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NASA NTRS1984

GD 323 - A white dwarf with a stratified H/He atmosphere?

New optical and ultraviolet (IUE) spectrophotometry of the hot white dwarf GD 323 are presented. The optical spectra exhibit broad, shallow Balmer lines, together with weaker He I lambda-4471, lambda-4921 lines. However, the effective temperature and helium abundance derived from the optical spectrum appear incompatible with parameters derived from the optical colors and the overall energy distribution - thus suggesting that GD 323 does not have a homogeneous, mixed composition atmosphere. The alternative interpretations of substantial reddening, a rapidly rotating subdwarf, an accreting short-period binary, and a detached DA + DB binary system are ruled out. It is suggested instead that GD 323 may have a stratified atmosphere, with an optically thin hydrogen layer overlying a helium envelope at T(e) about 30,000 K; however, this conclusion must be properly tested.

Liebert, J.↗
NASA NTRS1984

Spectrophotometry of extreme helium stars - Ultraviolet fluxes and effective temperatures

Ultraviolet flux distributions are presented for the extremely helium rich stars BD +10 deg 2179, HD 124448, LSS 3378, BD -9 deg 4395, LSE 78, HD 160641, LSIV -1 deg 2, BD 1 deg 3438, HD 168476, MV Sgr, LS IV-14 deg 109 (CD -35 deg 11760), LSII +33 deg 5 and BD +1 deg 4381 (LSIV +2 deg 13) obtained with the International Ultraviolet Explorer (IUE). Broadband photometry and a newly computed grid of line blanketed model atmospheres were used to determine accurate angular diameters and total stellar fluxes. The resultant effective temperatures are in most cases in satisfactory agreement with those based on broadband photometry and/or high resolution spectroscopy in the visible. For two objects, LSII +33 deg 5 and LSE 78, disagreement was found between the IUE observations and broadband photometry: the colors predict temperatures around 20,000 K, whereas the UV spectra indicate much lower photospheric temperatures of 14,000 to 15,000 K. The new temperature scale for extreme helium stars extends to lower effective temperatures than that of Heber and Schoenberner (1981) and covers the range from 8,500 K to 32,000 K. Previously announced in STAR as N83-24433

Heber, U.↗
NASA NTRS2003

5' modification of duplex DNA with a ruthenium electron donor-acceptor pair using solid-phase DNA synthesis

Incorporation of metalated nucleosides into DNA through covalent modification is crucial to measurement of thermal electron-transfer rates and the dependence of these rates with structure, distance, and position. Here, we report the first synthesis of an electron donor-acceptor pair of 5' metallonucleosides and their subsequent incorporation into oligonucleotides using solid-phase DNA synthesis techniques. Large-scale syntheses of metal-containing oligonucleotides are achieved using 5' modified phosporamidites containing [Ru(acac)(2)(IMPy)](2+) (acac is acetylacetonato; IMPy is 2'-iminomethylpyridyl-2'-deoxyuridine) (3) and [Ru(bpy)(2)(IMPy)](2+) (bpy is 2,2'-bipyridine; IMPy is 2'-iminomethylpyridyl-2'-deoxyuridine) (4). Duplexes formed with the metal-containing oligonucleotides exhibit thermal stability comparable to the corresponding unmetalated duplexes (T(m) of modified duplex = 49 degrees C vs T(m) of unmodified duplex = 47 degrees C). Electrochemical (3, E(1/2) = -0.04 V vs NHE; 4, E(1/2) = 1.12 V vs NHE), absorption (3, lambda(max) = 568, 369 nm; 4, lambda(max) = 480 nm), and emission (4, lambda(max) = 720 nm, tau = 55 ns, Phi = 1.2 x 10(-)(4)) data for the ruthenium-modified nucleosides and oligonucleotides indicate that incorporation into an oligonucleotide does not perturb the electronic properties of the ruthenium complex or the DNA significantly. In addition, the absence of any change in the emission properties upon metalated duplex formation suggests that the [Ru(bpy)(2)(IMPy)](2+)[Ru(acac)(2)(IMPy)](2+) pair will provide a valuable probe for DNA-mediated electron-transfer studies.

Non-NASA Center↗
NASA NTRS2002

Entrainment in solution of an oscillating NADH oxidase activity from the bovine milk fat globule membrane with a temperature-compensated period length suggestive of an ultradian time-keeping (clock) function

Entrainment in solution of an oscillating activity with a temperature compensated period of 24 min is described for a NADH oxidase (NOX) activity of the bovine milk fat globule membrane, a derivative of the mammary epithelial cell plasma membrane. The period of 24 min remained unchanged at 17 degrees C, 27 degrees C and 37 degrees C whereas the amplitude approximately doubled with each 10 degree C rise in temperature (Q(10)congruent with 2). The periodicity was observed with both intact milk fat globule membranes and with detergent-solubilized membranes, demonstrating that the oscillations did not require an association with membranes. The periodicity was not the result of instrument variation or of chemical interactions among reactants in solution. Preparations with different periodicities entrained (autosynchronized) when mixed. Upon mixing, the preparations exhibited two oscillatory patterns but eventually a single pattern representing the mean of the farthest separated maxima of the two preparations analyzed separately emerged. The cell surface NOX protein is the first reported example of an entrainable biochemical entity with a temperature-compensated periodicity potentially capable of functioning as an ultradian or circadian clock driver.

Non-NASA Center↗
NASA NTRS2002

2'-modified nucleosides for site-specific labeling of oligonucleotides

We report the synthesis of 2'-modified nucleosides designed specifically for incorporating labels into oligonucleotides. Conversion of these nucleosides to phosphoramidite and solid support-bound derivatives proceeds in good yield. Large-scale synthesis of 11-mer oligonucleotides possessing the 2'-modified nucleosides is achieved using these derivatives. Thermal denaturation studies indicate that the presence of 2'-modified nucleosides in 11-mer duplexes has minimal destabilizing effects on the duplex structure when the nucleosides are placed at the duplex termini. The powerful combination of phosphoramidite and support-bound derivatives of 2'-modified nucleosides affords the large-scale preparation of an entirely new class of oligonucleotides. The ability to synthesize oligonucleotides containing label attachment sites at 3', intervening, and 5' locations of a duplex is a significant advance in the development of oligonucleotide conjugates.

Non-NASA Center↗
NASA NTRS1995

Aqueous nitrite ion determination by selective reduction and gas phase nitric oxide chemiluminescence

An improved method of flow injection analysis for aqueous nitrite ion exploits the sensitivity and selectivity of the nitric oxide (NO) chemilluminescence detector. Trace analysis of nitrite ion in a small sample (5-160 microL) is accomplished by conversion of nitrite ion to NO by aqueous iodide in acid. The resulting NO is transported to the gas phase through a semipermeable membrane and subsequently detected by monitoring the photoemission of the reaction between NO and ozone (O3). Chemiluminescence detection is selective for measurement of NO, and, since the detection occurs in the gas-phase, neither sample coloration nor turbidity interfere. The detection limit for a 100-microL sample is 0.04 ppb of nitrite ion. The precision at the 10 ppb level is 2% relative standard deviation, and 60-180 samples can be analyzed per hour. Samples of human saliva and food extracts were analyzed; the results from a standard colorimetric measurement are compared with those from the new chemiluminescence method in order to further validate the latter method. A high degree of selectivity is obtained due to the three discriminating steps in the process: (1) the nitrite ion to NO conversion conditions are virtually specific for nitrite ion, (2) only volatile products of the conversion will be swept to the gas phase (avoiding turbidity or color in spectrophotometric methods), and (3) the NO chemiluminescence detector selectively detects the emission from the NO + O3 reaction. The method is free of interferences, offers detection limits of low parts per billion of nitrite ion, and allows the analysis of up to 180 microL-sized samples per hour, with little sample preparation and no chromatographic separation. Much smaller samples can be analyzed by this method than in previously reported batch analysis methods, which typically require 5 mL or more of sample and often need chromatographic separations as well.

NASA Discipline Environmental Health↗
NASA NTRS1993

A search for life on Earth from the Galileo spacecraft

In its December 1990 fly-by of Earth, the Galileo spacecraft found evidence of abundant gaseous oxygen, a widely distributed surface pigment with a sharp absorption edge in the red part of the visible spectrum, and atmospheric methane in extreme thermodynamic disequilibrium; together, these are strongly suggestive of life on Earth. Moreover, the presence of narrow-band, pulsed, amplitude-modulated radio transmission seems uniquely attributable to intelligence. These observations constitute a control experiment for the serach for extraterrestrial life by modern interplanetary spacecraft.

Flight Experiment↗
NASA NTRS1991

Synthesis of the coenzymes adenosine diphosphate glucose, guanosine diphosphate glucose, and cytidine diphosphoethanolamine under primitive Earth conditions

The nonenzymatic synthesis of the coenzymes adenosine diphosphate glucose (ADPG), guanosine diphosphate glucose (GDPG), and cytidine diphosphoethanolamine (CDP-ethanolamine) has been carried out under conditions considered to have been prevalent on the early Earth. The production of these compounds was performed by allowing simple precursor molecules to react under aqueous solutions, at moderate temperatures and short periods of time, with mediation by cyanamide or urea. These two condensing agents are considered to have been present in significant amounts on the primitive Earth and have been previously used in the nonenzymatic synthesis of several other important biochemical compounds. In our experiments, ADPG was obtained by heating glucose-1-phosphate (G1P) and ATP in the presence of cyanamide for 24 h at 70 degrees C. The reaction of G1P and GTP under the same conditions yielded GDPG. The cyanamide-mediated production of CDP-ethanolamine was carried out by reacting a mixture of ethanolamine phosphate and CTP for 24 h at 70 degrees C. The separation and identification of the reaction products was carried out by paper chromatography, thin-layer chromatography, high performance thin-layer chromatography, high performance liquid chromatography, both normal and reverse-phase, UV spectroscopy, enzymatic assays, and acid hydrolysis. Due to the mild conditions employed, and to the relative ease of these reactions, these studies offer a simple attractive system for the nonenzymatic synthesis of phosphorylated high-energy metabolic intermediates under conditions considered to have been prevalent on the ancient Earth.

NASA Discipline Exobiology↗
NASA NTRS2000

Peptide nucleic acids rather than RNA may have been the first genetic molecule

Numerous problems exist with the current thinking of RNA as the first genetic material. No plausible prebiotic processes have yet been demonstrated to produce the nucleosides or nucleotides or for efficient two-way nonenzymatic replication. Peptide nucleic acid (PNA) is a promising precursor to RNA, consisting of N-(2-aminoethyl)glycine (AEG) and the adenine, uracil, guanine, and cytosine-N-acetic acids. However, PNA has not yet been demonstrated to be prebiotic. We show here that AEG is produced directly in electric discharge reactions from CH(4), N(2), NH(3), and H(2)O. Electric discharges also produce ethylenediamine, as do NH(4)CN polymerizations. AEG is produced from the robust Strecker synthesis with ethylenediamine. The NH(4)CN polymerization in the presence of glycine leads to the adenine and guanine-N(9)-acetic acids, and the cytosine and uracil-N(1)-acetic acids are produced in high yield from the reaction of cyanoacetaldehyde with hydantoic acid, rather than urea. Preliminary experiments suggest that AEG may polymerize rapidly at 100 degrees C to give the polypeptide backbone of PNA. The ease of synthesis of the components of PNA and possibility of polymerization of AEG reinforce the possibility that PNA may have been the first genetic material.

NASA Discipline Exobiology↗
NASA NTRS1999

Boronate affinity adsorption of RNA: possible role of conformational changes

Batch equilibrium adsorption isotherm determination is used to characterize the adsorption of mixed yeast RNA on agarose-immobilized m-aminophenylboronic acid. It is shown that the affinity-enhancing influence of divalent cations depends strongly on the precise nature of the cation used, with barium being far more effective than the conventionally-used magnesium. This adsorption-promoting influence of barium is suggested to arise primarily from ionic influences on the structure and rigidity of the RNA molecule, as the adsorption of ribose-based small molecules is not similarly affected. The substitution of barium for the standard magnesium counterion does not greatly promote the adsorption of DNA, implying that the effect is specific to RNA and may be useful in boronate-based RNA separations. RNA adsorption isotherms exhibit sharp transitions as functions of temperature, and these transitions occur at different temperatures with Mg2+ and with Ba2+. Adsorption affinity and capacity were found to increase markedly at lower temperatures, suggestive of an enthalpically favored interaction process. The stoichiometric displacement parameter, Z, in Ba2+ buffer is three times the value in Mg2+ buffer, and is close to unity.

NASA Discipline Environmental Health↗
NASA NTRS1999

Transduction-like gene transfer in the methanogen Methanococcus voltae

Strain PS of Methanococcus voltae (a methanogenic, anaerobic archaebacterium) was shown to generate spontaneously 4.4-kbp chromosomal DNA fragments that are fully protected from DNase and that, upon contact with a cell, transform it genetically. This activity, here called VTA (voltae transfer agent), affects all markers tested: three different auxotrophies (histidine, purine, and cobalamin) and resistance to BES (2-bromoethanesulfonate, an inhibitor of methanogenesis). VTA was most effectively prepared by culture filtration. This process disrupted a fraction of the M. voltae cells (which have only an S-layer covering their cytoplasmic membrane). VTA was rapidly inactivated upon storage. VTA particles were present in cultures at concentrations of approximately two per cell. Gene transfer activity varied from a minimum of 2 x 10(-5) (BES resistance) to a maximum of 10(-3) (histidine independence) per donor cell. Very little VTA was found free in culture supernatants. The phenomenon is functionally similar to generalized transduction, but there is no evidence, for the time being, of intrinsically viral (i.e., containing a complete viral genome) particles. Consideration of VTA DNA size makes the existence of such viral particles unlikely. If they exist, they must be relatively few in number;perhaps they differ from VTA particles in size and other properties and thus escaped detection. Digestion of VTA DNA with the AluI restriction enzyme suggests that it is a random sample of the bacterial DNA, except for a 0.9-kbp sequence which is amplified relative to the rest of the bacterial chromosome. A VTA-sized DNA fraction was demonstrated in a few other isolates of M. voltae.

Methanococcus/genetics/growth & development/metabo↗
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 NTRS1992

Influence of the size and protonation state of acidic residue 85 on the absorption spectrum and photoreaction of the bacteriorhodopsin chromophore

The consequences of replacing Asp-85 with glutamate in bacteriorhodopsin, as expressed in Halobacterium sp. GRB, were investigated. Similarly to the in vitro mutated and in Escherichia coli expressed protein, the chromophore was found to exist as a mixture of blue (absorption maximum 615 nm) and red (532 nm) forms, depending on the pH. However, we found two widely separated pKa values (about 5.4 and 10.4 without added salt), arguing for two blue and two red forms in separate equilibria. Both blue and red forms of the protein are in the two-dimensional crystalline state. A single pKa, such as in the E. coli expressed protein, was observed only after solubilization with detergent. The photocycle of the blue forms was determined at pH 4.0 with 610 nm photoexcitation, and that of the red forms at pH 10.5 and with 520 nm photoexcitation, in the time-range of 100 ns to 1 s. The blue forms produced no M, but a K- and an L-like intermediate, whose spectra and kinetics resembled those of blue wild-type bacteriorhodopsin below pH 3. The red forms produced a K-like intermediate, as well as M and N. Only the red forms transported protons. Specific perturbation of the neighborhood of the Schiff base by the replacement of Asp-85 with glutamate was suggested by (1) the shift and splitting of the pKa for what is presumably the protonation of residue 85, (2) a 36 nm blue-shift in the absorption of the all-trans red chromophore and a 25 nm red-shift of the 13-cis N chromophore, as compared to wild-type bacteriorhodopsin and its N intermediate, and (3) significant acceleration of the deprotonation of the Schiff base at pH 7, but not of its reprotonation and the following steps in the photocycle.

Non-NASA Center↗
NASA NTRS1984

The rate of the reaction of OH with HCl

Using the techniques of laser/flash photolysis-resonance fluorescence, the absolute rate constant for the OH + HCl reaction is measured from 240 to 295 K. Ultraviolet and infrared spectrophotometry are used to continuously monitor the HCl concentrations. It is shown that the rate constant values obtained in the study are 20-30 percent larger than those recommended by Smith and Zellner (1974) and Ravishankara et al. (1977) for modeling of stratospheric chemistry.

Molina, M. J.↗
NASA NTRS1997

Physical Structure of Four Symbiotic Binaries

Disk accretion powers many astronomical objects, including pre-main sequence stars, interacting binary systems, and active galactic nuclei. Unfortunately, models developed to explain the behavior of disks and their surroundings - boundary layers, jets, and winds - lack much predictive power, because the physical mechanism driving disk evolution - the viscosity - is not understood. Observations of many types of accreting systems are needed to constrain the basic physics of disks and provide input for improved models. Symbiotic stars are an attractive laboratory for studying physical phenomena associated with disk accretion. These long period binaries (P(sub orb) approx. 2-3 yr) contain an evolved red giant star, a hot companion, and an ionized nebula. The secondary star usually is a white dwarf accreting material from the wind of its red giant companion. A good example of this type of symbiotic is BF Cygni: our analysis shows that disk accretion powers the nuclear burning shell of the hot white dwarf and also manages to eject material perpendicular to the orbital plane (Mikolajewska, Kenyon, and Mikolajewski 1989). The hot components in other symbiotic binaries appear powered by tidal overflow from a very evolved red giant companion. We recently completed a study of CI Cygni and demonstrated that the accreting secondary is a solar-type main sequence star, rather than a white dwarf (Kenyon et aL 1991). This project continued our study of symbiotic binary systems. Our general plan was to combine archival ultraviolet and optical spectrophotometry with high quality optical radial velocity observations to determine the variation of line and continuum sources as functions of orbital phase. We were very successful in generating orbital solutions and phasing UV+optical spectra for five systems: AG Dra, V443 Her, RW Hya, AG Peg, and AX Per. Summaries of our main results for these systems appear below. A second goal of our project was to consider general models for the outbursts of symbiotic stars, with an emphasis on understanding the differences between disk-driven and nuclear-powered eruptions.

Kenyon, Scott J.↗
NASA NTRS1995

Papers Presented at IAU Colloquium No. 152: Astrophysics in the Extreme Ultraviolet

This colloquium reviews findings in the Astrophysics in the Extreme Ultraviolet region. The papers presented are: (1) The Composition and Structure of White Dwarf Atmospheres Revealed by Extreme Ultraviolet Spectroscopy, (2) EUVE Spectrophotometry of QS Tel: The Second Pole Becomes Active, (3) Results from the ROSAT EUV Wide Field Camera, (4) Temporal Behaviour of Sources in the ROSAT Extreme-Ultraviolet All-Sky Survey, (5) A Search for the Signature of the Diffuse Soft X-ray Background in the ROSAT Wide-Field Camera All-Sky Survey, (6) Features of the Soft X-ray Background and Implications for the EUV Background.

Bowyer, S.↗
NASA NTRS1981

Spectrophotometry of planets, asteroids and satellites from the international ultraviolet explorer satellite

A total of 14 8 hour I.U.E. observing sessions resulted in 39 spectra of 11 asteroids and 9 solar type stars as well as 57 spectra at various locations on the disk of Jupiter. The Jupiter observations include a total of 5 center to limb series of spectra at various latitudes and a North South series along the central meridian. In the range from 2000-3000 A, the planet shows a striking decrease in brightness at latitudes greater than about 30 degrees, and exhibits limb brightening at low latitudes and limb darkening at high latitudes. Preliminary results indicate that about 6 km-amagats of clean hydrogen are required above a haze of absorbing aerosols to reproduce the limb brightening observed at 2500 A in the equatorial regions. At higher latitudes, the aerosols extend to even higher levels of the atmosphere. Comparison of the Jovian data with detailed model calculations and the analyses of the asteroid spectra are still in progress with other support.

Tomasko, M. G.↗