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All sourcesNASA NTRSDOE OSTI
NASA NTRS2003

Hypergravity Stimulates Osteoblast Proliferation Via Matrix-Integrin-Signaling Pathways

Extensive characterizations of the physiologic consequences of microgravity and gravity indicate that lack of weight-bearing may cause tissue atrophy through cellular and subcellular level mechanisms. We hypothesize that gravity is needed for the efficient transduction of cell growth and survival signals from the extra-cellular matrix (ECM) in mechanosensitive tissues. Recent work from our laboratory and from others shows that an increase of gravity increases bone cell growth and survival. We found that 50-g hypergravity stimulation increased osteoblast proliferation for cells grown on Collagen Type I and Fibronectin, but not on Laminin or uncoated plastic. This may be a tissue-specific response, because 50-g hypergravity stimulation caused no increase in proliferation for primary rat fibroblasts. These results combined with RT-PCR for all possible integrins indicate that beta1 integrin subunit may be involved. The osteoblast proliferation response on Collagen Type I was greater at 25-g than at 10-g or 50-g; 24-h duration of hypergravity was necessary to see an increase in proliferation. Survival was enhanced during hypergravity stimulation by the presence of matrix. Flow cytometry analysis indicated that cell cycle may be altered; BrdU incorporation in proliferating cells showed an increase in the number of actively dividing cells from about 60% at 1-g to over 90% at 25-g. To further investigate the molecular components involved, we applied fluorescence labeling of cytoskeletal and signaling molecules to cells after 2 to 30 minutes of hypergravity stimulation. While structural components did not appear to be altered, phosphorylation increased, indicating that signaling pathways may be activated. These data indicate that gravity mechanostimulation of osteoblast proliferation involves specific matrix-integrin signaling pathways which are sensitive to duration and g-level.

Vercoutere, W.↗
NASA NTRS2004

Surface Figure Measurement at 20K: Silicon Carbide and Cesic Mirrors

This report presents the facility, methods, and results of testing cryogenically-cooled spherical mirrors, using standard phase-shifting interferometry, at the Goddard Space Flight Center (GSFC). Two mirrors were supplied to GSFC by the European Space Technology Center, so that GSFC could render a second, independent cryo-measurement of their surface figures at 20K. These mirrors, produced by Galileo Avionica and its partners, demonstrate the technology of silicon carbide and SiC-composite lightweighted mirrors, designed for high accuracy (10 nm rms surface figure error) at both room temperature and 20K. The two mirrors provided for testing at GSFC include one made of sintered silicon carbide (mirror blank by Bettini), and a second made of the C/Si/SiC composite known as Cesic (supplied by ECM). Both mirrors are f/2 spheres with 150 mm clear aperture, and with integral mounts. At GSFC, the mirrors will be measured first at at room temperature, making use of standard techniques, with a predicted uncertainty of 2 nm. Then each mirror will be tested in the cryostat, down to 20K: first freely supported and, second, mounted to a Cesic plate that models a Cesic optical bench. The uncertainty of the resultant surface figure error is predicted to be about 5 nm rms. Details of the uncertainty budget are discussed in a related poster. GSFC's techniques and results can be usefully compared to the measurements performed on the same two mirrors by and for Galileo Avionica, who used a somewhat different test configuration and alignment approach.

Blake, Peter↗
NASA NTRS1993

Embryonic lung morphogenesis in organ culture: experimental evidence for a proteoglycan function in the extracellular matrix

The lung rudiment, isolated from mid-gestation (11 day) mouse embryos, can undergo morphogenesis in organ culture. Observation of living rudiments, in culture, reveals both growth and ongoing bronchiolar branching activity. To detect proteoglycan (PG) biosynthesis, and deposition in the extracellular matrix, rudiments were metabolically labeled with radioactive sulfate, then fixed, embedded, sectioned and processed for autoradiography. The sulfated glycosaminoglycan (GAG) types, composing the carbohydrate component of the proteoglycans, were evaluated by selective GAG degradative approaches that showed chondroitin sulfate PG principally associated with the interstitial matrix, and heparan sulfate PG principally associated with the basement membrane. Experiments using the proteoglycan biosynthesis disrupter, beta-xyloside, suggest that when chondroitin sulfate PG deposition into the ECM is perturbed, branching morphogenesis is compromised.

NASA Discipline Number 93-10↗
NASA NTRS2000

Particle irradiation induces FGF2 expression in normal human lens cells

Particle Irradiation Induces FGF2 Expression in Normal Human Lens Cells. Particle radiations, including both proton and helium-ion beams, have been used to successfully treat choroidal melanoma, but with the complication of radiation-induced cataract. We have investigated a role for radiation-induced changes in the expression of basic fibroblast growth factor (FGF2) gene expression as part of the mechanism(s) underlying lens cell injury associated with cataract. Normal human lens epithelial (HLE) cells were cultured in vitro on extracellular matrix (ECM) originated from bovine corneal endothelial cells. This study reports evidence for rapid but transient induction of FGF2 transcripts, an increase of between 5- and 8-fold, within 0.5 h after exposure to particle radiation, followed by another wave of increased transcription at 2-3 h postirradiation. Immunofluorescence results confirm the enhanced levels of FGF2 protein rapidly after exposure to protons or helium ions, followed by another wave of increased activity unique to helium at 6 h postirradiation. This second wave of increased immunoreactivity was not observed in the proton-irradiated samples. Total FGF2 protein analysis after helium-ion exposures shows induced expression of three FGF2 isoforms, with an increase of up to 2-fold in the 18-kDa low-molecular-weight species. Studies of the effects of protons on individual FGF2 protein isoforms are in progress. Several mechanisms involving a role for FGF2 in radiation-induced cataract are discussed.

NASA Discipline Radiation Health↗
NASA NTRS1995

Cell shape, cytoskeletal mechanics, and cell cycle control in angiogenesis

Capillary endothelial cells can be switched between growth and differentiation by altering cell-extracellular matrix interactions and thereby, modulating cell shape. Studies were carried out to determine when cell shape exerts its growth-regulatory influence during cell cycle progression and to explore the role of cytoskeletal structure and mechanics in this control mechanism. When G0-synchronized cells were cultured in basic fibroblast growth factor (FGF)-containing defined medium on dishes coated with increasing densities of fibronectin or a synthetic integrin ligand (RGD-containing peptide), cell spreading, nuclear extension, and DNA synthesis all increased in parallel. To determine the minimum time cells must be adherent and spread on extracellular matrix (ECM) to gain entry into S phase, cells were removed with trypsin or induced to retract using cytochalasin D at different times after plating. Both approaches revealed that cells must remain extended for approximately 12-15 h and hence, most of G1, in order to enter S phase. After this restriction point was passed, normally 'anchorage-dependent' endothelial cells turned on DNA synthesis even when round and in suspension. The importance of actin-containing microfilaments in shape-dependent growth control was confirmed by culturing cells in the presence of cytochalasin D (25-1000 ng ml-1): dose-dependent inhibition of cell spreading, nuclear extension, and DNA synthesis resulted. In contrast, induction of microtubule disassembly using nocodazole had little effect on cell or nuclear spreading and only partially inhibited DNA synthesis. Interestingly, combination of nocodazole with a suboptimal dose of cytochalasin D (100 ng ml-1) resulted in potent inhibition of both spreading and growth, suggesting that microtubules are redundant structural elements which can provide critical load-bearing functions when microfilaments are partially compromised. Similar synergism between nocodazole and cytochalasin D was observed when cytoskeletal stiffness was measured directly in living cells using magnetic twisting cytometry. These results emphasize the importance of matrix-dependent changes in cell and nuclear shape as well as higher order structural interactions between different cytoskeletal filament systems for control of capillary cell growth during angiogenesis.

Non-NASA Center↗
NASA NTRS1992

Altering the cellular mechanical force balance results in integrated changes in cell, cytoskeletal and nuclear shape

Studies were carried out with capillary endothelial cells cultured on fibronectin (FN)-coated dishes in order to analyze the mechanism of cell and nuclear shape control by extracellular matrix (ECM). To examine the role of the cytoskeleton in shape determination independent of changes in transmembrane osmotic pressure, membranes of adherent cells were permeabilized with saponin (25 micrograms/ml) using a buffer that maintains the functional integrity of contractile microfilaments. Real-time videomicroscopic studies revealed that addition of 250 microM ATP resulted in time-dependent retraction and rounding of permeabilized cells and nuclei in a manner similar to that observed in intact living cells following detachment using trypsin-EDTA. Computerized image analysis confirmed that permeabilized cells remained essentially rigid in the absence of ATP and that retraction was stimulated in a dose-dependent manner as the concentration of ATP was raised from 10 to 250 microM. Maximal rounding occurred by 30 min with projected cell and nuclear areas being reduced by 69 and 41%, respectively. ATP-induced rounding was also accompanied by a redistribution of microfilaments resulting in formation of a dense net of F-actin surrounding retracted nuclei. Importantly, ATP-stimulated changes in cell, cytoskeletal, and nuclear form were prevented in permeabilized cells using a synthetic myosin peptide (IRICRKG) that has been previously shown to inhibit actomyosin filament sliding in muscle. In contrast, both the rate and extent of cell and nuclear rounding were increased in permeabilized cells exposed to ATP when the soluble FN peptide, GRGDSP, was used to dislodge immobilized FN from cell surface integrin receptors.(ABSTRACT TRUNCATED AT 250 WORDS).

Non-NASA Center↗
NASA NTRS2007

Intelligent Entity Behavior Within Synthetic Environments

This paper describes some elements in the development of realistic performance and behavior in the synthetic entities (players) which support Modeling and Simulation (M&S) applications, particularly military training. Modern human-in-the-loop (virtual) training systems incorporate sophisticated synthetic environments, which provide: 1. The operational environment, including, for example, terrain databases; 2. Physical entity parameters which define performance in engineered systems, such as aircraft aerodynamics; 3. Platform/system characteristics such as acoustic, IR and radar signatures; 4. Behavioral entity parameters which define interactive performance, including knowledge/reasoning about terrain, tactics; and, 5. Doctrine, which combines knowledge and tactics into behavior rule sets. The resolution and fidelity of these model/database elements can vary substantially, but as synthetic environments are designed to be compose able, attributes may easily be added (e.g., adding a new radar to an aircraft) or enhanced (e.g. Amending or replacing missile seeker head/ Electronic Counter Measures (ECM) models to improve the realism of their interaction). To a human in the loop with synthetic entities, their observed veridicality is assessed via engagement responses (e.g. effect of countermeasures upon a closing missile), as seen on systems displays, and visual (image) behavior. The realism of visual models in a simulation (level of detail as well as motion fidelity) remains a challenge in realistic articulation of elements such as vehicle antennae and turrets, or, with human figures; posture, joint articulation, response to uneven ground. Currently the adequacy of visual representation is more dependant upon the quality and resolution of the physical models driving those entities than graphics processing power per Se. Synthetic entities in M&S applications traditionally have represented engineered systems (e.g. aircraft) with human-in-the-loop performance characteristics (e.g. visual acuity) included in the system behavioral specification. As well, performance affecting human parameters such as experience level, fatigue and stress are coming into wider use (via AI approaches) to incorporate more uncertainty as to response type as well as performance (e.g. Where an opposing entity might go and what it might do, as well as how well it might perform).

Kruk, R. V.↗
NASA NTRS2009

Evolutionary Computing Methods for Spectral Retrieval

A methodology for processing spectral images to retrieve information on underlying physical, chemical, and/or biological phenomena is based on evolutionary and related computational methods implemented in software. In a typical case, the solution (the information that one seeks to retrieve) consists of parameters of a mathematical model that represents one or more of the phenomena of interest. The methodology was developed for the initial purpose of retrieving the desired information from spectral image data acquired by remote-sensing instruments aimed at planets (including the Earth). Examples of information desired in such applications include trace gas concentrations, temperature profiles, surface types, day/night fractions, cloud/aerosol fractions, seasons, and viewing angles. The methodology is also potentially useful for retrieving information on chemical and/or biological hazards in terrestrial settings. In this methodology, one utilizes an iterative process that minimizes a fitness function indicative of the degree of dissimilarity between observed and synthetic spectral and angular data. The evolutionary computing methods that lie at the heart of this process yield a population of solutions (sets of the desired parameters) within an accuracy represented by a fitness-function value specified by the user. The evolutionary computing methods (ECM) used in this methodology are Genetic Algorithms and Simulated Annealing, both of which are well-established optimization techniques and have also been described in previous NASA Tech Briefs articles. These are embedded in a conceptual framework, represented in the architecture of the implementing software, that enables automatic retrieval of spectral and angular data and analysis of the retrieved solutions for uniqueness.

Terrile, Richard↗
NASA NTRS2007

Rapid Engineering of Three-Dimensional, Multicellular Tissues With Polymeric Scaffolds

A process has been developed for the rapid tissue engineering of multicellular-tissue-equivalent assemblies by the controlled enzymatic degradation of polymeric beads in a low-fluid-shear bioreactor. In this process, the porous polymeric beads serve as temporary scaffolds to support the assemblies of cells in a tissuelike 3D configuration during the critical initial growth phases of attachment of anchorage-dependent cells, aggregation of the cells, and formation of a 3D extracellular matrix. Once the cells are assembled into a 3D array and enmeshed in a structural supportive 3D extracellular matrix (ECM), the polymeric scaffolds can be degraded in the low-fluid-shear environment of the NASA-designed bioreactor. The natural 3D tissuelike assembly, devoid of any artificial support structure, is maintained in the low-shear bioreactor environment by the newly formed natural cellular/ECM. The elimination of the artificial scaffold allows normal tissue structure and function.

Gonda, Steve R.↗
NASA NTRS2012

SLR Station Recovery, Center of Frame Motion, and Time Varying Gravity

Weekly station position estimates, beginning with 1993, are derived from the ITRF2008-based SLR processing of up to four satellites: Lageos 1, Lageos2, Starlette, and Stella. Helmert parameters obtained from c omparison of weekly SLR station positions and the a-priori SLRF2008 station complement are evaluated for geocenter motion and scale. Two me thods for modeling time varying gravity are employed in the SLR satel lite POD processing, with GGM03S serving as the static gravity field. Both methods forward model atmosphere gravity derived from 6-hour ECM WF pressure data. The standard approach applies an annual 20x20 field estimated from 4 years of GRACE data, and the IERS2003 recommended linear rates for C20, C30, C40, C21, and S21. The alternate approach us es a new set of low-order/degree 4x4 coefficients estimated weekly fr om SLR & DORIS processing to 10 satellites from 1993-2012. This exper imental tvg4x4 model has been shown to improve the TOPEX, Jason-1, and Jason-2 altimeter satellite orbits,. In this paper we apply the more detailed time-variable gravity modeling to the SLR satellite POD pro cessing and subsequent reference frame analyses. For this study we will evaluate the orbit differences (periodic and secular) for the satel lites concerned, characterize the impact on the station coordinate solutions, and the impact on reference frame parameters (geocenter and s cale).

Zelensky, Nikita P.↗
NASA NTRS2010

Pathways and Challenges to Innovation in Aerospace

This paper explores impediments to innovation in aerospace and suggests how successful pathways from other industries can be adopted to facilitate greater innovation. Because of its nature, space exploration would seem to be a ripe field of technical innovation. However, engineering can also be a frustratingly conservative endeavor when the realities of cost and risk are included. Impediments like the "find the fault" engineering culture, the treatment of technical risk as almost always evaluated in terms of negative impact, the difficult to account for expansive Moore's Law growth when making predictions, and the stove-piped structural organization of most large aerospace companies and federally funded research laboratories tend to inhibit cross-cutting technical innovation. One successful example of a multi-use cross cutting application that can scale with Moore's Law is the Evolutionary Computational Methods (ECM) technique developed at the Jet Propulsion Lab for automated spectral retrieval. Future innovations like computational engineering and automated design optimization can potentially redefine space exploration, but will require learning lessons from successful innovators.

Terrile, Richard J.↗
NASA NTRS2009

Evolutionary Computation for the Identification of Emergent Behavior in Autonomous Systems

Over the past several years the Center for Evolutionary Computation and Automated Design at the Jet Propulsion Laboratory has developed a technique based on Evolutionary Computational Methods (ECM) that allows for the automated optimization of complex computationally modeled systems. An important application of this technique is for the identification of emergent behaviors in autonomous systems. Mobility platforms such as rovers or airborne vehicles are now being designed with autonomous mission controllers that can find trajectories over a solution space that is larger than can reasonably be tested. It is critical to identify control behaviors that are not predicted and can have surprising results (both good and bad). These emergent behaviors need to be identified, characterized and either incorporated into or isolated from the acceptable range of control characteristics. We use cluster analysis of automatically retrieved solutions to identify isolated populations of solutions with divergent behaviors.

Terrile, Richard J.↗
NASA NTRS2015

ARC EMCS Experiments (Seedling Growth-2) Experiment Status

Presentation of the status of the ARC ISS (International Space Station) Experiment, Seedling Growth-2 to the Payload Operations Investigator Working Group meeting at MSFC, Huntsville AL. The experiment employs the European Modular Cultivation System (ECMS).

EMCS↗
NASA NTRS2017

Modulation of Bone Marrow Primary Cell Osteoblastogenesis and Cell Senescence by Mechanical Stimulation

Cell and animal studies conducted onboard the International Space Station and formerly the Shuttle flights have provided groundbreaking data illuminating the deleterious biological response of bone to mechanical unloading. Specifically CDKN1A/p-21 a cell senescence protein, was found to be upregulated in osteoprecursor cells of the femur during 15-day spaceflight, leading to the working hypothesis that CDKN1A/p-21 plays a role in inhibition of bone formation via mechanical regulation. To evaluate this hypothesis, utilizing a p-21 knockout mouse-line and relevant wildtype control, we cultured femoral bone marrow primary cells under unloaded (static) and cyclically stretched loading through a 30 day osteoblastogenesis protocol. Morphologic evaluation of the cultures demonstrated that mechanical stretching aligned the cells and increased the presence of defined focal adhesion expressing talin, integrin αvβ3, and PTK2 protein tyrosine kinase 2, also known as focal adhesion kinase (FAK) in both mouse strains. In corroboration with previous investigations of cell survival signals relation to FAK, our study found that with greater concentration of focal adhesions via stretch stimulation the live cell percentage was significantly higher than the unloaded controls (p-21 knockout line: +49.70%, p*=0.009, wildtype control: +18.14%, p*=.01). Also evaluated was the mineralization and ECM secretion capability of the differentiating cells. Von Kossa staining has shown that in the p-21 knockout cells unloaded cells produce more matrix that the stretch stimulated, however the matrix is unorganized presenting in sporadic nodules covering approximately 30% of the culture area at day 14 (n=6 wells) while the stretch stimulated cultures have less mineralization content the surface area containing mineralized matrix is greater (~68% at day 14). Q-PCR evaluation of the p-21 knockout cells revealed that canonical (β-catenin cascade) and non-canonical wnt11 and downstream planar cell polarity (wnt/PCP) pathway molecule RAC1 are prevalently upregulated with mechanical stimulation. Immunofluorescence for β-catenin and RAC1 showed co-localization at the nuclear membrane of the p-21 knockout cells but not the wildtype (n=1) suggesting that molecular communication via the canonical and wnt/PCP pathway are initiated by mechanical loading and experience regulation along the signaling cascade by CDKN1A/p-21. Future investigations will further elucidate this relationship and provide causal data demonstrating mechanical loading’s modulatory effect on p-21 expression change.

Juran, Cassandra M.↗
NASA NTRSDate not supplied

Functional Volume Assessment of an Early Version of the Mars Transit Habitat

During the summer of 2020, the NASA Mars Architecture Team (MAT) conducted a functional volume assessment of the Transit Habitat (TH). The TH has evolved substantially since that time and the current TH does not share the same internal configuration, but the insights gained from the assessment remain relevant. In virtually all architectures involving chemical, electric, or nuclear propulsion, the majority of the crew mission is spent aboard the TH. The Earth to Mars transit durations may vary from architecture to architecture, but all are on the order of hundreds of days, regardless of whether an opposition or conjunction class trajectory is selected, and regardless of whether the propulsion system is chemical, electric, or nuclear. Thus, the TH must provide capabilities appropriate to a very long duration. So, despite significant changes in the current Mars architecture over the past few years, this evaluation still provides useful recommendations in the form of habitability guidance that can be applied to current and future TH concepts. This assessment had six primary objectives: provide a sanity check to the BOC-derived TH layout; understand if we can fit the hardware and functional tasks in the volume; provide a high-level assessment of how aggressive the layout is; generate a list of challenges or assumptions necessary to make it work; generate a list of future work to refine understanding; and identify proposed requirements. All of this information is critical to drive habitat sizing studies and key architecture decisions. It was clear that a human-in-the-loop (HITL) evaluation of some kind would be necessary, but several challenges were immediately identified. The most information could be gleaned from a Desert Research and Technology Studies (DRATS) type of mission operations test (MOT), but the TH concept was too low fidelity to construct the type of prototype necessary to conduct a MOT, nor were their financial resources to do so. The next best option – and really the one most appropriate for the BOC’s stage of maturity – is a Virtual Reality (VR) walk-through evaluation. However, NASA was in a shutdown state due to the COVID-19 pandemic and the VR labs were inaccessible. As a result, a tabletop evaluation was created, using .jpg imagery from the ECM CAD model along with Excel-based datasheets. High-level crew living and working functions within the habitat were identified for evaluation. Questionnaires using Likert scales assessed acceptability of habitat functions and sim quality – the degree to which the function was represented in CAD. This enabled the evaluation to be conducted by personnel working remotely. Each function was evaluated individually along with several overarching habitability parameters and vehicle subsystems. The results of this evaluation are discussed, including methodological challenges and rating challenges. Acceptability results are discussed for functions that the participants were able to rate and participant comments for functions that could not be related are also described. Final conclusions are described, including challenges or assumptions needed to make the TH design acceptable, future work needed to refine understanding of the TH, and proposed habitat requirements based on test data.

Transit Habitat↗