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At least 271 records · Page 15

Comparative Modeling Studies of Boreal Water and Carbon Balance

The coordination of the modeling and field efforts for an Intensive Field Campaign (IFC) may resemble the chicken and egg dilemma. This session's theme advocates that early and proactive involvement by modeling teams can produce a scientific and operational benefit for the IFC and Experiment. This talk will provide some examples and suggestions originating from the NASA funded IFC's of the FIFE First ISLSCP (International Satellite Land Surface Climatology Project) Field Experiment, Oregon Transect Ecosystem Research (OTTER) and predominately Boreal Ecosystem-Atmosphere Study (BOREAS) Experiments. In February 1994 and prior to the final selection of the BOREAS study sites, a group of funded BOREAS investigators agreed to run their models with data for five community types representing the proposed tower flux sites. All participating models were given identical initial values and boundary conditions and driven with identical climate data. The objectives of the intercomparison exercise were: 1) compare simulation results of participating terrestrial, hydrological, and atmospheric models over selected time frames; 2) learn about model behavior and sensitivity to estimated boreal site and vegetation definitions; 3) prioritize BOREAS field data collection efforts supporting modeling studies; 4) identify individual model deficiencies as early as possible. Out of these objectives evolved some important coordination and science issues for the BOREAS Experiment that can be generalized to IFCs and long term archiving of the data. Some problems are acceptable because they are endemic to maintaining fair and open competition prior to the peer review process. Others are logistical and addressable through application of planning, management, and information sciences. This investigator has identified one source of measurement and model incompatibility that is manifest in the IFC scaling approach. Although intuitively obvious, scaling problems are already more formally defined in the Geography literature. An example of the scaling problem will be demonstrated with Vegetation/Ecosystem Mapping and Analysis Project (VEMAP) and OTTER data.

Coughlan, J.↗

XCTK1: A Xenopus C-terminal Kinesin-like Protein

XCTK1 is 97kDa kinesin-like protein homologous to FKIF2 and KIFC3. XCTK1 is present at picomolar levels in eggs, embryos and cultured cells in a soluble high-molecular weight complex that is not associated with membranes. XCKT1 localizes to centrosomes in Xenopus A6 cells. Anti-XCTK1 antibodies also localize to spindle poles when injected into A6 cells or when added to extracts during in vitro spindle assembly reactions. XCTK1 is associated with the center of taxol-induced microtubule asters in extracts. Therefore its localization to poles is dependent on microtubule minus-ends and not on centrosomes per se. Overexpression of XCTK1 leads to centrosome destruction in cultured cells. XCTK1 was tagged at either the N- or C-terminus and transfected into Xenopus A6 cells At low expression levels, XCTK1 associated with centrosomes. At higher levels, the protein localized to insoluble cytoplasmic structures. Gamma-tubulin staining was dramatically decreased from centrosomes or altogether absent. The centrosomal SPJ antigen colocalized with XCTK1-containing structures. Upon nocodozole treatment, microtubules failed to regrow from the centrosomes indicating that overexpression of XCTK1 severely compromises centrosomal function. Current studies are aimed at determining whether XCTK1 interacts directly with centrosomal proteins and to determine the effects of XCTK1 depletion on oocyte maturation and embryogenesis.

Winfree, Seth↗

Heart Research

James Antaki and a group of researchers from the University of Pittsburgh School of Medicine used many elements of the Technology Utilization Program while looking for a way to visualize and track material points within the heart muscle. What they needed were tiny artificial "eggs" containing copper sulfate solution, small enough (about 2 mm in diameter) that they would not injure the heart, and large enough to be seen in Magnetic Resonance Imaging (MRI) images; they also had to be biocompatible and tough enough to withstand the beating of the muscle. The group could not make nor buy sufficient containers. After reading an article on microspheres in NASA Tech Briefs, and a complete set of reports on microencapsulation from the Jet Propulsion Laboratory (JPL), JPL put Antaki in touch with Dr.Taylor Wang of Vanderbilt University who helped construct the myocardial markers. The research is expected to lead to improved understanding of how the heart works and what takes place when it fails.

Source record↗

Fractals and Spatial Methods for Mining Remote Sensing Imagery

The rapid increase in digital remote sensing and GIS data raises a critical problem -- how can such an enormous amount of data be handled and analyzed so that useful information can be derived quickly? Efficient handling and analysis of large spatial data sets is central to environmental research, particularly in global change studies that employ time series. Advances in large-scale environmental monitoring and modeling require not only high-quality data, but also reliable tools to analyze the various types of data. A major difficulty facing geographers and environmental scientists in environmental assessment and monitoring is that spatial analytical tools are not easily accessible. Although many spatial techniques have been described recently in the literature, they are typically presented in an analytical form and are difficult to transform to a numerical algorithm. Moreover, these spatial techniques are not necessarily designed for remote sensing and GIS applications, and research must be conducted to examine their applicability and effectiveness in different types of environmental applications. This poses a chicken-and-egg problem: on one hand we need more research to examine the usability of the newer techniques and tools, yet on the other hand, this type of research is difficult to conduct if the tools to be explored are not accessible. Another problem that is fundamental to environmental research are issues related to spatial scale. The scale issue is especially acute in the context of global change studies because of the need to integrate remote-sensing and other spatial data that are collected at different scales and resolutions. Extrapolation of results across broad spatial scales remains the most difficult problem in global environmental research. There is a need for basic characterization of the effects of scale on image data, and the techniques used to measure these effects must be developed and implemented to allow for a multiple scale assessment of the data before any useful process-oriented modeling involving scale-dependent data can be conducted. Through the support of research grants from NASA, we have developed a software module called ICAMS (Image Characterization And Modeling System) to address the need to develop innovative spatial techniques and make them available to the broader scientific communities. ICAMS provides new spatial techniques, such as fractal analysis, geostatistical functions, and multiscale analysis that are not easily available in commercial GIS/image processing software. By bundling newer spatial methods in a user-friendly software module, researchers can begin to test and experiment with the new spatial analysis methods and they can gauge scale effects using a variety of remote sensing imagery. In the following, we describe briefly the development of ICAMS and present application examples.

Lam, Nina↗

AFM Studies of Salt Concentration Effects on the (110) Surface Structure of Tetragonal Lysozyme Crystals

Previous high resolution AFM studies of the (110) surface of tetragonal chicken egg white lysozyme crystals had shown that only one of two possible molecular surfaces is present, those constituting the completed 43 helices. These suggested that the crystal growth process was by the solution-phase assembly of the growth units, which then attach to the surface. However, the best fit for the imaged surfaces, vs. those predicted based upon the bulk crystallographic coordinates, were obtained when the packing about the 43 helices was "tightened up", while maintaining the underlying crystallographic unit cell spacing. This results in a widening of the gap between adjacent helices, and the top- most layer(s) may no longer be in contact. We postulated that the tightened packing about the helices is a result of the high salt concentrations in the bulk solution, used to crystallize the protein, driving hydrophobic interactions. Once the crystal surface is sufficiently buried by subsequent growth layers the ratio of salt to protein molecules decreases and the helices relax to their bulk crystallographic coordinates. The crystal surface helix structure is thus a reflection of the solution structure, and the tightness of the packing about the 43 helices would be a function of the bulk salt concentration. AFM images of the (110) surface of tetragonal lysozyme crystals grown under low (2%) and high (5%) NaCl concentrations reveal differences in the packing about the 43 helices consistent with the above proposal.

Pusey, Marc Lee↗

Changes in the Molar Ellipticities of HEWL Observed by Circular Dichroism and Quantitated by Time Resolved Fluorescence Anisotropy Under Crystallizing Conditions

Fluid models for simple colloids predict that as the protein concentration is increased, crystallization should occur at some sufficiently high concentration regardless of the strength of attraction. However, empirical measurements do not fully support this assertion. Measurements of the second virial coefficient (B22) indicate that protein crystallization occurs only over a discrete range of solution parameters. Furthermore, observations of a strong correlation between protein solubility and B22, has led to an ongoing debate regarding the relationship between the two. Experimental work in our lab, using Hen Egg White Lysozyme (HEWL), previously revealed that the rotational anisotropy of the protein under crystallizing conditions changes systematically with pH, ionic strength and temperature. These observations are now supported by recent work revealing that small changes in the molar ellipticity also occur systematically with changes in ionic strength and temperature. This work demonstrates that under crystallization conditions, the protein native state is characterized by a conformational heterogeneity that may prove fundamental to the relationship between protein crystallization and protein solubility.

Sumida, John↗

An Alternative Hypothesis for How Microgravity Improves Macromolecular Crystal Quality

There is a substantial body of experimental evidence, from this and other laboratories, that strongly suggests that for many proteins crystal nucleation and growth is by addition of associated species that are preformed by reversible concentration-driven self association processes in the bulk solution. We have developed a self-association model for the crystal nucleation and growth of the protein chicken egg lysozyme. The model accounts for the obtained crystal symmetry, explains the observed surface structures, and shows the importance of the symmetry obtained by self-association in solution to the process as a whole. This model also offers a possible mechanism for fluid flow effects on the growth process and how microgravity may affect it. While a single lysozyme molecule is relatively small an octamer in the 43 helix configuration (the proposed average sized growth unit) would have a M.W. approx. 115,000 and dimensions of 5.6 x 5.6 x 7.6 nm. Direct AFM measurements of growth unit incorporation indicate that units as wide as 11.2 nm and as long as 11.4 nm (a 24-mer) commonly attach to the crystal. AFM results from Weichmann et al. (Ultramicroscopy 86, 159-166, 2001) suggest that associated species of up to 40-mers in size add to the (101) faces. These measurements reflect the sizes of units that both added and desorbed from the crystal surface. The larger and less isotropic the associated species the more likely that it will be oriented to some degree in a flowing boundary layer, even at the low flow velocities measured about macromolecule crystals. On Earth, concentration gradient driven flow will maintain a high interfacial concentration, i.e., a high level (essentially that of the bulk solution) of solute association at the interface and higher growth rate. Higher growth rates mean an increased probability that misaligned growth units are trapped by subsequent growth layers before they can be desorbed and try again, or that the desorbing species is more likely to be smaller than the adsorbing species. In microgravity the extended diffusive boundary layer will lower the interfacial concentration. This results in a net dissociation of aggregated species that diffuse in from the bulk solution, i.e., smaller associated species, which are more likely able to make multiple attempts to correctly bind, yielding higher quality crystals.

Pusey, Marc↗

Early Development of Gravity-Sensing Organs in Microgravity

Most animals have organs that sense gravity. These organs use dense stones (called otoliths or statoconia), which rest on the sensitive hairs of specialized gravity- and motion-sensing cells. The weight of the stones bends the hairs in the direction of gravitational pull. The cells in turn send a coded representation of the gravity or motion stimulus to the central nervous system. Previous experiments, in which the eggs or larvae of a marine mollusk (Aplysia californica, the sea hare) were raised on a centrifuge, demonstrated that the size of the stones (or test mass) was reduced in a graded manner as the gravity field was increased. This suggests that some control mechanism was acting to normalize the weight of the stones. The experiments described here were designed to test the hypothesis that, during their initial development, the mass of the stones is regulated to achieve a desired weight. If this is the case, we would expect a larger-than-normal otolith would develop in animals reared in the weightlessness of space. To test this, freshwater snails and swordtail fish were studied after spaceflight. The snails mated in space, and the stones (statoconia) in their statocysts developed in microgravity. Pre-mated adult female swordtail fish were flown on the Space Shuttle, and the developing larvae were collected after landing. Juvenile fish, where the larval development had taken place on the ground, were also flown. In snails that developed in space, the total volume of statoconia forming the test mass was 50% greater than in size-matched snails reared in functionally identical equipment on the ground. In the swordtail fish, the size of otoliths was compared between ground- and flight-reared larvae of the same size. For later-stage larvae, the growth of the otolith was significantly greater in the flight-reared fish. However, juvenile fish showed no significant difference in otolith size between flight- and ground-reared fish. Thus, it appears that fish and snails reared in space do produce larger-than-normal otoliths (or their analogs), apparently in an attempt to compensate for the reduced weight of the stones in space. The fish data suggest that there is a critical period during which altered gravity can affect the size of the test mass, since the larval, but not the juvenile, fish showed the changes.

Wiederhold, Michael L.↗

Remotely Sensing Larval Population Dynamics of Rice Field Anophelines

The primary objective of both studies was to determine if RS and GIS techniques could be used to distinguish between high and low larval-producing rice fields in California. Results of the first study suggested that early-season green-up and proximity to livestock pastures were positively correlated with high larval abundance. Based on the early-season spectral differences between high and low larval-producing fields, it appeared that canopy development and tillering influenced mosquito habitat quality. At that time, rice fields consisted of a mixture of plants and water, a combination that allowed An. freeborni females to lay eggs in partial sunlight, protected from both predators and wind. This established a population earlier in the season than in other, 'less-green' fields where tillering and plant emergence was too minimal for ovipositioning. The study also indicated the importance of the distance that a mosquito would have to fly in order to take a bloodmeal prior to ovipositing. These associations were fully explored in an expanded study two years later. The second study confirmed the positive relationship between early season canopy development and larval abundance, and also demonstrated the relationship between abundance and distance-to-pasture. The association between greenness (as measured using NDVI), distance-to-pasture, and abundance is illustrated. The second study also indicated the siginificance of the landscape context of rice fields for larval production. Fields that included opportunities for feeding and resting within the flight range of the mosquito had higher abundances than did fields that were in a homogeneous rice area.

Beck, Louisa R.↗

Solution-Phase Processes of Macromolecular Crystallization

We have proposed, for the tetragonal form of chicken egg lysozyme, that solution phase assembly processes are needed to form the growth units for crystal nucleation and growth. The starting point for the self-association process is the monomeric protein, and the final crystallographic symmetry is defined by the initial dimerization interactions of the monomers and subsequent n-mers formed, which in turn are a function of the crystallization conditions. It has been suggested that multimeric proteins generally incorporate the underlying multimers symmetry into the final crystallographic symmetry. We posed the question of what happens to a protein that is known to grow as an n-mer when it is placed in solution conditions where it is monomeric. The trypsin-treated, or cut, form of the protein canavalin (CCAN) has been shown to nucleate and grow crystals as a trimer from neutral to slightly acidic solutions. Under these conditions the solution is composed almost wholly of trimers. The insoluble protein can be readily dissolved by weakly basic solution, which results in a solution that is monomeric. There are three possible outcomes to an attempt at crystallization of the protein under monomeric (high pH) conditions: 1) we will obtain the same crystals as under trimer conditions, but at different protein concentrations governed by the self association equilibria; 2) we will obtain crystals having a different symmetry, based upon a monomeric growth unit; 3) we will not obtain crystals. Obtaining the first result would be indicative that the solution-phase self-association process is critical to the crystal nucleation and growth process. The second result would be less clear, as it may also reflect a pH-dependent shift in the trimer-trimer molecular interactions. The third result, particularly for experiments in the transition pH's between trimeric and monomeric CCAN, would indicate that the monomer does not crystallize, and that solution phase self association is not part of the crystal nucleation and growth path. Results are presented for crystallization experiments of CCAN over the pH 6.8 to 9.6 range.

Pusey, Marc L.↗

The endomembrane requirement for cell surface repair

The capacity to reseal a plasma membrane disruption rapidly is required for cell survival in many physiological environments. Intracellular membrane (endomembrane) is thought to play a central role in the rapid resealing response. We here directly compare the resealing response of a cell that lacks endomembrane, the red blood cell, with that of several nucleated cells possessing an abundant endomembrane compartment. RBC membrane disruptions inflicted by a mode-locked Ti:sapphire laser, even those initially smaller than hemoglobin, failed to reseal rapidly. By contrast, much larger laser-induced disruptions made in sea urchin eggs, fibroblasts, and neurons exhibited rapid, Ca(2+)-dependent resealing. We conclude that rapid resealing is not mediated by simple physiochemical mechanisms; endomembrane is required.

NASA Discipline Cell Biology↗

Space Station Biological Research Project

To meet NASA's objective of using the unique aspects of the space environment to expand fundamental knowledge in the biological sciences, the Space Station Biological Research Project at Ames Research Center is developing, or providing oversight, for two major suites of hardware which will be installed on the International Space Station (ISS). The first, the Gravitational Biology Facility, consists of Habitats to support plants, rodents, cells, aquatic specimens, avian and reptilian eggs, and insects and the Habitat Holding Rack in which to house them at microgravity; the second, the Centrifuge Facility, consists of a 2.5 m diameter centrifuge that will provide acceleration levels between 0.01 g and 2.0 g and a Life Sciences Glovebox. These two facilities will support the conduct of experiments to: 1) investigate the effect of microgravity on living systems; 2) what level of gravity is required to maintain normal form and function, and 3) study the use of artificial gravity as a countermeasure to the deleterious effects of microgravity observed in the crew. Upon completion, the ISS will have three complementary laboratory modules provided by NASA, the European Space Agency and the Japanese space agency, NASDA. Use of all facilities in each of the modules will be available to investigators from participating space agencies. With the advent of the ISS, space-based gravitational biology research will transition from 10-16 day short-duration Space Shuttle flights to 90-day-or-longer ISS increments.

NASA Discipline General Space Life Sciences↗

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.

Non-NASA Center↗

Microtubules as key cytoskeletal elements in cellular transport and shape changes: their expected responses to space environments

Application of reference standard reagents to alternatively depolymerize or stabilize microtubules in a cell that undergoes very regular cytoskeleton-dependent shape changes provides a model system in which some expected components of the environments of spacecraft and space can be tested on Earth for their effects on the cytoskeleton. The fertilized eggs of Ilyanassa obsoleta undergo polar lobe formation by repeated, dramatic, constriction and relaxation of a microfilamentous band localized in the cortical cytoplasm and activated by microtubules.

NASA Discipline Number 93-10↗

Microgravity alters protein phosphorylation changes during initiation of sea urchin sperm motility

European Space Agency (ESA) studies demonstrated that bull sperm swim with higher velocity in microgravity (microG) than at 1 G. Coupling between protein phosphorylation and sperm motility during activation in microG and at 1 G was examined in the ESA Biorack on two space shuttle missions. Immotile sperm were activated to swim (86-90% motility) at launch +20 h by dilution into artificial seawater (ASW). Parallel ground controls were performed 2 h after the flight experiment. Activation after 0, 30, and 60 s was terminated with electrophoresis sample buffer and samples analyzed for phosphoamino acids by Western blotting. Phosphorylation of a 130-kDa phosphothreonine-containing protein (FP130) occurred three to four times faster in microG than at 1 G. A 32-kDa phosphoserine-containing protein was significantly stimulated at 30 s but returned to 1 G control levels at 60 s. The rate of FP130 phosphorylation in microG was attenuated by D2O, suggesting that changes in water properties participate in altering signal transduction. Changes in FP130 phosphorylation triggered by the egg peptide speract were delayed in microG. These results demonstrate that previously observed effects of microG on sperm motility are coupled to changes in phosphorylation of specific flagellar proteins and that early events of sperm activation and fertilization are altered in microG.

NASA Experiment Number 9403057 2/2↗

Blastomeres show differential fate changes in 8-cell Xenopus laevis embryos that are rotated 90 degrees before first cleavage

To study the mechanisms of dorsal axis specification, the alteration in dorsal cell fate of cleavage stage blastomeres in axis-respecified Xenopus laevis embryos was investigated. Fertilized eggs were rotated 90 degrees with the sperm entry point up or down with respect to the gravitational field. At the 8-cell stage, blastomeres were injected with the lineage tracers, Texas Red- or FITC-Dextran Amines. The distribution of the labeled progeny was mapped at the tail-bud stages (stages 35-38) and compared with the fate map of an 8-cell embryo raised in a normal orientation. As in the normal embryos, each blastomere in the rotated embryos has a characteristic and predictable cell fate. After 90 degrees rotation the blastomeres in the 8-cell stage embryo roughly switched their position by 90 degrees, but the fate of the blastomeres did not simply show a 90 degrees switch appropriate for their new location. Four types of fate change were observed: (i) the normal fate of the blastomere is conserved with little change; (ii) the normal fate is completely changed and a new fate is adopted according to the blastomere's new position: (iii) the normal fate is completely changed, but the new fate is not appropriate for its new position; and (4) the blastomere partially changed its fate and the new fate is a combination of its original fate and a fate appropriate to its new location. According to the changed fates, the blastomeres that adopt dorsal fates were identified in rotated embryos. This identification of dorsal blastomeres provides basic important information for further study of dorsal signaling in Xenopus embryos.

NASA Discipline Developmental Biology↗

Development of the otolith organs and semicircular canals in the Japanese red-bellied newt, Cynops pyrrhogaster

The sequence in which the otoliths and semicircular canals and their associated sensory epithelia appear and develop in the newt are described. Three-dimensional reconstruction of serial sections through the otic vesicle of newt embryos from stages 31 through 58 demonstrate the first appearance, relative position and growth of the otoliths. A single otolith is first seen in stage 33 embryos (approximately 9 days old); this splits into separate utricular and saccular otoliths at stage 40 (13 days). The lateral semicircular canal is the first to appear, at stage 41 (14 days). The anterior and posterior canals appear approximately one week later and the vestibular apparatus is essentially fully formed at stage 58 (approximately 5 weeks). The data reported here will serve as ground-based controls for fertilized newt eggs flown on the International Microgravity Laboratory-2 Space Shuttle flight, to investigate the influence of microgravity on the development of the gravity-sensing organs.

NASA Discipline Neuroscience↗

The chronic infusion of nicotine into the developing chick embryo does not alter the density of (-)-[3H]nicotine-binding sites or vestibular function

(-)-Nicotine (1.2 mg/day) or saline was infused into chick embryos (Gallus domesticus) for 10 days beginning 12 h beyond the eight day of incubation (E8 + 12 h). Twelve h beyond the eighteenth day of incubation (E18 + 12 h), the eggs were opened to access the embryos and subcutaneous skull electrodes placed. Short latency vestibular response thresholds and input/output functions were determined to assess neurophysiological consequences of chronic nicotine administration. Samples of serum and extraembryonic (amniotic and albumen) fluid were analyzed by gas chromatography-mass spectrometry to determine the levels of nicotine and its major metabolite, cotinine. The brains were removed and divided into diencephalon and mesencephalon and the density of (-)-[3H]nicotine binding sites in each brain area was measured. Nicotine and cotinine were found in the serum and extraembryonic fluid, but nicotinic receptors were not up-regulated in the brains of animals infused with nicotine in comparison to controls. Vestibular response thresholds also did not differ between nicotine-treated and control animals.

NASA Discipline Neuroscience↗