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At least 991 records · Page 55

Cloning and Characterization of an alpha-amylase Gene from the Hyperthermophilic Archaeon Thermococcus Thioreducens

The gene encoding an extracellular alpha-amylase, TTA, from the hyperthermophilic archaeon Thermococcus thioreducens was cloned and expressed in Escherichia coli. Primary structural analysis revealed high similarity with other a-amylases from the Thermococcus and Pyrococcus genera, as well as the four highly conserved regions typical for a-amylases. The 1374 bp gene encodes a protein of 457 amino acids, of which 435 constitute the mature protein preceded by a 22 amino acid signal peptide. The molecular weight of the purified recombinant enzyme was estimated to be 43 kDa by denaturing gel electrophoresis. Maximal enzymatic activity of recombinant TTA was observed at 90 C and pH 5.5 in the absence of exogenous Ca(2+), and the enzyme was considerably stable even after incubation at 90 C for 2 hours. The thermostability at 90 and 102 C was enhanced in the presence of 5 mM Ca(2+). The extraordinarily high specific activity (about 7.4 x 10(exp 3) U/mg protein at 90 C, pH 5.5 with soluble starch as substrate) together with its low pH optimum makes this enzyme an interesting candidate for starch processing applications.

Bernhardsdotter, Eva C. M. J.↗

Enzymatic Properties of an Alkaline and Chelator Resistant alpha-amylase from the Alkaliphilic Bacillus sp. Isolate L1711

An alkaliphilic amylase producing bacterium, Bacillus sp. strain L 711, was selected among 13 soda lakes isolates. When grown at pH 10.5 and 37 C, strain L711 produced multiple forms of amylases in the culture broth. One of these, BAA, was purified from the culture supernatant by QAE column chromatography and preparative native gel electrophoresis. The molecular weight of BAA was determined to be 51 kDa by denaturing gel electrophoresis. The pH optima for activity below and above 40 C were 9.5 - 10.0 and 7.0 - 7.5 respectively. BAA was stable in the pH range 6-11 and was completely inactivated at 55 C. The thermostability was not increased in the presence of Ca(2+). The enzyme was strongly inhibited by Ca(2+), Zn(2+), Mg(2+), Mn(2+), Ba(2+) and Cu(2+), whereas the presence of Na(+), Co(2+) and EDTA (10 mM) enhanced enzymatic activity. The K(sub m), and specific activity of BAA on soluble starch were 1.9 mg/ml and 18.5 U/mg respectively. The main end products of hydrolysis were maltotetraose, maltose and glucose.

Bernhardsdotter, Eva C. M. J.↗

Aqueous Chemical Modeling of Sedimentation on Early Mars with Application to Surface-Atmosphere Evolution

This project was to investigate models for aqueous sedimentation on early Mars from fluid evaporation. Results focused on three specific areas: (1) First, a fluid evaporation model incorporating iron minerals was developed to compute the evaporation of a likely solution on early Mars derived from the weathering of mafic rock. (2) Second, the fluid evaporation model was applied to salts within Martian meteorites, specifically salts in the nakhlites and ALH84001. Evaporation models were found to be consistent with the mineralogy of salt assemblages-anhydrite, gypsum, Fe-Mg-Ca carbonates, halite, clays-- and the concentric chemical fractionation of Ca-to Mg-rich carbonate rosettes in ALH84001. We made progress in further developing our models of fluid concentration by contributing to updating the FREZCHEM model. (3) Third, theoretical investigation was done to determine the thermodynamics and kinetics involved in the formation of gray, crystalline hematite. This mineral, of probable ancient aqueous origin, has been observed in several areas on the surface of Mars by the Thermal Emission Spectrometer on Mars Global Surveyor. The "Opportunity" Mars Exploration Rover has also detected gray hematite at its landing site in Meridiani Planum. We investigated how gray hematite can be formed via atmospheric oxidation, aqueous precipitation and subsequent diagenesis, or hydrothermal processes. We also studied the geomorphology of the Aram Chaos hematite region using Mars Orbiter Camera (MOC) images.

Catling, David C.↗

Chemical Weathering Records of Martian Soils Preserved in the Martian Meteorite EET79001

Impact-melt glasses, rich in Martian atmospheric gases, contain Martian soil fines (MSF) mixed with other coarse-grained regolith fractions which are produced during impact bombardment on Mars surface. An important characteristic of the MSF fraction is the simultaneous enrichment of felsic component accompanied by the depletion of mafic component relative to the host phase in these glasses. In addition, these glasses yield large sulfur abundances due to the occurrence of secondary mineral phases such as sulfates produced during acid-sulfate weathering of the regolith material near the Martian surface. Sulfurous gases released into atmosphere by volcanoes on Mars are oxidized to H2SO4 which deposit back on the surface of Mars as aerosol particles. Depending on the water availability, sulfuric acids dissolve into solutions which aggressively decompose the Fe-Mg silicates in the Martian regolith. During chemical weathering, structural elements such as Fe, Mg and Ca (among others) are released into the transgressing solutions. These solutions leach away the soluble components of Mg, Ca and Na, leaving behind insoluble iron as Fe3(+) hydroxysulfate mixed with poorly crystalline hydroxide- precipitates under oxidizing conditions. In this study, we focus on the elemental distribution of FeO and SO3 in the glass veins of EET79001, 507 sample, determined by Electron Microprobe and FE SEM measurements at JSC. This glass sample is an aliquot of a bigger glass inclusion ,104 analysed by where large concentrations of Martian atmospheric noble gases are found.

Rao, M. N.↗

Compositional Controls on the Formation of Kaersutite Amphibole in Shergottite Meteorites

The shergottite basalts, meteorites of Martian origin, contain rare small grains (approx. 10-100 microns diam.) of kaersutite, a Ca-amphibole rich in Al and Ti. Kaersutites have been used to estimate the water content of shergottites and the Martian mantle; however, questions remain about the original water content of the amphiboles and if they formed from magma. We investigated the petrographic settings of amphiboles in two shergottites and confirm that these amphiboles occur only in multiphase inclusions in pyroxene. In fact, kaersutite is found only in pigeonite. This suggests that the occurrence of amphibole is controlled in part by the composition of its host phase. Crystallization of host (cognate) pigeonite from a magmatic inclusion will enrich the remaining melt in Ca, Al, and Ti, supporting formation of kaersutite.

Pitman, K. M.↗

Enzymatic Properties of an Alkaline and Chelator Resistant Proportional to alpha-Amylase from the Alkaliphilic Bacillus sp. Isolate L1711

An alkaliphilic amylase producing bacterium, Bacillus sp. strain L1711, was selected among 13 soda lakes isolates. When grown at pH 10.5 and 370 C, strain L1711 produced multiple forms of amylases in the culture broth. One of these, BAA, was purified from the culture supernatant by QAE column chromatography and preparative native gel electrophoresis. The molecular weight of BAA was determined to be 51 kDa by denaturing gel electrophoresis. The pH optima for activity below and above 40 C were 9.5-10.0 and 7.0-7.5 respectively. BAA was stable in the pH range 6-11 and was completely inactivated at 55?C. The thermostability was not increased in the presence of Ca(2+). The enzyme was strongly inhibited by Ca(2+), Zn(2+), Mg(2+), Mn(2+), Ba(2+) and Cu(2+), whereas the presence of Na(+), Co2+ and EDTA (10 mM) enhanced enzymatic activity. The K(sub m) and specific activity of BAA on soluble starch were 1.9 mg/ml and 18.5 U/mg respectively. The main end products of hydrolysis were maltotetraose, maltose and glucose .

Bernhardsdotter, Eva C. M. J.↗

Cloning and Characterization of an Alpha-amylase Gene from the Hyperthermophilic Archaeon Thermococcus Thioreducens

The gene encoding an extracellular a-amylase, TTA, from the hyperthermophilic archaeon Thermococcus thioreducens was cloned and expressed in Escherichia coli. Primary structural analysis revealed high similarity with other a-amylases from the Thermococcus and Pyrococcus genera, as well as the four highly conserved regions typical for a-amylases. The 1374 bp gene encodes a protein of 457 amino acids, of which 435 constitute the mature protein preceded by a 22 amino acid signal peptide. The molecular weight of the purified recombinant enzyme was estimated to be 43 kDa by denaturing gel electrophoresis. Maximal enzymatic activity of recombinant TTA was observed at 90 C and pH 5.5 in the absence of exogenous Ca(2+), and the enzyme was considerably stable even after incubation at 90 C for 2 hours. The thermostability at 90 and 102 C was enhanced in the presence of 5 mM Ca(2+). The extraordinarily high specific activity (about 7.4 x 10(exp 3) U/mg protein at 90 C, pH 5.5 with soluble starch as substrate) together with its low pH optimum makes this enzyme an interesting candidate for starch processing applications.

Bernhardsdotter, Eva C. M. J.↗

Mineralogical and chemical characterization of iron-, manganese-, and copper-containing synthetic hydroxyapatites

The National Aeronautics and Space Administration's (NASA) Advanced Life Support (ALS) Program is evaluating the use of Fe-, Mn-, and Cu-containing synthetic hydroxyapatite (SHA) as a slow release fertilizer for crops that might be grown on the International Space Station or at Lunar and Martian outposts. Separate Fe-, Mn-, and Cu-containing SHA materials along with a transition-metal free SHA (pure-SHA) were synthesized using a precipitation method. Chemical and mineralogical analyses determined if and how Fe, Mn, and Cu were incorporated into the SHA structure. X-ray diffraction (XRD), Rietveld refinement, and transmission electron microscopy (TEM) confirmed that SHA materials with the apatite structure were produced. Chemical analyses indicated that the metal containing SHA materials were deficient in Ca relative to pure-SHA. The shift in the infrared PO4-mu 3 vibrations, smaller unit cell parameters, smaller particle size, and greater structural strain for Fe-, Mn-, and Cu-containing SHA compared with pure-SHA suggested that Fe, Mn, and Cu were incorporated into SHA structure. Rietveld analyses revealed that Fe, Mn, and Cu substituted into the Ca2 site of SHA. An Fe-rich phase was detected by TEM analyses and backscattered electron microscopy in the Fe-containing SHA material with the greatest Fe content. The substitution of metals into SHA suggests that metal-SHA materials are potential slow-release sources of micronutrients for plant uptake in addition to Ca and P.

NASA Center JSC↗

Calcium/calmodulin-mediated signal network in plants

Various extracellular stimuli elicit specific calcium signatures that can be recognized by different calcium sensors. Calmodulin, the predominant calcium receptor, is one of the best-characterized calcium sensors in eukaryotes. In recent years, completion of the Arabidopsis genome project and advances in functional genomics have helped to identify and characterize numerous calmodulin-binding proteins in plants. There are some similarities in Ca(2+)/calmodulin-mediated signaling in plants and animals. However, plants possess multiple calmodulin genes and many calmodulin target proteins, including unique protein kinases and transcription factors. Some of these proteins are likely to act as "hubs" during calcium signal transduction. Hence, a better understanding of the function of these calmodulin target proteins should help in deciphering the Ca(2+)/calmodulin-mediated signal network and its role in plant growth, development and response to environmental stimuli.

NASA Discipline Plant Biology↗

Plant growth during the Greenhouse II experiment on the Mir orbital station

We carried out three experiments with Super Dwarf wheat in the Bulgarian/Russian growth chamber Svet (0.1 m2 growing area) on the Space Station Mir. This paper mostly describes the first of these NASA-supported trials, began on Aug. 13, 1995. Plants were sampled five times and harvested on Nov. 9 after 90 days. Equipment failures led to low irradiance (3, then 4 of 6 lamp sets failed), instances of high temperatures (ca. 37 degrees C), and sometimes excessive substrate moisture. Although plants grew for the 90 d, no wheat heads were produced. Considering the low light levels, plants were surprisingly green, but of course biomass production was low. Plants were highly disoriented (low light, mirror walls?). Fixed and dried samples and the root module were returned on the U.S. Shuttle Atlantis on Nov. 20, 1995. Samples of the substrate, a nutrient-charged zeolite called Balkanine, were taken from the root module, carefully examined for roots, weighed, dried, and reweighed. The Svet control unit and the light bank were shipped to Moscow. An experiment validation test (EVT) of plant growth and experimental procedures, carried out in Moscow, was highly successful. Equipment built in Utah to measure CO2, H2O vapor, irradiance, air and leaf (IR) temperature, O2, pressure, and substrate moisture worked well in the EVT and in space. After this manuscript was first prepared, plants were grown in Mir with a new light bank and controller for 123 d in late 1996 and 39 days in 1996/1997. Plants grew exceptionally well with higher biomass production than in any previous space experiment, but the ca. 280 wheat heads that were produced in 1996 contained no seeds. Ethylene in the cabin atmosphere was responsible. c2002 Published by Elsevier Science Ltd on behalf of COSPAR.

Mir Project↗

Hypertonicity-induced transmitter release at Drosophila neuromuscular junctions is partly mediated by integrins and cAMP/protein kinase A

The frequency of quantal transmitter release increases upon application of hypertonic solutions. This effect bypasses the Ca(2+) triggering step, but requires the presence of key molecules involved in vesicle fusion, and hence could be a useful tool for dissecting the molecular process of vesicle fusion. We have examined the hypertonicity response at neuromuscular junctions of Drosophila embryos in Ca(2+)-free saline. Relative to wild-type, the response induced by puff application of hypertonic solution was enhanced in a mutant, dunce, in which the cAMP level is elevated, or in wild-type embryos treated with forskolin, an activator of adenylyl cyclase, while protein kinase A (PKA) inhibitors decreased it. The response was also smaller in a mutant, DC0, which lacks the major subunit of PKA. Thus the cAMP/PKA cascade is involved in the hypertonicity response. Peptides containing the sequence Arg-Gly-Asp (RGD), which inhibit binding of integrins to natural ligands, reduced the response, whereas a peptide containing the non-binding sequence Arg-Gly-Glu (RGE) did not. A reduced response persisted in a mutant, myospheroid, which expresses no integrins, and the response in DC0 was unaffected by RGD peptides. These data indicate that there are at lease two components in the hypertonicity response: one that is integrin mediated and involves the cAMP/PKA cascade, and another that is not integrin mediated and does not involve the cAMP/PKA cascade.

NASA Discipline Cell Biology↗

Gravity-dependent polarity of cytoplasmic streaming in Nitellopsis

The internodal cells of the characean alga Nitellopsis obtusa were chosen to investigate the effect of gravity on cytoplasmic streaming. Horizontal cells exhibit streaming with equal velocities in both directions, whereas in vertically oriented cells, the downward-streaming cytoplasm flows ca. 10% faster than the upward-streaming cytoplasm. These results are independent of the orientation of the morphological top and bottom of the cell. We define the ratio of the velocity of the downward- to the upward-streaming cytoplasm as the polar ratio (PR). The normal polarity of a cell can be reversed (PR < 1) by treatment with neutral red (NR). The NR effect may be the result of membrane hyperpolarization, caused by the opening of K+ channels. The K+ channel blocker TEA Cl- inhibits the NR effect. External Ca2+ is required for normal graviresponsiveness. The [Ca2+] of the medium determines the polarity of cytoplasmic streaming. Less than 1 micromole Ca2+ resulted in a PR < 1 while greater than 1 micromole Ca2+ resulted in the normal gravity response. The voltage-dependent Ca(2+)-channel blocker, nifedipine, inhibited the gravity response in a reversible manner, while treatment with LaCl3 resulted in a PR < 1, indicating the presence of two types of Ca2+ channels. A new model for graviperception is presented in which the whole cell acts as the gravity sensor, and the plasma membrane acts as the gravireceptor. This is supported by ligation and UV irradiation experiments which indicate that the membranes at both ends of the cell are required for graviperception. The density of the external medium also affects the PR of Nitellopsis. Calculations are presented that indicate that the weight of the protoplasm may provide enough potential energy to open ion channels.

NASA Discipline Plant Biology↗

Determination of cytoplasmic calcium concentration in Dryopteris spores: a developmentally non-disruptive technique for loading of the calcium indicator fura-2

Germination of Dryopteris spores is mediated by the physiologically active, far-red-absorbing form of phytochrome, Pfr, and external Ca2+ is necessary for the transduction of the light signal. Because knowledge about the cytoplasmic calcium ion concentration, [Ca2+]i, is of great importance for understanding the role of calcium during signal transduction, this value was measured using fura-2 in fern spores undergoing the normal developmental progression into germination. Fura-2 was loaded into the spores by electroporation, which does not disrupt the normal process of germination. The intensity of the fluorescence emission of the loaded fura-2 was analysed by a microspectrophotometric assay of single spores, and successful loading could be obtained by the application of ten electrical pulses (field strength 7.5 kV cm-1, half-life (time constant) 230 microseconds). Fura-2 was alternately excited by light of wavelengths 355 and 385 nm through an inverted fluorescence microscope, and the emitted fura-2 fluorescence was collected by a silicon-intensified video camera. The cytoplasmic calcium ion concentration was calculated from the ratio of the camera output obtained for both wavelengths and displayed by a pseudo-color technique. Spores responded to changes of the extracellular Ca2+ concentration, and this observation is considered as evidence that fura-2 is loaded into the cytoplasm. The substitution of a low external [Ca2+] (1 mM ethyleneglycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA)) by 1 mM CaCl2 caused a fast increase of [Ca2+]i from approx. 50 nM to above 500 nM. In contrast, the subsequent substitution of CaCl2 by EGTA decreased [Ca2+]i again below 100 nM within 0.5 h. Furthermore, the application of ionomycin could initiate a change in [Ca2+]i according to the Ca2+ gradient established between the extracellular medium and cytoplasm. In spores sown on a Ca(2+) -free medium, [Ca2+]i, analysed in a buffer containing EGTA, was found to be around 50 nM during the first days of cultivation, independent of the irradiation protocol. However, if spores were grown in darkness on a Ca(2+) -containing medium and analysed in EGTA, [Ca2+]i was significantly higher (> or = 500 nM). In red-light-irradiated spores, [Ca2+]i was found to decrease with increasing time after irradiation, and was determined to be less than 100 nM when analysis was done 44 h after germination was initiated by the light treatment.

NASA Discipline Plant Biology↗

Stimulation of root elongation and curvature by calcium

Ca2+ has been proposed to mediate inhibition of root elongation. However, exogenous Ca2+ at 10 or 20 millimolar, applied directly to the root cap, significantly stimulated root elongation in pea (Pisum sativum L.) and corn (Zea mays L.) seedlings. Furthermore, Ca2+ at 1 to 20 millimolar, applied unilaterally to the caps of Alaska pea roots, caused root curvature away from the Ca2+ source, which was caused by an acceleration of elongation growth on the convex side (Ca2+ side) of the roots. Roots of an agravitropic pea mutant, ageotropum, responded to a greater extent. Roots of Merit and Silver Queen corn also responded to Ca2+ in similar ways but required a higher Ca2+ concentration than that of pea roots. Roots of all other cultivars tested (additional four cultivars of pea and one of corn) curved away from the unilateral Ca2+ source as well. The Ca(2+)-stimulated curvature was substantially enhanced by light. A Ca2+ ionophore, A23187, at 20 micromolar or abscisic acid at 0.1 to 100 micromolar partially substituted for the light effect and enhanced the Ca(2+)-stimulated curvature in the dark. Unilateral application of Ca2+ to the elongation zone of intact roots or to the cut end of detipped roots caused either no curvature or very slight curvature toward the Ca2+. Thus, Ca2+ action on root elongation differs depending on its site of application. The stimulatory action of Ca2+ may involve an elevation of cytoplasmic Ca2+ in root cap cells and may partipate in root tropisms.

NASA Program Space Biology↗

Radiobiological experiments with plant seeds aboard the biosatellite Kosmos 1887

The effects of spaceflight factors on the seeds of Arabidopsis thaliana and Crepis capillaris were studied provided with various protective measures: the seeds were located inside the satellite and in open space, protected with aluminium foil and also exposed without the foil cover. When the seeds were in open space without any protection, their viability was found to be suppressed; the survival rate and fertility of plants grown from these seeds were also diminished. An increase in the frequency of chromosome aberrations (CA) and in the number of multiple injuries was registered in this case. Experiments with the aluminium foil shielding showed a decrease in the suppression of the seeds' viability, but mutational changes were found to be even more increased, while the survival and fertility of the plants decreased. An increase in the thickness of shielding resulted in a decrease in the effects up to the level of the control, except for the effects connected with CA and fertility of the plants. Analysis of the results shows that these impairments can be ascribed to the action of single heavy charged particles (HCP). The seeds can be thus regarded as an integral biological 'dosimeter' which allows estimation of the total effects of radiation, ecological and biological factors.

Cosmos 1887 Project↗

Growth, carbon dioxide exchange and mineral accumulation in potatoes grown at different magnesium concentrations

Plants of Norland potatoes (Solanum tuberosum L.) were maintained for 42 days at Mg concentrations of 0.05, 0.125, 0.25, 1, 2, and 4 mM in a nonrecirculating nutrient film system under controlled environment. With the increased Mg supply from 0.05 to 4 mM, Mg concentrations in the leaves of the 42-day old plants increased significantly from 1.1 to 11.2 mg g-1 dry weight. Plant leaf area and plant and tuber dry weights increased with increased Mg concentrations up to 1 mM in solution or 6.7 mg g-1 in leaves, and then decreased with further increases in Mg concentrations. Rates of CO2 assimilation measured on leaflets in situ at ambient and various intercellular CO2 concentrations were consistently lower at 0.05 and 4 mM Mg than at other Mg treatments, which may indicate decreased photosynthetic activity in mesophyll tissues at the lowest and highest Mg concentrations. Dark respiration rates in leaves were highest at 0.05 and 4 mM Mg, lowest at 0.25 and 1 mM Mg, and intermediate at 0.125 and 2 mM Mg. The different Mg treatments also influenced accumulation of other minerals in leaves. Leaf concentrations of Ca and Mn decreased with increased Mg supply except that Ca and Mn were lower at 0.05 mM than at 0.125 mM Mg. Leaf K concentrations were lower at 1, 2 and 4 mM Mg than at other Mg treatments. Foliar concentrations of P, Fe, Zn, and Cu had small but inconsistent variation with different Mg concentrations. Leaf concentrations of N, S, and B were similar at different Mg concentrations. This study demonstrates that various Mg nutrition, along with altered accumulation of other nutrients, could regulate dry matter production in potatoes by affecting not only leaf area but also leaf carbon dioxide assimilation and respiration.

NASA Discipline Number 61-10↗

An Overview of Recent Developments in Computational Aeroelasticity

The motivation for Computational Aeroelasticity (CA) and the elements of one type of the analysis or simulation process are briefly reviewed. The need for streamlining and improving the overall process to reduce elapsed time and improve overall accuracy is discussed. Further effort is needed to establish the credibility of the methodology, obtain experience, and to incorporate the experience base to simplify the method for future use. Experience with the application of a variety of Computational Aeroelasticity programs is summarized for the transonic flutter of two wings, the AGARD 445.6 wing and a typical business jet wing. There is a compelling need for a broad range of additional flutter test cases for further comparisons. Some existing data sets that may offer CA challenges are presented.

Bennett, Robert M.↗

A Review of Solar-Powered Aircraft Flight Activity at the Pacific Missile Range Test Facility, Kauai, Hawaii

A series of solar-powered aircraft have been designed and operated by AeroVironment, Inc. (Monrovia, CA) as a part of National Aeronautics and Space Administration (NASA) objectives to develop energy-efficient high-altitude long-endurance platforms for earth observations and communications applications. Flight operations have been conducted at NASA's Dryden Flight Research Center, Edwards CA and at the U.S. Navy Pacific Missile Range Facility (PMRF) at Barking Sands, Kauai, HI. These aircraft flown at PMRF are named Pathfinder , Pathfinder Plus and Helios . Sizes of these three aircraft range from 560 lb with a 99-ft wingspan to 2300 lb with a 247-ft wingspan. Available payload capacity reaches approximately 200 lb. Pathfinder uses six engines and propellers: Pathfinder Plus 8; and Helios 14. The 2003 Helios fuel cell configurations used 10 engines and propellers. The PMRF was selected as a base of operations because if offers optimal summertime solar exposure, low prevailing wind-speeds on the runway, modest upper-air wind-speeds and the availability of suitable airspace. Between 1997 and 2001, successive altitude records of 71,530 ft, 80,200 ft, and 96,863 ft were established. Flight durations extended to 18 hours.

Ehernberger, L. J.↗