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At least 1,585 records · Page 88
All sourcesNASA NTRSDOE OSTI
NASA NTRS2000

Aqueous two-phase partition applied to the isolation of plasma membranes and Golgi apparatus from cultured mammalian cells

Partitioning in dextran-poly(ethylene)glycol (PEG) aqueous-aqueous phase systems represents a mature technology with many applications to separations of cells and to the preparation of membranes from mammalian cells. Most applications to membrane isolation and purification have focused on plasma membranes, plasma membrane domains and separation of right side-out and inside-out plasma membrane vesicles. The method exploits a combination of membrane properties, including charge and hydrophobicity. Purification is based upon differential distributions of the constituents in a sample between the two principal compartments of the two phases (upper and lower) and at the interface. The order of affinity of animal cell membranes for the upper phase is: endoplasmic reticulum <mitochondria<Golgi apparatus<lysosomes and endosomes< plasma membranes. Salt concentrations and temperature affect partitioning behavior and must be precisely standardized. In some cases, it is more fortuitous to combine aqueous two-phase partition with other procedures to obtain a more highly purified preparation. A procedure is described for preparation of Golgi apparatus from transformed mammalian cells that combines aqueous two-phase partition and centrifugation. Also described is a periodic NADH oxidase, a new enzyme marker for right side-out plasma membrane vesicles not requiring detergent disruptions for measurement of activity.

Non-NASA Center↗
NASA NTRS2000

Rhodoferax antarcticus sp. nov., a moderately psychrophilic purple nonsulfur bacterium isolated from an Antarctic microbial mat

A new species of purple nonsulfur bacteria isolated from an Antarctic microbial mat is described. The organism, designated strain ANT.BR, was mildly psychrophilic, growing optimally at 15-18 degrees C with a growth temperature range of 0-25 degrees C. Cells of strain ANT.BR were highly motile curved rods and spirals, contained bacteriochlorophyll a, and showed a multicomponent in vivo absorption spectrum. A specific phylogenetic relationship was observed between strain ANT.BR and the purple bacterium Rhodoferax fermentans FR2T, and the two organisms shared several physiological and other phenotypic properties, with the notable exception of growth temperature optimum. Tests of genomic DNA hybridization, however, showed Rfx. fermentans FR2T and strain ANT.BR to be genetically distinct bacteria. Because of its unique set of properties, especially its requirement for low growth temperatures, we propose to recognize strain ANT.BR as a new species of the genus Rhodoferax, Rhodoferax antarcticus, named for its known habitat, the Antarctic.

NASA Discipline Exobiology↗
NASA NTRS2004

Isolated Liquid Droplet Combustion: Inhibition and Extinction Studies

Introduction of fire suppressants to the ambient environment surrounding a heterogeneous diffusion flame may be an inefficient technique for fire safety in systems without buoyant flows. Carbon dioxide substitution for nitrogen diluent leads to significant modifications of the sphero-symmetric burning behavior of isolated n-heptane droplets, partly through increased heat capacity within the gaseous diffusion flame, but mostly because of modifications in spectral radiative coupling in the gas phase. Effects of longer time scale phenomena such as sooting and slow gas-phase/droplet convection remain to be determined. Similar methodologies can be applied to evaluate the effects and efficacy of chemical inhibitors in the liquid and gas phases.

Dryer, F. L.↗
NASA NTRS1998

Isolation and characterization of Arabidopsis mutants defective in the induction of ethylene biosynthesis by cytokinin

Cytokinins elevate ethylene biosynthesis in etiolated Arabidopsis seedlings via a post-transcriptional modification of one isoform of the key biosynthetic enzyme ACC synthase. In order to begin to dissect the signaling events leading from cytokinin perception to this modification, we have isolated a series of mutants that lack the ethylene-mediated triple response in the presence of cytokinin due to their failure to increase ethylene biosynthesis. Analysis of genetic complementation and mapping revealed that these Cin mutants (cytokinin-insensitive) represent four distinct complementation groups, one of which, cin4, is allelic to the constitutive photomorphogenic mutant fus9/cop10. The Cin mutants have subtle effects on the morphology of adult plants. We further characterized the Cin mutants by analyzing ethylene biosynthesis in response to various other inducers and in adult tissues, as well as by assaying additional cytokinin responses. The cin3 mutant did not disrupt ethylene biosynthesis under any other conditions, nor did it disrupt any other cytokinin responses. Only cin2 disrupted ethylene biosynthesis in multiple circumstances. cin1 and cin2 made less anthocyanin in response to cytokinin. cin1 also displayed reduced shoot initiation in tissue culture in response to cytokinin, suggesting that it affects a cytokinin signaling element.

Non-NASA Center↗
NASA NTRS1995

Isolation, characterization, and primary structure of rubredoxin from the photosynthetic bacterium, Heliobacillus mobilis

Rubredoxin is a small nonheme iron protein that serves as an electron carrier in bacterial systems. Rubredoxin has now been isolated and characterized from the strictly anaerobic phototroph, Heliobacillus mobilis. THe molecular mass (5671.3 Da from the amino acid sequence) was confirmed and partial formylation of the N-terminal methionyl residue was established by matrix-assisted laser desorption mass spectroscopy. The complete 52-amino-acid sequence was determined by a combination of N-terminal sequencing by Edman degradation and C-terminal sequencing by a novel method using carboxypeptidase treatment in conjunction with amino acid analysis and laser desorption time of flight mass spectrometry. The molar absorption coefficient of Hc. mobilis rubredoxin at 490 nm is 6.9 mM-1 cm-1 and the midpoint redox potential at pH 8.0 is -46 mV. The EPR spectrum of the oxidized form shows resonances at g = 9.66 and 4.30 due to a high-spin ferric iron. The amino acid sequence is homologous to those of rubredoxins from other species, in particular, the gram-positive bacteria, and the phototrophic green sulfur bacteria, and the evolutionary implications of this are discussed.

NASA Discipline Number 52-30↗
NASA NTRS1992

Casein kinase II protein kinase is bound to lamina-matrix and phosphorylates lamin-like protein in isolated pea nuclei

A casein kinase II (CK II)-like protein kinase was identified and partially isolated from a purified envelope-matrix fraction of pea (Pisum sativum L.) nuclei. When [gamma-32P]ATP was directly added to the envelope-matrix preparation, the three most heavily labeled protein bands had molecular masses near 71, 48, and 46 kDa. Protein kinases were removed from the preparation by sequential extraction with Triton X-100, EGTA, 0.3 M NaCl, and a pH 10.5 buffer, but an active kinase still remained bound to the remaining lamina-matrix fraction after these treatments. This kinase had properties resembling CK II kinases previously characterized from animal and plant sources: it preferred casein as an artificial substrate, could use GTP as efficiently as ATP as the phosphoryl donor, was stimulated by spermine, was calcium independent, and had a catalytic subunit of 36 kDa. Some animal and plant CK II kinases have regulatory subunits near 29 kDa, and a lamina-matrix-bound protein of this molecular mass was recognized on immunoblot by anti-Drosophila CK II polyclonal antibodies. Also found associated with the envelope-matrix fraction of pea nuclei were p34cdc2-like and Ca(2+)-dependent protein kinases, but their properties could not account for the protein kinase activity bound to the lamina. The 71-kDa substrate of the CK II-like kinase was lamin A-like, both in its molecular mass and in its cross-reactivity with anti-intermediate filament antibodies. Lamin phosphorylation is considered a crucial early step in the entry of cells into mitosis, so lamina-bound CK II kinases may be important control points for cellular proliferation.

NASA Discipline Plant Biology↗
NASA NTRS1992

Isolation, characterization, and amino acid sequences of auracyanins, blue copper proteins from the green photosynthetic bacterium Chloroflexus aurantiacus

Three small blue copper proteins designated auracyanin A, auracyanin B-1, and auracyanin B-2 have been isolated from the thermophilic green gliding photosynthetic bacterium Chloroflexus aurantiacus. All three auracyanins are peripheral membrane proteins. Auracyanin A was described previously (Trost, J. T., McManus, J. D., Freeman, J. C., Ramakrishna, B. L., and Blankenship, R. E. (1988) Biochemistry 27, 7858-7863) and is not glycosylated. The two B forms are glycoproteins and have almost identical properties to each other, but are distinct from the A form. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis apparent monomer molecular masses are 14 (A), 18 (B-2), and 22 (B-1) kDa. The amino acid sequences of the B forms are presented. All three proteins have similar absorbance, circular dichroism, and resonance Raman spectra, but the electron spin resonance signals are quite different. Laser flash photolysis kinetic analysis of the reactions of the three forms of auracyanin with lumiflavin and flavin mononucleotide semiquinones indicates that the site of electron transfer is negatively charged and has an accessibility similar to that found in other blue copper proteins. Copper analysis indicates that all three proteins contain 1 mol of copper per mol of protein. All three auracyanins exhibit a midpoint redox potential of +240 mV. Light-induced absorbance changes and electron spin resonance signals suggest that auracyanin A may play a role in photosynthetic electron transfer. Kinetic data indicate that all three proteins can donate electrons to cytochrome c-554, the electron donor to the photosynthetic reaction center.

NASA Discipline Exobiology↗
NASA NTRS2004

Infrared Spectroscopy of Matrix-Isolated Polycyclic Aromatic Compounds and Their Ions: Phenazine, a Dual Substituted Polycyclic Aromatic Nitrogen Heterocycle - 7

The matrix-isolation technique has been employed to measure the mid-infrared spectra of phenazine (C12H8N2), a dual substituted Polycyclic Aromatic Nitrogen Heterocycle (PANH), in the neutral, cationic and anionic forms. The experimentally measured band frequencies and intensities are tabulated and compared with their calculated values as well as those of the non-substituted parent molecule, anthracene. The theoretical band positions and intensities were calculated using both the 3-31 G as well as the larger 6-3lG* Basis Sets. A comparison of the results can be found in the tables. The spectroscopic properties of phenazine and its cation are similar to those observed in mono-substituted PANHs, with one exception. The presence of a second nitrogen atom results in an additional enhancement of the cation's total integrated intensity, for the 1500-1000 cm(sup -1) (6.7 to 10 micron) region, over that observed for a mono-substituted PANH cation. The significance of this enhancement and the astrobiological implications of these results are discussed.

Mattioda, A. L.↗
NASA NTRS2003

Massive Stars and the Energy Balance of the Interstellar Medium: The Impact of an Isolated 60 M. Star - 1

We present results of numerical simulations carried out with a two-dimensional radiation hydrodynamics code in order to study the impact of massive stars on their surrounding interstellar medium. This first paper deals with the evolution of the circumstellar gas around an isolated 60 M. star. The interaction of the photo- ionized H II region with the stellar wind bubble forms a variety of interesting structures like shells, clouds, fingers, and spokes. These results demonstrate that complex structures found in H II regions are not necessarily relics from the time before the gas became ionized but may result from dynamical processes during the course of the H II region evolution. We have also analyzed the transfer and deposit of the stellar wind and radiation energy into the circumstellar medium until the star explodes as a supernova. Although the total mechanical wind energy supplied by the star is negligible compared to the accumulated energy of the Lyman continuum photons, the kinetic energy imparted to the circumstellar gas over the star s lifetime is 4 times higher than for a comparable windless simulation. Furthermore, the thermal energy of warm photoionized gas is lower by some 55%). Our results document the necessity to consider both ionizing radiation and stellar winds for an appropriate description of the interaction of OB stars with their circumstellar environment.

Freyer, Tim↗
NASA NTRS2004

Chondrules and Isolated Grains in the Fountain Hills Bencubbinite

The Fountain Hills (FH) meteorite was recently classified as a Bencubbin-like (CB(sub a)) chondrite, which are part of the CR clan [1]. The FH O-isotopic composition is indistinguishable from CB(sub a) chondrites. Metal and silicate compositions are consistent with the CB(sub a) classification. Significant differences between FH and the other CB(sub a) chondrites were noted. These include abundant porphyritic chondrules and complete lack of sulfide minerals. We are furthering this investigation by analyzing silicate chondrules and isolated grains in FH to determine more about its composition, thermal history, and implications for chondrule formation in the early solar system.

LaBlue, A. R.↗
NASA NTRS2005

High Resolution Spectroscopy And Timing Of The Isolated Neutron Star RBS 1774

The 2004 May 31 XMM-Newton observation was reprocessed using SASv6.0.0 and times of high background were filtered out. The net exposure time remaining was 23 ks. The source was clearly detected in MOS1, MOS2 and PN chips. We performed both timing and spectroscopic analysis on the data. We performed a spectral analysis by fitting data from the three EPIC detectors simultaneously, finding that the broadband spectrum can be represented by a single absorbed blackbody, with kT = 0.10 keV. The fitting revealed the presence of an absorption feature at 0:7 keV, but the data did have enough resolution to allow us to discriminate between an absorption line and an edge. We also tested magnetized models of Pavlov et a1 and Zavlin et al, but found that fits with these models were considerably worse than with a blackbody. For the timing analysis, we extracted the counts within a 3000 radius aperture in both PN and MOS 1 and MOS2 but with the aperture truncated by a chord where it approached the edge of the CCD window in each case: this maximized the counts while avoiding any edge effects. We analyzed PN, MOSl and MOS2 data both individually and combined using the Maximum Likelihood Periodogram technique of Zane et al. (2002) and Cropper et al. (2004). Periods from 10000 s to 30 ms were searched, ensuring that in each case the period grid was 2.5 times better sampled than the Nyquist frequency. The search revealed a significant period at 9.437s. Taken overall, we found the characteristics of RBS 1774 to be remarkably similar to those of another X-ray faint isolated neutron stars. These results were written up for the Astrophysical Journal, and the paper has recently been accepted for publication.

Mushotzky, Richard↗
NASA NTRS1999

Customized Hermetic Feedthrough Developed to Isolate Fluids

A common problem occurs when refrigerant fluids wick inside the insulation of thermocouple wires through a compressor's casing feedthrough and then leak into the adjacent disconnect box outside the casing. Leaking fluids create an unfavorable situation inside the disconnect box and may contaminate the fluids. To address this problem, NASA Lewis Research Center s Manufacturing Engineering Division developed a customized hermetic feedthrough for a bank of Worthington compressors. In these compressors, bearing temperatures are measured by internal thermocouples embedded in bearings located inside the compressor casings. The thermocouple wires need to be routed outside the casing and read at another location. These wires are short and are terminated to a disconnect strip inside the casing. The bearings operate at about 170 F, but because the casing is filled with R12 refrigerant oil, the casing has a maximum temperature of about 100 F. The operating conditions of these compressors permit the use of an epoxy that is compatible with the R12 fluid. The desired finished product is a stainless steel tube that has been filled solid with epoxy after thermocouple wires bonded and sealed by epoxy have been inserted through its length. Shrink tubing extends from both ends of the tube. The process that was developed to isolate the thermocouple wires from the R12 fluid follows. For this application, use an 8-in.-long piece of 0.500-in. 304 stainless steel tube with six pairs of 24-gauge stranded, PTFE-insulated (polytetrafluoroethylene) type "T" thermocouple wires for each feedthrough. Use shrink tubing to strain relief the insulated wires at their exit from the stainless steel tube. Cut the wire to length and identify the location of the stainless steel tube sleeve with masking tape. Then, remove the outer insulation from a 2-in. section of wire that will be inside the tube, and carefully strip to bare wire a 1-in. section in the middle of the section with the outer insulation removed. For an effective seal, the epoxy must penetrate between the strands when stranded conductors are used. Make the seal with epoxy bond on the bare wire. The bare wire must be encapsulated with a thin layer of the epoxy that leaves only a very low profile. These encapsulated wires must cure before the assembly can be continued. Then, inspect the cured wires for complete encapsulation before going to the next step. Insert the wires in the stainless steel tube and orient them so that the epoxied stripped sections are staggered within the tube; then, apply shrink tubing to one end of the cleaned wires, positioning it inside the edge of the tube. The small gaps between the wires on the other end will be used to inject the epoxy into the tube. Let the epoxy cure inside the tube, free of any voids. Then, continue to fill the tube until the entire 8-in. length is nearly filled, allowing room for the other strain-relieving shrink tubing. Since this first design, the process has been adjusted to fit many needs and situations. Customized feedthroughs have been assembled from various wire types, wire gauges, and/or stainless steel tube passages. The fittings selected to mount these feedthroughs allow their use in other areas, such as pressure or vacuum systems.

Meredith, Roger D.↗
NASA NTRS2002

Low-Loss, High-Isolation Microwave Microelectromechanical Systems (MEMS) Switches Being Developed

Switches, electrical components that either permit or prevent the flow of electricity, are the most important and widely used electrical devices in integrated circuits. In microwave systems, switches are required for switching between the transmitter and receiver; in communication systems, they are needed for phase shifters in phased-array antennas, for radar and communication systems, and for the new class of digital or software definable radios. Ideally, switches would be lossless devices that did not depend on the electrical signal's frequency or power, and they would not consume electrical power to change from OFF to ON or to maintain one of these two states. Reality is quite different, especially at microwave frequencies. Typical switches in microwave integrated circuits are pin diodes or gallium arsenide (GaAs) field-effect transistors that are nonlinear, with characteristics that depend on the power of the signal. In addition, they are frequency-dependent, lossy, and require electrical power to maintain a certain state. A new type of component has been developed that overcomes most of these technical difficulties. Microelectromechanical (MEMS) switches rely on mechanical movement as a response to an applied electrical force to either transmit or reflect electrical signal power. The NASA Glenn Research Center has been actively developing MEMS for microwave applications for over the last 5 years. Complete fabrication procedures have been developed so that the moving parts of the switch can be released with near 100-percent yield. Moreover, the switches fabricated at Glenn have demonstrated state-of-the-art performance. A typical MEMS switch is shown. The switch extends over the signal and ground lines of a finite ground coplanar waveguide, a commonly used microwave transmission line. In the state shown, the switch is in the UP state and all the microwave power traveling along the transmission line proceeds unimpeded. When a potential difference is applied between the cantilever and the transmission line, the cantilever is pulled downward until it connects the signal line to the ground planes, creating a short circuit. In this state, all the microwave power is reflected. The graph shows the measured performance of the switch, which has less than 0.1 dB of insertion loss and greater than 30dB of isolation. These switches consume negligible electrical power and are extremely linear. Additional research is required to address reliability and to increase the switching speed.

Ponchak, George E.↗
NASA NTRS2005

A "Kane's Dynamics" Model for the Active Rack Isolation System: Addition of Umbilicals to the Nonlinear Model - Part 3

In the late 1980s, microgravity researchers began to voice their concern that umbilical-transmitted energy could significantly degrade the acceleration environment of microgravity space science experiments onboard manned spacecraft. Since umbilicals are necessary for many experiments, control designers began to seek ways to compensate for these "indirect" disturbances. Hampton, et al., used the Kane s method to develop a model of the active rack isolation system (ARIS) that includes (1) actuator control forces, (2) direct disturbance forces, and (3) indirect, actuator-transmitted disturbances. Their model does not, however, include the indirect, umbilical-transmitted disturbances. Since the umbilical stiffnesses are not negligible, these indirect disturbances must be included in the model. Until the umbilicals have been appropriately included, the model will be incomplete. This Technical Memorandum presents a nonlinear model of ARIS with umbilicals included. Model verification was achieved by utilizing two commercial-off-the-shelf software tools. Various forces and moments were applied to the model to yield simulated responses of the system. Plots of the simulation results show how various critical points on an ARIS-outfitted international standard payload rack behave under the application of direct disturbances, indirect disturbances, and control forces. Simulations also show system response to a variety of initial conditions.

Rupert, J. K.↗
NASA NTRS2006

A Search for X-ray Emission in Isolated Compact Triplets

We describe preliminary results of an exploratory search for diffuse X-ray emission in a sample of the poorest galaxy groups, i.e., isolated compact triplets of galaxies. These systems represent the simplest forms of galaxy clustering while manifesting all the complexities inherent in other groups. We have selected 20 compact triplets for this initial study. The component galaxies are expected to interact with each other and with the group's intergalactic medium, if present, in complex ways that trigger high-energy processes.

Brown, Beth A.↗
NASA NTRS2002

Companions to isolated elliptical galaxies: revisiting the Bothun-Sullivan (1977) sample using the NASA/IPAC extragalactic database

We investigate the number of physical companion galaxies for a sample of relatively isolated elliptical galaxies. The NASA/IPAC Extragalactic Database (NED) has been usedto reinvestigate the incidence of satellite galaxies for a sample of 34 elliptical galaxies, firstinvestigated by Bothun & Sullivan (1977) using a visual inspection of Palomar Sky Survey prints out to a projected search radius of 75 kpc. We have repeated their original investigation usingdata cataloged data in NED. Nine of these ellipticals appear to be members of galaxy clusters:the remaining sample of 25 galaxies reveals an average of +1.0 f 0.5 apparent companions per galaxy within a projected search radius of 75 kpc, in excess of two equal-area comparisonregions displaced by 150-300 kpc. This is nearly an order of magnitude larger than the +0.12+/- 0.42 companions/galaxy found by Bothun & Sullivan for the identical sample. Making use of published radial velocities, mostly available since the completion of the Bothun-Sullivan study,identifies the physical companions and gives a somewhat lower estimate of +0.4 companions per elliptical. This is still a factor of 3x larger than the original statistical study, but giventhe incomplete and heterogeneous nature of the survey redshifts in NED, it still yields a firmlower limit on the number (and identity) of physical companions. An expansion of the searchradius out to 300 kpc, again restricted to sampling only those objects with known redshifts in NED, gives another lower limit of 4.3 physical companions per galaxy. (Excluding fiveelliptical galaxies in the Fornax cluster this average drops to 3.5 companions per elliptical.)These physical companions are individually identified and listed, and the ensemble-averagedradial density distribution of these associated galaxies is presented. For the ensemble, the radial density distribution is found to have a fall-off consistent with p c( R^-0.5 out to approximately150 kpc. For non-Fornax cluster companions the fall-off continues out to the 300-kpc limit of thesurvey. The velocity dispersion of these companions is found to be constant with projected radial distance from the central elliptical, holding at a value of approximately +/- 300-350 km/sec overall.

NED NASA/IPAC Extragalactic Database↗