced Aircraft

Environmental conditions, cyclic loading, and aging contribute to structural wear and degradation, and thus potentially catastrophic events. The challenge of health monitoring technology is to determine incipient changes accurately and efficiently. This project addresses this challenge by developing health monitoring techniques that depend only on sensor measurements. Since actively controlled excitation is not needed, sensor-to-sensor identification (S2SID) provides an in-flight diagnostic tool that exploits ambient excitation to provide advance warning of significant changes. S2SID can subsequently be followed up by ground testing to localize and quantify structural changes. The conceptual foundation of S2SID is the notion of a pseudo-transfer function, where one sensor is viewed as the pseudo-input and another is viewed as the pseudo-output, is approach is less restrictive than transmissibility identification and operational modal analysis since no assumption is made about the locations of the sensors relative to the excitation.

Kukreja, Sunil L.↗
NASA NTRS2012

In Flight Evaluation of Active Inceptor Force-Feel Characteristics and Handling Qualities

The effect of inceptor feel-system characteristics on piloted handling qualities has been a research topic of interest for many years. Most of the research efforts have focused on advanced fly-by-wire fixed-wing aircraft with only a few studies investigating the effects on rotorcraft. Consequently, only limited guidance is available on how cyclic force-feel characteristics should be set to obtain optimal handling qualities for rotorcraft. To study this effect, the U.S. Army Aeroflightdynamics Directorate working with the DLR Institute of Flight Systems in Germany under Task X of the U.S. German Memorandum of Understanding have been conducting flight test evaluations. In the U.S., five experimental test pilots have completed evaluations of two Mission Task Elements (MTEs) from ADS-33E-PRF and two command/response types for a matrix of center-stick cyclic force-feel characteristics at Moffett Field. In Germany, three experimental test Pilots have conducted initial evaluations of the two MTEs with two command/response types for a parallel matrix of side-stick cyclic force-feel characteristics at WTD-61 in Manching. The resulting data set is used to correlate the effect of changes in natural frequency and damping ratio of the cyclic inceptor on the piloted handling qualities. Existing criteria in ADS-33E and a proposed Handling Qualities Sensitivity Function that includes the effects of the cyclic force-feel characteristics are also evaluated against the data set and discussed.

Lusardi, Jeff A.↗
NASA NTRS1996

Modulation of adhesion-dependent cAMP signaling by echistatin and alendronate

We measured intracellular cAMP levels in cells during attachment and spreading on different extracellular matrix (ECM) proteins. Increases in cAMP were observed within minutes when cells attached to fibronectin, vitronectin, and a synthetic RGD-containing fibronectin peptide (Petite 2000), but not when they adhered to another integrin alpha nu beta 3 ligand, echistatin. Because echistatin also inhibits bone resorption, we measured the effects of adding another osteoporosis inhibitor, alendronate, in this system. Alendronate inhibited the cAMP increase induced by ligands that primarily utilize integrin alpha nu beta 3 (vitronectin, Peptite 2000), but not by fibronectin which can also use integrin alpha 5 beta 1. These results show that cell adhesion to ECM can increase intracellular cAPM levels and raise the possibility that inhibitors of osteoporosis may act, in part, by preventing activation of this pathway by integrins.

NASA Discipline Number 40-20↗
NASA NTRS1991

Evaluation of Fungal Metabolic Compounds Released to the Air in a Restricted Environment

The metabolic action of selected fungi species on common components of the interior of Space Station Freedom (SSF) will be tested. When present, volatile organic chemicals will be collected on porous polymer adsorbent columns. Using thermal desorption, the volatile compounds will be passed onto a gas chromatographic column for analysis. The Space Station Freedom (SSF) modular complex will largely be individually self contained and the established air environment will not be easily adjusted. The development and maintenance of a safe working environment offers a considerable challenge. Present plans for use of SSF acknowledge periods of manned activities and alternate times when the station is unmanned. The obvious necessity for clean and safe air and water during periods of use have been pursued as fundamental systems to SSF success. Somewhat less obvious, although perhaps of no less importance to the success of long term cyclic usage, are those periods of inactivity. It is during these periods when spores from microorganisms may be afforded the best conditions to germinate and in the vegetative form react with the complex synthetic chemical polymers which compose the furnishings and hardware of SSF nodes. Biodegradation could constitute a real hygiene problem, if the organisms form and release volatile organic chemicals. Similar problems have been documented in closed and improperly ventilated buildings and work spaces. Many of the metabolic products of fungi and bacterial growth create a variety of health problems. Analytical chemical techniques will first be used to document the growth of Aspergillus, Penicillium, and Cladosporium fungal species on the potential substrates Nomex and Kevlar. Any volatile organics that are released will be measured using the spectrum of gas adsorption chromatography. The level of microbial contamination that is necessary to produce such volatile compounds and the relative amounts expected to accumulate will be estimated.

Ferebee, Robert N.↗
NASA NTRS1993

Physical mechanism of comet outbursts - An experimental result

Attention is given to an experimental investigation of the physical mechanism of comet outbursts which is consistent with the general picture of mantle presence on comets and clarifies the relation of mantles to eruptive activity. The experiment and closeup observation of Comet P/Halley suggest a result different from most mathematical models in that the release of gas pressure does not occur only from uniform gas flow out of the entire surface. In some active comets near perihelion within a few AU of the sun, gas production rates and disturbance of the surface may be so high that the outflow is nearly continuous, with the regolith being entirely stripped away, as in many of the models. The present model provides a cyclic eruption and recharge mechanism which is lacking in most other models.

Hartmann, William K.↗
NASA NTRS2001

Integral Twist Actuation of Helicopter Rotor Blades for Vibration Reduction

Active integral twist control for vibration reduction of helicopter rotors during forward flight is investigated. The twist deformation is obtained using embedded anisotropic piezocomposite actuators. An analytical framework is developed to examine integrally-twisted blades and their aeroelastic response during different flight conditions: frequency domain analysis for hover, and time domain analysis for forward flight. Both stem from the same three-dimensional electroelastic beam formulation with geometrical-exactness, and axe coupled with a finite-state dynamic inflow aerodynamics model. A prototype Active Twist Rotor blade was designed with this framework using Active Fiber Composites as the actuator. The ATR prototype blade was successfully tested under non-rotating conditions. Hover testing was conducted to evaluate structural integrity and dynamic response. In both conditions, a very good correlation was obtained against the analysis. Finally, a four-bladed ATR system is built and tested to demonstrate its concept in forward flight. This experiment was conducted at NASA Langley T~ansonic Dynamics Tunnel and represents the first-of-a-kind Mach-scaled fully-active-twist rotor system to undergo forward flight test. In parallel, the impact upon the fixed- and rotating-system loads is estimated by the analysis. While discrepancies are found in the amplitude of the loads under actuation, the predicted trend of load variation with respect to its control phase correlates well. It was also shown, both experimentally and numerically, that the ATR blade design has the potential for hub vibratory load reduction of up to 90% using individual blade control actuation. Using the numerical framework, system identification is performed to estimate the harmonic transfer functions. The linear time-periodic system can be represented by a linear time-invariant system under the three modes of blade actuation: collective, longitudinal cyclic, and lateral cyclic. A vibration minimizing controller is designed based on this result, which implements classical disturbance rejection algorithm with some modifications. The controller is simulated numerically, and more than 90% of the 4P hub vibratory load is eliminated. By accomplishing the experimental and analytical steps described in this thesis, the present concept is found to be a viable candidate for future generation low-vibration helicopters. Also, the analytical framework is shown to be very appropriate for exploring active blade designs, aeroelastic behavior prediction, and as simulation tool for closed-loop controllers.

Shin, SangJoon↗
NASA NTRS1987

Turbulence control on an airborne laser platform

An active flow control device to generate large-scale, periodic structures in a turbulent shear flow is developed. Together with adaptive optics, the device may be used on airborne laser platforms to reduce or eliminate optical distortion caused by the turbulence in the aircraft's boundary layer. A cyclic jet issuing from a spanwise slot is used to collect the turbulent boundary layer for a finite time and then release all of the flow instantaneously in one large eddy that convects downstream. Flow visualization and hot-film probe measurements are used together with pattern recognition algorithms to demonstrate the viability of the flow control method. A flat plate towed in a water channel is used as a test bed. The instantaneous velocity signal is used to compute important statistical quantities of the random velocity field, such as the mean, the root-mean-square, the spectral distribution, and the probability density function. When optimized for a given boundary layer, it is shown that the cyclic jet will produce periodic structures that are similar to the random, naturally occurring ones. These structures seem to trigger the onset of bursting events near the wall of the plate. Thus, the present device generates periodic structures in both the outer and inner regions of a turbulent boundary layer.

Gad-El-hak, Mohamed↗
NASA NTRS1985

Raman structural studies of the nickel electrode

Raman spectroscopy is sensitive to empirically controlled nickel electrode structural variations, and has unique potential for structural characterization of these materials. How the structure relates to electrochemical properties is examined so that the latter can be more completely understood, controlled, and optimized. Electrodes were impregnated and cycled, and cyclic voltammetry is being used for electrochemical characterization. Structural variation was observed which has escaped detection using other methods. Structural changes are induced by: (1) cobalt doping, (2) the state of change or discharge, (3) the preparation conditions and type of buffer used, and (4) the formation process. Charged active mass has an NiOOH-type structure, agreeing with X-ray diffraction results. Discharged active mass, however, is not isostructural with beta-Ni(OH)2. Chemically prepared alpha phases are not isostructural either. A disordered structural model, containing point defects, is proposed for the cycled materials. This model explains K(+) incorporation. Band assignments were made and spectra interpreted for beta-Ni(OH)2, electrochemical NiOOH and chemically precipitated NiOOH.

Cornilsen, B. C.↗
NASA NTRS1999

Relative reactivity of ribosyl 2'-OH vs. 3'-OH in concentrated aqueous solutions of phosphoimidazolide activated nucleotides

Phosphoimidazolide activated ribomononucleotides (*pN, see structure) are useful substrates for the non-enzymatic synthesis of oligonucleotides. In the presence of metal ions, aqueous solutions of *pN yield primarily the two internucleotide-linked (pN2' pN and pN3' pN) and the pyrophosphate-linked (N5' ppN) dimers. Small amounts of cyclic dimers and higher oligomers are also produced. In this study the relative reactivity of 2'-OH vs. 3'-OH was determined from the ratio of the yields of pN2' pN vs. pN3' pN. Experiments were performed at 23 degrees C in the range 7.2 < or = pH < or = 8.4 with substrates that differ in nucleobase (guanosine (G), cytidine (C), uridine (U), and adenosine (A)) and leaving group (imidazole (Im), 2-methylimidazole (2-MeIm) and 2,4-dimethylimidazole (2,4-diMeIm)). Two metal ions (Mg2+ or Mn2+) were employed as catalysts. The conditions used here, i.e. a substrate concentration in the range 0.1 M to 1.0 M and metal ion concentration in the range 0.05 M to 0.2 M, favor base-stacking interactions. The ratio pN2' pN: pN3' pN = 2'-5': 3'-5' was found independent of nucleobase and typically varied between 2 to 3 indicating that the 2'-OH is about 2 to 3 times more reactive than the 3'-OH. *pN with Im, compared to 2-MeIm and 2,4-diMeIm leaving group, produce lower yields of internucleotide linked dimers, and a higher pN2' pN: pN3' pN ratio. Trends in the data, observed with all three leaving groups, suggest an increase in pN2' pN: pN3' pN ratio with decreasing substrate concentration (up to 5.47 with 0.051 M ImpG). The observations are in accord with earlier studies reporting a relative reactivity 2'-5': 3'-5' = 6 to 9 obtained with Im as the leaving group, in dilute nucleotide solutions and under conditions that disfavor stacking. It is speculated that the concentration induced change in the relative reactivity is the result of self-association via base-stacking that enhances selectively the proximity of the 3'-OH of one molecule to the reactive P-N bond of an other molecule. The implication of these conclusions for oligomerization/ligation reactions is discussed.

NASA Discipline Exobiology↗
NASA NTRS1984

Development and evaluation of a generic active helicopter vibration controller

A computerized generic active controller is developed, which alleviates helicopter vibration by closed-loop implementation of higher harmonic control (HHC). In the system, the higher harmonic blade pitch is input through a standard helicopter swashplate; for a four-blade helicopter rotor the 4/rev vibration in the rotorcraft is minimized by inducing cyclic pitch motions at 3, 4, and 5/rev in the rotating system. The controller employs the deterministic, cautious, and dual control approaches and two linear system models (local and global), as well as several methods of limiting control. Based on model testing, performed at moderate to high values of forward velocity and rotor thrust, reductions in the rotor test apparatus vibration from 75 to 95 percent are predicted, with HHC pitch amplitudes of less than one degree. Good performance is also noted for short-duration maneuvers.

Davis, M. W.↗
NASA NTRS1999

Expression of Novel Gene Products Upregulated by Disuse is Normalized by an Osteogenic Mechanical Stimulus: Evidence for the Molecular Basis of a Low Level Biomechanical Countermeasure for Osteoporosis?

The National Research Council's report entitled: A Strategy for Space Biology and Medical Science, highlighted several areas of fundamental scientific investigation which must be addressed to make long-term space exploration not only feasible, but safe. This "Goldberg Strategy," as well as several subsequent reports published by the NRC's Space Studies Board (e.g., Assessment of Programs in Space Biology and Medicine, Smith et. al., 1991), suggests that the principal hurdle to man's extended presence in space is the osteopenia which parallels reduced gravity. Ironically, the most significant risk to the skeleton may only be realized on return to normal gravitational fields, and full recovery of bone mass may never occur. Effective counter-measures to this microgravity induced bone loss are thus essential. Considering the similarities of space and aging induced osteopenia, an indisputable benefit of such a prophylaxis would be its potential as a treatment for the bone loss which plagues over 25 million people in the U.S. The osteogenic potential of mechanical strain is strongly frequency dependent, with sensitivity increasing up through at least 60 Hz (cycles per second). One hundred seconds per day of a 1 Hz cyclic loading will inhibit disuse osteopenia only if sufficient in magnitude to engender 1000 microstrain (mu(epsilon)) in the tissue. When loading is applied at 30 Hz, however, mechanical strains on the order of 5O mu(epsilon) (approx. 1% of the peak strains which occur in bone during vigorous functional activity), can stimulate bone formation in a duration dependent manner. In longer term animal studies, strains of less than 10 mu(epsilon), induced non-invasively via a whole body vibration, will stimulate bone formation on the surfaces of trabeculae, increase bone density, and improve strength. Finally, preliminary results from a double blind prospective clinical trial shows promise in inhibiting the bone loss which parallels the menopause. Based on these observations, we propose that these high frequency, low magnitude, mechanical strains effectively serve as a "surrogate" for musculoskeletal ground reaction forces, and thus represent an ideal countermeasure to the osteopenia which parallels microgravity conditions. The specific goal of this NASA funded work is to identify genes in bone upregulated by disuse, and to determine the efficacy of an osteogenic mechanical stimulus to downregulate their expression.

Rubin, C.↗
NASA NTRS2005

Early immune response and regulation of IL-2 receptor subunits

Affymetrix oligonucleotide arrays were used to monitor expression of 8796 genes and probe sets in activated T-cells; analysis revealed that 217 genes were significantly upregulated within 4 h. Induced genes included transcription factors, cytokines and their receptor genes. Analysis by semi-quantitative RT-PCR confirmed the significant induction of IL-2, IL-2R(gamma) and IL-2R(alpha). Forty-eight of the 217 induced genes are known to or predicted to be regulated by a CRE promoter/enhancer. We found that T-cell activation caused a significant increase in CREB phosphorylation furthermore, inhibition of the PKC pathway by GF109203 reduced CREB activation by 50% and inhibition of the PKA pathway caused a total block of CREB phosphorylation and significantly reduced IFN(gamma), IL-2 and IL-2R(alpha) gene expression by approximately 40% (p<0.001). PKC(theta) plays a major role in T-cell activation: inhibition of PKC significantly reduced the expression of IFN(gamma), IL-2 and IL-2R(alpha). Since PKC blocked activation of CREB, we studied potential cross-talk between the PKC and the PKA/MAPK pathways, PMA-stimulated Jurkat cells were studied with specific signal pathway inhibitors. Extracellular signal-regulated kinase-2 (ERK2) pathway was found to be significantly activated greater than seven-fold within 30 min; however, there was little activation of ERK-1 and no activation of JNK or p38 MAPK. Inhibition of the PKA pathway, but not the PKC pathway, resulted in inhibition of ERK1/2 activation at all time points, inhibition of MEK1 and 2 significantly blocked expression of IL-2 and IL-2R(alpha). Gene expression of IL-2R(alpha) and IFN(gamma) was dependent on PKA in S49 wt cells but not in kin- mutants. Using gel shift analysis, we found that forskolin activation of T-cells resulted in activation of AP1 sites; this increase in nuclear extract AP1 was significantly blocked by MEK1 inhibitor U0126. Taken together, these results suggest that the PKA in addition to PKC and MAPK pathways plays a role in early T-cell activation and induction of IL-2, IL-2R(alpha) and IFN(gamma) gene expression.

Non-NASA Center↗