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Bioreactor Development for CO2-Based In Situ Resource Utilization Manufacturing

Sustainable long-duration manned missions on both the Moon and Mars will require in situ resource utilization (ISRU). Carbon dioxide (CO2) has great potential as a harvestable resource, making up 95% of the atmosphere on Mars and being produced as respiratory waste in spacecraft and future planetary habitats. Through ISRU, biomanufacturing has the capability to produce a near limitless array of products from local space resources. Here, a CO2-based ISRU recombinant protein producing bioreactor and associated biomanufacturing organisms were designed to produce a highly stable carbonic anhydrase (CA). Candidate organisms were selected by growth characterization on acetate and formic acid, carbon substrates that are synthesized via electrochemical conversion of CO2. To improve growth on the CO2 producing substrate formic acid and for direct integration of ISRU CO2, a synthetic Calvin-Benson-Bassam cycle was designed for use in Escherichia coli. Multiplex genetic modification in E. coli was facilitated by a tailored CRISPR/Cas9 and λ red recombineering two-vector system. For expression of CA, a blue light regulated T7 promoter was employed for dynamic and small molecule free induction. Efficient bioproduction through a fed-batch exponential feeding strategy was determined via mass balance analysis from ISRU substrates to biomass and CA yield. Flux balance analysis was used to model ISRU substrate metabolism and metabolic pathway engineering in candidate organisms under cultivation strategy conditions for both metabolism reconstruction and pathway design optimization. Finally, a small-scale, disposable bag bioreactor for use in the NASA Bioculture System infrastructure was designed to enable CO2-based CA biomanufacturing in reduced-gravity environments.

biomanufacturing↗

Independent Verification of Mars-GRAM 2010 with Mars Climate Sounder Data

The Mars Global Reference Atmospheric Model (Mars-GRAM) is an engineering-level atmospheric model widely used for diverse mission and engineering applications. Applications of Mars-GRAM include systems design, performance analysis, and operations planning for aerobraking, entry, descent and landing, and aerocapture. Atmospheric influences on landing site selection and long-term mission conceptualization and development can also be addressed utilizing Mars-GRAM. Mars-GRAM's perturbation modeling capability is commonly used, in a Monte Carlo mode, to perform high-fidelity engineering end-to-end simulations for entry, descent, and landing. Mars-GRAM is an evolving software package resulting in improved accuracy and additional features. Mars-GRAM 2005 has been validated against Radio Science data, and both nadir and limb data from the Thermal Emission Spectrometer (TES). From the surface to 80 km altitude, Mars-GRAM is based on the NASA Ames Mars General Circulation Model (MGCM). Above 80 km, Mars-GRAM is based on the University of Michigan Mars Thermospheric General Circulation Model (MTGCM). The most recent release of Mars-GRAM 2010 includes an update to Fortran 90/95 and the addition of adjustment factors. These adjustment factors are applied to the input data from the MGCM and the MTGCM for the mapping year 0 user-controlled dust case. The adjustment factors are expressed as a function of height (z), latitude and areocentric solar longitude (Ls).

Justh, Hilary L.↗

Bioreactor Development for CO2-Based In Situ Resource Utilization Manufacturing

Sustainable long-duration manned missions on both the Moon and Mars will require in situ resource utilization (ISRU). Carbon dioxide (CO2) has great potential as a harvestable resource, making up 95% of the atmosphere on Mars and being produced as respiratory waste in spacecraft and future planetary habitats. Through ISRU, biomanufacturing has the capability to produce a near limitless array of products from local space resources, which include pharmaceuticals, bioplastics, chemical feedstocks, and industrial enzymes. Here, a CO2-based ISRU recombinant protein bioreactor and associated biomanufacturing organisms were designed to produce a highly stable carbonic anhydrase (CA). Initial work characterized candidate organisms for growth on acetate and formic acid, carbon substrates that can be synthesized via electrochemical conversion of CO2. To improve growth on the CO2 producing substrate formic acid and for direct integration of ISRU CO2, a synthetic Calvin-Benson-Bassam cycle was designed for use in Cyberlindnera jadinii and Escherichia coli. Genetic modifications in E. coli will be facilitated by a tailored CRISPR/Cas9 and λ red recombineering two-vector system. For expression of CA, a blue light regulated T7 promoter was employed for dynamic and small molecule free induction. Efficient bioproduction through a fed-batch exponential feeding strategy was determined via mass balance calculations from ISRU substrates to biomass and CA yield. Flux balance analysis was used to model ISRU substrate metabolism and metabolic pathway engineering in candidate organisms under cultivation strategy conditions for both metabolism reconstruction and pathway design optimization. Finally, a small-scale, disposable bag bioreactor concept for use in the NASA Bioculture System infrastructure was designed to enable CO2-based CA production in reduced-gravity environments.

Biomanufacturing, Pathway Engineering, Flux Balanc↗

Nuclear Thermal Rocket (NTR) Development Risk Communication

There are clear advantages of development of a Nuclear Thermal Rocket (NTR) for a crewed mission to Mars. NTR for in-space propulsion enables more ambitious space missions by providing high thrust at high specific impulse (approximately 900 sec) that is 2 times the best theoretical performance possible for chemical rockets. Missions can be optimized for maximum payload capability to take more payload with reduced total mass to orbit; saving cost on reduction of the number of launch vehicles needed. Or missions can be optimized to minimize trip time significantly to reduce the deep space radiation exposure to the crew. NTR propulsion technology is a game changer for space exploration. However, "NUCLEAR" is a word that is feared and vilified by some groups and the hostility towards development of any nuclear systems can meet great opposition by the public as well as from national leaders and people in authority. Communication of nuclear safety will be critical to the success of the development of the NTR. Why is there a fear of nuclear? A bomb that can level a city is a scary weapon. The first and only times the Nuclear Bomb was used in a war was on Hiroshima and Nagasaki during World War 2. The "Little Boy" atomic bomb was dropped on Hiroshima on August 6, 1945 and the "Fat Man" on Nagasaki 3 days later on August 9th. Within the first 4 months of bombings, 90- 166 thousand people died in Hiroshima and 60-80 thousand died in Nagasaki. It is important to note for comparison that over 500 thousand people died and 5 million made homeless due to strategic bombing (approximately 150 thousand tons) of Japanese cities and war assets with conventional non-nuclear weapons between 1942- 1945. A major bombing campaign of "firebombing" of Tokyo called "Operation Meetinghouse" on March 9 and 10 consisting of 334 B-29's dropped approximately1,700 tons of bombs around 16 square mile area and over 100 thousand people have been estimated to have died. The declaration of death is very clear for conventional weapons and then the declaration of death due to radiation becomes vague and unclear. This may have been due to people mis-understanding the dangers and effects of radiation when assessing the damage and harm to people initially, but it is also become insidious when expressing opposition to nuclear energy. A nuclear radiation accident can be scary due to the power involved and the fear of radiation release. The International Atomic Energy Agency defines a nuclear and radiation accident a "an event that has led to significant consequences to people, the environment or the facility." There have been 3 commercial nuclear reactor accidents (Chernobyl, Three Mile Island, and Fukushima) that stand out to the public and much of the information about the result and impact to workers, environment, and public can be misleading. Often information is presented without clear correlation with radiation and other pertinent information is left out presenting a very scary situation to affect the emotions of the reader. A very boring but "critically acclaimed" movie was made in 1979 called "The China Syndrome" starring Jane Fonda and Jack Lemmon. The film was released on March 16, 1979, 12 days before the 3-Mile Island nuclear accident in Pennsylvania. The basis from the movie was from a few nuclear plant incidents and in particular, the Brown's Ferry Alabama Power Plant fire. In one scene from the movie, a physicist Dr. Elliott Lowell played by Donald Hotton states that a China Syndrome event would make "an area the size of Pennsylvania" permanently uninhabitable. Real serious nuclear incidents like Chernobyl and Fukushima are often sited to make people fear the consequences of using nuclear power. However, the consequences are at best poorly communicated and at worst fictitiously inflated to instigate social unrest against nuclear power. There is an article being circulated on Facebook with a title "28 Signs that the (US) West Coast is being absolutely fried with nuclear radiation from Fukushima" which focus on mis-information and fear mongering. Nuclear power and NTR are powerful resources that can open many doors for future prosperity and capability. With great power comes great responsibility. Radiation and its effects need to be better understood, quantified, and communicated. A human mission to mars has its own risks of deep space radiation and is considered a considerable risk at 400 milli-Sieverts per year in deep space and 245 milli-Sieverts per year on the surface of Mars as measured by the Mars Curiosity mission. Although these quantities of ionizing radiation are within the astronaut career limit, it exceeds the yearly average amounts of ionizing radiation. Astronaut crews have experienced these levels of radiation before, but for durations shorter than a year, and a mission to Mars could possibly be 3 years in length. There is also evidence that people can comfortably handle higher levels of ionizing radiation where the radiation occurs naturally like Ramsar, Iran when people can experience 270 milli-Sieverts per year. A risk posture that the development, test, and flight of an NTR will meet opposition from groups who oppose nuclear energy must be likely and the impact can be sever to the effort. Active risk mitigation must be taken for an NTR full-scale development project. The NTR design must take into account safety for transport and off nominal conditions. Nuclear fuel element must consider containment of fission products and Low Enriched Uranium (LEU) that may meet less opposition should be considered for safety and security reasons. Even though testing was conducted on Rover/NERVA safely and successfully in the 60's with exhaust sent heavenward in to open air, modern testing of NTR must consider full containment and no release of ionizing radiation to the public and must meet the current requirement of no more than 0.1 milli-Sieverts per year to the public. 0.1 milli-Sieverts is equivalent to eating one banana or a 20 hour plane flight. Good communication with the public and regulatory agencies will be essential to show that all effort is applied toward protection to the public and astronauts. The inspiring endeavor to put humans on Mars to study the planet, search for life, and learn more about this Solar System will be full of risks but it will be worth it. NTR will be worth the development effort if it allows humans to explore our Solar System.

Kim, Tony↗

Bacillus Subtilis Engineered for Aerospace Medicine: A Platform for Off-Planet Production of Pharmaceutical Peptides

Biologics, such as pharmaceutical peptides, have notoriously short shelf lives, insufficient for long-duration space flight missions to the Moon or Mars. To enable the sustainable presence of humans on the Moon or Mars, we must develop methods for on-site production of pharmaceutical peptides in space, a concept we call the Astropharmacy. Here, we present a proof-of-concept for the first step needed: a low-mass system for pharmaceutical production designed to be stable in space. To demonstrate feasibility, we engineered strains of the space-hardy spore-forming bacterium, Bacillus subtilis, to secrete two pharmaceutical peptides important for astronaut health: teriparatide (an anabolic agent for combating osteoporosis) and filgrastim (an effective countermeasure for radiation-induced neutropenia). We found that the secretion peptides from the walM and yoqH genes of B. subtilis worked well for secreting teriparatide and filgrastim, respectively. In consideration of the Translational Research Institute for Space Health (TRISH) challenge to produce a dose equivalent in 24 h, dried spores of our engineered strains were used to produce 1 dose equivalent of teriparatide from a 2 mL culture and 1 dose equivalent of filgrastim from 52 mL of culture in 24 h. Further optimization of strain growth conditions, expression conditions, and promoter sequences should allow for higher production rates to be achieved. These strains provide the template for future optimization efforts and address the first step in the Astropharmacy, capable of on-site production, purification, and processing of biopharmaceutical compounds in platforms amenable for use in space.

Alec Vallota-Eastman↗

Metabolomic and Genomic Markers of Atherosclerosis as Related to Oxidative Stress, Inflammation, and Vascular Function in Twin Astronauts

Background: Future human space travel will consist primarily of long-duration missions onboard the International Space Station (ISS) or exploration-class missions to Mars, its moons, or nearby asteroids. Astronauts participating in long-duration missions may be at an increased risk of oxidative stress and inflammatory damage due to radiation, psychological stress, altered physical activity, nutritional insufficiency, and hyperoxia during extravehicular activity. By studying one identical twin during his 1-year ISS mission and one ground-based twin, this work extends a current NASA-funded investigation to determine whether these spaceflight factors contribute to an accelerated progression of atherosclerosis. This study of twins affords a unique opportunity to examine the spaceflight-related atherosclerosis risk independent of the confounding factors associated with different genotypes. Purpose: The purpose of this investigation is to determine whether biomarkers of oxidative and inflammatory stress are elevated during and after long-duration spaceflight and determine if a relation exists between levels of these biomarkers and structural and functional indices of atherosclerotic risk measured in the carotid and brachial arteries. These physiological and biochemical data will be extended by using an exploratory approach to investigate the relationship between intermediate phenotypes and risk factors for atherosclerosis and the metabolomic signature from plasma and urine samples. Since metabolites are often the indirect products of gene expression, we will simultaneously assess gene expression and DNA methylation in leukocytes. Hypothesis: We predict that the space-flown twin will experience elevated biomarkers of oxidative stress and inflammatory damage, altered arterial structure and function, accelerated telomere shortening, dysregulation of genes associated with oxidative stress and inflammation, and a metabolic profile shift that is associated with elevated atherosclerosis risk factors. Conversely, these will not be observed in the ground-based twin. Methods: We will measure blood and urine biomarkers of oxidative stress and inflammation as well as arterial structure and function (carotid intima-medial thickness and brachial artery flow-mediated dilation) in one twin astronaut before, during, and after long-duration spaceflight and in his twin serving as a ground-based control. Furthermore, we will measure metabolomics (targeted and untargeted approaches) and genomic markers (DNA methylation, mRNA gene expression, telomere length) to elucidate the molecular mechanisms involved. A panel of biomarkers of oxidative and inflammatory stress will be measured in venous blood samples and 24-hour (in-flight) and 48-hour (pre- and post-flight) urine pools twice before flight, early (flight days 15 and 60) and late (2 weeks before landing) during the mission, and early in the post-flight recovery phase (approximately 3-5 days after landing). Arterial structure, assessed from measures of intima-media thickness, will be measured at the same times. Arterial function will be assessed using brachial flow-mediated dilation, a well-validated measure used to assess endothelium-dependent vasodilation and a sensitive predictor of atherosclerotic risk, only before and after spaceflight. Discussion: Pre- and in-flight data collection is in progress for the space-flown twin, and similar data have been obtained from the ground-based twin. Blood and urine samples will be batch processed when received from ISS after the conclusion of the 1-year mission. Results from these individual subjects will be compared to the larger complement of subjects participating in the companion study currently ongoing in ISS astronauts.

Lee, Stuart M. C.↗

Metabolomic and Genomic Markers of Atherosclerosis as Related to Oxidative Stress, Inflammation, and Vascular Function in Twin Astronauts

BACKGROUND: Future human space travel will consist primarily of long-duration missions onboard the International Space Station (ISS) or exploration-class missions to Mars, its moons, or nearby asteroids. Astronauts participating in long-duration missions may be at an increased risk of oxidative stress and inflammatory damage due to radiation, psychological stress, altered physical activity, nutritional insufficiency, and hyperoxia during extravehicular activity. By studying one identical twin during his 1-year ISS mission and his ground-based twin, this work extends a current NASA-funded investigation to determine whether these spaceflight factors contribute to an accelerated progression of atherosclerosis. This study of twins affords a unique opportunity to examine spaceflight-related atherosclerosis risk that is independent of the confounding factors associated with different genotypes. PURPOSE: The purpose of this investigation was to determine whether biomarkers of oxidative and inflammatory stress are elevated during and after long-duration spaceflight and determine if a relation exists between levels of these biomarkers and structural and functional indices of atherosclerotic risk measured in the carotid and brachial arteries. These physiological and biochemical data will be extended by using an exploratory approach to investigate the relationship between intermediate phenotypes and risk factors for atherosclerosis and the metabolomic signature from plasma and urine samples. Since metabolites are often the indirect products of gene expression, we simultaneously assessed gene expression and DNA methylation in leukocytes. HYPOTHESIS: We predict that, compared to the ground-based twin, the space-flown twin will experience elevated biomarkers of oxidative stress and inflammatory damage, altered arterial structure and function, accelerated telomere shortening, dysregulation of genes associated with oxidative stress and inflammation, and a metabolic profile shift that is associated with elevated atherosclerosis risk factors. METHODS: In the space-flown twin, a panel of biomarkers of oxidative and inflammatory stress were measured in venous blood samples and in 24-h (in-flight) and 48-h (pre- and post-flight) urine pools collected twice before flight, six times during the mission (~FD15, 75, 180, 240, 300, 335), and early in the post-flight recovery phase (3-5 days after landing). We also measured metabolomic (targeted and untargeted approaches) and genomic markers (DNA methylation, mRNA gene expression, telomere length) in these samples. Arterial structure, assessed from measures of intima-media thickness, also were measured using standard clinical ultrasound at the same time points. Arterial function was assessed using brachial flow-mediated dilation, a well-validated measure used to assess endothelium-dependent vasodilation and a sensitive predictor of atherosclerotic risk, only before and after spaceflight. All of the same measures were obtained in the ground-based twin, but less frequently. DISCUSSION: All data collection has been completed for both the space-flown twin and the ground-based twin. Vascular structure and function measures have been analyzed, blood and urine samples have been batch-processed. Results from these individuals will be compared to each other, to data from other Twin Study investigations, and to the larger complement of subjects participating in the companion study currently ongoing in ISS astronauts.

Lee, Stuart M. C.↗

Drosophila Habitat Developed to Support Research on the International Space Station

The Fruit Fly Lab is a hardware suite being designed to support research on the International Space Station (ISS) for use by the entire Drosophila research community. A validation mission will launch and return on SpaceX-5 in late 2014, followed by Principal Investigator-lead science flights thereafter. Space flight experiments are selected via peer-reviewed proposals open to the Drosophila community. The cassettes (containers) that will house the Drosophila cultures were successfully used to conduct an immunity study on the Space Shuttle. Results showed that the innate immune system of Drosophila melanogaster was affected by space flight with a reduction in phagocytosis function of plasmatocytes, changes in antimicrobial peptides and other gene expression levels, as well as changes in development of the animals. Scientific research topics that are of interest to NASA will be presented. Each cassette used to house the Drosophila has a removable food tray that can be replaced to sustain the growth of the culture, or can be transferred to another cassette, along with embryos and burrowed larvae, enabling multi-generational studies. The cassette can be frozen in the Minus Eighty Laboratory Freezer for ISS to preserve samples until post-flight analysis, expanding the applications of the hardware. Utilization of a centrifuge allows for on-orbit 1g controls for microgravity experiments, as well as variable g-levels for lunar or Mars environment studies. The standard form factor used also allows for implementation of modular upgrades. An observation system, circadian rhythm lighting system, and fixation capability are upgrades currently in development for near-term implementation. This hardware suite, with its flight- proven design and ability to utilize existing on-board facilities, offers the whole Drosophila research community a platform to address several key areas of the National Research Councils decadal survey, supporting the utilization of ISS for science discovery.

Drosophila↗

Observations Regarding Small Eolian Dunes and Large Ripples on Mars

Eolian bedforms occur at the interface between a planetary surface and its atmosphere; they present a proxy record of the influence of climate, expressed in sediment transport, over that surface. High resolution images (1.5 - 12 m/pixel) from the Mars Global Surveyor (MGS) Mars Orbiter Camera provide glimpses of the most recent events shaping the martian landscape. Thousands of images exhibit small transverse dunes or large eolian ripples that have crest-to-crest spacings of 10 to 60 m, heights of a few to 10 m. Bedforms of the size and patterns seen in the Mars photographs are rarely described among Earth's eolian landforms; in terms of size and morphology, most of these fall between traditional definitions of "ripples" and "dunes". Dunes are composed chiefly of materials transported by saltation, ripples are smaller forms moved along by the impact of saltating grains (traction). The largest reported eolian ripples on Earth (granule ripples, megaripples) are typically smaller than the bedforms observed on Mars; likewise, most dunes are typically larger. The small dunes and large ripples on Mars come in a variety of relative albedos, despite an early MGS impression that they are all of high albedo. Some ripples occur on the surfaces of sand dunes; these are most likely true granule ripples. However, most of these bedforms occur in troughs, pits, craters, and on deflated plains. Despite impressions early in the MGS mission, they do not occur everywhere (e.g., they are rare on the northern plains) but they do occur at a range of elevations from the highest volcanoes to the deepest basins. Where they occur on a hard substrate among larger sand dunes, the big dunes have over-ridden the smaller bedforms, indicating that the smaller features are older and perhaps indurated or very coarse-grained. At other locales, the small bedforms have been mantled by material settled from suspension, in other cases they are being exhumed and may be lithified. Still other examples are peppered with small impact craters, implying considerable age. These bedforms present a complicated record of the geologically-recent past, one that has involved changes in climate, sediment transport capabilities, and sediment sources and sinks over time.

Edgett, Kenneth S.↗

So What's an RTG and Are They Safe?

When one considers space missions to the outer edges of our solar system and far beyond, our sun cannot be relied on to produce the required spacecraft (s/c) power. Solar energy diminishes as the square of the distance from the Sun. At Mars it is only 43% of that at earth. At Jupiter, it falls off to only 3.6% of Earth's. By the time we get out to Pluto, solar energy is only .066% what it is on Earth. Beyond the orbit of Mars, it is not practical to depend on solar power for a s/c. However, the farther out we go the more power we need to heat the s/c and to transmit data back to Earth over the long distances. On Earth, knowledge is power. In the outer solar system, power is knowledge. Solar arrays only operate at 19% efficiency, are very vulnerable to damage from radiation and temperature extremes, and cannot be used for even nearby missions that operate in extended darkness, or under the surface of a planet or moon. Twenty-six U.S. space missions, from the Transit to Cassini, have used radioisotope power systems and heater units to take s/c to the far reaches of our solar system and have demonstrated an outstanding record of safety and reliability. Radioisotope thermoelectric generators (RTG's) have proven to be safe, reliable, maintenance-free, and capable of providing both thermal and electrical power for decades under the harsh environments of deep space. RTG's have no problem operating in the high radiation belts of space, the extreme temperatures, or the severe dust storms of Mars, and they have proven to be the most reliable power source ever flown on U.S. s/c. For example, the two Pioneer s/c operated for more than two decades and the Voyager s/c may last for 40 years. RTG's are not nuclear reactors, they serve only as power generators and are not involved in the propulsion of the s/c. They operate on the principle of thermoelectric generation that converts heat directly into electricity, they have no moving parts, are extremely reliable, and have met or exceeded all safety and performance expectations. Federal laws and regulations require analysis and evaluation of the safety risks and any potential environmental impacts. Extensive safety testing of RTG's and RTG components has been performed by the U.S. Department of Energy (DOE) to demonstrate the ability to survive accidents related to Space Shuttle launches and assure that the systems would be safe under all accident conditions, including accidents at or near the launch pad or during orbital reentry. Many design improvements have been made over the four decades that RTG's have been flown on space missions. This paper outlines the operation and safety standards of RTG's and the advanced developments expected to be used on future deep space missions such as the Europa Orbiter, Pluto/Kuiper Express, Solar Probe, Europa Lander, and Titan Explorer missions.

Barret, Chris↗

MOLA-derived Roughness Data Used to Predict Surface Scattering for Mars Subsurface Radar Sounding

The Mars Express orbiter, to be launched by the European Space Agency in 2003, will carry a low-frequency radar sounding instrument, MARSIS (Mars Advanced Radar for Subsurface and Ionospheric Sounding). The primary goal of MARSIS is to map the distribution of water, both solid and (if present) liquid, in the upper several km of the martian crust. Detecting discontinuities in the crust, such as an ice-water transition, presents many challenges for a Mars orbital radar sounder. One challenge that must be overcome is the presence of radar scattering (echoes) from the surface of Mars, expected to be detected by the sounder antennas at the same time as any echoes aris-ing from subsurface interfaces. As the transmitted spherical wavefront spreads within the crust of Mars,. it also interacts with surface topography at off-nadir positions, creating a "clutter" signal that can mask the subsurface echoes. The MARSIS instrument will utilize Doppler filtering to limit the off-nadir clutter in the along-track direction, and a nadir-null secondary antenna to identify strong off-nadir clutter from the cross-track direction. To evaluate the effects of off-nadir surface clutter and the capability of these schemes to reduce the clutter, it is necessary to predict the range of scattering behavior that may be expected from martian surface topography. In this paper, we utilize Mars Orbital Laser Altimeter (MOLA) data from the current Mars Global Surveyor mission to characterize the topographic roughness of a variety of martian terrain types, at scales relevant to the MARSIS clutter problem. Segments of MOLA altimetry profiles are reduced to the topographic parameters rms slope and fractal dimension, which then are used as inputs to a near-nadir radar scattering model to predict the strength of the clutter signal. Additional information is contained in the original extended abstract.

Plaut, J. J.↗

The Role of Cis-Lunar Space in Future Global Space Exploration

Cis-lunar space offers affordable near-term opportunities to help pave the way for future global human exploration of deep space, acting as a bridge between present missions and future deep space missions. While missions in cis-lunar space have value unto themselves, they can also play an important role in enabling and reducing risk for future human missions to the Moon, Near-Earth Asteroids (NEAs), Mars, and other deep space destinations. The Cis-Lunar Destination Team of NASA's Human Spaceflight Architecture Team (HAT) has been analyzing cis-lunar destination activities and developing notional missions (or "destination Design Reference Missions" [DRMs]) for cis-lunar locations to inform roadmap and architecture development, transportation and destination elements definition, operations, and strategic knowledge gaps. The cis-lunar domain is defined as that area of deep space under the gravitational influence of the earth-moon system. This includes a set of earth-centered orbital locations in low earth orbit (LEO), geosynchronous earth orbit (GEO), highly elliptical and high earth orbits (HEO), earth-moon libration or "Lagrange" points (E-ML1 through E-ML5, and in particular, E-ML1 and E-ML2), and low lunar orbit (LLO). To help explore this large possibility space, we developed a set of high level cis-lunar mission concepts in the form of a large mission tree, defined primarily by mission duration, pre-deployment, type of mission, and location. The mission tree has provided an overall analytical context and has helped in developing more detailed design reference missions that are then intended to inform capabilities, operations, and architectures. With the mission tree as context, we will describe two destination DRMs to LEO and GEO, based on present human space exploration architectural considerations, as well as our recent work on defining mission activities that could be conducted with an EML1 or EML2 facility, the latter of which will be an emphasis of this paper, motivated in part by recent interest expressed at the Global Exploration Roadmap Stakeholder meeting. This paper will also explore the links between this HAT Cis-Lunar Destination Team analysis and the recently released ISECG Global Exploration Roadmap and other potential international considerations, such as preventing harmful interference to radio astronomy observations in the shielded zone of the moon.

Bobskill, Marianne R.↗

Science Goals in Radiation Protection for Exploration

Space radiation presents major challenges to future missions to the Earth s moon or Mars. Health risks of concern include cancer, degenerative and performance risks to the central nervous system, heart and lens, and the acute radiation syndromes. The galactic cosmic rays (GCR) contain high energy and charge (HZE) nuclei, which have been shown to cause qualitatively distinct biological damage compared to terresterial radiation, such as X-rays or gamma-rays, causing risk estimates to be highly uncertain. The biological effects of solar particle events (SPE) are similar to terresterial radiation except for their biological dose-rate modifiers; however the onset and size of SPEs are difficult to predict. The high energies of GCR reduce the effectiveness of shielding, while SPE s can be shielded however the current gap in radiobiological knowledge hinders optimization. Methods used to project risks on Earth must be modified because of the large uncertainties in projecting health risks from space radiation, and thus impact mission requirements and costs. We describe NASA s unique approach to radiation safety that applies probabilistic risk assessments and uncertainty based criteria within the occupational health program for astronauts and to mission design. The two terrestrial criteria of a point estimate of maximum acceptable level of risk and application of the principle of As Low As Reasonably Achievable (ALARA) are supplemented by a third requirement that protects against risk projection uncertainties using the upper 95% confidence level (CL) in radiation risk projection models. Exploration science goals in radiation protection are centered on ground-based research to achieve the necessary biological knowledge, and in the development of new technologies to improve SPE monitoring and optimize shielding. Radiobiology research is centered on a ground based program investigating the radiobiology of high-energy protons and HZE nuclei at the NASA Space Radiation Laboratory (NSRL) located at DoE s Brookhaven National Laboratory in Upton, NY. We describe recent NSRL results that are closing the knowledge gap in HZE radiobiology and improving exploration risk estimates. Linking probabilistic risk assessment to research goals makes it possible to express risk management objectives in terms of quantitative metrics, which include the number of days in space without exceeding a given risk level within well defined confidence limits, and probabilistic assessments of the effectiveness of design trade spaces such as material type, mass, solar cycle, crew selection criteria, and biological countermeasures. New research in SPE alert and risk assessment, individual radiation sensitivity, and biological countermeasure development are described.

Cucinotta, Francs A.↗

Bionutrients-1: Utilizing Genomics and Transcriptomics to Assess the Reliability of Microorganisms for In Situ Nutrient Production on Long Duration Missions

The resupply of current long-duration crewed missions to the ISS relies on ground-launched supplies. As NASA looks toward Mars, ground-based resupply will no longer be an option. Critical nutrients, including vitamin C, vitamin K, folate, and thiamin, degrade during long-term storage, and regular consumption of these nutrients is essential for astronaut health. Another challenge of current food systems is the difficulty of consuming sufficient calories when subsisting on the limited flavors of freeze-dried food, which can lead to weight loss. The inclusion of microorganism-based food systems could alleviate both concerns. For example, the fermentation of rehydrated milk into yogurt with microorganisms genetically incorporating genes to produce critical vitamins would allow for both in situ production of nutrients and a fresh food product with additional flavor profiles. In comparison to plant food production, microorganisms require less flight infrastructure. The BioNutrients-1 mission is demonstrating viability of microbial fermentation food production in microgravity and testing the reliability of this approach for long-duration missions lacking resupply. While the BioNutrients-1 mission includes the collection of multiple phenotypic measurements, this status update will focus on the processing of samples for genomics and transcriptomics analyses as well as the planned analysis pipelines. First, the BioNutrients-1 mission seeks to identify microorganisms capable of surviving long-duration storage at ambient temperatures while maintaining genetic fidelity. To achieve this, nine commonly employed microbial species were stored at ambient temperatures in Stasis Packs on the ISS for five years. The viability and mutation rates will be measured at multiple time points for both flown and ground control samples. From an omics perspective, the changes in the bulk rates of point mutations and genetic rearrangements across the Stasis Pack species during the five years of storage will be determined, providing valuable insights into the potential of these microorganisms for long-duration space missions. Second, the BioNutrients-1 mission is characterizing the impact of microgravity on fermentation. Two strains of the yeast Saccharomyces cerevisiae, each encoding antioxidants (β-carotene or zeaxanthin) were flown to ISS for storage and fermentation within simplified bioreactors (Production Packs). The impact of microgravity on the expression of the antioxidant production genes and general metabolic genes will be determined using RNA sequencing. Ultimately, the transcriptome data will be compared to phenotypic measurements, such as the antioxidant yield, end-state biomass, and the production of EtOH, to determine the impacts of microgravity and long-term storage on microbial fermentation. The findings from this research will be instrumental in understanding the challenges and opportunities of microorganism-based food systems in space missions.

BioNutrients↗

Venus Global Reference Atmospheric Model (Venus-GRAM) Upgrades

Introduction: The Venus Global Reference Atmospheric Model (Venus-GRAM) is one of the most widely used engineering models of Venus’ atmosphere. The Venus-GRAM upgrades are being developed by NASA Marshall Space Flight Center (MSFC) and NASA Langley Research Center (LaRC). This presentation will provide details regarding the upgrades that have been made to Venus-GRAM and the ongoing objectives, tasks, and milestones related to the GRAM upgrades funded by the NASA Science Mission Directorate (SMD). Venus-GRAM: Venus-GRAM is an engineering-oriented atmospheric model that estimates mean values and statistical variations of the atmospheric properties of Venus. Venus-GRAM provides mean values and variability for any point in the atmosphere as well as seasonal, geographic, and altitude variations. Venus-GRAM outputs include atmospheric density, temperature, pressure, winds, and chemical composition along a user-defined path. It is extensively used by the engineering community because of its ability to create realistic dispersions. GRAMs have been integrated into high fidelity flight dynamic simulations of launch, entry, descent and landing (EDL), aerobraking and aerocapture. GRAMs are currently available for Earth, Mars, Venus, Neptune, Titan, and Uranus. The lower atmosphere model in Venus-GRAM (up to 250 km) is based on the Venus International Reference Atmosphere (VIRA) [1]. The Venus-GRAM thermosphere (250 to 1000 km) is based on a MSFC-developed model [2] which assumes an isothermal temperature profile initialized using VIRA conditions at 250 km [3]. The VIRA version included in Venus-GRAM includes Pioneer Venus Orbiter and Probe data as well as Venera probe data, but it does not include a solid planet model or a high-resolution gravity model [4]. Venus-GRAM Upgrade Status: Code Modernization. Venus-GRAM has been rearchitected from Fortran to a common object-oriented C++ framework called the GRAM Suite. This new architecture creates a common GRAM library of data models and utilities. The first C++ release of the rearchitected Venus-GRAM is a straight conversion from the latest Fortran version. Model Upgrades. The focus of the model upgrade task is to improve the atmosphere models in the existing GRAMs and to establish a foundation for developing GRAMs for additional destinations. The GRAM ephemeris has been upgraded to the NASA Navigation and Ancillary Information Facility (NAIF) SPICE toolkit (version N0066). The calculation of the speed of sound has also been improved in the GRAMs. In FY20, the GRAM project established contracts to improve the model data within Venus-GRAM. Hampton University is developing an empirical global model for Venus. The University of Wisconsin is reanalyzing the Venus Express radio occultation observations and analyzing the Akatsuki thermal imaging data. Upgraded Venus-GRAM Release. GRAM Suite Version 1.3 will be released in September 2021 and will contain the rearchitected Venus-GRAM, including the common GRAM framework and planet–specific code. A User Guide and Programmer’s Manual are released with all GRAMs. Conclusions: GRAMs are frequently used toolsets and vital in assessing effects of atmospheres on interplanetary spacecraft during the program life cycle process. Releases of the GRAM Suite, upgrades of the existing planetary GRAMs, and development of new planetary GRAMs are ongoing. Venus-GRAM atmosphere model upgrades will be included in the next phase of GRAM tasks. References: [1] Kliore, A. J. et al. (1985) ASR, 5, 11, 1-304. [2] Justh, H. L. et al. (2006) AIAA/AAS Astrodynamics Specialist Conference & Exhibit, Abstract AIAA-2006-6394. [3] Guide to Reference and Standard Atmosphere Models, BSR/AIAA G-003-2010. [4] Limaye, S. S. (2012), LPSC VEXAG Townhall Meeting. Acknowledgments: The authors gratefully acknowledge support from the NASA SMD.

atmospheric models↗

Global Reference Atmospheric Model (GRAM) Advancements and Additions

Introduction:The Global Reference Atmospheric Model (GRAM) is one of the most widely used engineering models of the atmosphere. GRAM development and maintenance has been led by NASA Marshall Space Flight Center (MSFC). The NASA Science Mission Directorate (SMD) has provided funding support to upgrade the GRAMs since Fiscal Year 2018. NASA Langley Research Center has been working with MSFC on the upgrades.This presentation will provide details regarding the upgrades that have been made to the existing GRAMs, the development of new GRAMs, as well as the ongoing objectives, tasks, and milestones re-lated to the GRAM upgrades funded by NASA SMD. GRAM: The GRAMs are engineering-oriented atmospheric models that estimate mean values and statistical variations of the atmospheric properties for numerous planetary destinations.They provide mean values and variability for any point in the atmosphere as well as seasonal, geographic, and altitude variations. GRAM outputs include atmospheric density, temperature, pressure, winds, and chemical composition along a user-defined path.Theyhave been widely used by the engineering community because of their ability to create realistic dispersions. GRAMs have been integrated into high fidelity flight dynamic simulations of launch, entry, descent and landing (EDL), aerobraking and aerocapture. MSFC has been developing and updating GRAMs since 1974; GRAMs are currently available for Earth, Mars, Venus, Neptune, and Titan. GRAM Upgrade Status: Code Moderization.The planetary GRAMs are being rearchitected from Fortran to a common object-ori-entedC++ framework called the GRAM Suite. This new architecture creates a common GRAM library of data models and utilities. The first C++ releases of the existing planetary GRAMs (Mars, Venus, Neptune, and Titan-GRAM) in the GRAM Suite are straight conversions from the latest Fortran version. Model Upgrades.The focus of the model upgrade task is to improve the atmosphere models in the existing GRAMs and to establish a foundation for developing GRAMs for additional destinations. The GRAM ephemerishas been upgraded to the NASA Navigation and Ancillary Information Facility (NAIF) SPICE toolkit (version N0066). The calculation of the speed of sound has also been improved in the GRAMs. The GRAM team has received updated Mars General Circulation Model (MGCM) datasets from NASA Ames Research Center. Mars Global Ionosphere-Thermosphere Model (M-GITM) data is being obtained toreplace the Mars Thermospheric General Circulation Model (MTGCM) data in legacy Mars-GRAM. M-GITM and updated MGCM data will be incorporated into a future GRAM Suite release.Twoprojects that will improve the atmospheric model data in the GRAMs have been funded by the GRAM team since Fiscal Year 2020. Sanjay Limaye and Patrick Fry at the University of Wisconsin are reanalyzing the Venus Express radio occultation observations and analyzing the Akatsukiradio occultation observations.This will lead tothe calculation of number density, temperature, and pressure profiles for the 40-90 km altitude range. Kunio Sayanagi, Justin Garland, and Ryan McCabeat Hampton University are developing empirical global models for Venus,Jupiter, Saturn, Uranus, Neptune, and Titan that incorporates the latest data available for each of these planetary destinations. Upgraded GRAM Releases. GRAM Suite Version 1.0 was released in May 2020 and contains the rearchitected Neptune-GRAM, including the common GRAM framework and planet–specific code. GRAM Suite Version 1.1 was released in September 2020 and addedthe rearchitected Titan-GRAM to the GRAM Suite. A User Guide and Programmer’s Manualarereleased with all GRAMs. The rearchitected Mars and Venus-GRAMs will be released in upcomingversions of the GRAM Suite. New GRAM Releases. New GRAMs have been de-veloped for Uranus and Jupiter. Uranus-GRAM is based on an individual profile generated by Gary Allen (ARC) from Voyager 2 occultation data and will be released in GRAM Suite Version 1.2. Jupiter-GRAM is based on individual profile produced from Al Seiff’s Ju-piter model[1].Jupiter-GRAM will be released in GRAM Suite Version 1.3. Saturn-GRAM is currently under development and will be released in a future version of the GRAM Suite. Conclusions: GRAMs are vital and frequently used toolsets. Releases of the GRAM Suite, upgrades of the existing planetary GRAMs, and development of new planetary GRAMs are ongoing. NASA SMD funding has been essential to addressing current limitations and accomplishing GRAM developmental goals. Continua-tion ofSMD fundingwill ensure the development, up-grades, and maintenance of the GRAMs. References: [1] Seiff, A., et al. (1998) JGR, 103, 22,857-22,889. Acknowledgments: The authors gratefully acknowledge support from the NASA SMD

atmospheric models↗

Stirling Engine Controller

Stirling technology is being developed to replace RTG s (Radioisotope Thermoelectric Generators), more specifically a stirling convertor, which is a stirling engine coupled to a linear alternator. Over the past three decades, the stirling engine has been designed to perform different functions. Stirling convertors have been designed to decrease fuel consumption in automobiles. They have also been designed for terrestrial and space applications. Currently NASA Glenn is using the convertor for space based applications. A stiring converter is a better means of power for deep space mission and "dusty" mission, like the Mars Rovers, than solar panels because it is not affected by dust. Spirit and Opportunity, two Mars rovers currently navigating the planet, are losing their ability to generate electricity because dust is collecting on their solar panels. Opportunity is losing more energy because its robotic arm has a heater with a switch that can not be turned off. The heater is not needed at night, but yet still runs. This generates a greater loss of electricity and in turn diminishes the performance of the rover. The stirling cycle has the potential to provide very efficient conversion of heat energy to electric a1 energy, more so than RTG's. The stirling engine converts the thermal energy produced by the decaying radioisotope to mechanical energy; the linear alternator converts this into electricity. convertor. Since the early 1990's tests have been performed to maximize the efficiency of the stirling converter. Many months, even years, are dedicated to preparing and performing tests. Currently, two stirling convertors #'s 13 and 14, which were developed by Stirling Technology Company, are on an extended operation test. As of June 7th, the two convertors reached 7,500 hours each of operation. Before the convertors could run unattended, many safety precautions had to be examined. So, special instrumentation and circuits were developed to detect off nominal conditions and also safely shutdown the engines. The test will last for a period of 8000 to 9000 hours. Other types of tests that have been performed are: performance mapping, controller development, launch environment, and vibration emissions testing. Currently, the thermo-mechanical system branch is housing a RG-350, a stirling convertor. The convertor was used in previous tests such as a Hall Thruster test, world s first integrated test of a dynamic power system with electric propulsion. Another test performed was to conclude if free piston stirling convertors can be synchronized for vibration balancing, with no thermodynamic or electrical connections and not cause both to shutdown if one failed. The ability to reduce vibration by synchronizing convertor operation but still be able to operate when one partner fails is pertinent in space and terrestrial applications. The convertor is now being brought back into operation and a controller is in the process of being developed. This convertor will be used as a testbed for new controllers. I worked with Mary Ellen Roth on the electric engineering aspects of the RG-350. My main goal was to enhance the data collection process. I worked on different aspects of the RG-350, with a main focus on the engine controller. I drew a schematic of the wire connections in the engine controller, using PCB Express, so that a plan could be devised to connect the power meter properly between the output of the engine and the engine controller. I measured the power using two different instruments: Valhalla Scientific power meter and Ohio Semitronics power measurement device. The convertor is connected to an Agilent 34970A Data Acquisition/Switch Unit, which allows the user to measure, record, and monitor voltage, current, frequency, and temperature. I assisted in preparing the Data Acquisition for general operation. I also helped test a panel of transducers, which will be placed in the rack that powers and monitors the convertor.

Blaze, Gina M.↗

Exploration Atmosphere Demonstration of A Multi-Functional Integrated Medical Device (Tempus Pro)

INTRODUCTION The Exploration Medical Capability (ExMC) element within the Human Research Program and the Exploration Medical Integrated Product Team (XMIPT) within the Mars Campaign Office seek to advance medical system design and risk-informed decision-making for exploration missions. This includes assessment of candidate devices and their compatibility within a variety of increasingly Earth-independent medical scenarios. The Tempus Pro (Remote Diagnostic Technologies, Ltd., Philips Corp., Farnborough, UK) is a commercial-off-the-shelf (COTS), multi-functional integrated medical device capable of vital signs monitoring, with built-in procedure support (iAssist), patient record and telemedicine communication features, and medical imaging (e.g., camera, laryngoscope, ultrasound) that meets multiple exploration medical capability needs. One of several hypobaric atmospheres being considered for exploration vehicles is 8.2 psi with 34% oxygen. To assess the useability of the Tempus Pro in such environments by individuals without formal medical training, the unit was tested in May and June of 2023 at the Johnson Space Center 20-Foot Exploration Atmosphere Chamber in conjunction with the Exploration Atmospheres-2 (EA-2) study. TECHNOLOGY DEMONSTRATION Eight subjects in the EA-2 study were assigned roles as the caregiver or patient – or both in the case of a self-exam – and caregivers were asked to place sensors for 3-Lead ECG, non-invasive blood pressure (NIBP), pulse oximetry (SpO2), and temperature using Tempus Pro’s iAssist. Depending on the procedure, additional tasks included an oropharyngeal exam of the throat with the laryngoscope and capturing ultrasound images of a cardiac subxiphoid view or of the user’s choice from a pre-specified list. Scenarios ranged from remote-guided to fully autonomous operations and surveyed the effectiveness of support, amount of support desired, and importance of support by type (e.g., written crew procedures, iAssist and device guidance, remote console guidance, etc.). Feedback from the subjects and the support personnel was also gathered to inform future designs for training and support and to determine what operations are plausible given different levels of each. An unweighted NASA task load index (TLX) was used to profile the demands of using the device, however the sample size does not support statistics. The research was exploratory in nature and qualitative information was the main goal. Calibration checks on the Tempus Pro were conducted before and after chamber testing to ensure the device was in useable condition. RESULTS All subjects performed their tasks successfully and found the Tempus Pro easy to use with the support provided. The calibration checks outside the chamber showed that the Tempus Pro remained unchanged and measurements inside the chamber were within normal values. Caregivers taking the NASA TLX reported mean scores ≤ 41/100 showing the tasks to be undemanding and less demanding with repeated use (~20/100). Novice users were able to easily connect sensors and take vital signs with the guidance from Tempus Pro’s iAssist feature. For more complex tasks, such as the oropharyngeal exam and ultrasound image acquisition, guidance beyond iAssist was needed, primarily from a supporting physician. Feedback on the importance of support types varied by person and scenario but greater than 50% of the support used by each was non-native to the device. Overall opinions were positive, but the ultrasound users expressed a lack of confidence in their results and 3 out of 4 desired more time, training, or support. Subjects found the sensors to be comfortable and comparable to prior experiences with such devices, except for the blood pressure cuff, which squeezed too tightly for uncomfortably long periods. Many observations provoked discussion, especially regarding ultrasound, that will aide decisions about future demonstrations.

R. S. Miller↗