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Response of Two Legumes to Two Ultraviolet-B Radiation Regimes

Depletion of the stratospheric ozone layer has been directly linked to increased levels of UV radiation at the earth's surface. The purpose of this study was to evaluate the responses of soybean (Glycine max) and alfalfa (Medicago sativa) to increased UV-B radiation (280-320 nm). Soybean and alfalfa were grown successively in a growth chamber that provided UV-B intensities 45% above nominal summer field levels. Mylar-D (UVB opaque) and mono-acetate (UV-B transparent) films were used to establish the two UV-B treatments. Soybean grown under increased UV showed 21% smaller internodal lengths and higher concentrations of UV-B absorbing pigments (i.e. flavonoids) compared to plants grown under no UV. Significant results for alfalfa included 22% greater leaf flavonoid concentration under increased UV, 14% greater leaf chlorophyll concentration under no UV, and 32% greater above-ground biomass with no UV. These leguminous species possess mechanisms that protect against UV-B damage as indicated by increases in foliar concentrations of UV-B absorbing compounds. Alfalfa appears to be more sensitive to UV-B damage than soybean. Remote sensing of chlorophyll fluorescence may offer a means of monitoring UV-induced plant stress and damage.

Levy, Daniel L.↗

Reflective Self-Metallizing Polyimide Films

A silver organic complex, such as silver acetate, is solubilized in a polyamic acid resin or soluble polyimide solution using a suitable solvent such as hexafluoroacetyl acetone. The mixture is stable and can be applied to both flat and contoured surfaces. Application can be performed by casting, dip-coating, spraying, or other suitable techniques. In addition, the mixture can be cast or extruded as a polyimide film which is not applied to an underlying substrate. Upon curing, a flexible silver coated polyimide film is produced.

Thompson, David W.↗

Fatigue Damage Mechanisms in Advanced Hybrid Titanium Composite Laminates

Hybrid Titanium Composite Laminates (HTCL) are a type of hybrid composite laminate with promise for high-speed aerospace applications, specifically designed for improved damage tolerance and strength at high-temperature (350 F, 177 C). However, in previous testing, HTCL demonstrated a propensity to excessive delamination at the titanium/PMC interface following titanium cracking. An advanced HTCL has been constructed with an emphasis on strengthening this interface, combining a PETI-5/IM7 PMC with Ti-15-3 foils prepared with an alkaline-perborate surface treatment. This paper discusses how the fatigue capabilities of the "advanced" HTCL compare to the first generation HTCL which was not modified for interface optimization, in both tension-tension (R = 0.1) and tension-compression (R=-0.2). The advanced HTCL under did not demonstrate a significant improvement in fatigue life, in either tension-tension or tension-compression loading. However, the advanced HTCL proved much more damage tolerant. The R = 0.1 tests revealed the advanced HTCL to increase the fatigue life following initial titanium ply damage up to 10X that of the initial HTCL at certain stress levels. The damage progression following the initial ply damage demonstrated the effect of the strengthened PMC/titanium interface. Acetate film replication of the advanced HTCL edges showed a propensity for some fibers in the adjacent PMC layers to fail at the point of titanium crack formation, suppressing delamination at the Ti/PMC interface. The inspection of failure surfaces validated these findings, revealing PMC fibers bonded to the majority of the titanium surfaces. Tension compression fatigue (R = -0.2) demonstrated the same trends in cycles between initial damage and failure, damage progression, and failure surfaces. Moreover, in possessing a higher resistance to delamination, the advanced HTCL did not exhibit buckling following initial titanium ply cracking under compression unlike the initial HTCL.

Johnson, W. Steven↗

Phenylethynyl Containing Reactive Additives

Phenylethynyl containing reactive additives were prepared from aromatic diamines containing phenylethynyl groups and various ratios of phthalic anhydride and 4-phenylethynylphthalic anhydride in glacial acetic acid to form the imide in one step or in N-methyl-2-pyrrolidinone to form the amide acid intermediate. The reactive additives were mixed in various amounts (10% to 90%) with oligomers containing either terminal or pendent phenylethynyl groups (or both) to reduce the melt viscosity and thereby enhance processability. Upon thermal cure, the additives react and become chemically incorporated into the matrix and effect an increase in crosslink density relative to that of the host resin. This resultant increase in crosslink density has advantageous consequences on the cured resin properties such as higher glass transition temperature and higher modulus as compared to that of the host resin.

John W Connell↗

Comparison of Long-Term Storage in Chemical Fixatives on Morphology and Anatomy of Super-Dwarf Wheat

Wheat plants (Triticum aestivum L. cv Super-Dwarf) are grown in the microgravity of space and harvested for morphological and anatomical comparison with those exposed to gravity on earth. Such plants are subjected to relatively long periods of storage in chemical fixatives. Examination, evaluation and verification that the integrity of the vascular system is maintained for extended periods of storage in fixatives are required. McDowell and Trump's [4% Formaldehyde -- 1 % Glutaraldehyde (4F: 1 G)] or Variant I [(Russian Fixative): Formalin: Acetic Acid: Alcohol] fixatives, adjusted to pH 7.2, were placed in Aclam(TM), FilmORap(TM), or FilmOSun(TM) plastic bags on April 4, 1994. Wheat seedlings were harvested on days 9, 28, and 68 and preserved in these fixatives. Subsamples of leaves and/or seeds were taken from these stocks after various times in storage, dehydrated, and embedded in Spurr's, LR White's or Unicryl resin. Semithin (1 mm) and thin (50-70 nm) sections were examined by light and transmission electron microscopy. In a few sections, we have observed a slight plasmolysis of the cytostol in leaf tissue fixed with the Variant I, but overall there seem to be no major artifacts in the anatomical structure. The plasmalemma and other organelles appeared normal in the McDowell and Trump fixative. Use of differential chromophores suggests that LR White or Unicryl resins may give greater flexibility for enzyme localizations at both the light and electron microscopical levels.

Bubenheim, David L.↗

Buffer Effects in the Solubility, Nucleation and Growth of Chicken Egg White Lysozyme

The growth of protein crystals is important for determination of their three-dimensional structure, which relates to their biochemical functions and to the practical goal of designing pharmaceuticals to modify that function. While many proteins have been successfully crystallized by a variety of methods, there is still limited understanding of the process of nucleation and growth of even the simplest proteins. Chicken egg-white lysozyme (CEWL) is readily crystallized under a variety of conditions, and studies underway at MSFC are designed to elucidate the mechanisms by which the crystals nucleate and grow. We have investigated the effect of buffer choice on the solubility, nucleation and growth of CEWL. CEWL was purified by dialysis against a .05M phosphate buffer and chromatographic separation from contaminants in a sepharose column. Solubility studies were made as a function of buffer concentration for phosphate and formate buffers, and the nucleation and growth of crystals at 10 C was studied as a function of pH for oxalate, succinate, formate, butyrate, carbonate, phosphate and acetate buffer solutions. The solubility data support the conclusion that there is a solubility minimum as a function of buffer concentration for amphiphilic molecules, while no minimum is observed for a phosphate buffer. Nucleation is suppressed at pH greater than pKa for all buffers except phosphate. The aspect ratio of the (110) faces is shown to be a function of crystal size, rather than pH.

Gibson, Ursula J.↗

Microgravity Compatible Reagentless Instrumentation for Detection of Dissolved Organic Acids and Alcohols in Potable Water

The Organic Acid and Alcohol Monitor (OAAM) program has resulted in the successful development of a computer controlled prototype analyzer capable of accurately determining aqueous organic acids and primary alcohol concentrations over a large dynamic range with high sensitivity. Formic, acetic, and propionic acid were accurately determined at concentrations as low as 5 to 10 micrograms/L in under 20 minutes, or as high as 10 to 20 mg/L in under 30 minutes. Methanol, ethanol, and propanol were determined at concentrations as low as 20 to 100 micrograms/L, or as high as 10 mg/L in under 30 minutes. Importantly for space based application, the OAAM requires no reagents or hazardous chemicals to perform these analyses needing only power, water, and CO2 free purge gas. The OAAM utilized two membrane processes to segregate organic acids from interfering ions. The organic acid concentration was then determined based upon the conductiometric signal. Separation of individual organic acids was accomplished using a chromatographic column. Alcohols are determined in a similar manner after conversion to organic acids by sequential biocatalytic and catalytic oxidation steps. The OAAM was designed to allow the early diagnosis of under performing or failing sub-systems within the Water Recovery System (WRS) baselined for the International Space Station (ISS). To achieve this goal, several new technologies were developed over the course of the OAAM program.

Akse, James R.↗

Spirochaeta americana sp. nov.: A New Haloalkaliphilic, Obligately Anaerobic Spirochete Isolated from Soda Mono Lake, California

A novel obligately anaerobic, mesophilic, haloalkaliphilic spirochete, strain ASpG1, was isolated from sediments of the alkaline, hypersaline Mono Lake in California, U.S.A. The gram-negative cells are motile and spirochete-shaped with sizes of 0.22 x 10-15 micron. Growth was observed over the temperature range of 10 C to 44 C (optimum 37 C), NaCl concentration range of greater than 1 - 12 % (wt/vol) (optimum 3%), and pH range 7.5 - 10.5 (optimum pH 9.5). The novel isolate is strictly alkaliphilic, requires high concentrations of carbonate in the medium, and is capable of utilizing D-glucose, fructose, maltose, sucrose, starch, and D-mannitol. Main end products of glucose fermentation are: H2, acetate, ethanol, and formate. Strain AspG1 is resistant to kanamycin, but sensitive to chloramphenicol, gentamycin and tetracycline. The G+C content of its DNA is 58.5 mol%. On the basis of its physiological and molecular properties, the isolate appears to be a novel species among the genus Spirochaeta; and the name Spirochaeta americana sp. nov., is proposed for the taxon (type strain ASpG1(sup T) = ATCC BAA_392(sup T) = DSMZ 14872(sup T)).

Hoover, Richard B.↗

Tindallia Californiensis sp. nov.: A New Halo-Alkaliphilic Primary Anaerobe, Isolated from Meromictic soda Mono Lake in California and the Correction of Diagnosis for Genus Tindallia

A novel extremely halo-alkaliphilic, bacterium strain APO (sup T) was isolated from sediments of the athalassic, meromictic, soda Mono Lake in California. Gram positive, spore-forming, slightly curved rods with sizes 0.6-0.7x 2.5-4.0 micrometers which occur singly, in pairs or short curved chains. Cells, are motile by singular subcentral flagellum. Strain APO (sup T) is mesophilic: growth was observed over the temperature range of +10 C to +48 C (optimum +37 C), NaCl concentration range 1-20 %, wt/vol (optimum 3-5%, wt/vol) and pH range 8.0-11.0 (optimum pH 9.5). The novel isolate is strictly halo-alkaliphilic, requires sodium chloride in medium, obligately anaerobic and catalase-negative. Strain APO (sup T) is organo-heterotroph with fermentative type of metabolism, and uses as substrates: peptone, badotryptone, casamino acids, yeast extract, L-serine, L-lysine, L-histidine, L-arginine, and pyruvate. The main end products of growth on peptone medium were: lactate, acetate, propionate, and ethanol. Strain APO (sup T) is resistant to kanamycin, but sensitive to chloramphenicol, tetracycline, and gentamycin. The sum of G+C in DNA is 44.4 mol% (by HPLC method). On the bait of physiological and molecular properties, the isolate was considered as novel species of genus Tindallia; and the name Tindallia californiensis sp. nov., is proposed for new isolate (type strain APO (sup T) - ATCC BAA_393(sup T) = DSMZ 14871 (sup T)).

Pikuta, Elena↗

Spirochaeta Americana Sp. Nov., A new Haloalkaliphilic, Obligately Anaerobic Spirochete Isolated from Soda Mono Lake in California

A novel obligately anaerobic, mesophilic, haloalkaliphilic spirochete, strain ASpG1(sup T), was isolated from sediments of the alkaline, hypersaline Mono Lake in California, U.S.A. The Gram-negative cells are motile and spirochete-shaped with sizes of 0.2 - 0.22 X 8-15 microns. Growth was observed over the following ranges: temperature 10 C to 44 C; optimum +37 C; NaCl concentration 2 - 12 % (w/v); optimum NaCl3 % and pH 8 - 10.5; optimum pH 9.5. The novel isolate is strictly alkaliphilic, requires high concentrations of carbonate in the medium, and is capable of utilizing D-glucose, fructose, maltose, sucrose, starch, and D-mannitol. The main end products of glucose fermentation are: H2, acetate, ethanol, and formate. Strain ASpG(sup T) is resistant to kanamycin, and rifampin, but sensitive to chloramphenicol, gentamycin and tetracycline. The G+C content of its DNA is 58.5 mol%, genome size is 2.98 x l0(exp 9) Daltons, Tm of the genomic DNA is 68 +/- 2 C, and DNA-DNA hybridization with the most closely related species, Spirocheta alkalica Strain Z-7491(sup T), exhibited 48.7% homology. On the basis of its physiological and molecular properties, the isolate appears to be a novel species of the genus Spirochaeta; and the name Spirochaeta americana sp. nov., is proposed for the taxon (type strain ASpG1(sup T) = ATCC BAA-392(sup T) = DSMZ 14872(sup T)).

Hoover, Richard B.↗

Phenylethynyl Containing Reactive Additives

Phenylethynyl containing reactive additives were prepared from aromatic diamine, containing phenylethvnvl groups and various ratios of phthalic anhydride and 4-phenylethynviphthalic anhydride in glacial acetic acid to form the imide in one step or in N-methyl-2-pvrrolidinone to form the amide acid intermediate. The reactive additives were mixed in various amounts (10% to 90%) with oligomers containing either terminal or pendent phenylethynyl groups (or both) to reduce the melt viscosity and thereby enhance processability. Upon thermal cure, the additives react and become chemically incorporated into the matrix and effect an increase in crosslink density relative to that of the host resin. This resultant increase in crosslink density has advantageous consequences on the cured resin properties such as higher glass transition temperature and higher modulus as compared to that of the host resin.

Connell, John W.↗

Synthesis,and structural characterization of [(CH3(C5H4N))Ga(SCH2(CO)O)]-[(4-MepyH)]+, a novel Ga(III) five coordinate complex.

The synthesis and structural characterization of a novel ionic Ga(III) five coordinate complex [{CH3(C5H4N)}Ga(SCH2(CO)O)2]-[(4-MepyH)]+, (4-Mepy = CH3(C5H5N)) from the reaction between Ga2Cl4 with sodium mercapto-acetic acid in 4-methylpyridine is described. Under basic reaction conditions the mercapto ligand is found to behave as a 2e- bidentate ligand. Single crystal X-ray diffraction studies show the complex to have a distorted square pyramidal geometry with the [(-SCH2(CO)CO-)] ligands in a trans conformation. The compound crystallizes in the P2(sub 1)/c (No. 14) space group with a = 7.7413(6) A, b = 16.744(2) A, c = 14.459(2) A, V = 1987.1(6) A(sup 3), R(F) = 0.032 and R(sub w) = 0.038.

Banger, Kulbinder K.↗

Kinetics of Organic Transformations Under Mild Aqueous Conditions: Implications for the Origin of Life and Its Metabolism

The rates of thermal transformation of organic molecules containing carbon, hydrogen, and oxygen were systematically examined in order to identify the kinetic constraints that governed origin-of-life organic chemistry under mild aqueous conditions. Arrhenius plots of the kinetic data were used to estimate the reaction half-life at 50 C, and to reveal the effect of functional groups on reactivity. This survey showed that hydrocarbons and organic substances containing a single oxygenated group were kinetically the most stable (i. e. acetate decarboxylation half-life was l0(exp 18) years at 50 C); whereas, organic substances containing two oxygenated groups in which one group was a beta-positioned carbonyl group were the most reactive (i. e. acetoacetate decarboxylation half-life was l0(exp-2) years at 50 C). Of all functional groups the beta-positioned carbonyl group (aldehyde or ketone) was the strongest activating group, giving rates of reaction that were up to 10(exp 24)-times faster than rates of similar molecules lacking the beta-carbonyl group. From this knowledge of organic reactivity and the inherent constraints of autocatalytic processes, we concluded that an origins-of-life process based on autocatalytic transformation of C,H,O-substrates was constrained to using the most reactive organic molecules that contain alpha- or beta-carbonyl groups, since small autocatalytic domains of plausible catalytic power that used less reactive substrates could not carry out chemical transformations fast enough to prevent catastrophic efflux (escape) of reaction intermediates. Knowledge of the kinetics of organic transformations is useful, not only in constraining the chemistry of the earliest autocatalytic process related to the origin of life, but also in establishing the relative reactivity of organic molecules on the early Earth and other planets that may or may not be related to the origin of life.

Weber, Arthur L.↗

Macrovoid Defect Growth during Evaporative Casting of Polymeric Membranes

Macrovoid (MV) formation is a significant problem in evaporatively cast polymeric membranes. MVs are large, elongated or teardrop-shaped pores (~10-50 micron) that can impair membrane structural integrity. Although MVs have been extensively studied, there is no general agreement on the mechanisms governing MV growth. Recently, our research group has formulated the solutocapillary convection (SC) hypothesis, which contends that MV growth involves three principal forces: a Marangoni force generated by surface tension gradients within the MV interface, a viscous drag force, and a gravitationally induced body force. Two sets of complementary experiments were conducted to test the SC hypothesis. Ground-based videomicroscopy flow-visualization (VMFV) was utilized to measure the flow velocities at the MV-casting solution interface and deep within the casting solution. The measurements were performed with casting solutions containing 10 wt% cellulose acetate (CA), 30 wt% H2O, 60 wt% acetone, and 200- ppm TiO2 particles for flow visualization, and the surface tension was controlled by surfactant addition. Qualitatively, the experiments indicated that MV growth occurs in three distinct phases: (1) a very rapid initial growth period, (2) a much slower growth phase, and (3) absorption of selected MVs into the expanding demixed region. The presence of tracer particles inside the MVs suggests the presence of a convective flow, which transfers the particles from the bulk solution to the MV interior. Although the VMFV experiments did not establish any surfactant effect on the interfacial velocities, a statistically significant effect on the MV number density was observed. In the second set of experiments, membranes were cast aboard a KC-135 aircraft under 0-g and 2-g conditions. Despite careful attention to the design and fabrication of the membrane casting apparatus (MCA), several problems were encountered, the most significant of which was the contamination of the casting solution by the activated carbon particles used for solvent absorption.

Greenberg, A. R.↗

Gelidivirgula Patagoniensis Gen. Nov., Sp. Nov., A Novel Psychrotolerant, Sporeforming Anaerobe Isolated from Magellanic Penguin Guano in Patagonia, Chile

A novel obligately anaerobic, psychrotrophic bacterium, strain PPP2(sup T), was isolated from guano of the Magellanic penguin (Spheniscus magellanicus) in Patagonia, Chile. The Gram-positive, sporeforming, straight rods with sizes 0.6-0.9 x 3.0-5.0 microns, are motile by peritrichous flagella. Growth was observed to occur within the pH range 6.0-9.5 (optimum pH x), and temperature range 2-28 C (optimum 20 C). The novel isolate does not require NaCl for growth, but is halotolerant and growth was observed between 0 and 7 % NaCl (w/v) with optimum at 0.5 % (w/v). The new isolate is a catalase negative chemoorganohetherotroph with fermentative metabolism and uses as substrates: peptone, Bacto-tryptone, Casamino acids, and yeast extract. The major metabolic products are: acetate, butyrate, ethanol, and hydrogen is a minor gas product.. Strain PPP2 was sensitive to ampicillin, tetracycline, chloramphenicol, rifampin, kanamycin, and gentamycin. The G+C content of the DNA is 43.6 mol%. On the basis of 16S rDNA gene sequences and phenotypic characteristics, it is proposed that the strain PPP2(sup T) (= ATCC BAA-755(sup T) = JSM ...(sup T)) is assigned to the new genus Gelidivirgula gen. nov., as a representative of the new species, Gelidivirgula patagonensis sp. nov.

Pikuta, Elena V.↗

Purification, Crystallization, and Preliminary X-ray Analysis of Native Canavalin

The protein canavalin is a 7S vicilin, from the Jack Bean, Canavalis ensfomis. Canavalin is described as a seed storage protein, an energy source for a developing seed, as no other known activity or function has been found. The protein was first isolated and crystallized by Sumner and Howell (J. Biol. Chem. 113, 607-610, 1936). Canavalin spontaneously crystallizes after proteolytic cleavage at neutral pH, which removes residues 1-46, 224-245, and 325-330 and produces peptides of approximately 25, 13, and 12 kDa. Preliminary gel filtration experiments indicated the presence of nucleic acid with the uncleaved protein. We developed a dual column procedure, ion exchange followed by hydroxy apatite chromatography, that effectively removes the nucleic acid and yields an essentially pure uncut canavalin preparation with an OD 280/260 ratio of approximately 1.9-2.0. Standard crystallization screens using this material gave a number of positive results having a common requirement for alcohols and Mg(2+) ion, with crystals typically appearing within a day or less. Optimization experiments to date have shown that we can obtain crystals from pH 6.5 to pH 8.2, using MPD from 5 to 20% and 0.05 to 0.2M Mg(2+) (sulfate or acetate). The crystals are of space group P2(sub 1)2(sub 1)2(sub 1), unit cell dimensions, and a complete data set to 1.5 Angstroms, resolution has now been collected at a synchrotron source. Most importantly, the crystals are not twinned, a persistent problem with the most commonly obtained rhombohedral form of proteolytically cleaved canavalin.

Pusey, Marc↗

The Lifetimes of Nitriles (-C*N) and Acids (-COOH) during Ultraviolet Photolysis and Their Survival in Space

Nitriles are one of the most common classes of molecules observed in the gas phase in space, with over a dozen having been positively identified in interstellar and circumstellar environments through the detection of their rotational transitions. Acids, in contrast, are much less common. In this paper we present laboratory data comparing the stability of two structurally related acid-nitrile pairs to ultraviolet (UV) photolytic destruction: acetic acid vs. acetonitrile (CH3-COOH vs. CH3-CN) and glycine vs. aminoacetonitrile (H2N-CH2-COOH vs. H2N-CH2-CN). We find that the nitriles are destroyed ten and five times more slowly (respectively) by UV photolysis than are the corresponding acids. This suggests that whatever their relative formation rates, acids may be less abundant than nitriles in interstellar environments in part because they are more rapidly destroyed by photolysis. The results of this infrared (IR) spectral matrix isolation study indicate that during the lifetime of a typical interstellar cloud, even in its darkest regions, a population of acids in the gas phase will likely be diminished by at least half. Since aminoacetonitrile is a precursor to the amino acid glycine, and far more stable, presolar aminoacetonitrile may be a contributor to the deuterium enriched glycine detected in meteorites. It would clearly be informative to search for aminoacetonitrile (the nitrile corresponding to glycine) in the regions where the amino acid glycine has been reported.

Bernstein, Max P.↗

Tindallia californiensis sp. nov., a new anaerobic, haloalkaliphilic, spore-forming acetogen isolated from Mono Lake in California

A novel extremely haloalkaliphilic, strictly anaerobic, acetogenic bacterium strain APO was isolated from sediments of the athalassic, meromictic, alkaline Mono Lake in California. The Gram-positive, spore-forming, slightly curved rods with sizes 0.55- 0.7x1.7-3.0 microns were motile by a single laterally attached flagellum. Strain APO was mesophilic (range 10-48 C, optimum of 37 C); halophilic (NaCl range 1-20% (w/v) with optimum of 3-5% (w/v), and alkaliphilic (pH range 8.0-10.5, optimum 9.5). The novel isolate required sodium ions in the medium. Strain APO was an organotroph with a fermentative type of metabolism and used the substrates peptone, bacto-tryptone, casamino acid, yeast extract, L-serine, L-lysine, L-histidine, L-arginine, and pyruvate. The new isolate performed the Stickland reaction with the following amino acid pairs: proline + alanine, glycine + alanine, and tryptophan + valine. The main end product of growth was acetate. High activity of CO dehydrogenase and hydrogenase indicated the presence of a homoacetogenic, non-cycling acetyl-coA pathway. Strain APO was resistant to kanamycin but sensitive to chloramphenicol, tetracycline, and gentamycin. The G+C content of the genomic DNA was 44.4 mol% (by HPLC method). The sequence of the 16s rRNA gene of strain APO possessed 98.2% similarity with the sequence from Tindullia magadiensis Z-7934, but the DNA-DNA hybridization value between these organisms was only 55%. On the basis of these physiological and molecular properties, strain APO is proposed to be a novel species of the genus Tindallia with the name Tindallia californiensis sp. nov., (type strain APO = ATCC BAA-393 - DSM 14871).

Pikuta, E. V.↗