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Solid state 31phosphorus nuclear magnetic resonance of iron-, manganese-, and copper-containing synthetic hydroxyapatites.

The incorporation of micronutrients into synthetic hydroxyapatite (SHA) is proposed for slow release of these nutrients to crops in the National Aeronautics and Space Administration's (NASA's) Advanced Life Support (ALS) program for Lunar or Martian outposts. Solid state 31P nuclear magnetic resonance (NMR) was utilized to examine the paramagnetic effects of Fe3+, Mn2+, and Cu2+ to determine if they were incorporated into the SHA structure. Separate Fe3+, Mn2+, and Cu2+ containing SHA materials along with a transition metal free SHA (pure-SHA) were synthesized using a precipitation method. The proximity (<1 nm) of the transition metals to the 31P nuclei of SHA were apparent when comparing the integrated 31P signal intensities of the pure-SHA (87 arbitrary units g-1) with the Fe-, Mn-, and Cu-SHA materials (37-71 arbitrary units g-1). The lower integrated 31P signal intensities of the Fe-, Mn-, and Cu-SHA materials relative to the pure-SHA suggested that Fe3+, Mn2+, and Cu2+ were incorporated in the SHA structure. Further support for Fe3+, Mn2+, and Cu2+ incorporation was demonstrated by the reduced spin-lattice relaxation constants of the Fe-, Mn-, and Cu-SHA materials (T'=0.075-0.434s) relative to pure-SHA (T1=58.4s). Inversion recovery spectra indicated that Fe3+, Mn2+, and Cu2+ were not homogeneously distributed about the 31P nuclei in the SHA structure. Extraction with diethylene-triamine-penta-acetic acid (DTPA) suggested that between 50 and 80% of the total starting metal concentrations were incorporated in the SHA structure. Iron-, Mn-, and Cu-containing SHA are potential slow release sources of Fe, Mn, and Cu in the ALS cropping system.

Calcium↗

[Cyto-embryologic investigation of super dwarf wheat grown on board of the Mir orbital complex].

The cytoembryologic analysis of wet and dry samples of super dwarf wheat cultivated in greenhouse SVET aboard the MIR station over the whole cycle of vegetation was made with the use of light microscopy. Characteristic features of wheat development in space flight are plentiful early tillering, and formation and rapid growth of side shoots. Elementary spikelets in the composite head were more numerous but the top spikelets were rudimentary and, therefore, the ripe head contained less of these spikelets as compared with the ground controls (9-13 and 14, respectively). The number of florets in a spike was also higher reaching 14-16 vs. 7-8 in the control. Typically, no more than 4 to 5 florets vs. 3 in the control were fully differentiated while the others died off earlier in development. The fact that there were no caryopses found in the flight crop is explained by absolute male sterility appearing at different stages of staminal development: before archesporium formation, on the stage of differentiated archesporium, during meiosis, on the stage of microspores or uninucleate "pollen". The female generative system developed mainly without abnormalities. An assumption was made that elevated ethylene concentrations in the MIR atmosphere at the time of the wheat experiment were the cause for abnormal development of the male generative system which led to barrenness of the super dwarf wheat crop.

Animals↗

Effects of drying on nitrification activity in zeoponic medium used for long-term space missions.

One component of a proposed life support system is the use of zeoponic substrates, which slowly release NH4+ into "soil" solution, for the production of plants. Nitrifying bacteria that convert NH4+ to NO3- are among the important microbial components of these systems. Survival of nitrifying bacteria in dry zeoponic substrates is needed, because the substrate would likely be stored in an air-dry state between croppings. Substrate was enriched for nitrifying bacteria and allowed to air-dry in a laminar flow hood. Stored substrate was analyzed for nitrifier survivability by measuring nitrifier activity at the beginning, 3 days, 1, 2, and 3 weeks. After rewetting, activity was approximately 9 micrograms N g-1 h-1 regardless of storage time. Nitrification rates did not decrease during storage. It seems unlikely that drying between plantings would result in practical reductions in nitrification, and reinoculation with nitrifying bacteria would not be necessary.

Bacteria↗

[Growth of wheat from seed-to-seed in space flight].

In our earlier space experiment with super dwarf wheat we found the spikes developed in space to be barren. The cause of the full crop sterility was sensitivity of this wheat species to the ethylene concentration of 0.3-0.8 mg/m3 during the experiment. The follow-up ground experiments were made to identify species of dwarf wheat that could be raised in space greenhouse Svet and are distinguished by partial tolerance of their reproductive organs to elevated ethylene in air. The choice fell on the USU-Apogee cultivar specially developed for planting in growth chambers as an integral part of various bioregenerative life support systems, including the space ones. An experiment with wheat Apogee was performed in greenhouse Svet on board MIR. The period of the full crop vegetation cycle was not significantly altered under the spaceflight conditions. The experiment yielded 508 seeds from 12 plants, i.e. by 38% less than in laboratory experiments and by 69% more as compared with results of growing crops in ethylene-contaminated atmosphere (1 mg/m3). Mass of the space seeds was low if compared with the laboratory crops. This was the first time when the feasibility of gathering seeds from wheat that had passed the whole vegetation cycle in space flight was demonstrated. The experiment will give mightly impetus to the advancement of research on space biological LSS and gravitational biology.

Air Pollutants↗

Development of autonomous control in a closed microbial bioreactor.

Space-based life support systems which include ecological components will rely on sophisticated hardware and software to monitor and control key system parameters. Autonomous closed artificial ecosystems are useful for research in numerous fields. We are developing a bioreactor designed to study both microbe-environment interactions and autonomous control systems. Currently we are investigating N-cycling and N-mass balance in closed microbial systems. The design features of the system involve real-time monitoring of physical parameters (e.g. temperature, light), growth solution composition (e.g. pH, NOx, CO2), cell density and the status of important hardware components. Control of key system parameters is achieved by incorporation of artificial intelligence software tools that permit autonomous decision-making by the instrument. These developments provide a valuable research tool for terrestrial microbial ecology, as well as a testbed for implementation of artificial intelligence concepts. Autonomous instrumentation will be necessary for robust operation of space-based life support systems, and for use on robotic spacecraft. Sample data acquired from the system, important features of software components, and potential applications for terrestrial and space research will be presented.

Artificial Intelligence↗

CO2 capture by means of an enzyme-based reactor.

We report a means for efficient and selective extraction of carbon dioxide (CO(2)) at low to medium concentration from mixed gas streams. CO(2) capture was accomplished by use of a novel enzyme-based, facilitated transport contained liquid membrane (EBCLM) reactor. The parametric studies we report explore both structural and operational parameters of this design. The structural parameters include carbonic anhydrase (CA) concentration, buffer concentration and pH, and liquid membrane thickness. The operational parameters are temperature, humidity of the inlet gas stream, and CO(2) concentration in the feed stream. The data show that this system effectively captures CO(2) over the range 400 ppm to at least 100,000 ppm, at or around ambient temperature and pressure. In a single pass across this homogeneous catalyst design, given a feed of 0.1% CO(2), the selectivity of CO(2) versus N(2) is 1,090 : 1 and CO(2) versus O(2) is 790 :1. CO(2) permeance is 4.71 x 10(-8) molm(-2) Pa(-1) sec(-1). The CLM design results in a system that is very stable even in the presence of dry feed and sweep gases.

Animals↗

Anion-exchange high-performance liquid chromatography with post-column detection for the analysis of phytic acid and other inositol phosphates.

The use of gradient anion-exchange HPLC, with a simple post-column detection system, is described for the separation of myo-inositol phosphates, including "phytic acid" (myo-inositol hexaphosphate). Hexa-, penta-, tetra-, tri- and diphosphate members of this homologous series are clearly resolved within 30 min. This method should facilitate analysis and quantitation of "phytic acid" and other inositol phosphates in plant, food, and soil samples.

Chromatography, High Pressure Liquid↗

Recovery of resources for advanced life support space applications: effect of retention time on biodegradation of two crop residues in a fed-batch, continuous stirred tank reactor.

Bioreactor retention time is a key process variable that will influence costs that are relevant to long distance space travel or long duration space habitation. However. little is known about the effects of this parameter on the microbiological treatment options that are being proposed for Advanced Life Support (ALS) systems. Two bioreactor studies were designed to examine this variable. In the first one, six retention times ranging from 1.3 to 21.3 days--were run in duplicate, 81 working-volume continuous stirred tank reactors (CSTR) that were fed ALS wheat residues. Ash-free dry weight loss, carbon mineralization, soluble TOC reduction, changes in fiber content (cellulose, hemicellulose, and lignin), bacterial numbers, and mineral recoveries were monitored. At short retention times--1.33 days--biodegradation was poor (total: 16-20%, cellulose - 12%, hemicellulose - 28%) but soluble TOC was decreased by 75-80% and recovery of major crop inorganic nutrients was adequate, except for phosphorus. A high proportion of the total bacteria (ca. 83%) was actively respiring. At the longest retention time tested, 21.3 days, biodegradation was good (total: 55-60%, cellulose ca. 70%, hemicellulose - ca. 55%) and soluble TOC was decreased by 80%. Recovery of major nutrients, except phosphorus, remained adequate. A very low proportion of total bacteria was actively respiring (ca. 16%). The second bioreactor study used potato residue to determine if even shorter retention times could be used (range 0.25-2.0 days). Although overall biodegradation deteriorated, the degradation of soluble TOC continued to be ca. 75%. We conclude that if the goal of ALS bioprocessing is maximal degradation of crop residues, including cellulose, then retention times of 10 days or longer will be needed. If the goal is to provide inorganic nutrients with the smallest volume/weight bioreactor possible, then a retention time of 1 day (or less) is sufficient.

Aerobiosis↗

Super-optimal CO2 reduces seed yield but not vegetative growth in wheat.

Although terrestrial atmospheric CO2 levels will not reach 1000 micromoles mol-1 (0.1%) for decades, CO2 levels in growth chambers and greenhouses routinely exceed that concentration. CO2 levels in life support systems in space can exceed 10000 micromoles mol-1(1%). Numerous studies have examined CO2 effects up to 1000 micromoles mol-1, but biochemical measurements indicate that the beneficial effects of CO2 can continue beyond this concentration. We studied the effects of near-optimal (approximately 1200 micromoles mol-1) and super-optimal CO2 levels (2400 micromoles mol-1) on yield of two cultivars of hydroponically grown wheat (Triticum aestivum L.) in 12 trials in growth chambers. Increasing CO2 from sub-optimal to near-optimal (350-1200 micromoles mol-1) increased vegetative growth by 25% and seed yield by 15% in both cultivars. Yield increases were primarily the result of an increased number of heads per square meter. Further elevation of CO2 to 2500 micromoles mol-1 reduced seed yield by 22% (P < 0.001) in cv. Veery-10 and by 15% (P < 0.001) in cv. USU-Apogee. Super-optimal CO2 did not decrease the number of heads per square meter, but reduced seeds per head by 10% and mass per seed by 11%. The toxic effect of CO2 was similar over a range of light levels from half to full sunlight. Subsequent trials revealed that super-optimal CO2 during the interval between 2 wk before and after anthesis mimicked the effect of constant super-optimal CO2. Furthermore, near-optimal CO2 during the same interval mimicked the effect of constant near-optimal CO2. Nutrient concentration of leaves and heads was not affected by CO2. These results suggest that super-optimal CO2 inhibits some process that occurs near the time of seed set resulting in decreased seed set, seed mass, and yield.

Biomass↗

Rotating reverse osmosis: a dynamic model for flux and rejection.

Reverse osmosis (RO) is a compact process for the removal of ionic and organic pollutants from contaminated water. However, flux decline and rejection deterioration due to concentration polarization and membrane fouling hinders the application of RO technology. In this study, a rotating cylindrical RO membrane is theoretically investigated as a novel method to reduce polarization and fouling. A dynamic model based on RO membrane transport incorporating concentration polarization is used to predict the performance of rotating RO system. Operating parameters such as rotational speed and transmembrane pressure play an important role in determining the flux and rejection in rotating RO. For a given geometry, a rotational speed sufficient to generate Taylor vortices in the annulus is essential to maintain high flux as well as high rejection. The flux and rejection were calculated for wide range of operating pressures and rotational speeds.

Filtration↗

Electrochromatographic separation of proteins.

We have developed a modified electrochromatography system which minimizes Joule heating at electric field strengths up to 125 V/cm. A non-linear equilibrium model is described which incorporates electrophoretic mobility, hydrodynamic flow velocity, and an electrically induced concentration polarization at the surface of the stationary phase. This model is able to provide useful estimates of protein retention time and velocity in a column packed with Sephadex gel and subjected to an electric field. A correlation of electrophoretic mobility of peptide and proteins with respect to their charge, molecular mass, and asymmetry enables the selection of solute target molecules for electrochromatographic separations. Good separation of protein mixtures have been obtained.

Chromatography, High Pressure Liquid↗

A microwave-powered sterilizable interface for aseptic access to bioreactors that are vulnerable to microbial contamination.

Novel methods and apparatus that employ the rapid heating characteristics of microwave irradiation to facilitate the aseptic transfer of nutrients, products, and other materials between microbially sensitive systems and the external environment are described. The microwave-sterilizable access port (MSAP) consists of a 600-W magnetron emitting at a frequency of 2.45 GHz, a sterilization chamber with inlet and outlet flow lines, and a specimen transfer interface. Energy is routed to the sterilization chamber via a coaxial transmission line where small quantities of water couple strongly with the incident radiation to produce a superheated vapor phase. The efficiency of energy transfer is enhanced through the use of microwave susceptors within the sterilization chamber. Mating surfaces are thermally sterilized through direct contact with the hot gas. Efficacy has been demonstrated using the thermophile Bacillus stearothermophilus.

Bioreactors↗

Machine vision guided sensor positioning system for leaf temperature assessment.

A sensor positioning system was developed for monitoring plants' well-being using a non-contact sensor. Image processing algorithms were developed to identify a target region on a plant leaf. A novel algorithm to recover view depth was developed by using a camera equipped with a computer-controlled zoom lens. The methodology has improved depth recovery resolution over a conventional monocular imaging technique. An algorithm was also developed to find a maximum enclosed circle on a leaf surface so the conical field-of-view of an infrared temperature sensor could be filled by the target without peripheral noise. The center of the enclosed circle and the estimated depth were used to define the sensor 3-D location for accurate plant temperature measurement.

Algorithms↗

Canopy photosynthesis and transpiration in microgravity: gas exchange measurements aboard Mir.

The SVET Greenhouse on-board the Orbital Station Mir was used to measure canopy photosynthesis and transpiration rates for the first time in space. During the Greenhouse IIB experiment on Mir (June-January 1997), carbon and water vapor fluxes from two wheat (cv. Superdwarf) canopies were measured using the US developed Gas Exchange Measurement System (GEMS). Gas analyzers capable of resolving CO2 concentration differences of 5 micromoles mol-1 against a background of 0.9% CO2, are necessary to measure photosynthetic and respiratory rates on Mir. The ability of the GEMS gas analyzers to measure these CO2 concentration differences was determined during extensive ground calibrations. Similarly, the sensitivity of the analyzers to water vapor was sufficient to accurately measure canopy evapotranspiration. Evapotranspiration, which accounted for over 90% of the water added to the root zone, was estimated using gas exchange and used to estimate substrate moisture content. This paper presents canopy photosynthesis and transpiration data during the peak vegetative phase of development in microgravity.

Air Conditioning↗

Characterizing photosynthesis and transpiration of plant communities in controlled environments.

CO2 and water vapor fluxes of hydroponically grown wheat and soybean canopies were measured continuously in several environments with an open gas exchange system. Canopy CO2 fluxes reflect the photosynthetic efficiency of a plant community, and provide a record of plant growth and health. There were significant diurnal fluctuations in root and shoot CO2 fluxes, and in shoot water vapor fluxes. Canopy stomatal conductance (Gc) to water vapor was calculated from simultaneous measurements of canopy temperature (Tcan) and transpiration rates (Tr). Tr in the dark was substantial, and there were large diurnal fluctuations in both Gc and Tr. Canopy net Photosynthesis (Pnet), Tr, and Gc increased with increasing net radiation. Gc increased with Tr, suggesting that the stomata of plants in controlled environments (CEs) behave differently from field-grown plants. A transpiration model based on measurements of Gc was developed for CEs. The model accurately predicted Tr from a soybean canopy.

Carbon Dioxide↗

Low irradiances affect abscisic acid, indole-3-acidic acid, and cytokinin levels of wheat (Triticum aestivum L.) tissues.

Wheat (Triticum aestivum L.) plants were grown under four irradiance levels: 1,400, 400, 200, and 100 micromol m-2 s-1. Leaves and roots were sampled before, during, and after the boot stage, and levels of abscisic acid (ABA), indole-3-acetic acid (IAA), zeatin, zeatin riboside, dihydrozeatin, dihydrozeatin riboside, isopentenyl adenine, and isopentenyl adenosine were quantified using noncompetitive indirect ELISA systems. Levels of IAA in leaves and roots of plants exposed to 100 micromol m-2 s-1 of irradiance were 0.7 and 2.9 micromol kg-1 dry mass (DM), respectively. These levels were 0.2 and 1.0 micromol kg-1 DM, respectively, when plants were exposed to 1,400 micromol m-2 s-1. Levels of ABA in leaves and roots of plants exposed to 100 micromol m-2 s-1 were 0.65 and 0.55 micromol kg-1 DM, respectively. They were 0.24 micromol kg-1 DM (both leaves and roots) when plants were exposed to 1,400 micromol m-2 s-1. Levels of isopentenyl adenosine in leaves (24.3 nmol kg-1 DM) and roots (29.9 nmol kg-1 DM) were not affected by differences in the irradiance regime. Similar values were obtained in a second experiment. Other cytokinins could not be detected (<10 nmol kg 1 DM) in either experiment with the sample sizes used (150-600 mg DM for roots and shoots, respectively).

Abscisic Acid↗

Response of potatoes to nitrogen concentrations differ with nitrogen forms.

Two separate experiments were conducted to investigate plant growth and mineral composition of potatoes (Solanum tuberosum L.) at varied solution concentrations of nitrate (NO3-) and ammonium (NH4+). Each experiment evaluated five nitrogen (N) concentrations of 0.5, 2, 4, 8, and 12 mM, which were maintained with a non-recirculating nutrient film system in controlled environment. Plants were harvested on day 42 with NO3-; and day 35 with NH4+ after transplanting of tissue culture plantlets, and growth measurements were taken as leaf area, tuber number, and dry weights of different parts. With NO3-, plant growth was greatest and similar at 2, 4, and 8 mM of N whereas with NH4+, plant growth was best only at 2 and 4 mM of N. At 12 mM of N, plants exhibited interveinal ammonium toxicity with NH4+ nutrition, but healthy growth appearance with NO3- nutrition. With either N form, total N concentrations in tissues tended to increase with increased N concentrations, and tissue phosphorus (P) concentrations were reduced at 0.5 and 2 mM of N. Tissue concentrations of calcium (Ca), magnesium (Mg), and sulfur (S) changed only slightly at particular N concentrations, yet changed substantially with different N forms. The data indicate that the optimal ranges of N concentrations in both solution and tissues are wider and higher with NO3- than with NH4+ nutrition, and thus a careful control of NH4+ concentrations is necessary to minimize possible ammonium toxicity to potato plants.

Biomass↗

Yield and seed oil content response of dwarf, rapid-cycling Brassica to nitrogen treatments, planting density, and carbon dioxide enrichment.

Effects of N level (15 to 30 mM), time of N increase (14 to 28 days after planting), and planting density (1163 to 2093 plants/m2) were determined for crop yield responses of dwarf, rapid-cycling brassica (Brassica napus L., CrGC 5-2, Genome: ACaacc). Crops were grown in solid-matrix hydroponic systems and under controlled-environment conditions, including nonsupplemented (ambient) or elevated CO2 concentrations (998 +/- 12 micromoles mol-1). The highest seed yield rate obtained (4.4 g m-2 day-1) occurred with the lowest N level (15 mM) applied at the latest treatment time (day 28). In all trials, CO2 enrichment reduced seed yield rate and harvest index by delaying the onset of flowering and senescence and stimulating vegetative shoot growth. The highest shoot biomass accumulation rate (55.5 g m-2 day-1) occurred with the highest N level (30 mM) applied at the earliest time (day 14). Seed oil content was not significantly affected by CO2 enrichment. Maximum seed oil content (30% to 34%, dry weight basis) was obtained using the lowest N level (15 mM) initiated at the latest treatment time (day 28). In general, an increase in seed oil content was accompanied by a decrease in seed protein. Seed carbohydrate, moisture, and ash contents did not vary significantly in response to experimental treatments. Effects of N level and time of N increase were consistently significant for most crop responses. Planting density was significant only under elevated CO2 conditions.

Agriculture↗