The Pasteur probe - An assay for molecular asymmetry.
Pasteur Probe assay of asymmetry of D, L amino acids in detection of Martian life
LIFE DETECTOR↗
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The Pasteur probe - An assay for molecular asymmetry.
Pasteur Probe, assay of asymmetry of D, L amino acids in detection of Martian life
LIFE DETECTOR↗
BioNutrients-3: Precision Fermentation, Pasteurization, and Pathogen Detection - Towards Safe Fermentation and Production of Nutrients in Space
The BioNutrients (BN) project is developing a microbial manufacturing approach to supplement the NASA food system to address known nutrient degradation associated with long-term storage. BN uses synthetic biology to deliver high-value nutrients and therapeutics through genetic engineering of microbes and production of fermented food products like yogurt and kefir, in a fully dehydrated system. On-demand production of nutrients for human consumption requires rigorous safety protocols to ensure contaminants are not introduced during the fermentation process. The third iteration of the BN flight project, BN-3, further develops the BN project by investigating strategies for pathogen detection and pasteurization in microgravity. BN-3 will test the limits of pathogen detection using whole genome sequencing, standard microbial assays and the NASA Ames E-Nose, a volatile carbon nano tube-based sensor array, to detect unwanted microbes. BN-3 also expands on the number of nutrients produced in a single bioreactor to enhance efficiency of the system by combining production of B vitamins in conjunction with the carotenoids, beta-carotene and zeaxanthin. Serial production of food products through yogurt passaging, new updates to the fluorinated ethylene propylene bags to allow crew access via a straw, as well as use of a food safe pH indicator dye to indicate readiness of the food product, will further enhance the useability of this system. This presentation will provide status of the BN-3 flight project with the aim of advancing in-space biomanufacturing for on-demand microbially based food production for future space exploration.
Biomanufacturing↗
Miniaturized Single-Shot Valve and its Application to the ExoMars Pasteur Payload
Hermetically sealing a gas tank and opening it into tubing under telecommand control is a function required in various space instruments. There are a number of space valves that are power saving, withstand vibration, and do not contaminate the gas. But none of them combines these features with low mass and the ability to withstand temperatures of 130 C during the sterilization process mandatory for planetary missions. In this paper, a novel miniature valve is presented, which is particularly adapted to space applications. It is electrically actuated, utilizing a bimetallic snap-disc that pierces a metallic membrane by a needle pin, thereby opening the valve. The design of this single-shot valve is such that it allows it to withstand a temperature of 130 C and a pressure of 50 bars. The valve is also lightweight (6.62 g) and it requires only 9 W to operate.
Muller, Pierre↗
Effect of surfactants and temperature on the hyperfiltration performance of poly(ether/urea) membranes
The individual and combined effects of pasteurization temperature (347 K) and surfactants (anionic, cationic, and neutral) on a poly(ether/urea) thin-film hyperfiltration membrane were studied. Performance of this positively charged membrane was measured in terms of sodium chloride rejection and water flux. The observed effect was mostly on water flux and minimal on salt rejection. Pasteurization temperature caused an irreversible flux decline (flux decline slope of 0.09). The gradual flux reduction caused by neutral and cationic surfactants was reversible, whereas the flux reduction caused by anionic surfactant was irreversible and of similar magnitude to flux reduction caused by pasteurization temperature. The effects of anionic surfactant and pasteurization temperature were additive. Because of flux decline at elevated temperatures the poly(ether/urea) membrane is not very attractive for long-term spaceflight use.
Leban, M. I.↗
Health protection and food preservation by gamma irradiation
Results of several major studies on food systems for space missions beginning with Apollo 12 through Apollo-Soyuz and investigations of the application of irradiation to food for manned space flight are reported. The study of flight food systems involved the application of radurization (pasteurizing levels) doses of gamma irradiation to flour and bread supplied by Pepperidge Farms in advance of the missions. All flights from Apollo 12 through 17 carried irradiated fresh bread. On Apollo 17, cooperation with Natick Laboratories permitted the introduction of a ham sandwich using irradiated bread and irradiated sterile ham. Investigations centered on irradiated bread were conducted during the course of these missions. Studies were applied to the concept of improving fresh bread from the point of view of mold inhibition. The studies considered how irradiation could best be applied at what levels and on a variety of bread types. Throughout the studies of the application of gamma irradiation the emphasis was placed upon using low levels of irradiation in the pasteurizing or radurizing doses--under a Megarad. The primary goal was to determine if a public health benefit could be demonstrated using radurization along with food preservation and food quality improvements. The public health benefit would be parallel to that of pasteurization of milk as a concept. Publications are included providing the details of these observations, one dealing with the flour characteristics and the other dealing with the influence on fresh bread types. These demonstrate the major findings noted during the period of the studies examining bread.
Source record↗
A membrane-based subsystem for very high recoveries of spacecraft waste waters
This paper describes the continued development of a membrane-based subsystem designed to recover up to 99.5 percent of the water from various spacecraft waste waters. Specifically discussed are: (1) the design and fabrication of an energy-efficient reverse-osmosis (RO) breadboard subsystem; (2) data showing the performance of this subsystem when operated on a synthetic wash-water solution - including the results of a 92-day test; and (3) the results of pasteurization studies, including the design and operation of an in-line pasteurizer. Also included in this paper is a discussion of the design and performance of a second RO stage. This second stage results in higher-purity product water at a minimal energy requirement and provides a substantial redundancy factor to this subsystem.
Ray, Roderick J.↗
Hydrothermal organic synthesis experiments
The serious scientific debate about spontaneous generation which raged for centuries reached a climax in the nineteenth century with the work of Spallanzani, Schwann, Tyndall, and Pasteur. These investigators demonstrated that spontaneous generation from dead organic matter does not occur. Although no aspects of these experiments addressed the issue of whether organic compounds could be synthesized abiotically, the impact of the experiments was great enough to cause many investigators to assume that life and its organic compounds were somehow fundamentally different than inorganic compounds. Meanwhile, other nineteenth-century investigators were showing that organic compounds could indeed be synthesized from inorganic compounds. In 1828 Friedrich Wohler synthesized urea in an attempt to form ammonium cyanate by heating a solution containing ammonia and cyanic acid. This experiment is generally recognized to be the first to bridge the artificial gap between organic and inorganic chemistry, but it also showed the usefulness of heat in organic synthesis. Not only does an increase in temperature enhance the rate of urea synthesis, but Walker and Hambly showed that equilibrium between urea and ammonium cyanate was attainable and reversible at 100 C. Wohler's synthesis of urea, and subsequent syntheses of organic compounds from inorganic compounds over the next several decades dealt serious blows to the 'vital force' concept which held that: (1) organic compounds owe their formation to the action of a special force in living organisms; and (2) forces which determine the behavior of inorganic compounds play no part in living systems. Nevertheless, such progress was overshadowed by Pasteur's refutation of spontaneous generation which nearly extinguished experimental investigations into the origins of life for several decades. Vitalism was dealt a deadly blow in the 1950's with Miller's famous spark-discharge experiments which were undertaken in the framework of the Oparin and Haldane hypotheses concerning the origin of life. These hypotheses were constructed on some basic assumptions which included a reduced atmosphere, and a low surface temperature for the early Earth. These ideas meshed well with the prevailing hypothesis of the 1940's and 50's that the Earth had formed through heterogeneous accretion of dust from a condensing solar nebula. Miller's experiments were extremely successful, and were followed by numerous other experiments by various investigators who employed a wide variety of energy sources for abiotic synthesis including spark discharges, ultra-violet radiation, heat, shock waves, plasmas, gamma rays, and other forms of energy. The conclusion reached from this body of work is that energy inputs can drive organic synthesis from a variety of inorganic starting materials.
Shock, Everett L.↗
Construction of a Specialized Cloning Strain of E. Coli for the Nitrate Reductase Genes of Haloferax Denitrificans
This is the final report on Joint Research Interchange (NCC2-5011) "Construction of a Specialized Cloning Strain of E.. coli for the Nitrate Reductase Genes of Haloferax denitrificans." Originally the award was 11/l/93-10/31/95, but there were no-cost extensions made, because of a year Sabbatical at the Pasteur Institute in Paris and other leaves of 3 months each at the Pasteur Institute, during which work could not be done on this project, which extended the closing date to 10/30/98.
Johnson, Emmett↗
Irradiating of Bulk Soybeans: Influence on Their Functional and Sensory Properties for Soyfood Processing
Soybeans were chosen for lunar and planetary missions, where soybeans will be supplied in bulk or grown locally, due to their nutritive value and ability to produce oil and protein for further food applications. However, soybeans must be processed into foods prior to consumption. Radiation that soybeans would be exposed to during bulk storage prior to and during a Mars mission may influence their germination and functional properties. The influence of radiation includes the affect of surface pasteurization to ensure the astronauts safety from food-borne illnesses (HACCP, CCP), and the affect of the amount of radiation the soybeans receive during a Mars mission. Decreases in the amount of natural antioxidants free radical formation, and oxidation-induced changes in the soybean will influence the nutritional value, texture, color, and aroma of soyfoods. The objective of this study was to determine the influence of pasteurization and sterilization surface radiation on whole soybeans using gamma and electron beam radiation. The influence of 0, 1, 5, 10, and 30kGy on microbial load, germination rate, ease of processing, and quality of soymilk and tofu were determined. Surface radiation of whole dry soybeans using electron beam or gamma rays from 1-30kGy did provide microbial safety for the astronauts. However, the lower dose levels had surviving yeasts and molds. These doses caused oxidative changes that resulted in soymilk and tofu with rancid aromas. GC-MS of the aroma compounds using SPME Headspace confirmed the presence of lipid oxidation compounds. Soybean germination ability was reduced as radiation dosage increased. While lower doses may reduce these problems, the ability to insure microbial safety of bulk soybeans will be lost. Counter measures could include vacuum packaging, nitrogen flushing, added antioxidants, and radiating under freezing conditions. Doses below 1kGy need to be investigated further to determine the influence of the radiation encountered during Mars missions.
Chia, Chiew-Ling↗
Bacterial Utilization of L-sugars and D-amino Acids
The fact that organotrophic organisms on Earth use L-amino acids and D-sugars as an energy source is recognized as one of the universal features of life. The chirality of organic molecules with asymmetric location of group- radicals was described a relatively long time ago. In 1848, Louis Pasteur discovered chiral molecules when he investigated the way that crystals of sodium ammonium paratartrate rotated the plane of polarization of light. He found that the crystal structures represented the underlying asymmetry of molecules that existed in either lea-handed or right-handed forms (enantiomers). Pasteur observed that abiotic (chemical) processes produced mixtures with equal numbers (racemic) of the two forms but that living organisms possessed a molecular asymmetry that included only one of the enantiomers (homochirality). He speculated that the origin of the asymmetry of chiral biomolecules might hold the key to the nature of life. All of the amino acids in proteins (except for Glycine which is symmetrical) exhibit the same absolute steric configuration as L-glyceraldehyde. D-amino acids are never found in proteins, although they do exist in nature and are often found in polypeptide antibiotics. Constitutional sugars of cells, opposite to the amino acids, are the D-enantiomers, and the appearance of L-sugars in Nature is extremely rare. Notwithstanding this fact, the metabolism of some bacteria does have capability to use amino acids and sugars with alternative chirality. This property may be caused by the function of specific enzymes belonging to the class of isomerases (racemases, epimerases, isomerases, tautomerases). In our laboratory, we have investigated several anaerobic bacterial strains, and have found that some of these bacteria are capable of using D-amino acids and L-sugars. Strain BK1 is capable of growth on D-arginine, but its growth characteristics on L-arginine are approximately twice higher. Another alkaliphilic strain SCAT(sup T) (= ATCC BAA-1084(sup T)= JCM 12857(sup T) = DSM 17722(sup T) = CIP 107910(sup T)) was found to be capable of growth on L-ribose and L-arabinose. It is interesting that this strain was incapable of growth on D-arabinose, which suggests the involvement of some alternative mechanism of enzyme activity. In this paper, we describe the preliminary results of these microbiological studies and discuss some possible implications.
Pikuta, Elena↗
Studies on the energy metabolism of human leukocytes. II.
Glycolysis control by respiration in human leukocytes with and without Pasteur effect conditions
Baierlein, J. L.↗
Progress in the development of the reverse osmosis process for spacecraft wash water recovery.
Research work on ambient- and pasteurization-temperature reverse osmosis processes for wash water recovery in a spacecraft environment is reviewed, and the advantages and drawbacks of each are noted. A key requirement in each case is to provide a membrane of appropriate stability and semipermeability. Reverse osmosis systems intended for such use must also take into account the specific limitations and requirements imposed by the small volume of water to be processed and the high water recovery desired. The incorporation of advanced high-temperature membranes into specially designed modules is discussed.
Pecoraro, J. N.↗
Significant results from Apollo 14 lunar orbital photography.
Apollo 14 obtained 950 photographs from lunar orbit using the Hasselblad and Hycon cameras. The photographs reveal a number of new geologic features as well as previously unrecognized details of the morphology, structure, and stratigraphy of lunar surface units. The primary result is the verification of the extensive role of volcanism in the formation and modification of the lunar highlands, especially on the far side. Terra volcanism appears to be manifest in the formation of (1) constructional units of hilly and furrowed materials of regional extent as in the Kant Plateau in the central near-side highlands and northwest of the crater Pasteur near the eastern limb of the moon; (2) somewhat viscous lava flows and pools associated with fracture systems and/or what appear to be volcanic craters; (3) craters, crater chains, and irregular depressions, particularly on the lunar far side. The first photographs of a flow channel, a leveed sinuous rille that apparently originated by lava flowage on the surface, were obtained by Apollo 14. Another first is a high-resolution photograph of the interior of what appears to be the youngest lunar crater yet photographed in the 20- 40-km size range.
El-Baz, F.↗
Design strategies for the International Space University's variable gravity research facility
A variable gravity research facility named 'Newton' was designed by 58 students from 13 countries at the International Space University's 1989 summer session at the Universite Louis Pasteur, Strasbourge, France. The project was comprehensive in scope, including a political and legal foundation for international cooperation, development and financing; technical, science and engineering issues; architectural design; plausible schedules; and operations, crew issues and maintenance. Since log-term exposure to zero gravity is known to be harmful to the human body, the main goal was to design a unique variable gravity research facility which would find a practical solution to this problem, permitting a manned mission to Mars. The facility would not duplicate other space-based facilities and would provide the flexibility for examining a number of gravity levels, including lunar and Martian gravities. Major design alternatives included a truss versus a tether based system which also involved the question of docking while spinning or despinning to dock. These design issues are described. The relative advantages or disadvantages are discussed, including comments on the necessary research and technology development required for each.
Bailey, Sheila G.↗
Influence of Hydroponically Grown Hoyt Soybeans and Radiation Encountered on Mars Missions on the Yield and Quality of Soymilk and Tofu
Soybeans were chosen for hmar and planetary missions due to their nutritive value and ability to produce oil and protein for further food applications. However, soybeans must be processed into foods prior to crew consumption. Wilson et al. (2003) raised questions about (1) the influence of radiation (on germination and functional properties) that the soybeans would be exposed to during bulk storage for a Mars mission, and (2) the impact of using hydroponically grown versus field grown soybeans on the yield and quality of soyfoods. The influence of radiation can be broken down into two components: (A) affect of surface pasteurization to ensure the astronauts safety from food-borne illnesses (a Hazard Analysis Critical Control Point), and (B) affect of the amount of radiation the soybeans receive during a Mars mission. Decreases in the amount of natural antioxidants and free radical formation and oxidation induced changes in the soybean (lipid, protein, etc.) will influence the nutritional value, texture, quality, and safety of soyfoods made from them. The objectives of this project are to (1) evaluate the influence of gamma and electron beam radiation on bulk soybeans (HACCP, CCP) on the microbial load, germination, ease of processing, and quality of soymilk and tofu; (2) provide scale up and mass balance data for Advanced Life Support subsystems including Biomass, Solid Waste Processing, and Water Recovery Systems; and (3) to compare Hoyt field grown to hydroponically grown Hoyt soybeans for soymilk and tofu production. The soybean cultivar Hoyt, a small standing, high protein cultivar that could grow hydroponically in the AIMS facility on Mars) was evaluated for the production of soymilk and tofu. The quality and yield of the soymilk and tofu from hydroponic Hoyt, was compared to Vinton 81 (a soyfood industry standard), field Hoyt, IA 2032LS (lipoxygenase-free), and Proto (high protein and antioxidant potential). Soymilk and tofu were produced using the Japanese method. The soymilk was coagulated with calcium sulfate dihydrate. Soybeans and tofu were evaluated using chemical, microbial, and instrumental sensory methods. The surface radiation of whole dry soybeans using electron beam or gamma rays at 10 or 30 kGy did provide microbial safety for the astronauts. However, these doses caused oxidative changes that resulted in tofu with rancid aroma, darkening of the tofu, lower tofu yields, more solid waste, and loss of the ability of the seeds to germinate. While lower doses may reduce these problems, we lose the ability to insure microbial safety (cross-contamination) of bulk soybeans for the astronauts. Counter measures could include vacuum packaging, radiating under freezing conditions. A No Effect Dose for food quality, below 10 kGy needs to be determined. Better estimates of the radiation that the food will be exposed to need to determined and shared. Appropriate shielding for the food as well as the astronauts needs to be developed. The Hoyt soybean did not provide a high yielding, high quality tofu. A new small scale system for evaluating soybeans was developed using 50 g quantities of soybeans.
Wilson, Lester A.↗
Embossed Teflon AF Laminate Membrane Microfluidic Diaphragm Valves
A microfluidic system has been designed to survive spaceflight and to function autonomously on the Martian surface. It manipulates microscopic quantities of liquid water and performs chemical analyses on these samples to assay for the presence of molecules associated with past or present living processes. This technology lies at the core of the Urey Instrument, which is scheduled for inclusion on the Pasteur Payload of the ESA ExoMars rover mission in 2013. Fabrication processes have been developed to make the microfabricated Teflon-AF microfluidic diaphragm pumps capable of surviving extreme temperature excursions before and after exposure to liquid water. Two glass wafers are etched with features and a continuous Teflon membrane is sandwiched between them (see figure). Single valves are constructed using this geometry. The microfabricated devices are then post processed by heating the assembled device while applying pneumatic pressure to force the Teflon diaphragm against the valve seat while it is softened. After cooling the device, the embossed membrane retains this new shape. This solves previous problems with bubble introduction into the fluid flow where deformations of the membrane at the valve seat occurred during device bonding at elevated temperatures (100-150 C). The use of laminated membranes containing commercial Teflon AF 2400 sheet sandwiched between spun Teflon AF 1600 layers performed best, and were less gas permeable than Teflon AF 1600 membranes on their own. Spinning Teflon AF 1600 solution (6 percent in FLOURINERT(Registered TradeMark) FC40 solvent, 3M Company) at 500 rpm for 1.5 seconds, followed by 1,000 rpm for 3 seconds onto Borofloat glass wafers, results in a 10-micron-thick film of extremely smooth Teflon AF. This spinning process is repeated several times on flat, blank, glass wafers in order to gradually build a thick, smooth membrane. After running this process at least five times, the wafer and Teflon coating are heated under vacuum at 220 C for one hour in order to drive off any residual solvent present in the composite film. After this, a second blank, glass wafer is brought down from above and the stack is held under vacuum at 3 atm mechanical pressure for ten 10 hours.
Willis, Peter↗
Chiral Biomarkers in Meteorites
The chirality of organic molecules with the asymmetric location of group radicals was discovered in 1848 by Louis Pasteur during his investigations of the rotation of the plane of polarization of light by crystals of sodium ammonium paratartrate. It is well established that the amino acids in proteins are exclusively Levorotary (L-aminos) and the sugars in DNA and RNA are Dextrorotary (D-sugars). This phenomenon of homochirality of biological polymers is a fundamental property of all life known on Earth. Furthermore, abiotic production mechanisms typically yield recemic mixtures (i.e. equal amounts of the two enantiomers). When amino acids were first detected in carbonaceous meteorites, it was concluded that they were racemates. This conclusion was taken as evidence that they were extraterrestrial and produced by abiologically. Subsequent studies by numerous researchers have revealed that many of the amino acids in carbonaceous meteorites exhibit a significant L-excess. The observed chirality is much greater than that produced by any currently known abiotic processes (e.g. Linearly polarized light from neutron stars; Circularly polarized ultraviolet light from faint stars; optically active quartz powders; inclusion polymerization in clay minerals; Vester-Ulbricht hypothesis of parity violations, etc.). This paper compares the measured chirality detected in the amino acids of carbonaceous meteorites with the effect of these diverse abiotic processes. IT is concluded that the levels observed are inconsistent with post-arrival biological contamination or with any of the currently known abiotic production mechanisms. However, they are consistent with ancient biological processes on the meteorite parent body. This paper will consider these chiral biomarkers in view of the detection of possible microfossils found in the Orgueil and Murchison carbonaceous meteorites. Energy dispersive x-ray spectroscopy (EDS) data obtained on these morphological biomarkers will be presented to show that the elemental compositions of these morphological biomarkers are not consistent with the compositions expected for modern (i.e. post-arrival) biological contaminants.
Hoover, Richard B.↗