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At least 109 records · Page 6

Continuous flow electrophoresis system experiments on shuttle flights STS-6 and STS-7

The development of a space continuous flow electrophoresis system (CFES) is discussed. The objectives of the experiment were: (1) to use a model sample material at a high concentration to evaluate the continuous flow electrophoresis process in the McDonnell Douglass CFES instrument and compare its separation resolution and sample throughput with related devices on Earth, and (2) to expand the basic knowledge of the limitations imposed by fluid flows and particle concentration effects on the electrophoresis process by careful design and evaluation of the space experiment. Hemoglobin and polysaccharide were selected as samples of concentration effects. The results from space show a large band spread of the high concentration of the single species of hemoglobin that was principally due to the mismatch of electrical conductivity between the sample and buffer.

Snyder, Robert S.↗

Thermophysical properties of lysozyme (protein) solutions

Thermophysical properties of protein solutions composed of the lysozyme crystals with a 0.1 M sodium acetate and 5 percent NaCl solution as the buffer (pH = 4.0) are determined. The properties being measured include specific heat, thermal conductivity, dynamic viscosity, and surface tension. The protein concentrations are varied. Thermal diffusivity is calculated using the measured results. The purpose of the research is to measure thermophysical properties of lysozyme solutions which would serve as the data bank for controlling and modeling the crystal growth process on earth as well as in space.

Liu, Jiaching↗

Microscopic iron metal on glass and minerals - A tool for studying regolith maturity

A novel method of producing mixtures of glass or minerals with iron metal is presented. A portion of the Fe(2+) in basaltic glass and minerals can be reduced to metal in a few hours at 1100 C and an oxygen fugacity well below the iron-wustite buffer. Part of the iron metals forms rounded submicrometer blebs on the surfaces and in some cases within the grains. A concentration of such blebs equivalent to 20-30 percent of a grain's surface area can totally dominate the reflectance spectra of basaltic glass, pyroxene, and olivine. The production of optically opaque iron metal blebs, combined with the decline in Fe(2+), affects the glass and mineral reflectance spectra in three ways: by lowering the overall reflectivity, reducing the spectral contrast of absorption features, and producing a continuum with a general rise in reflectivity toward longer wavelengths.

Allen, C. C.↗

Bonding-Compatible Corrosion Inhibitor for Rinsing Metals

A corrosion-inhibiting mixture of compounds has been developed for addition to the water used to rinse metal parts that have been cleaned with aqueous solutions in preparation for adhesive bonding of the metals to rubber and rubber-like materials. Prior to the development of this corrosion inhibitor, the parts (made, variously, of D6AC steel and 7075-T73 aluminum) were rinsed by deionized water, which caused corrosion in some places on the steel parts especially in such occluded places as sealing surfaces and threaded blind holes. An integral part of the particular cleaning process is the deposition of a thin layer of silicates and silane primers that increase the strength of the adhesive bond. The corrosion inhibitor is formulated, not only to inhibit corrosion of both D6AC steel and 7075- T73 aluminum, but also to either increase or at least not reduce the strength of the adhesive bond to be formed subsequently. The corrosion inhibitor is a mixture of sodium silicate and sodium tetraborate. The sodium silicate functions as both a corrosion inhibitor and a bond-strength promoter in association with the silane primers. The sodium tetraborate buffers the rinse solution at the optimum pH and functions as a secondary corrosion inhibitor for the steel. For a given application, the concentrations of sodium silicate and sodium tetraborate must be chosen in a compromise among the needs to inhibit corrosion of steel, inhibit corrosion of aluminum, and minimize cosmetic staining of both steel and aluminum. Concentrations of sodium silicate in excess of 150 parts of silicon per million parts of solution (ppm Si) have been determined to enhance inhibition of corrosion; unfortunately, because of the alkalinity of sodium silicate, even a small concentration can raise the pH of the rinse solution to such a level that aluminum becomes corroded despite the inhibiting effect. The pH of a solution that contains a high concentration of sodium silicate can be decreased by adding sodium tetraborate. On the other hand, the addition of sodium tetraborate increases the concentration of dissolved solids to such a high level that cosmetic staining becomes an issue.

Saunders, C. R.↗

Buffering of stratospheric circulation by changing amounts of tropical ozone - A Pinatubo case study

Stratospheric aerosol from Mount Pinatubo heated the tropical lower stratosphere by about 0.3 K/day mainly due to absorption of terrestrial infrared radiation. This heating was dissipated by: (1) an observed increase in stratospheric temperatures, which enhanced the radiation cooling; (2) additional mean upward motion, observed for the aerosol cloud, which led to adiabatic cooling; and (3) reductions in ozone concentrations resulting from enhanced upward motions. Each of these processes operated on a different time scale: maximum temperatures were observed after about 90 days; maximum ozone losses of about -1.5 ppm occurred after 140 days when the enhanced vertical velocities effectively lifted the ozone profile by about 2 km. We believe this shows that ozone plays an important role in buffering vertical motion in the tropical lower stratosphere, and hence the residual Brewer Dobson circulation of the whole stratosphere.

Kinne, S.↗

Oxidation state of the atmosphere and crust of Venus from Pioneer Venus results

Recent spacecraft observations of Venus permit a detailed model of sulfur chemistry in the atmosphere-lithosphere system. Pioneer Venus experiments confirm that, as predicted, COS and H2S are dominant over SO2 in the lower atmosphere, and that the equilibrium concentrations of S2 and S3 are significant. Many criteria serve to bracket the oxidation state of the crust: it is nearly certain that the S2(2-)/SO4(2-) buffer regulates the oxygen fugacity, and that FeO is at least as abundant as F2O3 in crustal silicates. A highly oxidized crust (as, for example, would result from O2 absorption complementary to escape of vast amounts of H2) is incompatible with the gas-phase sulfur chemistry. If the Pioneer Venus mass spectrometer estimates of the abundance of sulfur gases are correct, earth-like models for the bulk composition of Venus are seriously in error, and a far lower FeO content is required for Venus.

Lewis, J. S.↗

DMA Modulus as a Screening Parameter for Compatibility of Polymeric Containment Materials with Various Solutions for use in Space Shuttle Microgravity Protein Crystal Growth (PCG) Experiments

Protein crystals are grown in microgravity experiments inside the Space Shuttle during orbit. Such crystals are basically grown in a five-component system containing a salt, buffer, polymer, organic and water. During these experiments, a number of different polymeric containment materials must be compatible with up to hundreds of different PCG solutions in various concentrations for durations up to 180 days. When such compatibility experiments are performed at NASA/MSFC (Marshall Space Flight Center) simultaneously on containment material samples immersed in various solutions in vials, the samples are rather small out of necessity. DMA4 modulus was often used as the primary screening parameter for such small samples as a pass/fail criterion for incompatibility issues. In particular, the TA Instruments DMA 2980 film tension clamp was used to test rubber O-rings as small in I.D. as 0.091 in. by cutting through the cross-section at one place, then clamping the stretched linear cord stock at each end. The film tension clamp was also used to successfully test short length samples of medical/surgical grade tubing with an O.D. of 0.125 in.

Wingard, Charles Doug↗

Administration of the non-steroidal anti-inflammatory drug ibuprofen increases macrophage concentrations but reduces necrosis during modified muscle use

OBJECTIVE: To test the hypothesis that ibuprofen administration during modified muscle use reduces muscle necrosis and invasion by select myeloid cell populations. METHODS: Rats were subjected to hindlimb unloading for 10 days, after which they experienced muscle reloading by normal weight-bearing to induce muscle inflammation and necrosis. Some animals received ibuprofen by intraperitoneal injection 8 h prior to the onset of muscle reloading, and then again at 8 and 16 h following the onset of reloading. Other animals received buffer injection at 8 h prior to reloading and then ibuprofen at 8 and 16 h following the onset of reloading. Control animals received buffer only at each time point. Quantitative immunohistochemical analysis was used to assess the presence of necrotic muscle fibers, total inflammatory infiltrate, neutrophils, ED1+ macrophages and ED2+ macrophages at 24 h following the onset of reloading. RESULT: Administration of ibuprofen beginning 8 h prior to reloading caused significant reduction in the concentration of necrotic fibers, but increased the concentration of inflammatory cells in muscle. The increase in inflammatory cells was attributable to a 2.6-fold increase in the concentration of ED2+ macrophages. Animals treated with ibuprofen 8 h following the onset of reloading showed no decrease in muscle necrosis or increase in ED2+ macrophage concentrations. CONCLUSION: Administration of ibuprofen prior to increased muscle loading reduces muscle damage, but increases the concentration of macrophages that express the ED2 antigen. The increase in ED2+ macrophage concentration and decrease in necrosis may be mechanistically related because ED2+ macrophages have been associated with muscle regeneration and repair.

Non-NASA Center↗

Geochemical Advances in Mercury Science Facilitated by a Landed Mission

The data from the MErcury Surface, Space ENvironment, GEochemistry, and Ranging (MESSENGER) spacecraft have revealed several surprising characteristics about the surface of Mercury, leading to its classification as a geochemical endmember among the terrestrial planets. Some of these features include elevated abundances of up to 3 wt% S, C enrichment as high as 4 wt% over the local mean in low reflectance materials (LRM), Na up to 5 wt% at high northern latitudes, and Fe abundances typically lower than 2 wt% [e.g., 1–4]. The S and Fe concentrations have been used to infer that Mercury’s igneous history evolved under highly reduced oxygen fugacity conditions between 2.6 and 7.3 log10 units below the iron-wüstite buffer [e.g., 5], which is more reducing than any other terrestrial planet in the solar system [e.g., 6]. This highly reduced nature has important consequences for the differentiation and thermal/magmatic evolution of Mercury. While the immense amount of data collected by MESSENGER revealed Mercury as a geochemical endmember, this new knowledge gained raised additional questions that necessitate continued exploration of the planet. Fortunately, BepiColombo launched in October of 2018, and this joint ESA/JAXA dual-orbiter spacecraft is the most ambitious effort yet attempted to explore Mercury [e.g., 7]. Looking beyond BepiColombo, there are major aspects of Mercury’s geochemical character and evolution for which significant knowledge gaps can be dramatically improved with data acquired from the planet’s surface via in situ landed science.

Vander Kaaden, K. E.↗

Size Exclusion Chromatography Studies of the Initial Self-Association Steps of Chicken Egg White Lysozyme Nucleation

Nucleation is one of the least understood aspects of crystallogenesis. In the case of macromolecule nucleation, this understanding is further hampered by uncertainty over what precisely is being discussed. We define the process of solute self-association (aggregation, oligomerization, interaction, clustering, etc.) whereby n-mers (n > or = 2) having a crystallographic or nascent crystallographic arrangement leading to the critical nucleus reversibly form in the solution, to be part of the nucleation process. This reversible self-association process is a fundamental part of the nucleation process, and occurs as a function of the solute concentration. In the case of chicken egg white lysozyme, a considerable body of experimental evidence leads us to the conclusion that it also forms the crystal growth units. Size exclusion chromatography is a simple and direct method for determining the equilibrium constants for the self-association process. A Pharmacia FPLC system was used to provide accurate solution flow rates. The column, injection valve, and sample loop were all mounted within a temperature-controlled chamber. Chromatographically re-purified lysozyme was first dialyzed against the column equilibration buffer, with injection onto the column after several hours pre-incubation at the running temperature. Preliminary experiments, were carried out using a Toyopearl HW-50F column (1 x 50cm), equilibrated with 0.1 M sodium acetate, 5% sodium chloride, pH 4.6, at 15C. Protein concentrations from 0.1 to 4 mg/ml were employed (C(sub sat) = 1.2 mg/ml). The data from several different protein preparations consistently shows a progressively decreasing elution volume with increasing protein concentration, indicating that reversible self-association is occurring. The dotted line indicates the monomeric lysozyme elution volume. However, lysozyme interacts with the column matrix in these experiments, which complicates data analysis.Accordingly, we are testing silica-based HPLC columns in an effort to eliminate this problem and substantially reduce the column volume and experimental run time. The results and data analysis from these and subsequent experiments will be presented.

Ewing, Felecia↗

Kinetics and Mechanism of the Reaction of Hydoxyl Radicals with Acetonitrile under Atmospheric Conditions

The pulsed laser photolysis-pulsed laser induced fluorescence technique has been employed to determine absolute rate coefficients for the reaction OH + CH3CN (1) and its isotopic variants, OH + CD3CN (2), OD + CH3CN (3), and OD + CD3CN (4). Reactions 1 and 2 were studied as a function of pressure and temperature in N2, N2/O2, and He buffer gases. In the absence of O2 all four reactions displayed well-behaved kinetics with exponential OH decays and pseudo-first rate constants which were proportional to substrate concentration. Data obtained in N2 over the range 50-700 Torr at 298 K are consistent with k(sub 1), showing a small pressure dependence. The Arrhenius expression obtained by averaging data at all pressures in k(sub 1)(T) = (1.1(sup +0.5)/(sub -0.3)) x 10(exp -12) exp[(-1130 +/- 90)/T] cu cm /(molecule s). The kinetics of reaction 2 are found to be pressure dependent with k(sub 2) (298 K) increasing from (1.21 +/- 0.12) x 10(exp -14) to (2.16 +/- 0.11) x 10(exp -14) cm(exp 3)/ (molecule s) over the pressure range 50-700 Torr of N2 at 298 K. Data at pressures greater than 600 Torr give k(sub 2)(T) = (9.4((sup +13.4)(sub -5.0))) x 10(exp -13) exp[(-1180 +/- 250)/T] cu cm/(molecule s). The rates of reactions 3 and 4 are found to be independent of pressure over the range 50-700 Torr of N2 with 298 K rate coefficient given by k(sub 3) =(3.18 +/- 0.40) x 10(exp -14) cu cm/(molecule s) and k(sub 4) = (2.25 +/-0.28) x 10(exp -14) cu cm/(molecule s). In the presence of O2 each reaction shows complex (non-pseudo-first-order) kinetic behavior and/or an apparent decrease in the observed rate constant with increasing [O2], indicating the presence of significant OH or OD regeneration. Observation of regeneration of OH in (2) and OD in (3) is indicative of a reaction channel which proceeds via addition followed by reaction of the adduct, or one of its decomposition products, with O2. The observed OH and OD decay profiles have been modeled by using a simple mechanistic scheme to extract kinetic information about the adduct reations with O2 and branching ratios for OH regeneration. A plausible mechanism for OH regeneration in (2) involves OH addition to the nitrogen atom followed by O2 addition to the cyano carbon atom, isomeriazation and decomposition to D2CO + DOCN + OH. Our results suggest that the OH + CH3CN reaction occurs via a complex mechanism involving both bimolecular and termolecular pathways, analogous to the mechanisms for the the important atmospheric reactions of OH with CO and HNO3.

Hynes, A. J.↗

Landing on Mercury: A Geochemical Perspective

The data from the MErcury Surface, Space ENvironment, GEochemistry, and Ranging (MESSENGER) spacecraft have revealed several surprising characteristics about the surface of Mercury, leading to its classification as a geochemical endmember among the terrestrial planets. Some of these features include elevated abundances of up to 3 wt% S, as much as 4 wt% C enrichment in low-reflectance materials (LRM) over the local mean, Na up to 5 wt% at high northern latitudes, and Fe abundances typically lower than 2 wt% [e.g., 1–4]. The S and Fe concentrations have been used to infer that Mercury’s igneous history evolved under highly reduced oxygen fugacity conditions between 2.6 and 7.3 log(sub 10) units below the iron–wüstite buffer [e.g., 5], which is more reducing than any other terrestrial planet in the solar system [e.g., 6]. This highly reduced nature has important con-sequences for the differentiation and thermal/magmatic evolution of Mercury. While the immense amount of data collected by MESSENGER revealed Mercury as a geochemical endmember, this new knowledge raised additional questions that necessitate continued exploration of the planet. Indeed, the joint ESA–JAXA dual-orbiter BepiColombo mission, launched in October 2018, is the most ambitious effort yet attempted to explore Mercury [e.g., 7]. Direct in situ elemental and mineralogical measurements on Mercury’s surface, however, are essential for addressing the new science questions that have arisen since MESSENGER.

K E Vander Kaaden↗

Evidence for the Synthesis of ATP by an F0F1 ATP Synthase in Membrane Vesicles from Halorubrum Saccharovorum

Vesicles prepared in a buffer containing ADP, Mg(2+) and Pi synthesized ATP at an initial rate of 2 nmols/min/mg protein after acidification of the bulk medium (pH 8 (right arrow) 4). The intravesicular ATP concentration reached a steady state after about 30 seconds and slowly declined thereafter. ATP synthesis was inhibited by low concentrations of dicyclohexylcarbodiimide and m-chlorophenylhydrazone indicating that synthesis took place in response to the proton gradient. NEM and PCMS, which inhibit vacuolar ATPases and the vacuolar-like ATPases of extreme halophiles, did not affect ATP synthesis, and, in fact, produced higher steady state levels of ATP. This suggested that two ATPase activities were present, one which catalyzed ATP synthesis and one that caused its hydrolysis. Azide, a specific inhibitor of F0F1 ATP Synthases, inhibited halobacterial ATP synthesis. The distribution of acridine orange as imposed by a delta pH demonstrated that azide inhibition was not due to the collapse of the proton gradient due to azide acting as a protonophore. Such an effect was observed, but only at azide concentrations higher than those that inhibited ATP synthesis. These results confirm the earler observations with cells of H. saccharovorum and other extreme halophiles that ATP synthesis is inconsistent with the operation of a vacuolar-like ATPase. Therefore, the observation that a vacuolar-like enzyme is responsible for ATP synthesis (and which serves as the basis for imputing ATP synthesis to the vacuolar-like ATPases of the extreme halophiles, and the Archaea in general) should be taken with some degree of caution.

Faguy, David↗

Human serum albumin crystals and method of preparation

Human serum albumin (HSA) crystals are provided in the form of tetragonal plates having the space groups P42(sub 1)2, the crystals being grown to sizes in excess of 0.5 mm in two dimensions and a thickness of 0.1 mm. Growth of the crystals is carried out by a hanging drop method wherein a precipitant solution containing polyethylene glycol (PEG) and a phosphate buffer is mixed with an HSA solution, and a droplet of mixed solution is suspended over a well of precipitant solution. Crystals grow to the desired size in 3 to 7 days. Concentration of reagents, pH and other parameters are controlled within prescribed limits. The resulting crystals exhibit a size and quality such as to allow performance of x ray diffraction studies and enable the conduct of drug binding studies as well as genetic engineering studies.

Carter, Daniel C.↗

Partitioning of Ni, Co and V between Spinel-Structured Oxides and Silicate Melts: Importance of Spinel Composition

Partitioning of Ni, Co and V between Cr-rich spinels and basaltic melt has been studied experimentally between 1150 and 1325 C, and at controlled oxygen fugacity from the Co-CoO buffer to slightly above the hematite magnetite buffer. These new results, together with new Ni, Co and V analyses of experimental run products from Leeman [Leeman, W.P., 1974. Experimental determination of the partitioning of divalent cations between olivine and basaltic liquid, Pt. II. PhD thesis, Univ. Oregon, 231 - 337.], show that experimentally determined spinel melt partition coefficients (D) are dependent upon temperature (T), oxygen fugacity (fO2) and spinel composition. In particular, partition coefficients determined on doped systems are higher than those in natural (undoped) systems, perhaps due to changing activity coefficients over the composition range defined by the experimental data. Using our new results and published runs (n =85), we obtain a multilinear regression equation that predicts experimental D(V) values as a function of T, fO2, concentration of V in melt and spinel composition. This equation allows prediction of D(V) spinel/melt values for natural mafic liquids at relevant crystallization conditions. Similarly, D(Ni) and D(Co) values can be inferred from our experiments at redox conditions approaching the QFM buffer, temperatures of 1150 to 1250 C and spinel composition (early Cr-bearing and later Ti-magnetite) appropriate for basic magma differentiation. When coupled with major element modelling of liquid lines of descent, these values (D(Ni) sp/melt=10 and D(Co) sp/melt=5) closely reproduce the compositional variation observed in komatiite, mid-ocean ridge basalt (MORB), ocean island basalt (OIB) and basalt to rhyolite suites.

Righter, K.↗

Fluorescence Studies of Protein Crystal Nucleation

Fluorescence can be used to study protein crystal nucleation through methods such as anisotropy, quenching, and resonance energy transfer (FRET), to follow pH and ionic strength changes, and follow events occurring at the growth interface. We have postulated, based upon a range of experimental evidence that the growth unit of tetragonal hen egg white lysozyme is an octamer. Several fluorescent derivatives of chicken egg white lysozyme have been prepared. The fluorescent probes lucifer yellow (LY), cascade blue, and 5-((2-aminoethyl)aminonapthalene-1-sulfonic acid (EDANS), have been covalently attached to ASP 101. All crystallize in the characteristic tetragonal form, indicating that the bound probes are likely laying within the active site cleft. Crystals of the LY and EDANS derivatives have been found to diffract to at least 1.7 A. A second group of derivatives is to the N-terminal amine group, and these do not crystallize as this site is part of the contact region between the adjacent 43 helix chains. However derivatives at these sites would not interfere with formation of the 43 helices in solution. Preliminary FRET studies have been carried out using N-terminal bound pyrene acetic acid (Ex 340 nm, Em 376 nm) lysozyme as a donor and LY (Ex -425 nm, Em 525 nm) labeled lysozyme as an acceptor. FRET data have been obtained at pH 4.6, 0.1 M NaAc buffer, at 5 and 7% NaCl, 4 C. The corresponding Csat values are 0.471 and 0.362 mg/ml (approximately 3.3 and approximately 2.5 x 10(exp -5) M respectively). The data at both salt concentrations show a consistent trend of decreasing fluorescence intensity of the donor species (PAA) with increasing total protein concentration. This decrease is more pronounced at 7% NaCl, consistent with the expected increased intermolecular interactions at higher salt concentrations reflected in the lower solubility. The calculated average distance between any two protein molecules at 5 x 10(exp -6) M is approximately 70nm, well beyond the range where any FRET can be expected. Results from these and ongoing studies will be presented.

Pusey, Marc L.↗

Two distinct regions of response drive differential growth in Vigna root electrotropism

Although exogenous electric fields have been reported to influence the orientation of plant root growth, reports of the ultimate direction of differential growth have been contradictory. Using a high-resolution image analysis approach, the kinetics of electrotropic curvature in Vigna mungo L. roots were investigated. It was found that curvature occurred in the same root toward both the anode and cathode. However, these two responses occurred in two different regions of the root, the central elongation zone (CEZ) and distal elongation zone (DEZ), respectively. These oppositely directed responses could be reproduced individually by a localized electric field application to the region of response. This indicates that both are true responses to the electric field, rather than one being a secondary response to an induced gravitropic stimulation. The individual responses differed in the type of differential growth giving rise to curvature. In the CEZ, curvature was driven by inhibition of elongation, whereas curvature in the DEZ was primarily due to stimulation of elongation. This stimulation of elongation is consistent with the growth response of the DEZ to other environmental stimuli.

Non-NASA Center↗

Effects of substrates and phosphate on INT (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl tetrazolium chloride) and CTC (5-cyano-2,3-ditolyl tetrazolium chloride) reduction in Escherichia coli

The effects of substrates of primary aerobic dehydrogenases, and inorganic phosphate on aerobic INT and CTC reduction in Escherichia coli were examined. In general, INT produced less formazan than CTC, but INT (+) cell counts remained near values of CTC (+) cells. INT and CTC (+) cell numbers were higher than plate counts on R2A medium using succinate, formate, lactate, casamino acids, glucose, glycerol (INT only) and no substrate. Formate resulted in the greatest amount of INT and CTC formazan. Reduction of both INT and CTC was inhibited above 10 mmol l-1 phosphate, and this appeared to be related to decreased rates of O2 consumption. Formation of fluorescent CTC (+), but not INT (+) cells was also inhibited in a concentration dependent manner by phosphate above 10 mmol l-1. From light microscopic observations it appeared CTC formed increasing amounts of poorly or non-fluorescent formazan with increasing phosphate. Therefore, use of phosphate buffer in excess of 10 mmol l-1 may not be appropriate in CTC and INT reduction assays.

NASA Program Environmental Health↗