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NASA NTRSDate not supplied

BioLEAD: BioLogical Exploration via Autonomous Detection – Using Dielectric Spectroscopy to Monitor Biological Changes in Response to Deep Space Radiation

Leveraging the bio-fluidic hardware developed for BioSentinel, we propose a new payload for autonomous missions: BioLEAD – BioLogical Exploration via Autonomous Detection. With NASA's renewed focus to continue human exploration of the Moon (Artemis Program), the BioLEAD payload aims to investigate the effects of the lunar environment on biology, whether onboard a lunar lander, the Lunar Gateway, or as a free-flying CubeSat orbiting the Moon. In place of an optical detection system, BioLEAD will employ a miniaturized, non-invasive dielectric spectroscopy sensor to enable real-time monitoring of biological activity. The sensor operates by relating capacitance measurements to the dielectric properties of the cell, such as cell morphology, doubling time, and cell cycle stage. The implementation of this new sensor technology will address limitations of the optical measurement system used on BioSentinel. It will also advance the use of autonomous bioanalytical microsystems and reveal new information regarding biological responses to the Moon’s radiation environment. Most importantly, BioLEAD’s technology will be adaptable for a wide array of future missions.

Biosensor↗

NASA NTRS2003

Mechanical injury and repair of cells

OBJECTIVE: To concisely review the field of cell plasma membrane disruption (torn cell surface) and repair. MAIN POINTS: Plasma membrane disruption is a common form of cell injury under physiologic conditions, after trauma, in certain muscular dystrophies, and during certain forms of clinical intervention. Rapid repair of a disruption is essential to cell survival and involves a complex and active cell response that includes membrane fusion and cytoskeletal activation. Tissues, such as cardiac and skeletal muscle, adapt to a disruption injury by hypertrophying. Cells adapt by increasing the efficiency of their resealing response. CONCLUSION: Plasma membrane disruption is an important cellular event in both health and disease. The disruption repair mechanism is now well understood at the cellular level, but much remains to be learned at the molecular level. Cell and tissue level adaptational responses to the disruption either prevent its further occurrence or facilitate future repairs. Therapeutically useful drugs might result if, using this accumulating knowledge, chemical agents can be developed that can enhance repair or adaptive responses.

Review, Tutorial↗

NASA NTRS1996

On the search for extant life on Mars

Proposals for continuing the search for extant life on Mars are primarily predicated on the assumption that specialized environmental niches that could support a biota may exist on the planet. Before attempting any critical tests for extant organisms, either in situ or on returned samples, it is imperative to determine whether any such sites actually exist. If, through remote sensing and landed instrumentation, sites of potential biological interest are discovered and characterized, biological tests can then more effectively be planned to elicit the presence of organisms that are adapted to living in these particular environments.

NASA Discipline Exobiology↗

NASA NTRS1987

Photosynthetic action spectra and adaptation to spectral light distribution in a benthic cyanobacterial mat

We studied adaptation to spectral light distribution in undisturbed benthic communities of cyanobacterial mats growing in hypersaline ponds at Guerrero Negro, Baja California, Mexico. Microscale measurements of oxygen photosynthesis and action spectra were performed with microelectrodes; spectral radiance was measured with fiber-optic microprobes. The spatial resolution of all measurements was 0.1 mm, and the spectral resolution was 10 to 15 nm. Light attenuation spectra showed absorption predominantly by chlorophyll a (Chl a) (430 and 670 nm), phycocyanin (620 nm), and carotenoids (440 to 500 nm). Blue light (450 nm) was attenuated 10-fold more strongly than red light (600 nm). The action spectra of the surface film of diatoms accordingly showed activity over the whole spectrum, with maxima for Chl a and carotenoids. The underlying dense Microcoleus population showed almost exclusively activity dependent upon light harvesting by phycobilins at 550 to 660 nm. Maximum activity was at 580 and 650 nm, indicating absorption by phycoerythrin and phycocyanin as well as by allophycocyanin. Very little Chl a-dependent activity could be detected in the cyanobacterial action spectrum, even with additional 600-nm light to excite photosystem II. The depth distribution of photosynthesis showed detectable activity down to a depth of 0.8 to 2.5 mm, where the downwelling radiant flux at 600 nm was reduced to 0.2 to 0.6% of the surface flux.

NASA Discipline Exobiology↗

NASA NTRS2010

Bird Migration Under Climate Change - A Mechanistic Approach Using Remote Sensing

The broad-scale reductions and shifts that may be expected under climate change in the availability and quality of stopover habitat for long-distance migrants is an area of increasing concern for conservation biologists. Researchers generally have taken two broad approaches to the modeling of migration behaviour to understand the impact of these changes on migratory bird populations. These include models based on causal processes and their response to environmental stimulation, "mechanistic models", or models that primarily are based on observed animal distribution patterns and the correlation of these patterns with environmental variables, i.e. "data driven" models. Investigators have applied the latter technique to forecast changes in migration patterns with changes in the environment, for example, as might be expected under climate change, by forecasting how the underlying environmental data layers upon which the relationships are built will change over time. The learned geostatstical correlations are then applied to the modified data layers.. However, this is problematic. Even if the projections of how the underlying data layers will change are correct, it is not evident that the statistical relationships will remain the same, i.e. that the animal organism may not adapt its' behaviour to the changing conditions. Mechanistic models that explicitly take into account the physical, biological, and behaviour responses of an organism as well as the underlying changes in the landscape offer an alternative to address these shortcomings. The availability of satellite remote sensing observations at multiple spatial and temporal scales, coupled with advances in climate modeling and information technologies enable the application of the mechanistic models to predict how continental bird migration patterns may change in response to environmental change. In earlier work, we simulated the impact of effects of wetland loss and inter-annual variability on the fitness of migratory shorebirds in the central fly ways of North America. We demonstrated the phenotypic plasticity of a migratory population of Pectoral sandpipers consisting of an ensemble of 10,000 individual birds in response to changes in stopover locations using an individual based migration model driven by remotely sensed land surface data, climate data and biological field data. With the advent of new computing capabilities enabled hy recent GPU-GP computing paradigms and commodity hardware, it now is possible to simulate both larger ensemble populations and to incorporate more realistic mechanistic factors into migration models. Here, we take our first steps use these tools to study the impact of long-term drought variability on shorebird survival.

Smith, James A.↗

NASA NTRS2019

Constraints on the Metabolic Activity of Microorganisms in Atacama Surface Soils Inferred from Refractory Biomarkers: Implications for Martian Habitability and Biomarker Detection

Dryness is one of the main environmental challenges to microbial survival. Understanding the threshold of microbial tolerance to extreme dryness is relevant to better constrain the environmental limits of life on Earth and critically evaluate long-term habitability models of Mars. Biomolecular proxies for microbial adaptation and growth were measured in Mars-like hyper-arid surface soils in the Atacama Desert that experience only a few millimeters of precipitation per decade, and in biologically active soils a few hundred kilometers away that experience two- to fivefold more precipitation. Diversity and abundance of lipids and other biomolecules decreased with increasing dryness. Cyclopropane fatty acids (CFAs), which are indicative of adaptive response to environmental stress and growth in bacteria, were only detected in the wetter surface soils. The ratio of trans to cis isomers of an unsaturated fatty acid, another bacterial stress indicator, decreased with increasingly dry conditions. Aspartic acid racemization ratios increased from 0.01 in the wetter soils to 0.1 in the driest soils, which is indicative of racemization rates comparable to de novo biosynthesis over long timescales (10,000 years). The content and integrity of stress proteins profiled by immunoassays were additional indicators that biomass in the driest soils is sin situ microbial growth in the driest surface soils of the Atacama, and any metabolic activity is likely to be basal for cellular repair and maintenance only. Our data add to a growing body of evidence that the driest Atacama surface soils represent a threshold for long-term habitability (i.e., growth and reproduction). These results place constraints on the potential for extant life on the surface of Mars, which is 100–1000 times drier than the driest regions in the Atacama.

Wilhelm, Mary Beth↗

NASA NTRS2001

Skeletal adaptation to external loads optimizes mechanical properties: fact or fiction

The skeleton adapts to a changing mechanical environment but the widely held concept that bone cells are programmed to respond to local mechanical loads to produce an optimal mechanical structure is not consistent with the high frequency of bone fractures. Instead, the author suggests that other important functions of bone compete with mechanical adaptation to determine structure. As a consequence of competing demands, bone architecture never achieves an optimal mechanical structure. c2001 Lippincott Williams & Wilkins, Inc.

Review, Tutorial↗

NASA NTRS1997

Intracellular distributions and putative functions of calcium-binding proteins in the bullfrog vestibular otolith organs

Hair cells in the bullfrog vestibular otolith organs were immunolabeled by monoclonal and polyclonal antisera against calbindin (CaB), calmodulin (CaM), calretinin (CaR), and parvalbumin (PA). S-100, previously shown to immunolabel striolar hair cells in fish vestibular organs, only weakly immunolabeled hair cells in the bullfrog vestibular otolith organs. Immunolabeling was not detected in supporting cells. With the exception of CaR, myelinated axons and unmyelinated nerve terminals were immunolabeled by all of the above antisera. Immunolabeling was seen in all saccular hair cells, although hair cells at the macular margins were immunolabeled more intensely for CaB, CaM, and PA than more centrally located hair cells. As the macula margins are known to be a growth zone, this labeling pattern suggests that marginal hair cells up-regulate their calcium-binding proteins during hair cell development. In the utriculus, immunolabeling for CaM and PA was generally restricted to striolar hair cells. CaR immunolabeling was restricted to the stereociliary array. Immunolabeling for other calcium-binding proteins was generally seen in both the cell body and hair bundles of hair cells, although this labeling was often localized to the stereociliary array and the apical portion of the cell body. CaM and PA immunolabeling in the stereociliary array in saccular and utricular striolar cells suggests a functional role for these proteins in mechanoelectric transduction and adaptation.

NASA Discipline Number 40-10↗

NASA NTRSDate not supplied

BioSentinel: To the Moon or Beyond?

BioSentinel, an Artemis-1 secondary spacecraft, will carry a biology experiment into deep space for the first time in 50 years. A 6U CubeSat form factor was utilized for the spacecraft and included technologies newly developed or adapted for operations beyond Earth orbit. This is the maiden deep-space voyage for the radio, propulsion system, electrical power system, and BioSensor payload. The spacecraft carries onboard budding yeast, Saccharomyces cerevisiae, as an analog to human cells to test the biological response to deep space radiation. Flying a secondary payload beyond LEO comes with unique challenges with respect to trajectory uncertainty and mission operations planning. BioSentinel does not carry propulsion for trajectory maneuvers, so plans for Comms and Power need to be developed for all trajectories. The nominal plan is a lunar flyby followed by an insertion into Heliocentric orbit. However, some possible scenarios include lunar eclipses that could severely impact the power budget during that phase of the mission, while others could result in a “Retrograde” hyperbola at swingby resulting in the spacecraft traveling inward toward Earth or even towards a collision with the lunar surface. BioSentinel’s final trajectory will not be known until after launch and deployment so possible scenarios need to be planned for ahead of time. This paper discusses the operational scenarios that were planned for as well as the actual execution of the mission operations including: command pass scheduling with the Deep Space Network, selecting bandwidth limits, medium gain antenna versus low gain antenna usage, and conserving power prior to a lunar eclipse. Note: Artemis-1 is planned to launch in the Spring of 2022

BioSentinel↗

NASA NTRSDate not supplied

BioSentinel SmallSat 2022 Presentation

BioSentinel↗

NASA NTRS2004

Detecting Organic Compounds in Martian Soil Analogues Using Gas Chromatography Mass Spectrometry

One of the primary objectives of the 1976 Viking missions was to determine whether organic compounds, possibly of biological origin, were present in the Martian surface soils. The Viking gas chromatography mass spectrometry (GCMS) instruments found no evidence for any organic compounds of Martian origin above a few parts per billion in the upper 10 cm of surface soil [l], suggesting the absence of a widely distributed Martian biota. However, Benner et d. have suggested that significant amounts of non-volatile organic compounds, possibly including oxidation products of bioorganic molecules (e.g. carboxylic acids) would not have been detected by the Viking GCMS [2]. Moreover, other key organic compounds important to biology, such as amino acids and nucleobases, would also likely have been missed by the Viking GCMS as these compounds require chemical derivatization to be stable in a GC column [3]. Recent pyrolysis experiments with a Mars soil analogue that had been innoculated with Escherichia coli bacteria have shown that amino acid decomposition products (amines) and nucleobases are among the most abundant products generated after pyrolysis of the bacterial cells [4,5]. At the part per billion level (Viking GCMS detection limit), these pyrolysis products generated from several million bacterial cells per gram of Martian soil would not have been detected by the Viking GCMS instruments [4]. Analytical protocols are under development for upcoming in situ lander opportunities to target several important biological compounds including amino acids and nucleobases. For example, extraction and chemical derivatization techniques [3] are being adapted for space flight use to transform reactive or fragile molecules that would not have been detected by the Viking GCMS instruments, into species that are sufficiently volatile to be detected by GCMS. Recent experiments carried out at NASA Goddard have shown that using this derivatization technique all of the targeted compounds mentioned above can be separated on a GC column and detected by MS at sub-picomole (< 10(exp -l2 mole) levels. With these methods, the detection limit for amino acids, carboxylic acids and nucleobases is several orders of magnitude more sensitive than the Viking GCMS instruments for these compounds. Preliminary results using this analytical technique on a variety of Martian soil analogues will be presented.

Glavin, D. P.↗

NASA NTRS2016

Synthetic Biological Membrane

The ultimate goal of the Synthetic Biological Membrane project is to develop a new type of membrane (using FOST - Forward Osmosis Secondary Treatment) that will enable the waste water treatment system required on exploration missions to operate for extended periods while requiring no maintenance, and using only available resources. Because current water treatment systems have a lifetime of less than 1 year, extending membrane life will benefit exploration missions by reducing upmass and crew time requirements associated with maintenance and resupply/replacement of membranes. The project will use synthetic biology to engineer organisms that protect and replenish the membrane, and this new technology can be easily adapted for use in terrestrial waste water treatment plants.

Biomimetic membrane↗

NASA NTRSDate not supplied

PLANNING FOR VEG-05 TOMATO CROP PRODUCTION ON THE INTERNATIONAL SPACE STATION

Growing fresh, nutritious, palatable produce for crew consumption during spaceflight may provide health-promoting, bioavailable nutrients and enhance the astronaut dietary experience as we move toward longer-duration missions. Tending plants may also serve as a countermeasure for crew psychological stresses associated with spaceflight. However, requirements to support consistent growth of a variety of nutritious crops under spaceflight environmental conditions remain unclear. This study explores the potential to grow crops for consumption on the International Space Station (ISS) using the Veggie vegetable-production system. Mizuna mustard was grown during VEG-04 studies in 2019, and human and plant data from those tests continue to be analyzed. Tomato plants will be grown for the VEG-05 experiment, and preflight definition studies are underway to ascertain the best hardware and operations for growing this fruiting crop. The variety of tomato selected for this study is ‘Red Robin’, a compact cherry tomato. Plants will be grown under two different red: blue lighting treatments using the Veggie units on ISS, and the impact of spectral quality on plant growth and yield, nutrient content, organoleptic acceptability, and microbial composition of the tomatoes will be assessed. Preliminary testing with this crop helped to identify responses to different analog types of growth hardware including both plant pillows and Passive Orbital Nutrient Delivery System (PONDS) units, and current testing is focusing on flight-like versions of this hardware. Challenges with fertilizer salts leaching out into the plant wicks and burning the stems of tomato were observed in earlier tests, so amended fertilizer and wick configurations have been developed. Crew procedures including plant thinning, watering, height adjustment with respect to lighting, and pollination are being validated. VEG-05 will be the first test of tomato fruit production on ISS. Lessons learned during preparation and implementation of the VEG-04 mizuna test for watering, and a separate ISS investigation with peppers (PH-04) in the Advanced Plant Habitat (APH) in terms of fertilizer, plant wicks, and pollination, will be adapted for this experiment. This research was co-funded by the Human Research Program and Space Biology (MTL#1075) in the ILSRA 2015 NRA call.

Veggie↗

NASA NTRSDate not supplied

Planning for VEG-05 Tomato Crop Production on the International Space Station

Veggie↗

NASA NTRSDate not supplied

The Sufficient Component Cause Model Explaining Low-Dose Radiobiology through an Epidemiologic Lens

One of the major challenges in understanding space radiation-induced carcinogenesis is the uncertainty from translating radiobiological research in cellular and animal models to humans, especially at doses below 100 mSv. Biological studies have shown a host of potential outcomes at low doses in animal and cellular models. The existence of non-targeted effects as bystander and abscopal effects is well-documented from clinical research and basic science1,2,3. While some studies have shown increased effects at low doses, others have shown evidence of hormetic bystander effects, where radiation exposure may be beneficial4,5,6. Despite these varied and diverse findings, epidemiologic studies largely support the linear-no threshold (LNT) assumption used by radiation protection guidance7,8. Translational animal-to-human models have predominantly considered ratio values such as the relative biological effectiveness (RBE) and dose and dose-rate effectiveness factor (DDREF) that rely on the assumption of LNT rather than implementing specific dose-response shapes in the low-dose region, and translational cell-to-human models are uncommon. The sufficient component cause model presents an opportunity to examine biological findings at low doses from an epidemiological lens9. In this model, exposures “sufficient” to cause an outcome of interest are presented in pie charts, such that when all slices of a pie chart are fulfilled, the outcome will occur. Multiple pie charts may exist for a single outcome, illustrating individual differences9. In 1988,Greenland and Poole adapted the sufficient component cause model to incorporate interaction with other exposures (such as genetics or lifestyle factors)10. They show that a range of biological processes are possible in a population, but that an epidemiologic study will only reveal the mean outcome from the population at large10,11. Using this construct, the multiple outcomes presented in radiobiological models to date can be explained in the context of epidemiologic studies. This presentation aims to demonstrate a causal framework that can integrate radiobiological and epidemiological models to date. It is intended as a conversation starter to spur future research.

C M Milder↗

NASA NTRSDate not supplied

The Potential Outcome Model: Explaining Low-Dose Radiobiology through an Epidemiologic Lens

One of the major challenges in understanding space radiation-induced carcinogenesis is the uncertainty from translating radiobiological research in cellular and animal models to humans, especially at doses below 100 mSv. Biological studies have shown a host of potential outcomes at lowdoses in animal and cellular models. The existence of non-targeted effects as bystander and abscopal effects is well-documented from clinical research and basic science1,2,3. While some studies have shown increased effects at low doses, others have shown evidence of hormetic bystander effects, where radiation exposure may be beneficial4,5,6. Despite these varied and diverse findings, epidemiologic studies largely support the linear-no threshold (LNT) assumption used by radiation protection guidance7,8. Translational animal-to-human models have predominantly considered ratio values such as the relative biological effectiveness (RBE) and dose and dose-rate effectiveness factor (DDREF) that rely on the assumption of LNT rather than implementing specific dose-response shapes in the low-dose region, and translational cell-to-human models are uncommon. The sufficient component cause model presents an opportunity to examine biological findings at low doses from an epidemiological lens9. In this model, exposures “sufficient” to cause an outcome of interest are presented in pie charts, such that when all slices of a pie chart are fulfilled, the outcome will occur. Multiple pie charts may exist for a single outcome, illustrating individual differences9. In 1988,Greenland and Poole adapted the sufficient component cause model to incorporate interaction with other exposures (such as genetics or lifestyle factors)10. They show that a range of biological processes are possible in a population, but that an epidemiologic study will only reveal the mean outcome from the population at large10,11. Using this construct, the multiple outcomes presented in radiobiological models to date can be explained in the context of epidemiologic studies. This presentation aims to demonstrate a causal framework that can integrate radiobiological and epidemiological models to date. It is intended as a conversation starter to spur future research.

C M Milder↗

NASA NTRS2003

Plasma membrane disruption: repair, prevention, adaptation

Many metazoan cells inhabit mechanically stressful environments and, consequently, their plasma membranes are frequently disrupted. Survival requires that the cell rapidly repair or reseal the disruption. Rapid resealing is an active and complex structural modification that employs endomembrane as its primary building block, and cytoskeletal and membrane fusion proteins as its catalysts. Endomembrane is delivered to the damaged plasma membrane through exocytosis, a ubiquitous Ca2+-triggered response to disruption. Tissue and cell level architecture prevent disruptions from occurring, either by shielding cells from damaging levels of force, or, when this is not possible, by promoting safe force transmission through the plasma membrane via protein-based cables and linkages. Prevention of disruption also can be a dynamic cell or tissue level adaptation triggered when a damaging level of mechanical stress is imposed. Disease results from failure of either the preventive or resealing mechanisms.

NASA Discipline Cell Biology↗

NASA NTRSDate not supplied

The BioSensor Instrument Beyond LEIA: a Versatile Platform for Lunar Biology

Introduction: The BioSensor is a deep-space-compatible automated microfluidic culturing instrument. While originally designed to measure the effects of deep space radiation on yeast growth for the BioSentinel mission, it has the potential to host a diverse range of life science experiments with single- and/or multi-celled organisms and can be adapted to interface with a diverse range of platforms in both crewed and uncrewed settings. It is therefore a leading candidate for hosting life sciences experimentation associated with a lunar surface habitat. BioSensor design: The function of the BioSensor is to monitor the growth and metabolic activity of samples in batch fluidic culture, without the need for crew involvement. The current configuration houses organisms in 16 wells within 16 microfluidic cards, accommodating a total of 256 samples, replicates, and controls. Each well has an optical system including three LEDs and a photodiode detector to measure absorbance at three wavelengths, enabling measurement of optical density, color change in dyes such as alamarBlue, and bioproduction of pigmented compounds. Organisms are loaded into fluidic wells and air-dried for storage during integration, launch, and transit, then activated by the introduction of culture medium from storage bags via manifolds that fill one card at a time. Temperature is controlled by individual card heaters, and timing of all activities (fluidics fills, optical measurements, temperatures) is directed by an experiment script. The self-contained BioSensor payload is roughly 4U in volume; with electrical/mechanical/thermal interface, e.g. for operation on ISS or a lunar lander, as well as a linear energy-transfer (LET) charged-particle radiation spectrometer, the entire system is closer to 6U. BioSentinel and LEIA: Flown on the ISS and in a deep-space free flyer for BioSentinel, the BioSensor has been modified for use in the LEIA mission, including improvements to reduce the sensitivity to lengthy launch delays. LEIA will monitor yeast growth in the radiation and reduced-gravity environment of the lunar surface no earlier than 2026, on a CLPS lander [4]. Changes include accommodating additional culture media and an additional LED color for a new biological assay (bioproduction of carotenoids-- dietary antioxidants), as well as modifications to the housing to allow late-load biology changeout and improved isolation between fluidics and electronics. Future prospects: Future work with the BioSensor, beyond LEIA, will include expanding the range and diversity of organisms and assays accommodated. Preliminary work has demonstrated the growth of Arabidopsis seedlings in BioSensor fluidic cards, including optical measurements of growth rate over time. Minor modifications could allow measurement of phenotypes related to photosynthetic capacity in both plants and cyanobacteria. The experimental capabilities of the BioSensor could be dramatically increased by introducing the capability for fluorescence measurements, and/or the design of novel biological assays using luminescence. The BioSensor can also be adapted for new platforms and experiment settings; in addition to free-flyer, ISS, and CLPS lander, a preliminary design concept has been developed for crewed deployment directly to the lunar surface. The instrument could be accommodated inside a lunar habitat, where its automated operation would make it an excellent candidate for experiments from fundamental investigations into the response of organisms to lunar surface conditions to applied-science purposes such as screening engineered strains of various organisms for bioproduction capability.

J A Lee↗