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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 37 records · Page 2

Automated Microbial Metabolism Laboratory

The effect of several environmental parameters on previously developed life detection systems is explored. Initial attempts were made to conduct all the experiments in a moist mode (high soil volume to water volume ratio). However, only labeled release and measurement of ATP were found to be feasible under conditions of low moisture. Therefore, these two life detection experiments were used for most of the environmental effects studies. Three soils, Mojave (California desert), Wyaconda (Maryland, sandy loam) and Victoria Valley (Antarctic desert) were generally used throughout. The environmental conditions studied included: incubation temperature 3 C to 80 C, ultraviolet irradiation of soils, variations in soil/liquid ratio, specific atmospheric gases, various antimetabolites, specific substrates, and variation in pH. An experiment designed to monitor nitrogen metabolism was also investigated.

Source record↗

Riveting-force gage

Gage monitors riveting forces applied when components are mounted on printed-circuit boards. Correct swaging pressures have been established for specific substrate materials such as phenolics and ceramics.

Rotta, J. W., Jr.↗

Microelectrophoretic apparatus and process

New gel tray and lid assemblies designed for use in conjunction with slotted electrophoretic membranes were developed to take advantage of recently improved microelectrophoretic accessories which include a multisample applicator capable of applying up to 10 samples consecutively or simultaneously, and a temperature control plate for dissipating the heat produced by electrophoresis in a gel. The trays and membranes can be marketed ready for use as electrophoretic media or impregnated with various specific substrates and dyes

Grunbaum, B. W.↗

Crystal Structure of the Catalytic Domain of a Serine Threonine Protein Phosphatase

Reversible phosphorylation of serine and threonine residues is a well-recognized mechanism in eukaryotic cells for the regulation of cell-cycle progression, cell growth and metabolism. Human serine/threonine phosphatases can be placed into two major families, PPP and PPM. To date the structure on one PPP family member (PPl) has been determined. Here we present the structure of a 323-residue catalytic domain of a second phosphatase belonging to the PPP family of enzyme. catalytic domain of the enzyme has been determined to 1.60Angstrom resolution and refined to R=17.5 and Rfree = 20.8%. The catalytic domain possesses a unique fold consisting of a largely monolithic structure, divisible into closely-associated helical and sheet regions. The catalytic site contains two manganese ions that are involved in substrate binding and catalysis. The enzyme crystallizes as a dimer that completely buries catalytic surfaces of both monomers, Also, the structure shows evidence of some flexibility around the active site cleft that may be related to substrate specificity of this enzyme.

Swinglel, Mark↗

Structure, Function, Self-Assembly and Origin of Simple Membrane Proteins

Integral membrane proteins perform such essential cellular functions as transport of ions, nutrients and waste products across cell walls, transduction of environmental signals, regulation of cell fusion, recognition of other cells, energy capture and its conversion into high-energy compounds. In fact, 30-40% of genes in modem organisms codes for membrane proteins. Although contemporary membrane proteins or their functional assemblies can be quite complex, their transmembrane fragments are usually remarkably simple. The most common structural motif for these fragments is a bundle of alpha-helices, but occasionally it could be a beta-barrel. In a series of molecular dynamics computer simulations we investigated self-organizing properties of simple membrane proteins based on these structural motifs. Specifically, we studied folding and insertion into membranes of short, nonpolar or amphiphatic peptides. We also investigated glycophorin A, a peptide that forms sequence-specific dimers, and a transmembrane aggregate of four identical alpha-helices that forms an efficient and selective voltage-gated proton channel was investigated. Many peptides are attracted to water-membrane interfaces. Once at the interface, nonpolar peptides spontaneously fold to a-helices. Whenever the sequence permits, peptides that contain both polar and nonpolar amino also adopt helical structures, in which polar and nonpolar amino acid side chains are immersed in water and membrane, respectively. Specific identity of side chains is less important. Helical peptides at the interface could insert into the membrane and adopt a transmembrane conformation. However, insertion of a single helix is unfavorable because polar groups in the peptide become completely dehydrated upon insertion. The unfavorable free energy of insertion can be regained by spontaneous association of peptides in the membrane. The first step in this process is the formation of dimers, although the most common are aggregates of 4-7 helices. The helices could arrange themselves such that they formed pores capable of transporting ions and small molecules across membranes. Stability of transmembrane aggregates of simple proteins is often only marginal and, therefore, it can be regulated by environmental signals or small sequence modifications in the region of interhelical interactions. A key step in the earliest evolution of membrane proteins was the emergence of selectivity for specific substrates. Many channels could become selective if one or only a few properly chosen amino acids are properly placed along the channel, acting as filters or gates. This is a convenient evolutionary solution because it does not require imposing conditions on the whole sequence.

Pohorille, Andrew↗

Purification and characterization of soluble (cytosolic) and bound (cell wall) isoforms of invertases in barley (Hordeum vulgare) elongating stem tissue

Three different isoforms of invertases have been detected in the developing internodes of barley (Hordeum vulgare). Based on substrate specificities, the isoforms have been identified to be invertases (beta-fructosidases EC 3.2.1.26). The soluble (cytosolic) invertase isoform can be purified to apparent homogeneity by diethylaminoethyl cellulose, Concanavalin-A Sepharose, organo-mercurial Sepharose, and Sephacryl S-300 chromatography. A bound (cell wall) invertase isoform can be released by 1 molar salt and purified further by the same procedures as above except omitting the organo-mercurial Sepharose affinity chromatography step. A third isoform of invertase, which is apparently tightly associated with the cell wall, cannot be isolated yet. The soluble and bound invertase isoforms were purified by factors of 60- and 7-fold, respectively. The native enzymes have an apparent molecular weight of 120 kilodaltons as estimated by gel filtration. They have been identified to be dimers under denaturing and nondenaturing conditions. The soluble enzyme has a pH optimum of 5.5, Km of 12 millimolar, and a Vmax of 80 micromole per minute per milligram of protein compared with cell wall isozyme which has a pH optimum of 4.5, Km of millimolar, and a Vmax of 9 micromole per minute per milligram of protein.

NASA Program Space Biology↗

Immunoassay readout method using extrinsic Raman labels adsorbed on immunogold colloids

An immunoassay readout method based on surface-enhanced Raman scattering (SERS) is described. The method exploits the SERS-derived signal from reporter molecules that are coimmobilized with biospecific species on gold colloids. This concept is demonstrated in a dualanalyte sandwich assay, in which two different antibodies covalently bound to a solid substrate specifically capture two different antigens from an aqueous sample. The captured antigens in turn bind selectively to their corresponding detection antibodies. The detection antibodies are conjugated with gold colloids that are labeled with different Raman reporter molecules, which serve as extrinsic labels for each type of antibody. The presence of a specific antigen is established by the characteristic SERS spectrum of the reporter molecule. A near-infrared diode laser was used to excite efficiently the SERS signal while minimizing fluorescence interference. We show that, by using different labels with little spectral overlap, two different antigenic species can be detected simultaneously. The potential of this concept to function as a readout strategy for multiple analytes is briefly discussed.

NASA Program Advanced Human Support Technology↗

Ubiquitin conjugation by the N-end rule pathway and mRNAs for its components increase in muscles of diabetic rats

Insulin deficiency (e.g., in acute diabetes or fasting) is associated with enhanced protein breakdown in skeletal muscle leading to muscle wasting. Because recent studies have suggested that this increased proteolysis is due to activation of the ubiquitin-proteasome (Ub-proteasome) pathway, we investigated whether diabetes is associated with an increased rate of Ub conjugation to muscle protein. Muscle extracts from streptozotocin-induced insulin-deficient rats contained greater amounts of Ub-conjugated proteins than extracts from control animals and also 40-50% greater rates of conjugation of (125)I-Ub to endogenous muscle proteins. This enhanced Ub-conjugation occurred mainly through the N-end rule pathway that involves E2(14k) and E3alpha. A specific substrate of this pathway, alpha-lactalbumin, was ubiquitinated faster in the diabetic extracts, and a dominant negative form of E2(14k) inhibited this increase in ubiquitination rates. Both E2(14k) and E3alpha were shown to be rate-limiting for Ub conjugation because adding small amounts of either to extracts stimulated Ub conjugation. Furthermore, mRNA for E2(14k) and E3alpha (but not E1) were elevated 2-fold in muscles from diabetic rats, although no significant increase in E2(14k) and E3alpha content could be detected by immunoblot or activity assays. The simplest interpretation of these results is that small increases in both E2(14k) and E3alpha in muscles of insulin-deficient animals together accelerate Ub conjugation and protein degradation by the N-end rule pathway, the same pathway activated in cancer cachexia, sepsis, and hyperthyroidism.

Non-NASA Center↗

Interactions of the C-terminal Domain of Human Ku70 with DNA Substrate: A Molecular Dynamics Study

NASA is developing a systems biology approach to improve the assessment of health risks associated with space radiation. The primary toxic and mutagenic lesion following radiation exposure is the DNA double strand break (DSB), thus a model incorporating proteins and pathways important in response and repair of this lesion is critical. One key protein heterodimer for systems models of radiation effects is the Ku(sub 70/80) complex. The Ku70/80 complex is important in the initial binding of DSB ends following DNA damage, and is a component of nonhomologous end joining repair, the primary pathway for DSB repair in mammalian cells. The C-terminal domain of Ku70 (Ku70c, residues 559-609), contains an helix-extended strand-helix motif and similar motifs have been found in other nucleic acid-binding proteins critical for DNA repair. However, the exact mechanism of damage recognition and substrate specificity for the Ku heterodimer remains unclear in part due to the absence of a high-resolution structure of the Ku70c/DNA complex. We performed a series of molecular dynamics (MD) simulations on a system with the subunit Ku70c and a 14 base pairs DNA duplex, whose starting structures are designed to be variable so as to mimic their different binding modes. By analyzing conformational changes and energetic properties of the complex during MD simulations, we found that interactions are preferred at DNA ends, and within the major groove, which is consistent with previous experimental investigations. In addition, the results indicate that cooperation of Ku70c with other subunits of Ku(sub 70/80) is necessary to explain the high affinity of binding as observed in experiments.

Hu, Shaowen↗

Interactions of the SAP Domain of Human Ku70 with DNA Substrate: A Molecular Dynamics Study

NASA is developing a systems biology approach to improve the assessment of health risks associated with space radiation. The primary toxic and mutagenic lesion following radiation exposure is the DNA double strand break (DSB), thus a model incorporating proteins and pathways important in response and repair of this lesion is critical. One key protein heterodimer for systems models of radiation effects is the Ku70/80 complex. The Ku70/80 complex is important in the initial binding of DSB ends following DNA damage, and is a component of nonhomologous end joining repair, the primary pathway for DSB repair in mammalian cells. The SAP domain of Ku70 (residues 556-609), contains an a helix-extended strand-helix motif and similar motifs have been found in other nucleic acid-binding proteins critical for DNA repair. However, the exact mechanism of damage recognition and substrate specificity for the Ku heterodimer remains unclear in part due to the absence of a high-resolution structure of the SAP/DNA complex. We performed a series of molecular dynamics (MD) simulations on a system with the SAP domain of Ku70 and a 10 base pairs DNA duplex. Large-scale conformational changes were observed and some putative binding modes were suggested based on energetic analysis. These modes are consistent with previous experimental investigations. In addition, the results indicate that cooperation of SAP with other domains of Ku70/80 is necessary to explain the high affinity of binding as observed in experiments.

Hu, Shaowen↗

Structurally altered capsular polysaccharides produced by mutant bacteria

Structurally altered capsular polysaccharides are produced by mutant bacteria. These polysaccharides are isolated by selecting a wild type bacterial strain and a phage producing degradative enzymes that have substrate specificity for the capsular polysaccharides produced by the wild type bacteria. Phage-resistant mutants producing capsular polysaccharides are selected and the structurally altered capsular polysaccharide is isolated therefrom.

Kern, Roger G.↗

Method for producing capsular polysaccharides

Structurally altered capsular polysaccharides are produced by mutant bacteria. These polysaccharides are isolated by selecting a wild type bacterial strain and a phage producing degradative enzymes that have substrate specificity for the capsular polysaccharides produced by the wild type bacteria. Phage-resistant mutants producing capsular polysaccharides are selected and the structurally altered capsular polysaccharide is isolated therefrom.

Kern, Roger G.↗

Demonstration of Fluid Dynamics for Plant Growing Systems in Varied Gravity Environments Through Scaled Capillary Models

The development of reliable and bioregenerative crop growth production systems is vital for human exploration into deep space. As humans prepare for space travel beyond LEO, scientists need to find a way to provide reliable and adequate water delivery for all stages of the plant's life cycle. Past production systems struggled with this and often led to overwatering in the system. To better understand this issue, NASA's Plant Water Management (PWM) experiments were able to model fluid flow through granular substrates, specifically a clay-based material arcilite, in 0-G. The PWM study can help researchers to predict fluid flow through the systems, however their model only works for an arcilite based system and was unable to account for different stages of plant growth. For future missions, payload requirements to support crop production system will need to be limited, leading to the use of in situ resources. This project aims to validate the PWM experiments as well as incorporate various materials into the design for growth systems. To meet these objectives, a series of terrestrial experiments will be deployed to mimic all gravities. By modifying the material, fluid, and size of the test subjects, the effect of Earth's gravity can be minimized. This project will aid researchers in the design of future crop production systems for surface missions by creating a refined model that can be used at all stages of plant growth and utilize in situ resources.

Plant Biology↗

Evolution in vitro of an RNA enzyme with altered metal dependence

The Tetrahymena group I ribozyme catalyses a sequence-specific phosphodiester cleavage reaction on an external RNA oligonucleotide substrate in the presence of a divalent metal cation cofactor. This reaction proceeds readily with either Mg2+ or Mn2+, but no detectable reaction has been reported when other divalent cations are used as the sole cofactor. Cations such as Ca2+, Sr2+ and Ba2+ can stabilize the correct folded conformation of the ribozyme, thereby partially alleviating the Mg2+ or Mn2+ requirement. But catalysis by the ribozyme involves coordination of either Mg2+ or Mn2+ at the active site, resulting in an overall requirement for one of these two cations. Here we use an in vitro evolution process to obtain variants of the Tetrahymena ribozyme that are capable of cleaving an RNA substrate in reaction mixtures containing Ca2+ as the divalent cation. These findings extend the range of different chemical environments available to RNA enzymes and illustrate the power of in vitro evolution in generating macromolecular catalysts with desired properties.

NASA Discipline Exobiology↗

Automated microbial metabolism laboratory

The design and rationale of an advanced labeled release experiment based on single addition of soil and multiple sequential additions of media into each of four test chambers are outlined. The feasibility for multiple addition tests was established and various details of the methodology were studied. The four chamber battery of tests include: (1) determination of the effect of various atmospheric gases and selection of that gas which produces an optimum response; (2) determination of the effect of incubation temperature and selection of the optimum temperature for performing Martian biochemical tests; (3) sterile soil is dosed with a battery of C-14 labeled substrates and subjected to experimental temperature range; and (4) determination of the possible inhibitory effects of water on Martian organisms is performed initially by dosing with 0.01 ml and 0.5 ml of medium, respectively. A series of specifically labeled substrates are then added to obtain patterns in metabolic 14CO2 (C-14)O2 evolution.

Source record↗

Microbial ecology of extreme environments: Antarctic dry valley yeasts and growth in substrate-limited habitats

The success of the Antarctic Dry Valley yeasts presumeably results from adaptations to multiple stresses, to low temperatures and substrate-limitation as well as prolonged resting periods enforced by low water availability. Previous investigations have suggested that the crucial stress is substrate limitation. Specific adaptations may be pinpointed by comparing the physiology of the Cryptococcus vishniacii complex, the yeasts of the Tyrol Valley, with their congeners from other habitats. Progress was made in methods of isolation and definition of ecological niches, in the design of experiments in competition for limited substrate, and in establishing the relationships of the Cryptococcus vishniacii complex with other yeasts. In the course of investigating relationships, a new method for 25SrRNA homology was developed. For the first time it appears that 25SrRNA homology may reflect parallel or convergent evolution.

Vishniac, H. S.↗

Numerical Solution of Light Scattered from and Transmitted through a Rough Dielectric Surface with Applications to Periodic Roughness and Isolated Structures

Light scattering and transmission by rough surfaces are of considerable interest in a variety of applications including remote sensing and characterization of surfaces. In this work, the finite-difference time domain technique is applied to calculate the scattered and transmitted electromagnetic fields of an infinite periodic rough surface. The elements of Mueller matrix for scattered light are calculated by an integral of the near fields over a significant number of periods of the surface. The normalized Mueller matrix elements of the scattered light and the spatial distribution of the transmitted flux for a monolayer of micron-sized dielectric spheres on a silicon substrate are presented. The numerical results show that the nonzero Mueller matrix elements of the system of the monolayer of dielectric spheres on a silicon substrate have specific maxima at some scattering angles. These maxima may be used in characterization of the feature of the system. For light transmitted through the monolayer of spheres, our results show that the transmitted energy focuses around the ray passing through centers of the spheres. At other locations, the transmitted flux is very small. The technique also may be used to calculate the perturbance of the electromagnetic field due to the presence of an isolated structure on the substrate.

Sun, Wenbo↗

Amorphous metallic films in silicon metallization systems

The general objective was to determine the potential of amorphous metallic thin films as a means of improving the stability of metallic contacts to a silicon substrate. The specific objective pursued was to determine the role of nitrogen in the formation and the resulting properties of amorphous thin-film diffusion barriers. Amorphous metallic films are attractive as diffusion barriers because of the low atomic diffusivity in these materials. Previous investigations revealed that in meeting this condition alone, good diffusion barriers are not necessarily obtained, because amorphous films can react with an adjacent medium (e.g., Si, Al) before they recrystallize. In the case of a silicon single-crystalline substrate, correlation exists between the temperature at which an amorphous metallic binary thin film reacts and the temperatures at which the films made of the same two metallic elements react individually. Amorphous binary films made of Zr and W were investigated. Both react with Si individually only at elevated temperatures. It was confirmed that such films react with Si only above 700 C when annealed in vacuum for 30 min. Amorphous W-N films were also investigated. They are more stable as barriers between Al and Si than polycrystalline W. Nitrogen effectively prevents the W-Al reaction that sets in at 500 C with polycrystalline W.

Nicolet, M. A.↗