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At least 757 records · Page 42

Transduction-like gene transfer in the methanogen Methanococcus voltae

Strain PS of Methanococcus voltae (a methanogenic, anaerobic archaebacterium) was shown to generate spontaneously 4.4-kbp chromosomal DNA fragments that are fully protected from DNase and that, upon contact with a cell, transform it genetically. This activity, here called VTA (voltae transfer agent), affects all markers tested: three different auxotrophies (histidine, purine, and cobalamin) and resistance to BES (2-bromoethanesulfonate, an inhibitor of methanogenesis). VTA was most effectively prepared by culture filtration. This process disrupted a fraction of the M. voltae cells (which have only an S-layer covering their cytoplasmic membrane). VTA was rapidly inactivated upon storage. VTA particles were present in cultures at concentrations of approximately two per cell. Gene transfer activity varied from a minimum of 2 x 10(-5) (BES resistance) to a maximum of 10(-3) (histidine independence) per donor cell. Very little VTA was found free in culture supernatants. The phenomenon is functionally similar to generalized transduction, but there is no evidence, for the time being, of intrinsically viral (i.e., containing a complete viral genome) particles. Consideration of VTA DNA size makes the existence of such viral particles unlikely. If they exist, they must be relatively few in number;perhaps they differ from VTA particles in size and other properties and thus escaped detection. Digestion of VTA DNA with the AluI restriction enzyme suggests that it is a random sample of the bacterial DNA, except for a 0.9-kbp sequence which is amplified relative to the rest of the bacterial chromosome. A VTA-sized DNA fraction was demonstrated in a few other isolates of M. voltae.

Methanococcus/genetics/growth & development/metabo↗

Alteration of renal function of rats following spaceflight

Following spaceflight, changes in renal function of humans have been suggested. To assess the effects of readaptation on renal function, urine was collected from male rats ( approximately 245 g) over a 2-wk period following a 14-day spaceflight. Rats were assigned to three groups: flight animals (n = 6), flight controls (n = 6) housed in the flight cages on the ground, and vivarium controls (n = 5) housed in standard shoe box cages. Animals were placed into individual metabolic cages for urine collection. Urine output was significantly increased for 3 days following flight. Excretion rates of Na+ and K+ were increased, resulting in an increased osmotic excretion rate. Creatinine excretion rate increased over the first two postflight days. Glomerular filtration rate increased immediately following spaceflight without changes in plasma creatinine, Na+, K+, or osmolality. Increased excretion of solute was thus the result of increased delivery and a decreased percent reabsorption of the filtered load. Osmolal clearance was increased immediately postflight while free water clearance was decreased. In growing rats, the diuresis after short-duration spaceflight is the result of an increase in solute excretion with an accompanying reduction in free water clearance.

STS-58 Shuttle Project↗

Site-specific regulatory interaction between spinach leaf sucrose-phosphate synthase and 14-3-3 proteins

We report an Mg2+-dependent interaction between spinach leaf sucrose-phosphate synthase (SPS) and endogenous 14-3-3 proteins, as evidenced by co-elution during gel filtration and co-immunoprecipitation. The content of 14-3-3s associated with an SPS immunoprecipitate was inversely related to activity, and was specifically reduced when tissue was pretreated with 5-aminoimidazole-4-carboxamide riboside, suggesting metabolite control in vivo. A synthetic phosphopeptide based on Ser-229 was shown by surface plasmon resonance to bind a recombinant plant 14-3-3, and addition of the phosphorylated SPS-229 peptide was found to stimulate the SPS activity of an SPS:14-3-3 complex. Taken together, the results suggest a regulatory interaction of 14-3-3 proteins with Ser-229 of SPS.

Non-NASA Center↗

Use of an adaptable cell culture kit for performing lymphocyte and monocyte cell cultures in microgravity

The results of experiments performed in recent years on board facilities such as the Space Shuttle/Spacelab have demonstrated that many cell systems, ranging from simple bacteria to mammalian cells, are sensitive to the microgravity environment, suggesting gravity affects fundamental cellular processes. However, performing well-controlled experiments aboard spacecraft offers unique challenges to the cell biologist. Although systems such as the European 'Biorack' provide generic experiment facilities including an incubator, on-board 1-g reference centrifuge, and contained area for manipulations, the experimenter must still establish a system for performing cell culture experiments that is compatible with the constraints of spaceflight. Two different cell culture kits developed by the French Space Agency, CNES, were recently used to perform a series of experiments during four flights of the 'Biorack' facility aboard the Space Shuttle. The first unit, Generic Cell Activation Kit 1 (GCAK-1), contains six separate culture units per cassette, each consisting of a culture chamber, activator chamber, filtration system (permitting separation of cells from supernatant in-flight), injection port, and supernatant collection chamber. The second unit (GCAK-2) also contains six separate culture units, including a culture, activator, and fixation chambers. Both hardware units permit relatively complex cell culture manipulations without extensive use of spacecraft resources (crew time, volume, mass, power), or the need for excessive safety measures. Possible operations include stimulation of cultures with activators, separation of cells from supernatant, fixation/lysis, manipulation of radiolabelled reagents, and medium exchange. Investigations performed aboard the Space Shuttle in six different experiments used Jurkat, purified T-cells or U937 cells, the results of which are reported separately. We report here the behaviour of Jurkat and U937 cells in the GCAK hardware in ground-based investigations simulating the conditions expected in the flight experiment. Several parameters including cell concentration, time between cell loading and activation, and storage temperature on cell survival were examined to characterise cell response and optimise the experiments to be flown aboard the Space Shuttle. Results indicate that the objectives of the experiments could be met with delays up to 5 days between cell loading into the hardware and initial in flight experiment activation, without the need for medium exchange. Experiment hardware of this kind, which is adaptable to a wide range of cell types and can be easily interfaced to different spacecraft facilities, offers the possibility for a wide range of experimenters successfully and easily to utilise future flight opportunities.

short duration↗

Regulation of body fluid volume and electrolyte concentrations in spaceflight

Despite a number of difficulties in performing experiments during weightlessness, a great deal of information has been obtained concerning the effects of spaceflight on the regulation of body fluid and electrolytes. Many paradoxes and questions remain, however. Although body mass, extracellular fluid volume, and plasma volume are reduced during spaceflight and remain so at landing, the changes in total body water are comparatively small. Serum or plasma sodium and osmolality have generally been unchanged or reduced during the spaceflight, and fluid intake is substantially reduced, especially during the first of flight. The diuresis that was predicted to be caused by weightlessness, has only rarely been observed as an increased urine volume. What has been well established by now, is the occurrence of a relative diuresis, where fluid intake decreases more than urine volume does. Urinary excretion of electrolytes has been variable during spaceflight, but retention of fluid and electrolytes at landing has been consistently observed. The glomerular filtration rate was significantly elevated during the SLS missions, and water and electrolyte loading tests have indicated that renal function is altered during readaptation to Earth's gravity. Endocrine control of fluid volumes and electrolyte concentrations may be altered during weightlessness, but levels of hormones in body fluids do not conform to predictions based on early hypotheses. Antidiuretic hormone is not suppressed, though its level is highly variable and its secretion may be affected by space motion sickness and environmental factors. Plasma renin activity and aldosterone are generally elevated at landing, consistent with sodium retention, but inflight levels have been variable. Salt intake may be an important factor influencing the levels of these hormones. The circadian rhythm of cortisol has undoubtedly contributed to its variability, and little is known yet about the influence of spaceflight on circadian rhythms. Atrial natriuretic peptide does not seem to play an important role in the control of natriuresis during spaceflight. Inflight activity of the sympathetic nervous system, assessed by measuring catecholamines and their metabolites and precursors in body fluids, generally seems to be no greater than on Earth, but this system is usually activated at landing. Collaborative experiments on the Mir and the International Space Station should provide more of the data needed from long-term flights, and perhaps help to resolve some of the discrepancies between U.S. and Russian data. The use of alternative methods that are easier to execute during spaceflight, such as collection of saliva instead of blood and urine, should permit more thorough study of circadian rhythms and rapid hormone changes in weightlessness. More investigations of dietary intake of fluid and electrolytes must be performed to understand regulatory processes. Additional hormones that may participate in these processes, such as other natriuretic hormones, should be determined during and after spaceflight. Alterations in body fluid volume and blood electrolyte concentrations during spaceflight have important consequences for readaptation to the 1-G environment. The current assessment of fluid and electrolyte status during weightlessness and at landing and our still incomplete understanding of the processes of adaptation to weightlessness and readaptation to Earth's gravity have resulted in the development of countermeasures that are only partly successful in reducing the postflight orthostatic intolerance experienced by astronauts and cosmonauts. More complete knowledge of these processes can be expected to produce countermeasures that are even more successful, as well as expand our comprehension of the range of adaptability of human physiologic processes.

NASA Discipline Number 18-10↗

Multiple forms of ADP-glucose pyrophosphorylase from tomato fruit

ADP-glucose pyrophosphorylase (AGP) was purified from tomato (Lycopersicon esculentum Mill.) fruit to apparent homogeneity. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis the enzyme migrated as two close bands with molecular weights of 50,000 and 51,000. Two-dimensional polyacrylamide gel electrophoresis analysis of the purified enzyme, however, revealed at least five major protein spots that could be distinguished by their slight differences in net charge and molecular weight. Whereas all of the spots were recognized by the antiserum raised against tomato fruit AGP holoenzyme, only three of them reacted strongly with antiserum raised against the potato tuber AGP large subunit, and the other two spots (with lower molecular weights) reacted specifically with antisera raised against spinach leaf AGP holoenzyme and the potato tuber AGP small subunit. The results suggest the existence of at least three isoforms of the AGP large subunit and two isoforms of the small subunit in tomato fruit in vivo. The native molecular mass of the enzyme determined by gel filtration was 220 +/- 10 kD, indicating a tetrameric structure for AGP from tomato fruit. The purified enzyme is very sensitive to 3-phosphoglycerate/inorganic phosphate regulation.

NASA Discipline Life Support Systems↗

Interaction between alpha 2-adrenergic and angiotensin II systems in the control of glomerular hemodynamics as assessed by renal micropuncture in the rat

The hypothesis that renal alpha 2 adrenoceptors influence nephron filtration rate (SNGFR) via interaction with angiotensin II (AII) was tested by renal micropuncture. The physical determinants of SNGFR were assessed in adult male Munich Wistar rats 5-7 d after ipsilateral surgical renal denervation (DNX). DNX was performed to isolate inhibitory central and presynaptic alpha 2 adrenoceptors from end-organ receptors within the kidney. Two experimental protocols were employed: one to test whether prior AII receptor blockade with saralasin would alter the glomerular hemodynamic response to alpha 2 adrenoceptor stimulation with the selective agonist B-HT 933 under euvolemic conditions, and the other to test whether B-HT 933 would alter the response to exogenous AII under conditions of plasma volume expansion. In euvolemic rats, B-HT 933 caused SNGFR to decline as the result of a decrease in glomerular ultrafiltration coefficient (LpA), an effect that was blocked by saralasin. After plasma volume expansion, B-HT 933 showed no primary effect on LpA but heightened the response of arterial blood pressure, glomerular transcapillary pressure gradient, and LpA to AII. The parallel results of these converse experiments suggest a complementary interaction between renal alpha 2-adrenergic and AII systems in the control of LpA.

NASA Discipline Cardiopulmonary↗

Freeing Water from Viruses and Bacteria

Four years ago, Argonide Corporation, a company focused on the research, production, and marketing of specialty nano materials, was seeking to develop applications for its NanoCeram[R] fibers. Only 2 nanometers in diameter, these nano aluminum oxide fibers possessed unusual bio-adhesive properties. When formulated into a filter material, the electropositive fibers attracted and retained electro-negative particles such as bacteria and viruses in water-based solutions. This technology caught the interest of NASA as a possible solution for improved water filtration in space cabins. NASA's Johnson Space Center awarded Sanford, Florida-based Argonide a Phase I Small Business Innovation Research (SBIR) contract to determine the feasibility of using the company's filter for purifying recycled space cabin water. Since viruses and bacteria can be carried aboard space cabins by space crews, the ability to detect and remove these harmful substances is a concern for NASA. The Space Agency also desired an improved filter to polish the effluent from condensed and waste water, producing potable drinking water. During its Phase I partnership with NASA, Argonide developed a laboratory-size filter capable of removing greater than 99.9999 percent of bacteria and viruses from water at flow rates more than 200 times faster than virus-rated membranes that remove particles by sieving. Since the new filter s pore size is rather large compared to other membranes, it is also less susceptible to clogging by small particles. In September 2002, Argonide began a Phase II SBIR project with Johnson to develop a full-size cartridge capable of serving a full space crew. This effort, which is still ongoing, enabled the company to demonstrate that its filter media is an efficient absorbent for DNA and RNA.

Source record↗

The Chemical Composition and Gas-to-Dust Mass Ratio of Nearby Interstellar Matter

We use recent results on interstellar gas toward nearby stars and interstellar by-products within the solar system to select among the equilibrium radiative transfer models of the nearest interstellar material presented in Slavin & Frisch. For the assumption that O/H - 400 parts per million, models 2 and 8 are found to yield good fits to available data on interstellar material inside and outside of the heliosphere, with the exception of the Ne abundance in the pickup ion and anomalous cosmic-ray populations. For these models, the interstellar medium (ISM) at the entry point to the heliosphere has n(H(sup 0)) = 0.202-0.208/cu cm, n(He(sup 0) = 0.0137-0.0152/cu cm, and ionizations X(H) = 0.29-0.30, X(He) = 0.47-0.51. These best models suggest that the chemical composition of the nearby ISM is approx.60%-70% subsolar if S is undepleted. Both H(0) and H(+) need to be included when evaluating abundances of ions found in warm diffuse clouds. Models 2 and 8 yield an H filtration factor of approx.0.46. Gas-to-dust mass ratios for the ISM toward epsilon CMa are R(sub gd) = 178-183 for solar abundances of Holweger or R(sub gd) = 611-657 for an interstellar abundance standard 70% solar. Direct observations of dust grains in the solar system by Ulysses and Galileo yield R(sub gd) appr0x. 115 for models 2 and 8, supporting earlier results (Frisch and coworkers). If the local ISM abundances are subsolar, then gas and dust are decoupled over small spatial scales. The inferred variation in R(sub gd) over parsec length scales is consistent with the fact that the ISM near the Sun is part of a dynamically active cluster of cloudlets flowing away from the Sco-Cen association. Observations toward stars within approx.500 pc show that R(sub gd) correlates with the percentage of the dust mass that is carried by iron, suggesting that an Fe-rich grain core (by mass) remains after grain destruction. Evidently large dust grains (>10(exp -13) g) and small dust grains (<10(exp -13) g) are not well mixed over parsec length spatial scales in the ISM. It also appears that very small C-dominated dust grains have been destroyed in the ISM within several parsecs of the Sun, since C appears to be essentially undepleted. However, if gas-dust coupling breaks down over the cloud lifetime, the missing mass arguments applied here to determine R(sub gd) and dust grain mineralogy are not appropriate.

Frisch, Priscilla C.↗

Near-Net-Shape Processing of Sintered Fibrous Ceramics Achieved

A variety of sintered fibrous ceramic (SFC) materials have been developed over the last 50 years as thermal barrier materials for reentry applications. SFC materials typically exhibit very low thermal conductivities combined with low densities and good thermal stability up to 2500 F. These materials have flown successfully on the space shuttle orbiters since the 1960's. More recently, the McDonnell Douglas Corporation successfully used SFC tiles as a heat shield on the underside of its DC X test vehicle. For both of these applications, tiles are machined from blocks of a specific type of SFC called an alumina-enhanced thermal barrier (AETB). The sizes of these blocks have been limited by the manufacturing process. In addition, as much as 80 to 90 percent of the material can be lost during the machining of tiles with significant amounts of curvature. To address these problems, the NASA Glenn Research Center at Lewis Field entered a cooperative contract with the Boeing Company to develop a vacuum-assisted forming process that can produce large (approximately 4 square feet), severely contoured panels of AETB while saving costs in comparison to the conventional cast-and-machine billet process. For shuttle use, AETB is slurry cast, drained, and fired to form square billets conforming to the shape of the filtration box. The billets are then cut into tiles of the appropriate size for thermally protecting the space shuttle. Processing techniques have limited the maximum size of AETB billets to 21.5 square inches by 6.5-in. thick, but the space shuttles use discrete heat shield tiles no more than 8 to 12 square inches. However, in other applications, large, complex shapes are needed, and the tiling approach is undesirable. For such applications, vacuum-assisted forming can produce large parts with complex shapes while reducing machining waste and eliminating cemented joints between bonded billets. Because it allows contoured shapes to be formed, material utilization is inherently high. Initial estimates show that the amount of material lost during machining can be reduced by 50 percent or more. In addition, a fiber alignment favorable for minimum heat transfer is maintained for all panel shapes since the fibers are aligned parallel to the contoured surface of the forming tool or mold. The vacuum-assisted forming process can complete the entire forming operation in a matter of minutes and can produce multiple parts whose size is limited only by the size of the forming tool. To date, panels as large as 2 square feet have been demonstrated The vacuum-assisted forming process starts with the fabrication of a permeable forming tool, or mold, with the proper part contour. This reusable tool is mounted over an internal rib support structure, as depicted in the diagram, such that a vacuum can be pulled on the bottom portion of the tool. AETB slurry is then poured over and around the tool, liquid is drawn from the slurry, and the part forms over the tool surface. The part is then dried, fired, and finished machined. Future plans include an evaluation of the need for additional coatings and surface-toughness treatments to extend the durability and performance of this material.

Angel, Paul W.↗

Operation of magnetically assisted fluidized beds in microgravity and variable gravity: experiment and theory

The conversion of solid waste into useful resources in support of long duration manned missions in space presents serious technological challenges. Several technologies, including supercritical water oxidation, microwave powered combustion and fluidized bed incineration, have been tested for the conversion of solid waste. However, none of these technologies are compatible with microgravity or hypogravity operating conditions. In this paper, we present the gradient magnetically assisted fluidized bed (G-MAFB) as a promising operating platform for fluidized bed operations in the space environment. Our experimental and theoretical work has resulted in both the development of a theoretical model based on fundamental principles for the design of the G-MAFB, and also the practical implementation of the G-MAFB in the filtration and destruction of solid biomass waste particles from liquid streams. c2004 COSPAR. Published by Elsevier Ltd. All rights reserved.

manned↗

Novel Biomedical Device Utilizing Light-Emitting Nanostructures Developed

Sketches and chemical diagrams of state-of-the-art device and novel proposed device are presented. Current device uses a diode laser that emits into a fluorescent fluid only one wavelength and a photodetector diode that detects only one wavelength. Only one type of bacteria can be detected. The proposed device uses a quantum dot array that emits into a fluorescent fluid multiple wavelengths and an NIR 512 spectrometer that scans 0.8- to 1.7-mm wavelengths. Hundreds of different bacteria and viruses can be detected. A novel biomedical device is being developed at the NASA Glenn Research Center in cooperation with the University of Michigan. This device uses nano-structured quantum dots that emit light in the near-infrared (IR) region. The nanostructured quantum dots are used as a source and excite fluorochrome polymers coupled with antibodies that seek out and attach to specific bacteria and viruses. The fluorochrome polymers/antibodies fluoresce at specific wavelengths in the near-IR spectrum, but these wavelengths are offset from the excitation wavelength and can be detected with a tunable spectrometer. The device will be used to detect the presence of viruses and bacteria in simple fluids and eventually in more complex fluids, such as blood. Current state-of-the-art devices are limited to single bacteria or virus detection and a considerable amount of time and effort is required to prepare samples for analysis. Most importantly, the devices are quite large and cumbersome, which prohibits them from being used on the International Space Station and the space shuttles. This novel device uses nanostructured quantum dots which, through molecular beam epitaxy and highly selective annealing processes, can be developed into an illumination source that could potentially generate hundreds of specific wavelengths. As a result, this device will be able to excite hundreds of antibody/fluorochrome polymer combinations, which in turn could be used to detect hundreds of bacteria and viruses in fluids. A novel sample preparation technique that exploits micromembrane filtration and centrifugation methods has been developed for this device. The technique greatly reduces the time required to prepare the sample and the amount of sample needed to perform an accurate and comprehensive analysis. Last, and probably most important, because of the nano-light-emitting source and the novel sample preparation technique, the overall size of the device could be reduced dramatically. This device will serve as a nanoscale lab-on-a-chip for in situ microorganism detection and will enable tests to be performed on a time scale of minutes rather than days. Thus, it is ideally suited for monitoring the environmental conditions onboard the International Space Station and the space shuttles, thereby enhancing the safety of the astronauts. In addition, the device has important commercial applications, such as detecting the presence of bacteria and viruses in water at food- and beverage-processing centers, water treatment plants, and restaurants. Also, this technology has the potential to be used to detect bacteria and viruses in more complex fluids, such as blood--which in all likelihood would revolutionize blood analysis as it is performed today. This project was made possible through the Director's Discretionary Fund and is ongoing. In addition, this project provides funding to Dr. Rachel Goldman of the University of Michigan for the research and development of nanostructured quantum dots.

Scardelletti, Maximilian C.↗

Microscale Particulate Classifiers (MiPAC) Being Developed

The NASA Glenn Research Center is developing microscale sensors to characterize atmospheric-borne particulates. The devices are fabricated using MEMS (microelectromechanical systems) technologies. These technologies are derived from those originally developed in support of the semiconductor processing industry. The resulting microsensors can characterize a wide range of particles and are, therefore, suitable to a broad range of applications. This project is supported under a collaborative program called the Glennan Microsystems Initiative. The initiative comprises members of NASA Glenn Research Center, various university affiliates from the State of Ohio, and a number of participating industrial partners. Funding is jointly provided by NASA, the State of Ohio, and industrial members. The work described here is a collaborative arrangement between researchers at Glenn, the University of Minnesota, The National Institute of Standards and Technology (NIST), and the Cleveland State University. Actual device fabrication is conducted at Glenn and at the laboratories of Case Western Reserve University. Case Western is also located in Cleveland, Ohio, and is a participating member of the initiative. The principal investigator for this project is Paul S. Greenberg of Glenn. Two basic types of devices are being developed, and target different ranges of particle sizes. The first class of devices, which is used to measure nanoparticles (i.e., particles in the range of 0.002 to 1 mm), is based on the technique of Electrical Mobility Classification. This technique also affords the valuable ability of measuring the electrical charge state of the particles. Such information is important in the understanding of agglomeration mechanisms and is useful in the development of methods for particle repulsion. The second type of device being developed, which utilizes optical scattering, is suitable for particles larger than 1 mm. This technique also provides information on particle shape and composition. Applications for these sensors include fundamental planetary climatology, monitoring and filtration in spacecraft, human habitation modules and related systems, characterization of particulate emissions from propulsion and power systems, and as early warning sensors for both space-based and ter-restrial fire detection. These devices are also suitable for characterizing biological compounds such as allergens, infectious agents, and biotoxic agents.

Greenberg, Paul S.↗

Novel Shapes of Miscible Interfaces Observed

The dynamics of miscible displacements in a cylindrical tube are being investigated experimentally and numerically, with a view to understand the complex processes that occur, for example, in enhanced oil recovery, hydrology, and filtration. We have observed complex shapes of the interface between two liquids that mix with each other when the less viscous liquid is displaced by the more viscous one in a tube. A less viscous fluid that displaces a more viscous fluid is known to propagate in the form of a "finger," and a flight experiment proposed by Maxworthy et al. to investigate the miscible-interface dynamics is currently being developed by NASA. From the current theory of miscible displacements, which was developed for a porous medium satisfying Darcy's law, it can be shown that in the absence of gravity the interface between the fluids is destabilized and thus susceptible to fingering only when a more viscous fluid is displaced by a less viscous one. Therefore, if the interface is initially flat and the more viscous fluid displaces the less viscous fluid, the interface ought to be stable and remain flat. However, numerical simulations by Chen and Meiburg for such displacement in a cylindrical tube show that the interface is unstable and a finger of the more viscous fluid is indeed formed. Preliminary experiments performed at the NASA Glenn Research Center show that not only can fingering occur when the more viscous fluid displaces a less viscous one in a cylindrical tube, but also that under certain conditions the advancing finger achieves a sinuous or snakelike shape. These experiments were performed using silicone oils in a vertical pipette of small diameter. In the initial configuration, the more viscous fluid rested on top of the less viscous one, and the interface was nominally flat. A dye was added to the upper liquid for ease of observation of the interface between the fluids. The flow was initiated by draining the lower fluid from the bottom of the pipette, at speeds less than 0.1 mm/sec.

Balasubramaniam, Ramaswamy↗

Performance Qualification Test of the ISS Water Processor Assembly (WPA) Expendables

The Water Processor Assembly (WPA) for use on the International Space Station (ISS) includes various technologies for the treatment of waste water. These technologies include filtration, ion exchange, adsorption, catalytic oxidation, and iodination. The WPA hardware implementing portions of these technologies, including the Particulate Filter, Multifiltration Bed, Ion Exchange Bed, and Microbial Check Valve, was recently qualified for chemical performance at the Marshall Space Flight Center. Waste water representing the quality of that produced on the ISS was generated by test subjects and processed by the WPA. Water quality analysis and instrumentation data was acquired throughout the test to monitor hardware performance. This paper documents operation of the test and the assessment of the hardware performance.

Carter, Layne↗

Autonomous Medical Care for Exploration

The goal of Autonomous Medical Care (AMC) is to ensure a healthy, well-performing crew which is a primary need for exploration. The end result of this effort will be the requirements and design for medical systems for the CEV, lunar operations, and Martian operations as well as a ground-based crew health optimization plan. Without such systems, we increase the risk of medical events occurring during a mission and we risk being unable to deal with contingencies of illness and injury, potentially threatening mission success. AMC has two major components: 1) pre-flight crew health optimization and 2) in-flight medical care. The goal of pre-flight crew health optimization is to reduce the risk of illness occurring during a mission by primary prevention and prophylactic measures. In-flight autonomous medical care is the capability to provide medical care during a mission with little or no real-time support from Earth. Crew medical officers or other crew members provide routine medical care as well as medical care to ill or injured crew members using resources available in their location. Ground support becomes telemedical consultation on-board systems/people collect relevant data for ground support to review. The AMC system provides capabilities to incorporate new procedures and training and advice as required. The on-board resources in an autonomous system should be as intelligent and integrated as is feasible, but autonomous does not mean that no human will be involved. The medical field is changing rapidly, and so a challenge is to determine which items to pursue now, which to leverage other efforts (e.g. military), and which to wait for commercial forces to mature. Given that what is used for the CEV or the Moon will likely be updated before going to Mars, a critical piece of the system design will be an architecture that provides for easy incorporation of new technologies into the system. Another challenge is to determine the level of care to provide for each mission type. The level of care refers to the amount and type of care one will render based on perceived need and ability. This is in contrast to the standard of care which is the benchmark by which that care is provided. There are certainly some devices and procedures that have unique microgravity or partial gravity requirements such that terrestrial methods will not work. For example, performing CPR on Mars cannot be done in exactly the same way as on Earth because the reduced gravity causes too large a reduction in the forces available for effective compression of the chest. Likewise, fluid behavior in microgravity may require a specialized water filtration and mixing system for the creation of intravenous fluids. This paper will outline the drivers for the design of the medical care systems, prioritization and planning techniques, key system components, and long term goals.

Johnson-Throop, Kathy A.↗

Antigenic Protein In Microgravity-Grown Human Mixed Mullerian Tumor (LN1) Cells Preserved In RNA Stabilizing Agent

Cells treated with RNAlater(TradeMark) have previously been shown to contain antigenic proteins that can be visualized using Western blot analysis. These proteins seem to be stable for several months when stored in RNA stabilizer at 4 C. Antigenic protein can be recovered from cells that have been processed using an Ambion RNAqueous(Registered TradeMark) kit to remove RNA. In this set of experiments, human mixed Mullerian tumor (LNI) cells grown on the International Space Station during Expedition 3 were examined for antigenic stability after removal of RNA. The cells were stored for three months in RNAlater(TradeMark) and RNA was extracted. The RNA filtrate containing the protein was precipitated, washed, and suspended in buffer containing sodium dodecyl sulfate (SDS). Samples containing equal concentrations of protein were loaded onto SDS-polyacrylamide gels. Proteins were separated by electrophoresis and transferred by Western blot to polyvinylidene fluoride (PVDF) membrane. The Western blots were stained with an enhanced chemiluminescent ECL(Registered Trademark) Plus detection kit (Amersham) and scanned using a Storm 840 gel image analyzer (Amersham, Molecular Dynamics). ImageQuant(Registered TradeMark) software was used to quantify the densities of the protein bands. The ground control and flight LN1 cell samples showed a similar staining pattern over time with antibodies to vimentin, glyceraldehyde-3-phosphate dehydrogenase, and epithelial membrane antigens.

Hammond, Dianne K.↗

Antigenic Protein In Microgravity-Grown Human Mixed Mullerian Tumor (LN1) Cells Preserved In RNA Stabilizing Agent

Cells treated with RNAlater(TradeMark) have previously been shown to contain antigenic proteins that can be visualized using Western blot analysis. These proteins seem to be stable for several months when stored in RNA stabilizer at 4 C. Antigenic protein can be recovered from cells that have been processed using an Ambion RNAqueous(Registered TradeMark) kit to remove RNA. In this set of experiments, human mixed Mullerian tumor (LN1) cells grown on the International Space Station during Expedition 3 were examined for antigenic stability after removal of RNA. The cells were stored for three months in RNAlater(TradeMark) and RNA was extracted. The RNA filtrate Containing the protein was precipitated, washed, and suspended in buffer containing sodium dodecyl sulfate (SDS). Samples containing equal concentrations of protein were loaded onto SDS-polyacrylamide gels. Proteins were separated by electrophoresis and transferred by Western blot to polyvinylidene fluoride (PVDF) membrane. The Western blots were stained with an enhanced chemiluminescent ECL(Registered TradeMark)Plus detection kit (Amersham) and scanned using a Storm 840 gel image analyzer (Amersham, Molecular Dynamics). ImageQuant(Registered TradeMark)a software was used to quantify the densities of the protein bands. The ground control and flight LN1 cell samples showed a similar staining pattern over time with antibodies to vimentin, glyceraldehyde-3-phosphate dehydrogenase, and epithelial membrane antigens.

Hammond, Dianne K.↗