Effect of microgravity on stress ethylene and carbon dioxide production in sweet clover (Melilotus alba L.).
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Carbon Dioxide↗
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Plasmid acquisition in microgravity.
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Ampicillin Resistance↗
The Fluid Processing Apparatus: from flight hardware to electron micrographs.
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Calcium↗
Physical, chemical, biochemical and biological techniques and processes. Proceedings of the F2.2, F2.3, F2.5, F4.3, F4.4, F4.9, F4.10 and F3.5 Meetings of COSPAR Scientific Commission F which were held during the Thirtieth COSPAR Scientific Assembly, Hamburg, Germany, 11-21 July 1994.
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Animals↗
Scanning electron microscope observations of brine shrimp larvae from space shuttle experiments.
Brine shrimp are encysted as gastrula stage embryos, and may remain dehydrated and encysted for years without compromising their viability. This aspect of brine shrimp biology is desirable for studying development of animals during space shuttle flight, as cysts placed aboard a spacecraft may be rehydrated at the convenience of an astronaut, guaranteeing that subsequent brine shrimp development occurs only on orbit and not on the pad during launch delays. Brine shrimp cysts placed in 5 ml syringes were rehydrated with salt water and hatched during a 9 day space shuttle mission. Subsequent larvae developed to the 8th larval stage in the sealed syringes. We studied the morphogenesis of the brine shrimp larvae and found the larvae from the space shuttle experiments similar in rate of growth and extent of development, to larvae grown in sealed syringes on the ground. Extensive differentiation and development of embryos and larvae can occur in a microgravity environment.
Animals↗
Preliminary observations on the effects of vector-averaged gravity on the embryonic and larval development of the gastropod mollusk, Ilyanassa obsoleta Stimpson.
Fertilized eggs of Ilyanassa obsoleta Stimpson were collected immediately after their deposition in egg capsules. Unopened egg capsules then were affixed to glass slides, and incubated either statically (controls) or on a clinostat (experimentals). After incubation for 9-14 days, hatching occurred sooner and in a higher percentage of clinostated capsules than in controls. Embryos that hatched while undergoing clinostat incubation were abnormal in morphology, whereas other embryos present in non-hatched capsules in the same tubes appeared normal, as did embryos in the control tubes. Although the results are compatible with a conclusion that vector-averaged gravity in the experimental tubes caused the altered development, some other aspects of how the incubations were done may have contributed to the differences between the control and experimental results.
Animals↗
Preliminary observations on the effects of selenate on the development of the embryonic skate, Raja eglanteria.
Morphogenesis of the clearnose skate, Raja eglanteria, was not significantly inhibited as a result of 7 days of exposure to 1-2 mM selenate in the sea water during Days 59-69 of embryonic development (hatching would normally have occurred at 82 +/- 4 days of incubation). Although corneal transparency appeared normal in the eye, preliminary measurements of the thickness of Bowman's layer of the cornea suggested that it was significantly thinner in the corneas of embryos exposed to 1-2 mM selenate. Selenate is an ion reported to inhibit sulfation of glycosaminoglycans in connective tissue.
Animals↗
The use of centrifugation to study early Drosophila embryogenesis.
By the end of 10th nuclear cycle, the somatic nuclei of the Drosophila embryo have migrated to the periphery of the egg. Centrifugation of embryos did not result in the displacement of these nuclei, since cytoskeletal elements anchor them to the cortex. But, mild centrifugal forces displace the centrally located, nascent yolk nuclei. If this increased sensitivity to hypergravity occurs before the beginning of nuclear differentiation during cycle 8, when the nascent yolk and somatic nuclei physically separate, then it would mark the earliest functional difference between these two lineages.
Animals↗
A role for the diazotrophic cyanobacterium, Cyanothece sp. strain ATCC 51142, in nitrogen cycling for CELSS applications.
Simple calculations show that fixed nitrogen regeneration in a CELSS may not be as efficient as stowage and resupply of fixed nitrogen compounds. However, fixed nitrogen regeneration may be important for the sustainability and safety of a deployed CELSS. Cyanothece sp. strain ATCC 51142, a unicellular, aerobic, diazotrophic cyanobacterium, with high growth rates and a robust metabolism, is a reasonable candidate organism for a biological, fixed nitrogen regeneration system. In addition, Cyanothece sp. cultures may be used to balance gas exchange ratio imparities between plants and humans. The regeneration of fixed nitrogen compounds by cyanobacterial cultures was examined in the context of a broad computer model/simulation (called CELSS-3D). When cyanothece sp. cultures were used to balance gas exchange imparities, the biomass harvested could supply as much as half of the total fixed nitrogen needed for plant biomass production.
Biomass↗
Effect of microgravity and hypergravity on embryo axis alignment during postencystment embryogenesis in Artemia franciscana (Anostraca).
Cysts of brine shrimp attached with a liquid adhesive to 12-mm diameter glass coverslips in a syringe-type fluid processing apparatus were flown aboard the NASA space shuttle Discovery, flight STS-60, from 3-11 February 1994, and were allowed to undergo postencystment embryogenesis and to hatch in microgravity. The shuttle flight and the ground-based control coverslips with attached cysts were parallel to the earth's surface during incubation in salt water. Based on the position of the cyst shell crack in the attached cyst population, the ground-control nauplii emerged mostly upward. On the shuttle in microgravity, although our method of detection of orientation would not reveal emergence toward the coverslip, the ratio of the position of the cyst shell crack in the population after hatching best fit the predicted values of a random direction for nauplii emergence. Centrifugation on earth was then used to create hypergravity forces of up to 73 g during postencystment embryogenesis and hatching. The upward orientation of emerging nauplii showed a high degree of correlation (r(2) =98.8%) with a linear relationship to the log of g, with 78.2% of the total hatching upward at 1 g and 91.0% hatching upward at 73 g.
Animals↗
Identifying yield-optimizing environments for two cowpea breeding lines by manipulating photoperiod and harvest scenario.
Photoperiod and harvest scenario of cowpea (Vigna unguiculata L. Walp) canopies were manipulated to optimize productivity for use in future controlled ecological life-support systems. Productivity was measured by edible yield rate (EYR:g m-2 day-1), shoot harvest index (SHI: g edible biomass [g total shoot dry weight]), and yield-efficiency rate (YER:g edible biomass m-2 day-1 per[g nonedible shoot dry weight]). Breeding lines 'IT84S-2246' (S-2246) and "IT82D-889' (D-889) were grown in a greenhouse under 8-, 12-, or 24-h photoperiods. S-2246 was short-day and D-889 was day-neutral for flowering. Under each photoperiod, cowpeas were harvested either for leaves only, seeds only, or leaves plus seeds (mixed harvest). Photoperiod did not affect EYR of either breeding line for any harvest scenario tested. Averaged over both breeding lines, seed harvest gave the highest EYR at 6.7 g m-2 day-1. The highest SHI (65%) and YER (94 mg m-2 day-1 g-1) were achieved for leaf-only harvest of D-889 under an 8-h photoperiod. For leaf-only harvest of S-2246, both SHI and YER increased with increasing photoperiod, but declined for seed-only and mixed harvests. However, photoperiod had no effect on SHI or YER for D-889 for any harvest scenario. A second experiment utilized the short-day cowpea breeding line 'IT89KD-288' (D-288) and the day-neutral breeding line 'IT87D-941-1' (D-941) to compare yield parameters using photoperiod extension under differing lamp types. This experiment confirmed the photoperiod responses of D-889 and S-2246 to a mixed-harvest scenario and indicated that daylength extension with higher irradiance from high pressure sodium lamps further suppressed EYR, SHI, and YER of the short-day breeding line D-288.
Agriculture↗
Varying plant density and harvest time to optimize cowpea leaf yield and nutrient content.
Plant density and harvest time were manipulated to optimize vegetative (foliar) productivity of cowpea [Vigna unguiculata (L.) Walp.] canopies for future dietary use in controlled ecological life-support systems as vegetables or salad greens. Productivity was measured as total shoot and edible dry weights (DW), edible yield rate [(EYR) grams DW per square meter per day], shoot harvest index [(SHI) grams DW per edible gram DW total shoot], and yield-efficiency rate [(YER) grams DW edible per square meter per day per grams DW nonedible]. Cowpeas were grown in a greenhouse for leaf-only harvest at 14, 28, 42, 56, 84, or 99 plants/m2 and were harvested 20, 30, 40, or 50 days after planting (DAP). Shoot and edible dry weights increased as plant density and time to harvest increased. A maximum of 1189 g shoot DW/m2 and 594 g edible DW/m2 were achieved at an estimated plant density of 85 plants/m2 and harvest 50 DAP. EYR also increased as plant density and time to harvest increased. An EYR of 11 g m-2 day-1 was predicted to occur at 86 plants/m2 and harvest 50 DAP. SHI and YER were not affected by plant density. However, the highest values of SHI (64%) and YER (1.3 g m-2 day-1 g-1) were attained when cowpeas were harvested 20 DAP. The average fat and ash contents [dry-weight basis (dwb)] of harvested leaves remained constant regardless of harvest time. Average protein content increased from 25% DW at 30 DAP to 45% DW at 50 DAP. Carbohydrate content declined from 50% DW at 30 DAP to 45% DW at 50 DAP. Total dietary fiber content (dwb) of the leaves increased from 19% to 26% as time to harvest increased from 20 to 50 days.
Biomass↗
Clusters of DNA damage induced by ionizing radiation: formation of short DNA fragments. II. Experimental detection.
The basic 30-nm chromatin fiber in the mammalian cell consists of an unknown (possibly helical) arrangement of nucleosomes, with about 1.2 kb of DNA per 10-nm length of fiber. Track-structure considerations suggest that interactions of single delta rays or high-LET particles with the chromatin fiber might result in the formation of multiple lesions spread over a few kilobases of DNA (see the accompanying paper: W.R. Holley and A. Chatterjee, Radiat. Res. 145, 188-199, 1996). In particular, multiple DNA double-strand breaks and single-strand breaks may form. To test this experimentally, primary human fibroblasts were labeled with [3H]thymidine and exposed at 0 degrees C to X rays or accelerated nitrogen or iron ions in the LET range of 97-440 keV/microns. DNA was isolated inside agarose plugs and subjected to agarose gel electrophoresis under conditions that allowed good separation of 0.1-2 kb size DNA. The bulk of DNA remained in the well or migrated only a small distance into the gel. It was found that DNA fragments in the expected size range were formed linearly with dose with an efficiency that increased with LET. A comparison of the yield of such fragments with the yield of total DNA double-strand breaks suggests that for the high-LET ions a substantial proportion (20-90%) of DNA double-strand breaks are accompanied within 0.1-2 kb by at least one additional DNA double-strand break. It is shown that these results are in good agreement with theoretical calculations based on treating the 30-nm chromatin fiber as the target for ionizing particles. Theoretical considerations also predict that the clusters will contain numerous single-strand breaks and base damages. It is proposed that such clusters be designated "regionally multiply damaged sites." Postirradiation incubation at 37 degrees C resulted in a decline in the number of short DNA fragments, suggesting a repair activity. The biological significance of regionally multiply damaged sites is presently unknown.
Cells, Cultured↗
A mutation in protein phosphatase 2A regulatory subunit A affects auxin transport in Arabidopsis.
The phytohormone auxin controls processes such as cell elongation, root hair development and root branching. Tropisms, growth curvatures triggered by gravity, light and touch, are also auxin-mediated responses. Auxin is synthesized in the shoot apex and transported through the stem, but the molecular mechanism of auxin transport is not well understood. Naphthylphthalamic acid (NPA) and other inhibitors of auxin transport block tropic curvature responses and inhibit root and shoot elongation. We have isolated a novel Arabidopsis thaliana mutant designated roots curl in NPA (rcn1). Mutant seedlings exhibit altered responses to NPA in root curling and hypocotyl elongation. Auxin efflux in mutant seedlings displays increased sensitivity to NPA. The rcn1 mutation was transferred-DNA (T-DNA) tagged and sequences flanking the T-DNA insert were cloned. Analysis of the RCN1 cDNA reveals that the T-DNA insertion disrupts a gene for the regulatory A subunit of protein phosphatase 2A (PP2A-A). The RCN1 gene rescues the rcn1 mutant phenotype and also complements the temperature-sensitive phenotype of the Saccharomyces cerevisiae PP2A-A mutation, tpd3-1. These data implicate protein phosphatase 2A in the regulation of auxin transport in Arabidopsis.
Amino Acid Sequence↗
Composition of the carbohydrate granules of the cyanobacterium, Cyanothece sp. strain ATCC 51142.
Cyanothece sp. strain ATCC 51142 is an aerobic, unicellular, diazotrophic cyanobacterium that temporally separates O2-sensitive N2 fixation from oxygenic photosynthesis. The energy and reducing power needed for N2 fixation appears to be generated by an active respiratory apparatus that utilizes the contents of large interthylakoidal carbohydrate granules. We report here on the carbohydrate and protein composition of the granules of Cyanothece sp. strain ATCC 51142. The carbohydrate component is a glucose homopolymer with branches every nine residues and is chemically identical to glycogen. Granule-associated protein fractions showed temporal changes in the number of proteins and their abundance during the metabolic oscillations observed under diazotrophic conditions. There also were temporal changes in the protein pattern of the granule-depleted supernatant fractions from diazotrophic cultures. None of the granule-associated proteins crossreacted with antisera directed against several glycogen-metabolizing enzymes or nitrogenase, although these proteins were tentatively identified in supernatant fractions. It is suggested that the granule-associated proteins are structural proteins required to maintain a complex granule architecture.
1,4-alpha-Glucan Branching Enzyme↗
Creating webservers on the Internet to advance CELSS research.
Effective integration of CELSS design concepts depends on the transfer of research results from studies performed in several disciplines and at many locations to all members of the CELSS community. Some of these appear in sources outside the mainstream scientific journals and are often difficult to acquire. The Internet offers a way to make these research results more accessible. The NSCORT in Bioregenerative Life Support at Purdue University has created a webserver on the Internet that provides information about the Purdue NSCORT, the NSCORT program, and CELSS design and development. Other CELSS-related webservers are needed, and a tutorial on webserver development is presented here. The Purdue NSCORT webserver is discussed in relation to other CELSS-related Internet projects. Future applications of the Internet in promoting CELSS research also are discussed.
Computer Communication Networks↗
Four educational programs in Space Life Sciences.
Four different educational programs impacting Space Life Sciences are described: the NASA/USRA Advanced Design Program, the NASA Specialized Center of Research and Training (NSCORT) Program, the Centers for the Commercial Development of Space (CCDS) Program, and the NASA Graduate Research Fellow Program. Each program makes somewhat different demands on the students engaged in them. Each program, at the University of Colorado, involves Space Life Sciences training. While the Graduate Student Research Fellow and NSCORT Programs are discipline oriented, the Advanced Design and CCDS Programs are focused on design, technologies and applications. Clearly, the "training paradigms" differ for these educational endeavors. But, these paradigms can be made to mutually facilitate enthusiasm and motivation. Discipline-oriented academic programs, ideally, must be flexible enough to accommodate the emergent cross-disciplinary needs of Space Life Sciences students. Models for such flexibility and resultant student performance levels are discussed based upon actual academic and professional records.
Biological Science Disciplines↗
Physical and biological studies with protons and HZE particles in a NASA supported research center in radiation health.
NASA has established and supports a specialized center for research and training (NSCORT) to specifically address the potential deleterious effects of HZE particles on human health. The NSCORT in radiation health is a joint effort between Lawrence Berkeley National Laboratory (LBNL) and Colorado State University (CSU). The overall scope of research encompasses a broad range of subjects from microdosimetric studies to cellular and tissue responses to initial damage produced by highly energetic protons and heavy charged particles of the type found in galactic cosmic rays (GCR) spectrum. The objectives of the microdosimetry studies are to determine the response of Tissue Equivalent Proportional Counter (TEPC) to cosmic rays using ground based accelerators. This includes evaluation of energy loss due to the escape of high-energy delta rays and increased energy deposition due to the enhanced delta ray production in the wall of the detector. In this report major results are presented for 56Fe at 1000, 740, 600 and 400 MeV/nucleon. An assessment of DNA repair and early development of related chromosomal changes is extremely important to our overall understanding of enhanced biological effectiveness of high LET particle radiation. Results are presented with respect to the fidelity of the rejoining of double strand breaks and the implications of misrejoining. The relationship between molecular and cytogenetic measurements is presented by studying damage processing in highly heterochromatic supernumerary (correction of sypernumerary) X chromosomes and the active X-chromosome. One of the important consequences of cell's inability to handle DNA damage can be evaluated through mutation studies. Part of our goal is the assessment of potential radioprotectors to reduce the mutation yield following HZE exposures, and some promising results are presented on one compound. A second goal is the integration of DNA repair and mutation studies. Results are presented on a direct comparison of initial double strand breaks induction, the time course and fidelity of double strand break rejoining, cell killing and mutation induction in the same human model system. In order to understand the carcinogenic potential of protons and HZE particles, the role of damaged microenvironment in this process must be understood. In this project it has been postulated that radiation affects the microenvironment, which then modifies cell interactions in a manner conducive to neoplastic progression. Both TGF-beta and FGF-2 are important components of microenvironment. A recent result on the assessment of the role of FGF-2 and its cross-talk with TGF-beta as a function of radiation quality is presented. Theoretical modeling has so far played a central role in analyzing and integrating experimental data on repair and mutation studies and predicting new phenomena. The integrated NSCORT program also provides a broad training experience for students and postdoctoral fellows in space radiation health.
Animals↗