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NASA NTRS1992

The rhodopsins: structure and function. Introduction

Nature makes use of the propensity of retinal for light-dependent double-bond isomerization in a number of systems and in a variety of ways. The common theme for light receptors based on this kind of chemistry is that (1) the retinal is bound in most cases to a small membrane protein via a protonated lysine-retinal Schiff base, (2) the absorption maximum in the visible is tuned to a suitable wavelength largely by electrostatic interaction with polar protein residues, and (3) the light-induced bond rotations and strains in the retinal set off reaction chains during which at least part of the excess free energy acquired is transferred to the protein and causes pK shifts of acidic residues and/or backbone conformational changes. The physiological consequence of the process initiated by absorption of light is either the activation of an information transfer chain (sensory and visual rhodopsins) or energy transduction which drives the electrogenic movement of ions across the membrane (ion-motive rhodopsins). Rhodopsins with these functions occur in bacteria and in higher organisms; from an evolutionary standpoint they are not related to one another. Nevertheless, all of these proteins are remarkably similar and form a distinct family.

NASA Discipline Number 52-30↗

NASA NTRS1993

Design and testing of a unique randomized gravity, continuous flow bioreactor

A rotating, null gravity simulator, or Couette bioreactor was successfully used for the culture of mammalian cells in a simulated microgravity environment. Two limited studies using Lipomyces starkeyi and Streptomyces clavuligerus were also conducted under conditions of simulated weightlessness. Although these studies with microorganisms showed promising preliminary results, oxygen limitations presented significant limitations in studying the biochemical and cultural characteristics of these cell types. Microbial cell systems such as bacteria and yeast promise significant potential as investigative models to study the effects of microgravity on membrane transport, as well as substrate induction of inactive enzyme systems. Additionally, the smaller size of the microorganisms should further reduce the gravity induced oscillatory particle motion and thereby improve the microgravity simulation on earth. Focus is on the unique conceptual design, and subsequent development of a rotating bioreactor that is compatible with the culture and investigation of microgravity effects on microbial systems. The new reactor design will allow testing of highly aerobic cell types under simulated microgravity conditions. The described reactor affords a mechanism for investigating the long term effects of reduced gravity on cellular respiration, membrane transfer, ion exchange, and substrate conversions. It offers the capability of dynamically altering nutrients, oxygenation, pH, carbon dioxide, and substrate concentration without disturbing the microgravity simulation, or Couette flow, of the reactor. All progeny of the original cell inoculum may be acclimated to the simulated microgravity in the absence of a substrate or nutrient. The reactor has the promise of allowing scientists to probe the long term effects of weightlessness on cell interactions in plants, bacteria, yeast, and fungi. The reactor is designed to have a flow field growth chamber with uniform shear stress, yet transfer high concentrations of oxygen into the culture medium. The system described allows for continuous, on line sampling for production of product without disturbing fluid and particle dynamics in the reaction chamber. It provides for the introduction of substrate, or control substances after cell adaptation to simulated microgravity has been accomplished. The reactor system provides for the nondisruptive, continuous flow replacement of nutrient and removal of product. On line monitoring and control of growth conditions such as pH and nutrient status are provided. A rotating distribution valve allows cessation of growth chamber rotation, thereby preserving the simulated microgravity conditions over longer periods of time.

Lassiter, Carroll B.↗

NASA NTRS2004

Integrity of Ceramic Parts Predicted When Loads and Temperatures Fluctuate Over Time

Brittle materials are being used, and being considered for use, for a wide variety of high performance applications that operate in harsh environments, including static and rotating turbine parts for unmanned aerial vehicles, auxiliary power units, and distributed power generation. Other applications include thermal protection systems, dental prosthetics, fuel cells, oxygen transport membranes, radomes, and microelectromechanical systems (MEMS). In order for these high-technology ceramics to be used successfully for structural applications that push the envelope of materials capabilities, design engineers must consider that brittle materials are designed and analyzed differently than metallic materials. Unlike ductile metals, brittle materials display a stochastic strength response because of the combination of low fracture toughness and the random nature of the size, orientation, and distribution of inherent microscopic flaws. This plus the fact that the strength of a component under load may degrade over time because of slow crack growth means that a probabilistic-based life-prediction methodology must be used when the tradeoffs of failure probability, performance, and useful life are being optimized. The CARES/Life code (which was developed at the NASA Glenn Research Center) predicts the probability of ceramic components failing from spontaneous catastrophic rupture when these components are subjected to multiaxial loading and slow crack growth conditions. Enhancements to CARES/Life now allow for the component survival probability to be calculated when loading and temperature vary over time.

Nemeth, Noel N.↗

NASA NTRS1990

Direct formulation of a 4-node hybrid shell element with rotational degrees of freedom

A simple 4-node assumed-stress hybrid quadrilateral shell element with rotational or 'drilling' degrees of freedom is formulated. The element formulation is based directly on a 4-node element rather than being derived from an 'internal' 8-node isoparametric element. The formulation is based on the principle of minimum complementary energy. The membrane part of the element has 12 degrees of freedom. The bending part of the element also has 12 degrees of freedom. The bending part of the element uses the Reissner-Mindlin plate theory which takes into account the transverse shear effects. Quadratic variations for both in-plane and out-of-plane displacement fields and linear variations for both in-plane and out-of-plane rotation fields are assumed along the edges of the element. The element passes the patch test, is nearly insensitive to mesh distortion, has no spurious modes, and is invariant with respect to node numbering.

Aminpour, Mohammad A.↗

NASA NTRS2000

Primate translational vestibuloocular reflexes. III. Effects of bilateral labyrinthine electrical stimulation

The effects of functional, reversible ablation and potential recruitment of the most irregular otolith afferents on the dynamics and sensitivity of the translational vestibuloocular reflexes (trVORs) were investigated in rhesus monkeys trained to fixate near and far targets. Translational motion stimuli consisted of either steady-state lateral and fore-aft sinusoidal oscillations or short-lasting transient lateral head displacements. Short-duration (usually <2 s) anodal (inhibitory) and cathodal (excitatory) currents (50-100 microA) were delivered bilaterally during motion. In the presence of anodal labyrinthine stimulation, trVOR sensitivity and its dependence on viewing distance were significantly decreased. In addition, anodal currents significantly increased phase lags. During transient motion, anodal stimulation resulted in significantly lower initial eye acceleration and more sluggish responses. Cathodal currents tended to have opposite effects. The main characteristics of these results were simulated by a simple model where both regularly and irregularly discharging afferents contribute to the trVORs. Anodal labyrinthine currents also were found to decrease eye velocity during long-duration, constant velocity rotations, although results were generally more variable compared with those during translational motion.

NASA Program Biomedical Research and Countermeasur↗

NASA NTRS1989

The sensory transduction pathways in bacterial chemotaxis

Bacterial chemotaxis is a useful model for investigating in molecular detail the behavioral response of cells to changes in their environment. Peritrichously flagellated bacteria such as coli and typhimurium swim by rotating helical flagella in a counterclockwise direction. If flagellar rotation is briefly reversed, the bacteria tumble and change the direction of swimming. The bacteria continuously sample the environment and use a temporal sensing mechanism to compare the present and immediate past environments. Bacteria respond to a broad range of stimuli including changes in temperature, oxygen concentration, pH and osmotic strength. Bacteria are attracted to potential sources of nutrition such as sugars and amino acids and are repelled by other chemicals. In the methylation-dependent pathways for sensory transduction and adaptation in E. coli and S. typhimurium, chemoeffectors bind to transducing proteins that span the plasma membrane. The transducing proteins are postulated to control the rate of autophosphorylation of the CheA protein, which in turn phosphorylates the CheY protein. The phospho-CheY protein binds to the switch on the flagellar motor and is the signal for clockwise rotation of the motor. Adaptation to an attractant is achieved by increasing methylation of the transducing protein until the attractant stimulus is cancelled. Responses to oxygen and certain sugars involve methylation-independent pathways in which adaption occurs without methylation of a transducing protein. Taxis toward oxygen is mediated by the electron transport system and changes in the proton motive force. Recent studies have shown that the methylation-independent pathway converges with the methylation-dependent pathway at or before the CheA protein.

Taylor, Barry L.↗

NASA NTRS2017

Mapping Vinyl Cyanide and Other Nitriles in Titan's Atmosphere Using ALMA

Vinyl cyanide (C2H3CN) is theorized to form in Titan's atmosphere via high-altitude photochemistry and is of interest regarding the astrobiology of cold planetary surfaces due to its predicted ability to form cell membrane-like structures (azotosomes) in liquid methane. In this work, we follow up on the initial spectroscopic detection of C2H3CN on Titan by Palmer et al. with the detection of three new C2H3CN rotational emission lines at submillimeter frequencies. These new, high-resolution detections have allowed for the first spatial distribution mapping of C2H3CN on Titan. We present simultaneous observations of C2H5CN, HC3N, and CH3CN emission, and obtain the first (tentative) detection of C3H8 (propane) at radio wavelengths. We present disk-averaged vertical abundance profiles, two-dimensional spatial maps, and latitudinal flux profiles for the observed nitriles. Similarly to HC3N and C2H5CN, which are theorized to be short-lived in Titan's atmosphere, C2H3CN is most abundant over the southern (winter) pole, whereas the longer-lived CH3CN is more concentrated in the north. This abundance pattern is consistent with the combined effects of high-altitude photochemical production, poleward advection, and the subsequent reversal of Titan's atmospheric circulation system following the recent transition from northern to southern winter. We confirm that C2H3CN and C2H5CN are most abundant at altitudes above 200 km. Using a 300 km step model, the average abundance of C2H3CN is found to be 3.03 +/- 0.29ppb, with a C2H5CN/C2H3CN abundance ratio of 2.43 +/- 0.26. Our HC3N and CH3CN spectra can be accurately modeled using abundance gradients above the tropopause, with fractional scale-heights of 2.05 +/- 0.16 and 1.63 +/- 0.02, respectively.

Lai, J. C.-Y.↗

NASA NTRS2006

A Cassegrain Offset-Fed Dual-Band Reflectarray

An X/Ka dual-band microstrip reflectarray with circular polarization (CP) has been constructed using thin membranes and Cassegrain offset-fed configuration. It is believed that this is the first Cassegrain reflectarray ever been developed. This antenna has a 75-cm-diameter aperture and uses metallic subreflector and angular-rotated annular ring elements. It achieved a measured -3dB-gain bandwidth of 700 MHz at X-band and 1.5 GHz at Ka-band, as well as a CP bandwidth (3dB axial ratio) of more than 700 MHz at X-band and more than 2 GHz at Ka-band. The measured peak efficiencies are 49.8% at X-band and 48.2% at Ka-band.

offset fed↗

NASA NTRS2011

Low Fluid Shear Culture of Staphylococcus Aureus Represses hfq Expression and Induces an Attachment-Independent Biofilm Phenotype

Background: The opportunistic pathogen, Staphylococcus aureus, experiences fluctuations in fluid shear during infection and colonization of a human host. Colonization frequently occurs at mucus membrane sites such as in the gastrointestinal tract where the bacterium may experience low levels of fluid shear. The response of S. aureus to low fluid shear remains unclear. Methods: S. aureus was cultured to stationary phase using Rotating-Wall Vessel (RWV) bioreactors which produce a physiologically relevant low fluid shear environment. The bacterial aggregates that developed in the RWV were evaluated by electron microscopy as well as for antibiotic resistance and other virulence-associated stressors. Genetic expression profiles for the low-shear cultured S. aureus were determined by microarray analysis and quantitative real-time PCR. Results: Planktonic S. aureus cultures in the low-shear environment formed aggregates completely encased in high amounts of extracellular polymeric substances. In addition, these aggregates demonstrated increased antibiotic resistance indicating attachment-independent biofilm formation. Carotenoid production in the low-shear cultured S. aureus was significantly decreased, and these cultures displayed an increased susceptibility to oxidative stress and killing by whole blood. The hfq gene, associated with low-shear growth in Gram negative organisms, was also found to be down-regulated in S. aureus. Conclusions: Collectively, this data suggests that S. aureus decreases virulence characteristics in favor of a biofilm-dwelling colonization phenotype in response to a low fluid shear environment. Furthermore, the identification of an Hfq response to low-shear culture in S. aureus, in addition to the previously reported responses in Gram negative organisms, strongly suggests an evolutionarily conserved response to mechanical stimuli among structurally diverse prokaryotes.

Ott, C. Mark↗

NASA NTRS2009

Low-Pressure, Field-Ionizing Mass Spectrometer

A small mass spectrometer utilizing a miniature field ionization source is now undergoing development. It is designed for use in a variety of applications in which there are requirements for a lightweight, low-power-consumption instrument that can analyze the masses of a wide variety of molecules and ions. The device can operate without need for a high-vacuum, carrier-gas feed radioactive ionizing source, or thermal ionizer. This mass spectrometer can operate either in the natural vacuum of outer space or on Earth at any ambient pressure below 50 torr (below about 6.7 kPa) - a partial vacuum that can easily be reached by use of a small sampling pump. This mass spectrometer also has a large dynamic range - from singly charged small gas ions to deoxyribonucleic acid (DNA) fragments larger than 104 atomic mass units - with sensitivity adequate for detecting some molecules and ions at relative abundances of less than one part per billion. This instrument (see figure) includes a field ionizer integrated with a rotating-field mass spectrometer (RFMS). The field ionizer effects ionization of a type characterized as "soft" in the art because it does not fragment molecules or initiate avalanche arcing. What makes the "soft" ionization mode possible is that the distance between the ionizing electrodes is less than mean free path for ions at the maximum anticipated operating pressure, so that the ionizer always operates on the non-breakdown side of the applicable Paschen curve (a standard plot of breakdown potential on the ordinate and pressure electrode separation on the abscissa). The field ionizer in this instrument is fabricated by micromachining a submicron-thick membrane out of an electrically nonconductive substrate, coating the membrane on both sides to form electrodes, then micromachining small holes through the electrodes and membrane. Because of the submicron electrode separation, even a potential of only 1 V applied between the electrodes gives rise to an electric field with a strength of in excess of a megavolt per meter strong enough to ionize any gas molecules passing through the holes. An accelerator grid and an electrostatic deflector focus the ions from the field ionizer into the rotating-field cell of the RFMS. The potentials applied to the electrodes of the cell to generate the rotating electric field typically range from 1 to 13 V. The ions travel in well-defined helices within this cell, after which they are collected in a Faraday cup. The mass of most of the molecules reaching the Faraday cup decreases with increasing frequency of rotation of the electric field in the cell. Therefore, the frequency of rotation of the electric field is made to vary in order to scan through a desired range of ion masses: For example, lightweight gas molecules are scanned at frequencies in the megahertz range, while DNA and other large organic molecules are scanned at kilohertz frequencies.

Hartley, Frank↗

NASA NTRS1989

High density tape casting system

A system is provided for casting thin sheets (or tapes) of particles bound together, that are used for oxygen membranes and other applications, which enables the particles to be cast at a high packing density in a tape of uniform thickness. A slurry contains the particles, a binder, and a solvent, and is cast against the inside walls of a rotating chamber. Prior to spraying the slurry against the chamber walls, a solvent is applied to a container. The solvent evaporates to saturate the chamber with solvent vapor. Only then is the slurry cast. As a result, the slurry remains fluid long enough to spread evenly over the casting surface formed by the chamber, and for the slurry particles to become densely packed. Only then is the chamber vented to remove solvent, so the slurry can dry. The major novel feature is applying solvent vapor to a rotating chamber before casting slurry against the chamber walls.

Collins, Earl R., Jr.↗

NASA NTRS2004

Microgravity and Cellular Consequences in Lymphocyte Function

Mammalian cells adapt to the environment of low gravity and express a series of responses, some possibly from direct effects on cells and others based on environmental conditions created by microgravity. Human lymphocytes in microgravity culture are functionally diminished in activation and locomotion. Both processes are integral to optimal immune response to fight pathogens. The NASA Rotating-wall vessel (RWV) is a well-accepted analog for microgravity culture on the ground. Gene array experiments and immunoblotting identified upstream events in human lymphocytes adapting to microgravity analog culture. Microgravity induces selective changes, many of which are cell membrane related. Results showed that upstream of PKC in the T cell activation cascade, PLC-gamma and LAT are significantly diminished. ZAP 70 which controls LAT activation is also down regulated in modeled microgravity. Thus events governing cell shape might warrant attention in microgravity conditions. The goal of this study is to delineate response suites that are consequential, direct or indirect effects of the microgravity environment and which of these are essential to lymphocytes

Pellis, Neal R.↗

NASA NTRS2007

Telescope Formation at L2 for Observing Earth's Atmosphere

Two documents describe a proposed Earth-atmosphere observatory to orbit the Sun at the Sun-Earth L2 Lagrange point -- a point of unstable equilibrium in the shadow of the Earth, about 1.5 million km from the Earth along an outward projection of the Earth-Sun axis. The observatory would comprise two spacecraft flying in precision formation: (1) a primary-aperture spacecraft, from which would be deployed a 25-m diameter membrane primary mirror aimed at the Earth, and (2) a secondary-telescope spacecraft at the focal plane of the primary mirror, 125-m distant along the axis towards the Earth. The secondary telescope would be aimed at the primary mirror and slowly rotated to scan the focused annular image of the visible illuminated portion of the Earth's atmosphere during continuous occultation of the Sun.

Mettler, Edward↗

NASA NTRS2010

Ultrasonic Monitoring of Ply Crack and Delamination Formation in Composite Tube Under Torsion Load

As a simple model of a rotor spar, a circular graphite-epoxy composite laminate cylinder was subjected to cyclic torsional load. The test section of the cylindrical specimen varied from four to six plies of plus or minus 45 degree fibers, due to intentional ply overlaps and gaps. A layer of 13-micrometer Teflon film was inserted between plies at three locations to serve as delamination initiators. A commercial X-Y scanner was mounted to the load frame to enable ultrasonic inspection without removing the specimen. A focused immersion probe was mounted in a captive water column with a rugged Nitrile membrane tip, which was coupled to the cylinder using a mist of soapy water. The transducer was aligned normal to the cylinder surface using the X-axis. Scanning was performed along the length of the specimen with the Y-axis and the specimen was incrementally rotated by the torsion head of the load frame. After 350k cycles of torsion, several linear 45 degree diagonal indications appeared as 5-40% attenuation of the back wall echo, with no apparent echoes from the interior of the composite, suggesting through-ply cracks in the innermost ply. Crack indications grew and new cracks appeared as torsion cycling continued. Internal reflections from delaminations associated with the growing ply cracks appeared after 500k cycles. Three areas of extensive multi-layer delaminations appeared after 1150k cycles. Failure of the specimen occurred at 1600k cycles. The observed progressive damage was not associated with the Teflon inclusions. Concurrent thermographic measurements provided lower resolution confirmation of the damage observed.

Johnston, P. H.↗

NASA NTRS1993

Use of microgravity bioreactors for development of an in vitro rat salivary gland cell culture model

During development, salivary gland (SG) cells both secrete factors which modulate cellular behavior and express specific hormone receptors. Whether SG cell growth is modulated by an autocrine epidermal growth factor (EGF) receptor-mediated signal transduction pathway is not clearly understood. SG tissue is the synthesis site for functionally distinct products including growth factors, digestive enzymes, and homeostasis maintaining factors. Historically, SG cells have proven difficult to grow and may be only maintained as limited three-dimensional ductal-type structures in collagen gels or on reconstituted basement membrane gels. A novel approach to establishing primary rat SG cultures is use of microgravity bioreactors originally designed by NASA as low-shear culture systems for predicting cell growth and differentiation in the microgravity environment of space. These completely fluid-filled bioreactors, which are oriented horizontally and rotate, have proven advantageous for Earth-based culture of three-dimensional cell assemblies, tissue-like aggregates, and glandular structures. Use of microgravity bioreactors for establishing in vitro models to investigate steroid-mediated secretion of EGF by normal SG cells may also prove useful for the investigation of cancer and other salivary gland disorders. These microgravity bioreactors promise challenging opportunities for future applications in basic and applied cell research.

NASA Program Space Biology↗

NASA NTRS2008

Elastic Distribution of Microshutters, Measurements Obtainable on James Web Space Telescope

Spectrographic astronomy measurements in the near-infrared region will be done by functional two-dimensional microshutter arrays that are being fabricated at the NASA Goddard Space Flight Center for the James Webb Space Telescope (JWST). These micro-shutter arrays will represent the first mission-critical MEMS devices to be flown in space. JWST will use microshutter arrays to select focal plane object. 2-D programmable aperture masks of more than 200,000 elements select such space object. The use of silicon wafer material promises high efficiency and high contrast. Microshutter operation temperature is around 35K. Microshutter arrays are fabricated as close-packed silicon nitride membranes with a unit cell size of 105 x 204 micrometers. A layer of magnetic material is deposited onto each shutter. Individual shutters are equipped with a torsion flexure. Reactive ion etching (RIE) releases the shutters so they can open up to 90 degrees using the torsion flexure. Shutter rotation is initiated into a silicon support structure via an external magnetic field. Two electrically independent aluminum electrodes are deposited, one onto each shutter and another onto the support structure side-wall, permitting electrostatic latching and 2-D addressing to hold specific shutters open via external electronics.

Kletetschka, Gunther↗

NASA NTRS1994

Simulated microgravity does not alter epithelial cell adhesion to matrix and other molecules

Microgravity has advantages for the cultivation of tissues with high fidelity; however, tissue formation requires cellular recognition and adhesion. We tested the hypothesis that simulated microgravity does not affect cell adhesion. Human colorectal carcinoma cells were cultured in the NASA Rotating Wall Vessel (RWV) under low shear stress with randomization of the gravity vector that simulates microgravity. After 6 - 7 days, cells were assayed for binding to various substrates and compared to cells grown in standard tissue culture flasks and static suspension cultures. The RWV cultures bound as well to basement membrane proteins and to Carcinoembryonic Antigen (CEA), an intercellular adhesion molecule, as control cultures did. Thus, microgravity does not alter epithelial cell adhesion and may be useful for tissue engineering.

Jessup, J. M.↗

NASA NTRS1995

Effects of clinorotation and microgravity on sweet clover columella cells treated with cytochalasin D

The cytoskeleton of columella cells is believed to be involved in maintaining the developmental polarity of cells observed as a reproducible positioning of cellular organelles. It is also implicated in the transduction of gravitropic signals. Roots of sweet clover (Melilotus alba L.) seedlings were treated with a microfilament disrupter, cytochalasin D, on a slowly rotating horizontal clinostat (2 rpm). Electron micrographs of treated columella cells revealed several ultrastructural effects including repositioning of the nucleus and the amyloplasts and the formation of endoplasmic reticulum (ER) whorls. However, experiments performed during fast clinorotation (55 rpm) showed an accumulation (but no whorling) of a disorganized ER network at the proximal and distal pole and a random distribution of the amyloplasts. Therefore, formation of whorls depends upon the speed of clinorotation, and the overall impact of cytochalasin D suggests the necessity of microfilaments in organelle positioning. Interestingly, a similar drug treatment performed in microgravity aboard the US Space Shuttle Endeavour (STS-54, January 1993) caused a displacement of ER membranes and amyloplasts away from the distal plasma membrane. In the present study, we discuss the role of microfilaments in maintaining columella cell polarity and the utility of clinostats to simulate microgravity.

manned↗