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Formation and Control of Fluidic Species

This invention generally relates to systems and methods for the formation and/or control of fluidic species, and articles produced by such systems and methods. In some cases, the invention involves unique fluid channels, systems, controls, and/or restrictions, and combinations thereof. In certain embodiments, the invention allows fluidic streams (which can be continuous or discontinuous, i.e., droplets) to be formed and/or combined, at a variety of scales, including microfluidic scales. In one set of embodiments, a fluidic stream may be produced from a channel, where a cross-sectional dimension of the fluidic stream is smaller than that of the channel, for example, through the use of structural elements, other fluids, and/or applied external fields, etc. In some cases, a Taylor cone may be produced. In another set of embodiments, a fluidic stream may be manipulated in some fashion, for example, to create tubes (which may be hollow or solid), droplets, nested tubes or droplets, arrays of tubes or droplets, meshes of tubes, etc. In some cases, droplets produced using certain embodiments of the invention may be charged or substantially charged, which may allow their further manipulation, for instance, using applied external fields. Non-limiting examples of such manipulations include producing charged droplets, coalescing droplets (especially at the microscale), synchronizing droplet formation, aligning molecules within the droplet, etc. In some cases, the droplets and/or the fluidic streams may include colloids, cells, therapeutic agents, and the like.

Link, Darren Roy↗

Parameterizing Grid-Averaged Longwave Fluxes for Inhomogeneous Marine Boundary Layer Clouds

This paper examines the relative impacts on grid-averaged longwave flux transmittance (emittance) for Marine Boundary Layer (MBL) cloud fields arising from horizontal variability of optical depth tau and cloud sides, First, using fields of Landsat-inferred tau and a Monte Carlo photon transport algorithm, it is demonstrated that mean all-sky transmittances for 3D variable MBL clouds can be computed accurately by the conventional method of linearly weighting clear and cloudy transmittances by their respective sky fractions. Then, the approximations of decoupling cloud and radiative properties and assuming independent columns are shown to be adequate for computation of mean flux transmittance. Since real clouds have nonzero geometric thicknesses, cloud fractions A'(sub c) presented to isotropic beams usually exceed the more familiar vertically projected cloud fractions A(sub c). It is shown, however, that when A(sub c)less than or equal to 0.9, biases for all-sky transmittance stemming from use of A(sub c) as opposed to A'(sub c) are roughly 2-5 times smaller than, and opposite in sign to, biases due to neglect of horizontal variability of tau. By neglecting variable tau, all-sky transmittances are underestimated often by more than 0.1 for A(sub c) near 0.75 and this translates into relative errors that can exceed 40% (corresponding errors for all-sky emittance are about 20% for most values of A(sub c). Thus, priority should be given to development of General Circulation Model (GCM) parameterizations that account for the effects of horizontal variations in unresolved tau, effects of cloud sides are of secondary importance. On this note, an efficient stochastic model for computing grid-averaged cloudy-sky flux transmittances is furnished that assumes that distributions of tau, for regions comparable in size to GCM grid cells, can be described adequately by gamma distribution functions. While the plane-parallel, homogeneous model underestimates cloud transmittance by about an order of magnitude when 3D variable cloud transmittances are less than or equal to 0.2 and by approx. 20% to 100% otherwise, the stochastic model reduces these biases often by more than 80%.

Barker, Howard W.↗

Integrated Computational-Experimental Development of Lithium-Air Batteries for Electric Aircraft

The primary obstacle to enable NASA's vision of Green Aviation is the extraordinary energy storage requirements for electric aircraft. Significant advances in high energy, rechargeable, safe batteries are required to enable electric aviation. Boeing's SUGAR and NASA studies have identified 400 Wh/kg as the threshold energy density for general aviation and 750 Wh/kg for commercial regional air service. State of the Art Lithium Ion Battery (LIB) technology currently has a density of 200 Wh/kg and is expected to plateau at 300 Wh/kg due to fundamental chemistry limitations making it unsuitable for future electric aircraft. Additional demanding requirements include high power, rechargeability, and high safety. Such battery technology does not currently exist. The recent considerable activity in battery research (DOE, Tesla Gigafactory, etc) overwhelmingly has been geared towards reducing cost and improving safety of LIB technology in order to promote the adoption of electric automobiles; and thus it is expected to have little impact on electric aviation development. New battery materials will be needed for the "Beyond Li Ion" (BLI) technologies required for high energy, safe electric aviation. Li-Air batteries have the highest known theoretical energy density (3400 Wh/kg) and therefore and if realized promises to transform the global transportation system. These high energy batteries have the potential to meet the energy storage challenges of current and future NASA aeronautics and space missions in addition to many terrestrial transportation applications as well. However, this technology requires significant components development and integration, as it is currently unable to achieve aircraft requirements. The objective of this project is to leverage modern computational materials methods combined with battery multiphysics tools to develop radically advanced compatible cathode and electrolyte materials, build several Li-Air cells, and flight-demonstrate the corresponding Li-Air battery packs. A significant problem for current Lithium-Air batteries is large scale decomposition of the battery electrolyte during operation leading to battery failure after a handful of charge/discharge cycles. Therefore, development of large scale, ultra-high energy, rechargeable, and safe Lithium-Air batteries require highly stable electrolytes that are resistant to decomposition under operating conditions. A NASA-based cross-organizational "dream team" of high-powered experts combined integrated supercomputer modeling, fundamental chemistry analysis, advanced material science, and battery cell development to tackle this very challenging, multidisciplinary problem. The ultimate goal for the team is to develop an integrated experimental/computational infrastructure to produce a reliable predictive capability for the selection of optimal components, their fabrication parameters, and "design rules" of novel cell components for advanced ultra-high energy batteries that can meet energy storage challenges of NASA missions and many terrestrial transportation applications.

Li-air battery↗

Expression of Enzymes that Metabolize Medications

INTRODUCTION: Increased exposure to radiation is one physiological stressor associated with spaceflight and it is feasible to conduct ground experiments using known radiation exposures. The health of the liver, especially the activity rate of its metabolic enzymes, determines the concentration of circulating drugs as well as the duration of their efficacy. While radiation is known to alter normal physiological function, how radiation affects liver metabolism of administered medications is unclear. Crew health could be affected if the actions of medications used in spaceflight deviated from expectations formed during terrestrial medication use. This study is an effort to identify liver metabolic enzymes whose expression is altered by spaceflight or by radiation exposures that mimic features of the spaceflight environment. METHODS: Using procedures approved by the Animal Care and Use Committee, mice were exposed to either 137Cs (controls, 50 mGy, 6Gy, or 50 mGy + 6Gy separated by 24 hours) or 13 days of spaceflight on STS 135. Animals were anesthetized and sacrificed at several time points (4 hours, 24 hours or 7 days) after their last radiation exposure, or within 6 hours of return to Earth for the STS 135 animals. Livers were removed immediately and flash-frozen in liquid nitrogen. Tissue was homogenized, RNA extracted, purified and quality-tested. Complementary DNA was prepared from high-quality RNA samples, and used in RT-qPCR experiments to determine relative expression of a wide variety of genes involved in general metabolism and drug metabolism. RESULTS: Results of the ground radiation exposure experiments indicated ~65 genes of the 190 tested were significantly affected by at least one of the radiation doses. Many of the affected genes are involved in the metabolism of drugs with hydrophobic or steroid-like structures, maintenance of redox homeostasis and repair of DNA damage. Most affected genes returned to near control expression levels by 7 days post-treatment. Not all recovered completely by the final time point tested: with 6 Gy exposure, metallothionein expression was 132-fold more than control at the 4 hr time point, and fell at each later time point (11-fold at 24 hrs, and 8-fold at 7 days). In contrast, there were other genes whose expression was altered and remained relatively constant through the 7 day period we tested. One examples is Cyp17a1, which showed a 4-fold elevation at 4 hrs after exposure and remained constant for 7 days after the last treatment. Spaceflight samples evaluated with similar methods and comparisons will be made between the radiation-treated groups and the spaceflight samples. CONCLUSION It seems likely that radiation exposure triggers homeostatic mechanisms, which could include alterations of gene expression. Better understanding of these pathways could aid in optimizing medications doses given to crewmembers who require treatment and eventually, to development of new countermeasures to ameliorate or prevent radiation-induced damage to cells and tissues.

Wotring, V. E.↗

Neonatal rat heart cells cultured in simulated microgravity

In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit gravity constrain cells to propagate, differentiate, and interact in a two-dimensional (2D) plane. Neonatal rat cardiac cells in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by nonmyocyte cell types. Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic microgravity. The NASA-designed High-Aspect Ratio Vessel (HARV) bioreactors provide a low shear environment that allows cells to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited typical 2D organization. Cells cultured in HARVs adhered to microcarrier beads, the beads aggregated into defined clusters containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARVs using alternative support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV and control cultures were similar; however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel for the generation of tissuelike organization of cardiac cells in vitro.

Non-NASA Center↗

Neonatal rat heart cells cultured in simulated microgravity

In vitro characteristics of cardiac cells cultured in simulated microgravity are reported. Tissue culture methods performed at unit gravity constrain cells to propagate, differentiate, and interact in a two dimensional (2D) plane. Neonatal rat cardiac cells in 2D culture organize predominantly as bundles of cardiomyocytes with the intervening areas filled by non-myocyte cell types. Such cardiac cell cultures respond predictably to the addition of exogenous compounds, and in many ways they represent an excellent in vitro model system. The gravity-induced 2D organization of the cells, however, does not accurately reflect the distribution of cells in the intact tissue. We have begun characterizations of a three-dimensional (3D) culturing system designed to mimic microgravity. The NASA designed High-Aspect-Ratio-Vessel (HARV) bioreactors provide a low shear environment which allows cells to be cultured in static suspension. HARV-3D cultures were prepared on microcarrier beads and compared to control-2D cultures using a combination of microscopic and biochemical techniques. Both systems were uniformly inoculated and medium exchanged at standard intervals. Cells in control cultures adhered to the polystyrene surface of the tissue culture dishes and exhibited typical 2D organization. Cells in cultured in HARV's adhered to microcarrier beads, the beads aggregated into defined clusters containing 8 to 15 beads per cluster, and the clusters exhibited distinct 3D layers: myocytes and fibroblasts appeared attached to the surfaces of beads and were overlaid by an outer cell type. In addition, cultures prepared in HARV's using alternative support matrices also displayed morphological formations not seen in control cultures. Generally, the cells prepared in HARV and control cultures were similar, however, the dramatic alterations in 3D organization recommend the HARV as an ideal vessel for the generation of tissue-like organizations of cardiac cells in simulated microgravity.

Akins, Robert E.↗

Countermeasures to microgravity

Biological systems ranging from the most simple to the most complex generally survive exposure to microgravity. Changes in many characteristics of biological systems are well documented as a consequence of space flight. Attempts to devise countermeasures to microgravity may have direct pragmatic consequences for crew protection and may provide additional insights into the nature of microgravity influences on biological systems. Some of the most well documented changes occur in humans who have experienced space flight. Changes appear to be transient. Space adaption syndrome occurs relatively briefly whereas bone deterioration may require months of postflight time for restoration. It seems critical to recognize that these changes and others may derive from rather passive, active or even reactive changes in the biological systems that are hosts to them. For example, hydrostatic fluid redistributions may be quite passive occurrences that are realized through extensive fluid channels. Changes occur in cell metabolism because of fluid, nutrient and gas redistributions. Equally important are the misconstrued messages likely to be carried by fluid redistributions. These reactive events can trigger, for example, loss of fluids and electrolytes through altered kidney function. Each of these considerations must be evaluated in regard to the biological site affected. Countermeasures to the vast range of biological changes and sites are difficult to envision. The most obvious countermeasure is the restoration of gravity-like influences. Some options are discussed. Recent work has focussed on the use of magnetic fields. Pulsed electromagnetic fields (PEMF) are shown to alleviate bone deterioration produced in rodents exposed to tail suspension. Methods of PEMF exposure are consistent with human use in space. Related methods may provide muscular and neural benefits.

Luttges, Marvin W.↗

Application of a Threshold Method to the TRMM Radar for the Estimation of Space-Time Rain Rate Statistics

One of the TRMM radar products of interest is the monthly-averaged rain rates over 5 x 5 degree cells. Clearly, the most directly way of calculating these and similar statistics is to compute them from the individual estimates made over the instantaneous field of view of the Instrument (4.3 km horizontal resolution). An alternative approach is the use of a threshold method. It has been established that over sufficiently large regions the fractional area above a rain rate threshold and the area-average rain rate are well correlated for particular choices of the threshold [e.g., Kedem et al., 19901]. A straightforward application of this method to the TRMM data would consist of the conversion of the individual reflectivity factors to rain rates followed by a calculation of the fraction of these that exceed a particular threshold. Previous results indicate that for thresholds near or at 5 mm/h, the correlation between this fractional area and the area-average rain rate is high. There are several drawbacks to this approach, however. At the TRMM radar frequency of 13.8 GHz the signal suffers attenuation so that the negative bias of the high resolution rain rate estimates will increase as the path attenuation increases. To establish a quantitative relationship between fractional area and area-average rain rate, an independent means of calculating the area-average rain rate is needed such as an array of rain gauges. This type of calibration procedure, however, is difficult for a spaceborne radar such as TRMM. To estimate a statistic other than the mean of the distribution requires, in general, a different choice of threshold and a different set of tuning parameters.

Meneghini, Robert↗

Countering Solutal Buoyant Convection with High Magnetic Fields

An important component in biotechnology, particularly in the area of protein engineering and rational drug design is the knowledge of the precise three-dimensional molecular structure of proteins. The quality of structural information obtained from X-ray diffraction methods is directly dependent on the degree of perfection of the protein crystals. As a consequence, the growth of high quality macromolecular crystals for diffraction analyses has been the central focus for biochemist, biologists, and bioengineers. Macromolecular crystals are obtained from solutions that contain the crystallizing species in equilibrium with higher aggregates, ions, precipitant, other possible phases of the protein, foreign particles, the walls of the container, and a likely host of other impurities. By changing transport modes in general, i.e., reduction of convection and sedimentation, as is achieved in microgravity, we have been able to dramatically effect the movement and distribution of macromolecules in the fluid, and thus their transport, formation of crystal nuclei, and adsorption to the crystal surface. While a limited number of high quality crystals from space flights have been obtained, as the recent National Research Council (NRC) review of the NASA microgravity crystallization program pointed out, the scientific approach and research in crystallization of proteins has been mainly empirical yielding inconclusive results. We postulate that we can reduce convection in ground-based experiments and we can understand the different aspects of convection control through the use of strong magnetic fields and field gradients. We postulate that limited convection in a magnetic field will provide the environment for the growth of high quality crystals. The approach exploits the variation of fluid magnetic susceptibility with concentration for this purpose and the convective damping is realized by appropriately positioning the crystal growth cell so that the magnetic susceptibility force counteracts terrestrial gravity. The general objective is to test the hypothesis of convective control using a strong magnetic field and magnetic field gradient and to understand the nature of the various forces that come into play. Specifically we aim to delineate causative factors and to quantify them through experiments, analysis and numerical modeling. The paper will report on the current status of the investigation and discuss results from the experimental and modeling efforts.

Ramachandran, N.↗

Using Strong Magnetic Fields to Control Solutal Convection

An important component in biotechnology, particularly in the area of protein engineering and rational drug design is the knowledge of the precise three-dimensional molecular structure of proteins. The quality of structural information obtained from X-ray diffraction methods is directly dependent on the degree of perfection of the protein crystals. As a consequence, the growth of high quality macromolecular crystals for diffraction analyses has been the central focus for biochemists, biologists, and bioengineers. Macromolecular crystals are obtained from solutions that contain the crystallizing species in equilibrium with higher aggregates, ions, precipitants, other possible phases of the protein, foreign particles, the walls of the container, and a likely host of other impurities. By changing transport modes in general, i.e., reduction of convection and sedimentation, as is achieved in microgravity , we have been able to dramatically affect the movement and distribution of macromolecules in the fluid, and thus their transport, formation of crystal nuclei, and adsorption to the crystal surface. While a limited number of high quality crystals from space flights have been obtained, as the recent National Research Council (NRC) review of the NASA microgravity crystallization program pointed out, the scientific approach and research in crystallization of proteins has been mainly empirical yielding inconclusive results. We postulate that we can reduce convection in ground-based experiments and we can understand the different aspects of convection control through the use of strong magnetic fields and field gradients. We postulate that limited convection in a magnetic field will provide the environment for the growth of high quality crystals. The approach exploits the variation of fluid magnetic susceptibility with concentration for this purpose and the convective damping is realized by appropriately positioning the crystal growth cell so that the magnetic susceptibility force counteracts terrestrial gravity. The general objective is to test the hypothesis of convective control using a strong magnetic field and magnetic field gradient and to understand the nature of the various forces that come into play. Specifically we aim to delineate causative factors and to quantify them through experiments, analysis and numerical modeling. The paper will report on the experimental results using paramagnetic salts and solutions in magnetic fields and compare them to analytical predictions.

Ramachandran, N.↗

Magnetic Control of Convection during Protein Crystallization

An important component in biotechnology, particularly in the area of protein engineering and rational drug design is the knowledge of the precise three-dimensional molecular structure of proteins. The quality of structural information obtained from X-ray diffraction methods is directly dependent on the degree of perfection of the protein crystals. As a consequence, the growth of high quality macromolecular Crystals for diffraction analyses has been the central focus for bio-chemists, biologists, and bioengineers. Macromolecular crystals are obtained from solutions that contain the crystallizing species in equilibrium with higher aggregates, ions, precipitants, other possible phases of the protein, foreign particles, the walls of container, and a likely host of other impurities. By changing transport modes in general, i.e., reduction of convection and Sedimentation as is achieved in "microgravity", we have been able to dramatically affect the movement and distribution of macromolecules in the fluid, and thus their transport, f o d o n of crystal nuclei, and adsorption to the crystal surface. While a limited number of high quality crystals from space flights have been obtained, as the recent National Research Council (NRC) review of the NASA microgravity crystallization program pointed out, the scientific approach and research in crystallization of proteins has been mainly empirical yielding inconclusive results. We postulate that we can reduce convection in ground-based experiments and we can understand the different aspects of convection control through the use of strong magnetic fields and field gradients. We postulate that limited convection in a magnetic field will provide the environment for the growth of high quality crystals. The approach exploits the variation of fluid magnetic susceptibility with counteracts on for this purpose and the convective damping is realized by appropriately positioning the crystal growth cell so that the magnetic susceptibility force counteract terrestrial gravity. The genera1 objective is to test the hypothesis of convective control using a strong magnetic field and magnetic field gradient and to understand the nature of the various forces that come into play. Specifically we aim to delineate causative factors and to quantify them through experiments, analysis and numerical modeling. The paper will report on the experimental results using paramagentic salts and solutions in magnetic fields and compare them to analyticalpr~ctions.

Ramachandran, N.↗

Spurious Behavior of Shock-Capturing Methods: Problems Containing Stiff Source Terms and Discontinuities

The goal of this paper is to relate numerical dissipations that are inherited in high order shock-capturing schemes with the onset of wrong propagation speed of discontinuities. For pointwise evaluation of the source term, previous studies indicated that the phenomenon of wrong propagation speed of discontinuities is connected with the smearing of the discontinuity caused by the discretization of the advection term. The smearing introduces a nonequilibrium state into the calculation. Thus as soon as a nonequilibrium value is introduced in this manner, the source term turns on and immediately restores equilibrium, while at the same time shifting the discontinuity to a cell boundary. The present study is to show that the degree of wrong propagation speed of discontinuities is highly dependent on the accuracy of the numerical method. The manner in which the smearing of discontinuities is contained by the numerical method and the overall amount of numerical dissipation being employed play major roles. Moreover, employing finite time steps and grid spacings that are below the standard Courant-Friedrich-Levy (CFL) limit on shockcapturing methods for compressible Euler and Navier-Stokes equations containing stiff reacting source terms and discontinuities reveals surprising counter-intuitive results. Unlike non-reacting flows, for stiff reactions with discontinuities, employing a time step and grid spacing that are below the CFL limit (based on the homogeneous part or non-reacting part of the governing equations) does not guarantee a correct solution of the chosen governing equations. Instead, depending on the numerical method, time step and grid spacing, the numerical simulation may lead to (a) the correct solution (within the truncation error of the scheme), (b) a divergent solution, (c) a wrong propagation speed of discontinuities solution or (d) other spurious solutions that are solutions of the discretized counterparts but are not solutions of the governing equations. The present investigation for three very different stiff system cases confirms some of the findings of Lafon & Yee (1996) and LeVeque & Yee (1990) for a model scalar PDE. The findings might shed some light on the reported difficulties in numerical combustion and problems with stiff nonlinear (homogeneous) source terms and discontinuities in general.

Yee, Helen M. C.↗

Study of Fluid Flow Control In Protein Crystallization Using Strong Magnetic Fields

An important component in biotechnology, particularly in the area of protein engineering and rational drug design is the knowledge of the precise three-dimensional molecular structure of proteins. The quality of structural information obtained from X-ray diffraction methods is directly dependent on the degree of perfection of the protein crystals. As a consequence, the growth of high quality macromolecular crystals for diffraction analyses has been the central focus for biochemists, biologists, and bioengineers. Macromolecular crystals are obtained from solutions that contain the crystallizing species in equilibrium with higher aggregates, ions, precipitants, other possible phases of the protein, foreign particles, the walls of the container, and a likely host of other impurities. By changing transport modes in general, i.e., reduction of convection and sedimentation, as is achieved in 'microgravity', researchers have been able to dramatically affect the movement and distribution of macromolecules in the fluid, and thus their transport, formation of crystal nuclei, and adsorption to the crystal surface. While a limited number of high quality crystals from space flights have been obtained, as the recent National Research Council (NRC) review of the NASA microgravity crystallization program pointed out, the scientific approach and research in crystallization of proteins has been mainly empirical yielding inconclusive results. We postulate that we can reduce convection in ground-based experiments and we can understand the different aspects of convection control through the use of strong magnetic fields and field gradients. Whether this limited convection in a magnetic field will provide the environment for the growth of high quality crystals is still a matter of conjecture that our research will address. The approach exploits the variation of fluid magnetic susceptibility with concentration for this purpose and the convective damping is realized by appropriately positioning the crystal growth cell so that the magnetic susceptibility force counteracts terrestrial gravity. The general objective is to test the hypothesis of convective control using a strong magnetic field and magnetic field gradient and to understand the nature of the various forces that come into play. Specifically we aim to delineate causative factors and to quantify them through experiments, analysis and numerical modeling. Once the basic understanding is obtained, the study will focus on testing the hypothesis on proteins of pyruvate dehydrogenase complex (PDC), proteins E1 and E3. Obtaining high crystal quality of these proteins is of great importance to structural biologists since their structures need to be determined.

Ramachandran, N.↗

Study of Fluid Flow Control in Protein Crystallization using Strong Magnetic Fields

An important component in biotechnology, particularly in the area of protein engineering and rational drug design is the knowledge of the precise three-dimensional molecular structure of proteins. The quality of structural information obtained from X-ray diffraction methods is directly dependent on the degree of perfection of the protein crystals. As a consequence, the growth of high quality macromolecular crystals for diffraction analyses has been the central focus for biochemists, biologists, and bioengineers. Macromolecular crystals are obtained from solutions that contain the crystallizing species in equilibrium with higher aggregates, ions, precipitants, other possible phases of the protein, foreign particles, the walls of the container, and a likely host of other impurities. By changing transport modes in general, i.e., reduction of convection and sedimentation, as is achieved in "microgravity", researchers have been able to dramatically affect the movement and distribution of macromolecules in the fluid, and thus their transport, formation of crystal nuclei, and adsorption to the crystal surface. While a limited number of high quality crystals from space flights have been obtained, as the recent National Research Council (NRC) review of the NASA microgravity crystallization program pointed out, the scientific approach and research in crystallization of proteins has been mainly empirical yielding inconclusive results. We postulate that we can reduce convection in ground-based experiments and we can understand the different aspects of convection control through the use of strong magnetic fields and field gradients. Whether this limited convection in a magnetic field will provide the environment for the growth of high quality crystals is still a matter of conjecture that our research will address. The approach exploits the variation of fluid magnetic susceptibility with concentration for this purpose and the convective damping is realized by appropriately positioning the crystal growth cell so that the magnetic susceptibility force counteracts terrestrial gravity. The general objective is to test the hypothesis of convective control using a strong magnetic field and magnetic field gradient and to understand the nature of the various forces that come into play. Specifically we aim to delineate causative factors and to quantify them through experiments, analysis and numerical modeling. Once the basic understanding is obtained, the study will focus on testing the hypothesis on proteins of pyruvate dehydrogenase complex (PDC), proteins E1 and E3. Obtaining high crystal quality of these proteins is of great importance to structural biologists since their structures need to be determined. Specific goals for the investigation are: 1. To develop an understanding of convection control in diamagnetic fluids with concentration gradients through experimentation and numerical modeling. Specifically solutal buoyancy driven convection due to crystal growth will be considered. 2. To develop predictive measures for successful crystallization in a magnetic field using analyses and numerical modeling for use in future protein crystal growth experiments. This will establish criteria that can be used to estimate the efficacy of magnetic field flow damping on crystallization of candidate proteins. 3. To demonstrate the understanding of convection damping by high magnetic fields to a class of proteins that is of interest and whose structure is as yet not determined. 4. To compare quantitatively, the quality of the grown crystals with and without a magnetic field. X-ray diffraction techniques will be used for the comparative studies. In a preliminary set of experiments, we studied crystal dissolution effects in a 5 Tesla magnet available at NASA Marshall Space Flight Center (MSFC). Using a Schlieren setup, a 1mm crystal of Alum (Aluminum-Potassium Sulfate) was introduced in a 75% saturated solution and the resulting dissolution plume was observed. The experiment was conducted both in the presence and absence of a magnetic field gradient. The magnet produces a gradient field of approx. 1 Tesla2/cm. Image analysis of the recorded images indicated an enhanced plume velocity that was of the order of the measurement limit. For this experiment, both the gradient and gravity fields are in the same direction resulting in an enhanced effective gravity that tends to accelerate the observed plume velocity. While the results are not conclusive, pending further tests, it clearly points out the inadequacy of the MSFC magnet for conducting protein crystallization experiments and the need for a stronger magnet. In spacebased experiments, however, where the gravitational effects are small, only a weak magnetic field will be required to control or mitigate the effects of convective contamination.

Ramachandran, Narayanan↗

Elucidating Microbial Adaptation Dynamics via Autonomous Exposure and Sampling

The adaptation of micro-organisms to their environments is a complex process of interaction between the pressures of the environment and of competition. Reducing this multifactorial process to environmental exposure in the laboratory is a common tool for elucidating individual mechanisms of evolution, such as mutation rates. Although such studies inform fundamental questions about the way adaptation and even speciation occur, they are often limited by labor-intensive manual techniques. Current methods for controlled study of microbial adaptation limit the length of time, the depth of collected data, and the breadth of applied environmental conditions. Small idiosyncrasies in manual techniques can have large effects on outcomes; for example, there are significant variations in induced radiation resistances following similar repeated exposure protocols. We describe here a project under development to allow rapid cycling of multiple types of microbial environmental exposure. The system allows continuous autonomous monitoring and data collection of both single species and sampled communities, independently and concurrently providing multiple types of controlled environmental pressure (temperature, radiation, chemical presence or absence, and so on) to a microbial community in dynamic response to the ecosystem's current status. When combined with DNA sequencing and extraction, such a controlled environment can cast light on microbial functional development, population dynamics, inter- and intra-species competition, and microbe-environment interaction. The project's goal is to allow rapid, repeatable iteration of studies of both natural and artificial microbial adaptation. As an example, the same system can be used both to increase the pH of a wet soil aliquot over time while periodically sampling it for genetic activity analysis, or to repeatedly expose a culture of bacteria to the presence of a toxic metal, automatically adjusting the level of toxicity based on the number or growth rate of surviving cells. We are on our second prototype iteration, with demonstrated functions of microbial growth monitoring and dynamic exposure to UV-C radiation and temperature. We plan to add functionality for general chemical presence or absence by Nov. 2013. By making the project low-cost and open-source, we hope to encourage others to use it as a basis for future development of a common microbial environmental adaptation testbed.

Microbiology↗

An Experimental Study of Swirling Flows as Applied to Annular Combustors

This thesis presents an experimental study of swirling flows with direct applications to gas turbine combustors. Two separate flowfields were investigated: a round, swirling jet and a non-combusting annular combustor model. These studies were intended to allow both a further understanding of the behavior of general swirling flow characteristics, such as the recirculation zone, as well as to provide a base for the development of computational models. In order to determine the characteristics of swirling flows the concentration fields of a round, swirling jet were analyzed for varying amount of swirl. The experimental method used was a light scattering concentration measurement technique known as marker nephelometry. Results indicated the formation of a zone of recirculating fluid for swirl ratios (rotational speed x jet radius over mass average axial velocity) above a certain critical value. The size of this recirculation zone, as well as the spread angle of the jet, was found to increase with increase in the amount of applied swirl. The annular combustor model flowfield simulated the cold-flow characteristics of typical current annular combustors: swirl, recirculation, primary air cross jets and high levels of turbulence. The measurements in the combustor model made by the Laser Doppler Velocimetry technique, allowed the evaluation of the mean and rms velocities in the three coordinate directions, one Reynold's shear stress component and the turbulence kinetic energy: The primary cross jets were found to have a very strong effect on both the mean and turbulence flowfields. These cross jets, along with a large step change in area and wall jet inlet flow pattern, reduced the overall swirl in the test section to negligible levels. The formation of the strong recirculation zone is due mainly to the cross jets and the large step change in area. The cross jets were also found to drive a four-celled vortex-type motion (parallel to the combustor longitudinal axis) near the cross jet injection plane.

Seal, Michael Damian, II↗

Spatial Variability in Column CO2 Inferred from High Resolution GEOS-5 Global Model Simulations: Implications for Remote Sensing and Inversions

Column CO2 observations from current and future remote sensing missions represent a major advancement in our understanding of the carbon cycle and are expected to help constrain source and sink distributions. However, data assimilation and inversion methods are challenged by the difference in scale of models and observations. OCO-2 footprints represent an area of several square kilometers while NASA s future ASCENDS lidar mission is likely to have an even smaller footprint. In contrast, the resolution of models used in global inversions are typically hundreds of kilometers wide and often cover areas that include combinations of land, ocean and coastal areas and areas of significant topographic, land cover, and population density variations. To improve understanding of scales of atmospheric CO2 variability and representativeness of satellite observations, we will present results from a global, 10-km simulation of meteorology and atmospheric CO2 distributions performed using NASA s GEOS-5 general circulation model. This resolution, typical of mesoscale atmospheric models, represents an order of magnitude increase in resolution over typical global simulations of atmospheric composition allowing new insight into small scale CO2 variations across a wide range of surface flux and meteorological conditions. The simulation includes high resolution flux datasets provided by NASA s Carbon Monitoring System Flux Pilot Project at half degree resolution that have been down-scaled to 10-km using remote sensing datasets. Probability distribution functions are calculated over larger areas more typical of global models (100-400 km) to characterize subgrid-scale variability in these models. Particular emphasis is placed on coastal regions and regions containing megacities and fires to evaluate the ability of coarse resolution models to represent these small scale features. Additionally, model output are sampled using averaging kernels characteristic of OCO-2 and ASCENDS measurement concepts to create realistic pseudo-datasets. Pseudo-data are averaged over coarse model grid cell areas to better understand the ability of measurements to characterize CO2 distributions and spatial gradients on both short (daily to weekly) and long (monthly to seasonal) time scales

Ott, L.↗

Flow Cytometry Methods to Monitor Immune System Dysregulation in Astronauts

Persistent immune system dysregulation has been documented to occur in astronauts participating in orbital spaceflight onboard the International Space Station. The phenomenon consists of reductions in T and NK cell function, altered cytokine profiles, persistent inflammation, and the subclinical reactivation of latent herpesviruses. In select crewmembers the dysregulation does actually lead to clinical symptoms, primarily atypical allergy or atopic dermatitis/zoster. Flow cytometry has served a central role defining the ‘immune assessment’ panel of assays that allow monitoring of astronauts. The cytometry assays which have been utilized include: 1. Peripheral leukocyte subsets 2. T cell function 3. Monocyte function 4. NK cell function 5. Intracellular cytokine analysis 6. Virus-specific T cell number (tetramer assay) 7. Virus-specific T cell function (peptide stimulation) 8. Leukocyte-bacterial challenge cytometry 9. Cytometric bead/multiplex array (soluble proteins) The use of these assays has been validated through various ISS flight investigations to define, to varying degrees, both in-flight, and post-flight immune system alterations. The kinetics of the dysregulation through the various phases of spaceflight, as well as post-flight recovery, have also been documented. To allow the science to occur within the orbital constraints of a spaceflight investigation, particular sample collection and processing techniques were developed compatible with the delays associated with terrestrial processing of in-flight samples. A subset of the assays has been adapted to routine monitoring of astronauts via a NASA ‘ISS Standard Measures’ activity, with the data from a specific crew then provided to all science investigators for that particular mission. This battery of cytometry assays has also been applied, through ground investigations, to several terrestrial ‘spaceflight analog’ populations. The purpose was to validate the analog which most closely replicates the in-flight observed pattern of alterations, generally believed to be winterover at an Antarctica station. To assist in determination of clinical risk, the assay panel has also been applied to investigations of various terrestrial patient populations, particularly zoster patients. As NASA is initiating crewed lunar missions via the ‘Artemis’ program, deployment of a miniaturized, microgravity-compatible flow cytometer, would be extremely beneficial to allow real time monitoring of crewmembers. Real time medical data could influence use of several countermeasures options during deep space missions. Several such instruments have been developed and validated to varying degrees of success. Assay details and summary findings across the various flight and ground platforms will be presented, as will current status in developing such technology for in-flight use.

Brian Crucian↗