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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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At least 811 records · Page 45
All sourcesNASA NTRSDOE OSTI
DOE OSTI2010

Crystallographic Identification and Functional Characterization of Phospholipids as Ligands for the Orphan Nuclear Receptor Steroidogenic Factor-1

The orphan nuclear receptor steroidogenic factor 1 (SF-1) regulates the differentiation and function of endocrine glands. Although SF-1 is constitutively active in cell-based assays, it is not known whether this transcriptional activity is modulated by ligands. Here, we describe the 1.5 {angstrom} crystal structure of the SF-1 ligand binding domain in complex with an LXXLL motif from a coregulator protein. The structure reveals the presence of a phospholipid ligand in a surprisingly large pocket ({approx}1600 {angstrom}{sup 3}), with the receptor adopting the canonical active conformation. The bound phospholipid is readily exchanged and modulates SF-1 interactions with coactivators. Mutations designed to reduce the size of the SF-1 pocket or to disrupt hydrogen bonds with the phospholipid abolish SF-1/coactivator interactions and significantly reduce SF-1 transcriptional activity. These findings provide evidence that SF-1 is regulated by endogenous ligands and suggest an unexpected relationship between phospholipids and endocrine development and function.

08 HYDROGEN↗
DOE OSTI2010

The Crystal Structure of Coxsackievirus A21 and Its Interaction with ICAM-1

CVA21 and polioviruses both belong to the Enterovirus genus in the family of Picornaviridae, whereas rhinoviruses form a distinct picornavirus genus. Nevertheless, CVA21 and the major group of human rhinoviruses recognize intercellular adhesion molecule-1 (ICAM-1) as their cellular receptor, whereas polioviruses use poliovirus receptor. The crystal structure of CVA21 has been determined to 3.2 {angstrom} resolution. Its structure has greater similarity to poliovirus structures than to other known picornavirus structures. Cryo-electron microscopy (cryo-EM) was used to determine an 8.0 {angstrom} resolution structure of CVA21 complexed with an ICAM-1 variant, ICAM-1{sup Kilifi}. The cryo-EM map was fitted with the crystal structures of ICAM-1 and CVA21. Significant differences in the structure of CVA21 with respect to the poliovirus structures account for the inability of ICAM-1 to bind polioviruses. The interface between CVA21 and ICAM-1 has shape and electrostatic complementarity with many residues being conserved among those CVAs that bind ICAM-1.

36 MATERIALS SCIENCE↗
DOE OSTI2016

Implementation of Enhanced Attenuation at the DOE Mound Site OU-1 Landfill: Accelerating Progress and Reducing Costs - 16270

A field demonstration of enhanced attenuation (EA) was initiated in 2014 to expedite the remediation of industrial solvents in the groundwater impacted by the former landfill Operable Unit 1 (OU-1) at the US Department of Energy (DOE) Mound, Ohio, Site. EA uses active engineering solutions to alter the target site so that the contaminant plume will passively stabilize and shrink. For OU-1, the EA was implemented by targeted injection of an electron donor (vegetable oil) to create 'structured geochemical zones'. Volatile organic compounds (VOCs) in the altered subsurface system encounter alternating anaerobic and aerobic environments along the plume flow path. The effect of these structured geochemical zones is effective and rapid degradation of parent compounds such as trichloroethene (TCE) and tetrachloroethene (PCE), and daughter compounds such as cis-1,2-dichloroethene and vinyl chloride. EA at Mound OU-1 provides a transition to natural attenuation and is an alternative to the baseline pump-and-treat (P and T) system. Industrial solvents (primarily TCE) and other VOCs that originated from the former solid-waste landfill contaminated the groundwater in the Buried Valley Aquifer beneath the Mound site. A groundwater P and T system was initiated in 1996 to control the plume, and a soil vapor extraction system was installed and operated from 1997 to 2003 to remove the bulk of the VOC sources from the vadose zone. Physical removal of the landfill waste and contaminated soil was performed between 2006 and 2010. Approximately 76,100 cubic meters (99,500 cubic yards) of material were removed from the OU-1 landfill area. Operation of the groundwater P and T system continued throughout this period. The principles and technical basis of the ongoing EA field demonstration were developed by the Interstate Technology and Regulatory Council. The goal of the EA implementation is to provide a bridge between active groundwater treatments and monitored natural attenuation. EA relies on understanding and manipulating the contaminant mass balance. Technologies are applied to sustainably reduce the contaminant loading from the source and/or increase the rate of natural attenuation processes within the plume. In 2014, a core team that included DOE, the US Environmental Protection Agency (EPA), the Ohio EPA, and technical support organizations evaluated remedial options for OU-1. During the 2014 evaluation, the team projected a cleanup time frame for the baseline P and T system of 26 additional years and a cleanup time frame for the EA remedy of 13 years. On the basis of that evaluation, EPA approved the implementation of the EA remedy that required the shutdown of the P and T system. The combination of technologies implemented for OU-1 EA included (a) neat vegetable oil in the vadose zone in the former landfill and (b) emulsified vegetable oil within the footprint of the groundwater plume. In the first part of the deployment, neat oil spreads laterally and forms a thin layer on the water table beneath residual soil sources to intercept and reduce future VOC loading and to reduce oxygen inputs to the local groundwater. In the second part of the deployment, emulsified oil forms active bioremediation anaerobic treatment zones within the plume footprint to degrade existing groundwater contaminants (via reductive dechlorination, co-metabolism, or both) and stimulates long-term attenuation capacity in the down-gradient aerobic plume (via co-metabolism). The results from the first year of the field demonstration indicate that the dissolved TCE and PCE plumes have decreased in size and mass. Statistical tests indicate that the VOC concentrations in the majority of the source-area wells are decreasing. The VOC concentrations throughout the plume are projected to be below the target maximum contaminant level of 2 to 5 μg/L within 5 to 10 years-sooner than the design projection of 13 years. The biogeochemistry and microbial community within the structured geochemical zones are consistent with the design and appear to be stable. At Mound OU-1, EA has accelerated progress toward the remedial action objectives and reduced costs. Ongoing monitoring will determine whether the EA remedy has transitioned the site into monitored natural attenuation status and whether the EA will continue to be effective, timely, and sustainable. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2018

Crystal structure of the m4-1BB/4-1BBL complex reveals an unusual dimeric ligand that undergoes structural changes upon 4-1BB receptor binding

The interaction between the receptor 4-1BB and its ligand 4-1BBL provides co-stimulatory signals for T-cell activation and proliferation. However, differences in the mouse and human molecules might result in differential engagement of this pathway. Here, we report the crystal structure of mouse 4-1BBL and of the mouse 4-1BB/4-1BBL complex, which together provided insights into the molecular mechanism by which m4-1BBL and its cognate receptor recognize each other. Unlike all human or mouse tumor necrosis factor ligands that form noncovalent and mostly trimeric assemblies, the m4-1BBL structure formed a disulfide-linked dimeric assembly. The structure disclosed that certain differences in the amino acid composition along the intramolecular interface, together with two specific residues (Cys-246 and Ser-256) present exclusively in m4-1BBL, are responsible for this unique dimerization. Unexpectedly, upon m4-1BB binding, m4-1BBL undergoes structural changes within each protomer; moreover, the individual m4-1BBL protomers rotate relative to each other, yielding a dimerization interface with more inter-subunit interactions. We also observed that in the m4-1BB/4-1BBL complex, each receptor monomer binds exclusively to a single ligand subunit with contributions of cysteine-rich domain 1 (CRD1), CRD2, and CRD3. Furthermore, structure-guided mutagenesis of the binding interface revealed that novel binding interactions with the GH loop, rather than the DE loop, are energetically critical and define the m4-1BB receptor selectivity for m4-1BBL. As a result, a comparison with the human 4-1BB/4-1BBL complex highlighted several differences between the ligand- and receptor-binding interfaces, providing an explanation for the absence of inter-species cross-reactivity between human and mouse 4-1BB and 4-1BBL molecules.

4-1BB↗
DOE OSTI2018

An insoluble iron complex coated cathode enhances direct electron uptake by Rhodopseudomonas palustris TIE-1

Microbial electrosynthesis (MES) is a promising bioelectrochemical approach to produce biochemicals. A previous study showed that Rhodopseudomonas palustris TIE-1 can directly use poised electrodes as electron donors for photoautotrophic growth at cathodic potentials that avoid electrolytic H2 production (photoelectroautotrophy). To make TIE-1 an effective biocatalyst for MES, we need to improve its electron uptake ability and growth under photoelectroautotrophic conditions. Because TIE-1 interacts with various forms of iron while using it as a source of electrons for photoautotrophy (photoferroautotrophy), here we tested the ability of iron-based redox mediators to enhance direct electron uptake. Our data show that soluble iron cannot act as a redox mediator for electron uptake by TIE-1 from a cathode poised at +100 mV vs. Standard Hydrogen electrode. We then tested whether an immobilized iron-based redox mediator Prussian blue (PB) can enhance electron uptake by TIE-1. Chronoamperometry indicates that cathodic current uptake by TIE-1 increased from 1.47 ± 0.04 to 5.6 ± 0.09 μA/cm 2 (3.8 times). Overall, our data show that immobilized PB can enhance direct electron uptake by TIE-1.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Identification of a Covalent Molecular Inhibitor of Anti-apoptotic BFL-1 by Disulfide Tethering

The BCL-2 family is composed of anti- and pro-apoptotic members that respectively protect or disrupt mitochondrial integrity. Anti-apoptotic overexpression can promote oncogenesis by trapping the BCL-2 homology 3 (BH3) “killer domains” of pro-apoptotic proteins in a surface groove, blocking apoptosis. Groove inhibitors, such as the relatively large BCL-2 drug venetoclax (868 Da), have emerged as cancer therapies. BFL-1 remains an undrugged oncogenic protein and can cause venetoclax resistance. Having identified a unique C55 residue in the BFL-1 groove, we performed a disulfide tethering screen to determine if C55 reactivity could enable smaller molecules to block BFL-1's BH3-binding functionality. We found that a disulfide-bearing N-acetyltryptophan analog (304 Da adduct) effectively targeted BFL-1 C55 and reversed BFL-1-mediated suppression of mitochondrial apoptosis. Structural analyses implicated the conserved leucine-binding pocket of BFL-1 as the interaction site, resulting in conformational remodeling. Thus, therapeutic targeting of BFL-1 may be achievable through the design of small, cysteine-reactive drugs.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Dendritic cells focus CTL responses toward highly conserved and topologically important HIV-1 epitopes

During early HIV-1 infection, immunodominant T cell responses to highly variable epitopes lead to the establishment of immune escape virus variants. Here we assessed a type 1-polarized monocyte-derived dendritic cell (MDC1)-based approach to selectively elicit cytotoxic T lymphocyte (CTL) responses against highly conserved and topologically important HIV-1 epitopes in HIV-1-infected individuals from the Thailand RV254/SEARCH 010 cohort who initiated antiretroviral therapy (ART) during early infection (Fiebig stages I-IV). Autologous MDC1 were used as antigen presenting cells to induce in vitro CTL responses against HIV-1 Gag, Pol, Env, and Nef as determined by flow cytometry and ELISpot assay. Ultra-conserved or topologically important antigens were respectively identified using the Epigraph tool and a structure-based network analysis approach and compared to overlapping peptides spanning the Gag proteome. MDC1 presenting either the overlapping Gag, Epigraph, or Network 14-21mer peptide pools consistently activated and expanded HIV-1-specific T cells to epitopes identified at the 9-13mer peptide level. Interestingly, some CTL responses occurred outside known or expected HLA associations, providing evidence of new HLA-associated CTL epitopes. Comparative analyses demonstrated more sequence conservation among Epigraph antigens but a higher magnitude of CTL responses to Network and Gag peptide groups. Importantly, CTL responses against topologically constrained Gag epitopes contained in both the Network and Gag peptide pools were selectively enhanced in the Network pool-initiated cultures. Our study supports the use of MDC1 as a therapeutic strategy to induce and focus CTL responses toward putative fitness-constrained regions of HIV-1 to prevent immune escape and control HIV-1 infection.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2010

Fission Product Monitoring of TRISO Coated Fuel For The Advanced Gas Reactor -1 Experiment

The US Department of Energy has embarked on a series of tests of TRISO-coated particle reactor fuel intended for use in the Very High Temperature Reactor (VHTR) as part of the Advanced Gas Reactor (AGR) program. The AGR-1 TRISO fuel experiment, currently underway, is the first in a series of eight fuel tests planned for irradiation in the Advanced Test Reactor (ATR) located at the Idaho National Laboratory (INL). The AGR-1 experiment reached a peak compact averaged burn up of 9% FIMA with no known TRISO fuel particle failures in March 2008. The burnup goal for the majority of the fuel compacts is to have a compact averaged burnup greater than 18% FIMA and a minimum compact averaged burnup of 14% FIMA. At the INL the TRISO fuel in the AGR-1 experiment is closely monitored while it is being irradiated in the ATR. The effluent monitoring system used for the AGR-1 fuel is the Fission Product Monitoring System (FPMS). The FPMS is a valuable tool that provides near real-time data indicative of the AGR-1 test fuel performance and incorporates both high-purity germanium (HPGe) gamma-ray spectrometers and sodium iodide [NaI(Tl)] scintillation detector-based gross radiation monitors. To quantify the fuel performance, release-to-birth ratios (R/B’s) of radioactive fission gases are computed. The gamma-ray spectra acquired by the AGR-1 FPMS are analyzed and used to determine the released activities of specific fission gases, while a dedicated detector provides near-real time count rate information. Isotopic build up and depletion calculations provide the associated isotopic birth rates. This paper highlights the features of the FPMS, encompassing the equipment, methods and measures that enable the calculation of the release-to-birth ratios. Some preliminary results from the AGR-1 experiment are also presented.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
DOE OSTI2025

Creation of Self-Semi-Interpenetrating Network Structures in PIM-1 Membranes for Enhanced Physical Aging Resistance

A series of self-semi-interpenetrating network (ssIPN) thin films based on PIM-1 structure were developed by end-cross-linking telechelic PIM-1 oligomers end-capped with curable carboxylic acid groups to form model networks, which are penetrated by linear high-molecular-weight PIM-1 chains. ssIPN films with systematically varied network content ranging from 10 to 30 wt % were comprehensively examined on their microstructure and gas permeation properties. Fresh PIM-1 ssIPN films exhibited gas separation performances close to those of as-cast linear PIM-1, where the films closely followed the upper bound trade-off line, gaining as much selectivity as they lose gas permeability as the network content increases. This indicates that gas permeability is largely preserved despite cross-linking. Wide-angle X-ray scattering supported this observation, with peaks shifting toward lower d-spacing as oligomer content increased, suggesting tighter chain packing with higher cross-linkable oligomer loading. In physical aging studies over two months, all ssIPN films outperform linear PIM-1 for H 2 /CH 4 and O 2 /N 2 separations. Notably, the PIM-1 ssIPN with 10 wt % network content showed the best physical aging resistance, with negligible permeability loss even after one month. Finally, these findings highlight a promising macromolecular strategy for enhancing the physical aging resistance of microporous polymer membranes for gas separation.

PIM-1↗
DOE OSTI2016

Structural basis for human PECAM-1-mediated trans-homophilic cell adhesion

Cell adhesion involved in signal transduction, tissue integrity and pathogen infection is mainly mediated by cell adhesion molecules (CAM). One CAM member, platelet–endothelial-cell adhesion molecule-1 (PECAM-1), plays an important role in tight junction among endothelia cells, leukocyte trafficking, and immune response through its homophilic and heterophilic binding patterns. Both kinds of interactions, which lead to endogenous and exogenous signal transmission, are derived from extracellular immunoglobulin-like (IgL) domains and cytoplasmic immunoreceptor tyrosine-based inhibitory motifs (ITIMs) of PECAM-1. To date, the mechanism of trans-homophilic interaction of PECAM-1 remains unclear. Here, we present the crystal structure of PECAM-1 IgL1-2 trans-homo dimer. Both IgL 1 and 2 adopt the classical Ig domain conformation comprised of two layers of β-sheets possessing antiparallel β-strands with each being anchored by a pair of cysteines forming a disulfide bond. The dimer interface includes hydrophobic and hydrophilic interactions. The Small-Angle X-ray Scattering (SAXS) envelope of PECAM-1 IgL1-6 supported such a dimer formation in solution. As a result, cell adhesion assays on wildtype and mutant PECAM-1 further characterized the structural determinants in cell junction and communication.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2013

Plasma IgG to Linear Epitopes in the V2 and V3 Regions of HIV-1 gp120 Correlate with a Reduced Risk of Infection in the RV144 Vaccine Efficacy Trial

Neutralizing and non-neutralizing antibodies to linear epitopes on HIV-1 envelope glycoproteins have potential to mediate antiviral effector functions that could be beneficial to vaccine-induced protection. Here, plasma IgG responses were assessed in three HIV-1 gp120 vaccine efficacy trials (RV144, Vax003, Vax004) and in HIV-1- infected individuals by using arrays of overlapping peptides spanning the entire consensus gp160 of all major genetic subtypes and circulating recombinant forms (CRFs) of the virus. In RV144, where 31.2% efficacy against HIV-1 infection was seen, dominant responses targeted the C1, V2, V3 and C5 regions of gp120. An analysis of RV144 case-control samples showed that IgG to V2 CRF01_AE significantly inversely correlated with infection risk (OR= 0.54, p=0.0042), as did the response to other V2 subtypes (OR=0.60-0.63, p=0.016-0.025). The response to V3 CRF01_AE also inversely correlated with infection risk but only in vaccine recipients who had lower levels of other antibodies, especially Env-specific plasma IgA (OR=0.49, p=0.007) and neutralizing antibodies (OR=0.5, p=0.008). Responses to C1 and C5 showed no significant correlation with infection risk. In Vax003 and Vax004, where no significant protection was seen, serum IgG responses targeted the same epitopes as in RV144 with the exception of an additional C1 reactivity in Vax003 and infrequent V2 reactivity in Vax004. In HIV-1 infected subjects, dominant responses targeted the V3 and C5 regions of gp120, as well as the immunodominant domain, heptad repeat 1 (HR-1) and membrane proximal external region (MPER) of gp41. These results highlight the presence of several dominant linear B cell epitopes on the HIV-1 envelope glycoproteins. They also generate the hypothesis that IgG to linear epitopes in the V2 and V3 regions of gp120 are part of a complex interplay of immune responses that contributed to protection in RV144.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

ARM Aerial Facility (AAF) Merged Value-Added Product Report for Historical G-1 Field Campaigns

For 30 years, the U.S. Department of Energy (DOE) Office of Science supported an instrumented Grumman Gulfstream-1 (G-1) aircraft for atmospheric field campaigns. Data from the final decade of G-1 operations were archived by the Atmospheric Radiation Measurement (ARM) user facility Data Center (ADC) and made publicly available at no cost to all registered users. To ensure a consistent data format and to improve the accessibility of the ARM airborne data, an integrated data set was recently developed covering the final six years of G-1 operations (2013 to 2018). The integrated data set includes data collected from 236 flights (766.4 hours). Four of the seven field campaigns were based in the U.S. One campaign collected data from the wildfires in the U.S. Pacific Northwest and agricultural burns in the lower Mississippi River valley as part of the Biomass Burning Observation Project (BBOP) in 2013. In 2015, the ARM Cloud Aerosol Precipitation Experiment provided data on atmospheric rivers and associated aerosol-cloud interactions that produce heavy precipitation on the U.S. west coast during the early spring. Research data from Airborne Carbon Measurements-V (ACME-V), collected during the summer of 2015, gave scientists insight into trends and variability of trace gases in the atmosphere over the North Slope of Alaska to improve arctic climate models. In the early summer and autumn of 2016, the Holistic Interactions of Shallow Clouds, Aerosols, and Land-Ecosystems (HI-SCALE) campaign provided an extensive data set geared toward coupled processes that affect the life cycle of shallow clouds through the interaction among aerosol, cloud, land surface, and ecosystems. In 2014 (March and October), the airborne sampling moved outside of the U.S. to the city of Manaus in central Amazonia, Brazil, where residential and industrial emissions were extensively characterized by flights of the G-1. The GoAmazon2014/15 aircraft campaign data are being integrated with aquatic and terrestrial ecosystem measurements to quantify anthropogenic perturbations to a usually pristine tropical environment. Another international airborne mission was carried out in the Eastern North Atlantic region. The Aerosol and Cloud Experiments in the Eastern North Atlantic (ACE-ENA) campaign saw the G-1 aircraft fly from Terceira Island in the Azores during the summer of 2017 and the winter of 2018. The campaign studied both seasons to measure key aerosol and cloud processes under various meteorological and cloud conditions with different aerosol sources. Then the G-1 deployed to the Sierras de Córdoba range in central Argentina from October to November 2018 for the Cloud, Aerosol, and Complex Terrain Interactions (CACTI) campaign to study orographic convective cloud interactions with their surrounding environment. These comprehensive datastreams provide much-needed insight into spatiotemporal variability of thermodynamic quantities, aerosol and cloud states, and properties for addressing essential science questions in Earth system process studies.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Molecular Epidemiology of HIV-1 in Ghana: Subtype Distribution, Drug Resistance and Coreceptor Usage

The greatest HIV-1 genetic diversity is found in West/Central Africa due to the pandemic’s origins in this region, but this diversity remains understudied. We characterized HIV-1 subtype diversity (from both sub-genomic and full-genome viral sequences), drug resistance and coreceptor usage in 103 predominantly (90%) antiretroviral-naive individuals living with HIV-1 in Ghana. Full-genome HIV-1 subtyping confirmed the circulating recombinant form CRF02_AG as the dominant (53.9%) subtype in the region, with the complex recombinant 06_cpx (4%) present as well. Unique recombinants, most of which were mosaics containing CRF02_AG and/or 06_cpx, made up 37% of sequences, while “pure” subtypes were rare (<6%). Pretreatment resistance to at least one drug class was observed in 17% of the cohort, with NNRTI resistance being the most common (12%) and INSTI resistance being relatively rare (2%). CXCR4-using HIV-1 sequences were identified in 23% of participants. Overall, our findings advance our understanding of HIV-1 molecular epidemiology in Ghana. Extensive HIV-1 genetic diversity in the region appears to be fueling the ongoing creation of novel recombinants, the majority CRF02_AG-containing, in the region. The relatively high prevalence of pretreatment NNRTI resistance but low prevalence of INSTI resistance supports the use of INSTI-based first-line regimens in Ghana.

59 BASIC BIOLOGICAL SCIENCES↗
NASA NTRS2002

Physical interaction of the activator protein-1 factors c-Fos and c-Jun with Cbfa1 for collagenase-3 promoter activation

Previously, we determined that the activator protein-1 (AP-1)-binding site and the runt domain (RD)-binding site and their binding proteins, c-Fos.c-Jun and Cbfa, regulate the collagenase-3 promoter in parathyroid hormone-treated and differentiating osteoblasts. Here we show that Cbfa1 and c-Fos.c-Jun appear to cooperatively bind the RD- and AP-1-binding sites and form ternary structures in vitro. Both in vitro and in vivo co-immunoprecipitation and yeast two-hybrid studies further demonstrate interaction between Cbfa1 with c-Fos and c-Jun in the absence of phosphorylation and without binding to DNA. Additionally, only the runt domain of Cbfa1 was required for interaction with c-Jun and c-Fos. In mammalian cells, overexpression of Cbfa1 enhanced c-Jun activation of AP-1-binding site promoter activity, demonstrating functional interaction. Finally, insertion of base pairs that disrupted the helical phasing between the AP-1- and RD-binding sites also inhibited collagenase-3 promoter activation. Thus, we provide direct evidence that Cbfa1 and c-Fos.c-Jun physically interact and cooperatively bind the AP-1- and RD-binding sites in the collagenase-3 promoter. Moreover, the AP-1- and RD-binding sites appear to be organized in a specific required helical arrangement that facilitates transcription factor interaction and enables promoter activation.

Non-NASA Center↗
NASA NTRS2001

Twisting integrin receptors increases endothelin-1 gene expression in endothelial cells

A magnetic twisting stimulator was developed based on the previously published technique of magnetic twisting cytometry. Using ligand-coated ferromagnetic microbeads, this device can apply mechanical stresses with varying amplitudes, duration, frequencies, and waveforms to specific cell surface receptors. Biochemical and biological responses of the cells to the mechanical stimulation can be assayed. Twisting integrin receptors with RGD (Arg-Gly-Asp)-containing peptide-coated beads increased endothelin-1 (ET-1) gene expression by >100%. In contrast, twisting scavenger receptors with acetylated low-density lipoprotein-coated beads or twisting HLA antigen with anti-HLA antibody-coated beads did not lead to alterations in ET-1 gene expression. In situ hybridization showed that the increase in ET-1 mRNA was localized in the cells that were stressed with the RGD-coated beads. Blocking stretch-activated ion channels with gadolinium, chelating Ca2+ with EGTA, or inhibiting tyrosine phosphorylation with genistein abolished twist-induced ET-1 mRNA elevation. Abolishing cytoskeletal tension with an inhibitor of the myosin ATPase, with an inhibitor of myosin light chain kinase, or with an actin microfilament disrupter blocked twisted-induced increases in ET-1 expression. Our results are consistent with the hypothesis that the molecular structural linkage of integrin-cytoskeleton is an important pathway for stress-induced ET-1 gene expression.

Non-NASA Center↗
NASA NTRS2000

Developmental regulation of collagenase-3 mRNA in normal, differentiating osteoblasts through the activator protein-1 and the runt domain binding sites

Collagenase-3 mRNA is initially detectable when osteoblasts cease proliferation, increasing during differentiation and mineralization. We showed that this developmental expression is due to an increase in collagenase-3 gene transcription. Mutation of either the activator protein-1 or the runt domain binding site decreased collagenase-3 promoter activity, demonstrating that these sites are responsible for collagenase-3 gene transcription. The activator protein-1 and runt domain binding sites bind members of the activator protein-1 and core-binding factor family of transcription factors, respectively. We identified core-binding factor a1 binding to the runt domain binding site and JunD in addition to a Fos-related antigen binding to the activator protein-1 site. Overexpression of both c-Fos and c-Jun in osteoblasts or core-binding factor a1 increased collagenase-3 promoter activity. Furthermore, overexpression of c-Fos, c-Jun, and core-binding factor a1 synergistically increased collagenase-3 promoter activity. Mutation of either the activator protein-1 or the runt domain binding site resulted in the inability of c-Fos and c-Jun or core-binding factor a1 to increase collagenase-3 promoter activity, suggesting that there is cooperative interaction between the sites and the proteins. Overexpression of Fra-2 and JunD repressed core-binding factor a1-induced collagenase-3 promoter activity. Our results suggest that members of the activator protein-1 and core-binding factor families, binding to the activator protein-1 and runt domain binding sites are responsible for the developmental regulation of collagenase-3 gene expression in osteoblasts.

NASA Discipline Musculoskeletal↗
NASA NTRS2015

The Orion Pad Abort 1 (PA-1) Flight Test: A Propulsion Success

This poster provides a concise overview of the highly successful Orion Pad Abort 1 (PA-1) flight test, and the three rocket motors that contributed to this success. The primary purpose of the Orion PA-1 flight was to help certify the Orion Launch Abort System (LAS), which can be utilized in the unlikely event of an emergency on the launchpad or during mission vehicle ascent. The PA-1 test was the first fully integrated flight test of the Orion LAS, one of the primary systems within the Orion Multi-Purpose Crew Vehicle (MPCV). The Orion MPCV is part of the architecture within the Space Launch System (SLS), which is being designed to transport astronauts beyond low-Earth orbit for future exploration missions. Had the Orion PA-1 flight abort occurred during launch preparations for a real human spaceflight mission, the PA-1 LAS would have saved the lives of the crew. The PA-1 flight test was largely successful due to the three solid rocket motors of the LAS: the Attitude Control Motor (ACM); the Jettison Motor (JM); and the Abort Motor (AM). All three rocket motors successfully performed their required functions during the Orion PA-1 flight test, flown on May 6, 2010 at the White Sands Missile Range in New Mexico, culminating in a successful demonstration of an abort capability from the launchpad.

JM↗
NASA NTRS2015

Orion Launch Abort System (LAS) Propulsion on Pad Abort 1 (PA-1)

This presentation provides a concise overview of the highly successful Orion Pad Abort 1 (PA-1) flight test, and the three rocket motors that contributed to this success. The primary purpose of the Orion PA-1 flight was to help certify the Orion Launch Abort System (LAS), which can be utilized in the unlikely event of an emergency on the launchpad or during mission vehicle ascent. The PA-1 test was the first fully integrated flight test of the Orion LAS, one of the primary systems within the Orion Multi-Purpose Crew Vehicle (MPCV). The Orion MPCV is part of the architecture within the Space Launch System (SLS), which is being designed to transport astronauts beyond low-Earth orbit for future exploration missions. Had the Orion PA-1 flight abort occurred during launch preparations for a real human spaceflight mission, the PA-1 LAS would have saved the lives of the crew. The PA-1 flight test was largely successful due to the three solid rocket motors of the LAS: the Attitude Control Motor (ACM); the Jettison Motor (JM); and the Abort Motor (AM). All three rocket motors successfully performed their required functions during the Orion PA-1 flight test, flown on May 6, 2010 at the White Sands Missile Range in New Mexico, culminating in a successful demonstration of an abort capability from the launchpad.

SLS↗