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NASA NTRS2018

BioSentinel: Optimizing Growth Conditions for Improved Yeast Cell Viability After Long-Term Desiccation

NASA's BioSentinel mission is one of thirteen secondary payloads to be deployed on the Space Launch System Exploration Mission-1 (SLS EM-1). The BioSentinel nanosatellite will be sent into a heliocentric orbit beyond Low Earth Orbit (LEO), to study the effects of deep space radiation on the budding yeast, Saccharomyces cerevisiae. Ionizing radiation encountered in deep space can create damaging lesions in DNA, including double strand breaks (DSBs). Budding yeast is suitable as a biological model to study these effects, as it is eukaryotic, and can be desiccated for prolonged periods while retaining viability, thus serving as a robust analog for human cells. On the ground, yeast cells are grown in liquid medium, then loaded into the wells of microfluidic cards and air dried prior to integration into the payload. Once the spacecraft reaches its target heliocentric orbit, a mixture of growth medium and metabolic indicator dye will be pumped into the microwells at specific time points to rehydrate the cells and allow them to grow. A 3-color LED detection system will measure changes in growth and metabolism resulting from ionizing radiation exposure. BioSentinel contains a wild type control strain and a rad51 mutant that is defective for DNA damage repair. In this study, we will determine the optimal amount of time to grow diploid yeast cells in liquid culture before they are desiccated for space flight. After an extended time in stationary phase, they become more tolerant to desiccation due to stress caused by nitrogen starvation. However, excessive exposure can lead to loss of viability and to a heterogeneous cell population due to sporulation. Since viability loss during desiccation poses a risk to mission success, a stress preconditioning process during initial growth may increase long-term cell viability. To determine the growth period that improves desiccation tolerance but allows for retention of uniform radiation sensitivity, we will grow both strains in liquid medium for a varying number of days (4 to 7), desiccate the cells, and then observe changes to cell viability and ionizing radiation sensitivity over time. Supported by the Space Life Sciences Training Program at NASA Ames Research Center.

Cui, Kaixin↗

NASA NTRS2016

Measurements of Oxychlorine species on Mars

Mars landed and orbiter missions have instrumentation capable of detecting oxychlorine phases (e.g. perchlorate, chlorate) on the surface. Perchlorate (~0.6 wt%) was first detected by the Wet Chemistry Laboratory in the surface material at the Phoenix Mars Landing site. Subsequent analyses by the Thermal Evolved Gas Analyser aboard the same lander detected an oxygen release (~465°C) consistent with the thermal decomposition of perchlorate. Recent thermal analysis by the Mars Science Laboratory’s Sample Analysis at Mars instrument has also indicated the presence of oxychlorine phases (up to 1.2 wt%) in Gale Crater materials. Despite being at detectable concentrations, the Chemistry and Mineralogy (CheMin) X-ray diffractometer has not detected oxychlorine phases. This suggests that Gale Crater oxychlorine may exist as poorly crystalline phases or that perchlorate/chlorate mixtures exist, so that individual oxychlorine concentrations are below CheMin detection limits (~1 wt%). Although not initially designed to detect oxychlorine phases, reinterpretation of Viking Gas Chromatography/Mass Spectrometer data also suggest that oxychlorine phases are present in the Viking surface materials. Remote near-infrared spectral analyses by the Compact Reconnaissance Imaging Spectrometer for Mars (CRISM) instrument indicate that at least some martian recurring slope lineae (RSL) have spectral signatures consistent with the presence of hydrated perchlorates or chlorates during the seasons when RSL are most extensive. Despite the thermal emission spectrometer, Thermal Emission Imaging System, Observatoire pour la Minéralogie, l’Eau, les Glaces et l’Activité and CRISM detection of hundreds of anhydrous chloride (~10–25 vol%) deposits, expected associated oxychlorine phases (>5–10 vol%) have not been detected. Total Cl and oxychlorine data sets from the Phoenix Lander and the Mars Science Laboratory missions could be used to develop oxychlorine versus total Cl correlations, which may constrain oxychlorine concentrations at other locations on Mars by using total Cl determined by other missions (e.g. Viking, Pathfinder, MER and Odyssey). Development of microfluidic or ‘lab-on-a-chip’ instrumentation has the potential to be the next generation analytical capability used to identify and quantify individual oxychlorine species on future landed robotic missions to Mars.

Perchlorate↗

NASA NTRS2020

ExCALiBR: An Instrument for Uncovering the Origin of the Moonâ€™s Organics.

Lipids are organic molecules of great astrobiological interest for future NASA missions to the Moon, Mars and Icy Worlds. Lipids are essential for life as we know it, and likely also required for putative extraterrestrial organisms. These robust organics can be synthesized abiotically and survive for long periods of geological time. Laboratory characterization techniques are well established but are laborious, operator dependent, and require large volumes of consumables, precluding in situ analysis. We are developing an autonomous, miniaturized fluidic system, integrating lab techniques for lipid analysis while minimizing rea-gent volumes and concentrating organics for analysis, thereby increasing signal-to-noise ratios by orders of magnitude. This system, the Extractor for Chemical Analysis of Lipid Biomarkers in Regolith (ExCALiBR), will enable future organic surveys by extracting and concentrating lipids from approximately 50 grams of regolith using a fluidic and microfluidic sample processor made of materials compatible with non-aqueous solvents required for extraction. ExCAL-iBR can be deployed on a lander or used on a crewed mission.

M B Wilhelm↗

NASA NTRS2019

Gene Expression Measurement Module (GEMM) for space application: Design and validation

In order to facilitate studies on the impact of the space environment on biological systems, we have developed a prototype of GEMM (Gene Expression Measurement Module) - an automated, miniaturized, integrated fluidic system for in-situ measurements of gene expression in microbial samples. The GEMM instrument is capable of (1) lysing bacterial cell walls, (2) extracting and purifying RNA released from cells, (3) hybridizing the RNA to probes attached to a microarray and (4) providing electrochemical readout, all in a microfluidics cartridge. To function on small, uncrewed spacecraft, the conventional, laboratory protocols for both sample preparation and hybridization required significant modifications. Biological validation of the instrument was carried out on Synechococcus elongatus, a photosynthetic cyanobacterium known for its metabolic diversity and resilience to adverse conditions. It was demonstrated that GEMM yielded reliable, reproducible gene expression profiles. GEMM is the only high throughput instrument that can be deployed in near future on space platforms other than the ISS to advance biological research in space. It can also prove useful for numerous terrestrial applications in the field.

Kianoosh Peyvan↗

NASA NTRS2020

BioSentinel ISS Control Experiment Final Investigation Summary Form

Each payload has 18 fluidic cards and each fluidic card has 16 microfluidic wells, for a total of 288 wells. Card activation occurs at different time points throughout the 6-month mission and following ground commands. At each timepoint, two fluidic cards are filled with growth medium containing nutrients and a metabolic indicator dye. In addition to card filling, the temperature of the cards increases from ~4°C (stasis mode) to 23°C to allow for cell growth. The metabolic dye changes color from blue to pink, then pink to clearin the presence of metabolically active yeast cells. The colorimetric changes and cell growth are monitored using a 3-light-emitting diode(LED)optical detection system. The optical data determines how fast the cells are able to recover after accumulating radiation damage over time.After ground command initiation, all the fluidics, thermal and optical detection steps in BioSentinel are fully automated.The biosensor response data is compared to physical dosimetry data collected onboard the same payload. This dosimeter or linear energy transfer (LET)spectrometer is based on TimePix chip technology. This chip measures the total ionizing radiation dose in addition to calculating the LET of each particle that traverses its sensor, thus allowing researchers to characterize the radiation environment around the payload.The biological response (and physical dosimetry) on ISS over time is compared to the deep space mission and to data obtained on the ground.This analysis provides insight on the biological effects of the space radiation exposure in preparation for future long-duration missions to space and inhabitation beyond low-Earth orbit (LEO).

BioSentinel↗

NASA NTRS2020

Encealdus Orbilander: A Flagship Mission Concept for Astrobiology

"Whether life exists beyond Earth remains a fundamental question driving our exploration of the Solar System. At Saturn’s moon Enceladus, plumes of oceanic material vented into space allow the investigation of the astrobiological potential of an ocean world, hinted at by Cassini, without the necessity of drilling through kilometers of ice crust. The Enceladus Orbilander is a flagship ($2.56B in fiscal year 2025 dollars) mission concept created for the 2023–2033 Planetary Science Decadal Survey. Orbilander takes full advantage of the opportunity provided by Enceladus’ plumes to search for signs of life. A single spacecraft both orbits and lands, capturing samples from four distinct reservoirs offered by the plumes. These samples, both particulate and vapor, are then analyzed by the Life Detection Suite (LDS), a set of five instruments conducting complementary and orthogonal biosignature-seeking measurements. To provide the context that specifically enhances interpretation of LDS measurements, geochemical and geophysical investigations are conducted both in orbit and on the surface. These reveal the physio-chemical state of the ocean and core as well as the processes involved in ejection of plume material and how these affect the ocean material analyzed by the LDS. The Orbilander can be delivered to the Saturn system via several launch vehicle and trajectory options, including a direct trajectory (7-year cruise), a ∆V-EGA trajectory (9-year cruise) and several options using an inner cruise with Venus and Earth flybys (10-year cruise). Upon Saturn Orbit Insertion, a 4-year moon tour pumps down the Orbilander’s orbit to intercept Enceladus. The most optimal arrival times balance the Jupiter flyby opportunities of the late 2030s and solar illumination at the Enceladus high southern latitudes where plume material is most abundant. This mission concept therefore targets project start in 2030. Upon Enceladus Orbit Insertion, the Orbilander begins a 1.5-year-long campaign of landing site reconnaissance, remote sensing science, and collecting sufficient plume sample to run all but one of the LDS measurements. After successful landing, the Orbilander spends 2 years on the surface conducting multiple LDS measurements with all five instruments on actively and passively collected plume material, as well as seismic investigations. The schedule laid out here is well-defined, but the mission also has operational and resource flexibility should additional reconnaissance be needed. As part of the design study, mission and development risks were identified and mitigation strategies proposed. Technologies key to achieving the life detection science objectives include instrumentation matured under programs like COLDTech and ICEE-2, such as aspects of the sampling system and microfluidic devices, as well as well-known techniques like high-resolution and separation-capable mass spectrometers. Autonomous onboard navigation is planned to maintain a halo orbit around Enceladus to enable passive sampling from orbit as well as reconnaissance measurements for use in site selection and landing. Terrain relative navigation is included to ensure safe landing, given that targeted areas may contain landing hazards. Continued development of radioisotope thermoelectric generator (RTG) technology and long-life batteries is essential for this long duration mission. The Enceladus Orbilander represents an optimal point in the trade space of science value versus cost, taking advantage of the extensive knowledge of Enceladus provided by Cassini, how well Enceladus lends itself to a search for life in material from its ocean, and the flexibility afforded by the innovative design developed by the APL team. By taking full advantage of Enceladus’ plumes both in orbit and on the surface, Orbilander represents a robust search for life with complementary and orthogonal biosignatures as well as contextual geophysical and geochemical measurements, determining not only whether Enceladus is inhabited (at levels up to 500,000× scarcer than in Earth’s oceans) but also why. "

Exobiology↗

NASA NTRSDate not supplied

ELM: Europa Luminescence Microscope

The Europa Luminescence Microscope (ELM) is an automated fluorescence and bright-field microscope designed to meet key objectives defined in the 2016 NASA Europa Lander Study Report, including the identification and characterization of morphological biosignatures. ELM’s heritage stems from a 2U cubesat fluorescence microscope, the Fluorescence Analysis for In situ Research imager, designed and built at NASA Ames Research Center, for the autonomous study of microbial biology in low Earth orbit. For ELM implementation, a sample is autonomously manipulated with a microfluidic system using in-line 10, 1.0, and 0.2 μm pore-size filters to capture successively smaller particles for imaging. For bright-field imaging, ELM uses deep-ultraviolet, ultraviolet and visible light to image organic and inorganic structures with submicron resolution. The ability to detect biosignatures as small as 0.2 μm in size is achieved by imaging native fluorescence and using fluorescence microscopy stains to identify key structural and functional indicators of microbial life (proteins, lipids, nucleic acids). For fluorescence imaging, ELM uses 265, 370, 470, and 530 nm LEDs with five emission bands. The use of multiple excitation and emission wavelengths for native fluorescence imaging enables the detection of a wide range of molecular species and their rough classification. Excitation at 265 nm allows for the detection of smaller polycyclic aromatic hydrocarbons (PAH), aromatic amino acids, and proteins with little to no interference from mineral fluorescence, given proper emission band selection. 370 and 470 nm light excites increasingly larger PAH structures and larger aromatic biomolecules that may be present (e.g., protective pigments). Similarly, inorganic fluorescence can be characterized and separated from organic fluorescence, allowing the recognition and in some cases classification, of minerals and other abiotic particles. ELM is based upon work supported by the NASA COLDTech and ICEE-2 programs.

Richard C Quinn↗

NASA NTRS2012

Design and Operation of the Planetary In-Situ Capillary Electrophoresis System (PISCES)

The PISCES instrument is a portable, miniature, modular instrument designed to analyze organic molecular signatures of life. PISCES uses microcapillary electrophoresis (μCE) with laser-induced fluorescence detection (LIFD) to analyze fluorescently-labeled analytes prepared on a multi-layer microfluidic chip. As a modular instrument composed of five independently functioning units, each module can be easily detached from the system and used for subsystem testing in different environments, e.g. low temperature. The pneumatic module contains mechanical and electrical components necessary to control pneumatically-actuated valves for on-chip sample processing. A prototype LabVIEW program and two custom printed circuit boards (PCBs) operate the pneumatics module using an Arduino microcontroller, and a tentative design for the module’s housing has been developed. The high-voltage module contains circuitry required to apply varying electrical potentials to the electrophoretic channel of the microdevice. Work is currently being done to design the circuit and PCB for this module in collaboration with our SBIR partners at Los Gatos Research. The optical electronics module contains the spectrometer, laser, and laser power supply. The mounting components for the electronics and housing for the module have been designed, assembled, and tested. The optical module contains optical components to focus the laser to a 20μm spot in the center of the electrophoretic channel.The housing for this module has been designed and assembled with the aligned optical components inside.

Willis, Peter A.↗

NASA NTRSDate not supplied

Development of Electrolytic Oxygen Recovery System for Advanced Life Support

The oxygen (O2) recovery system for the International Space Station (ISS) can recover approximately 50 percent of O2 from metabolic carbon dioxide (CO2). Increasing the O2 recovery rate and closing the open loop for future long duration crewed missions in space beyond Low Earth Orbit (LEO) is essential. There are several developmental efforts underway to increase the recovery rate. However, most of these technologies result in a complex, heavy, and power consuming system. The desired exploration O2 recovery system would be reliable and efficient with maximum O2 recovery. Marshall Space Flight Center (MSFC) is currently investigating an electrolytic O2 recovery approach that will increase the O2 recovery to greater than 70 percent as well as lowering the complexity, mass, and power consumption than most other technologies currently under development. The electrolytic O2 recovery system consists of a Microfluidic Electrochemical Reactor (MFECR) that is based on the electrochemical reduction of CO2 to O2 and ethylene (C2H4) using water (H2O) as precursor and operates at standard condition with a theoretical recovery rate of 73 percent. In 2016, NASA’s Game Changing Development Program awarded the University of Texas Arlington (UTA) a grant to initiate the development of the MFECR. Since 2019, MSFC and UTA have been collaborating with the current goals of increasing the O2 recovery efficiency, advancing the technology readiness to a Technology Readiness Level (TRL) 4, and maturing the system to process CO2 of one crew-member. Based on the results from UTA’s initial efforts, the following were identified as key areas of improvement in order to maximize O2 recovery for the system: further development of the anode material and cathode catalyst, model-based cell design optimization, and the addition of a separation system and fuel cell. This paper will present the current developmental efforts of the electrolytic system including MFECR design and overall system enhancements as well as results from single cell stack testing.

Brittany R Brown↗

NASA NTRSDate not supplied

Rapid Nucleic Acid Isolation Method and Fluid Handling Devices Overview

Sample preparation is a common bottleneck in genetic analysis. Tools that reduce time and effort are of value in a laboratory setting. There is a need for a genetic analysis/detection system that is not only mobile, ruggedized, easy-to-use but also provides an enclosed fluid handling that diminishes the risks of Ribonucleic acid (RNA) contamination and degradation during processing. NASA has developed a novel assay methodology and suite of devices to isolate nucleic acids and prepare samples for real-time (quantitative) polymerase chain reaction (PCR) analysis that solves the problem of contamination and degradation because of the advanced microfluidics which provides better fluid handling. The assay employs an aqueous-based non-alcohol method that yields robust RNA quality. The suite of ready-to-use devices provide pre-loaded reagents in liquid and lyophilized formats to enable rapid, cost effective, and portable manual operation in the laboratory or remote field environments.

Travis Boone↗

NASA NTRSDate not supplied

Europa Luminescence Microscope

The Europa Luminescence Microscope (ELM) is an automated fluorescence and dark-field mi-croscope designed to meet key objectives defined in the NASA Europa Lander Study Report, includ-ing the identification and characterization of morphological biosignatures. ELM’s heritage stems from a 2U cubesat fluorescence microscope, the Fluorescence Analysis for In-situ Research (FLAIR) im-ager, designed and built at NASA Ames Research Center, for the autonomous study of microbial bi-ology in low Earth orbit. For the ELM implementation, a sample is autonomously manipulated with a microfluidic system using in-line filter sets to capture successively smaller particles on 10, 1.0, and 0.1 µm pore-size filters for imaging. For darkfield imaging, ELM uses ultraviolet and visible light to image organic and inorganic structures with submicron resolution. The ability to detect structural and chemical biosignatures as small as 0.2 µm in size is achieved by imaging native fluorescence and us-ing fluorescence microscopy stains to identify key molecular and structural indicators of microbial life (proteins, lipids, nucleic acids). To excite fluorescence, ELM uses LEDs with wavelengths centered near 265, 370, 470, and 530 nm and five emission bands. The use of multiple excitation and emission wavelengths for native fluorescence imaging not only enables the detection of different molecular species, but also their rough classification. Excitation at 265 nm allows for the detection of smaller polyaromatic hydrocarbons (PAHs; 1-5 rings), aromatic amino acids, and proteins with little to no interference from mineral fluorescence, given proper emission band selection. 370 and 470 nm light excites increasingly larger PAH structures (e.g., coronene) and larger aromatic biomolecules that may be present (e.g., protective pigments). Similarly, inorganic fluorescence can be characterized and sep-arated from organic fluorescence, allowing the recognition and in some cases classification, of miner-als and other abiotic particles. ELM is based upon work supported by the NASA COLDTech and ICEE-2 programs.

Microscope↗

NASA NTRSDate not supplied

AMMPER: Agent-based Model for Microbial Populations Exposed to Radiation

Exposure of microbial populations to galactic cosmic radiation (GCR) in the deep-space radiation environment may lead to intracellular damage that compromises the ability of cells to repair and replicate. While simulation programs of deep-space radiation do exist, they focus primarily on single-cell damage, rather than population-wide effects. In this work, a new application and graphical user interface, Agent-Based Model for Microbial Populations Exposed to Radiation (AMMPER) is presented, which simulates the effects of proton-based GCR on Saccharomyces cerevisiae population growth. AMMPER consists of a 50x50x50 µm simulation space, analogous to an aqueous culture medium with non-limiting nutrient and pH buffering, in a microwell plate or microfluidic culture card. This model utilizes Relativistic Ion Tracks (RITRACKS) to create detailed track structures of the radiation traversals. AMMPER then calculates the radiation dose present at each cell, and subsequently determines the damage (chromosomal aberrations, oxidative stress, etc.) and resulting loss of cell viability from both primary and secondary radiative effects. Through implementing cell replication, repair, damage, and death, the effect of radiation exposure on the population growth can be determined. With AMMPER, long-duration effects of the deep space environment on entire populations can be determined and used to assess the feasibility of sustaining life in space.

Amrita Singh↗

NASA NTRSDate not supplied

PowerCell aboard the EuCROPIS satellite - Results from the first synthetic biology experiment in space

Synthetic biology is the design and construction of new biological parts and systems not found in nature, a technology that we believe will be a game-changing technology for space exploration. The PowerCell concept is an ecology based on photosynthetic microbes taking advantage of in situ materials and energy to generate, on-demand, useful products (food, fuel, cloths, drugs, etc.) that satisfy specific needs of long-term human presence away from Earth. This first PowerCell experiment is testing this concept as well as several key mechanisms of engineering biology in a 48-well plate microfluidics system. NASA's PowerCell experiment was loaded in the fluidics system at NASA Ames in May 2016, and subsequently integrated on the DLR's compact satellite Eu:CROPIS (Euglena Combined Regenerative Organic food Production In Space), as a secondary payload. Launched into Earth’s orbit from the Vandenberg Air Force Base, California, USA, in December 3, 2019, the Eu:CROPIS platform has provided an artificial gravity through rotational acceleration, simulating gravity ranges compatible to those found on Mars and on the Moon, as well as microgravity in the absence of acceleration. Each set of biological experiments performed at Earth's orbit was compared to ground controls. Over 400 days' worth of mission data showed that all hardware and software worked as planned, and microbial growth was observed in all four gravity regimes, including growth on cyanobacterial cell extract in three out of four gravity regimes. The genetic transformation experiment was successful in microgravity and provided valuable long-term reagent stability data, including antibiotic efficacy. The demonstrated ability of hardy bacterial spores to recover from a long-term stasis period will be fundamental for deep human space missions. Lessons learned from hardware design to experimental operations are leading to improved designs and capabilities for future missions.

Ivan Glaucio Paulino Lima↗

NASA NTRSDate not supplied

BioSentinel: Mission Summary and Lessons Learned From the First Deep Space Biology CubeSat Mission

Launched on Artemis-1, BioSentinel carries a biology experiment into deep space for the first time in 50 years. A 6U CubeSat form factor was utilized for the spacecraft which included technologies newly developed or adapted for operations beyond Earth orbit. The spacecraft carries onboard budding yeast, Saccharomyces cerevisiae, as an analog to human cells to test the biological response to deep space radiation. This was the maiden deep-space voyage for many of the subsystems, and the first time to evaluate their performance in flight operation. Flying a CubeSat beyond LEO comes with unique challenges with respect to trajectory uncertainty and mission operations planning. The nominal plan was a lunar fly-by, followed by an insertion into Heliocentric orbit. However, some possible scenarios included lunar eclipses that could have severely impacted the power budget during that phase of the mission, while others could have resulted in a “Retrograde” hyperbola at swing-by resulting in the spacecraft traveling inward toward Earth or even towards a collision with the lunar surface. The commissioning phase of the mission was successful and completed a week ahead of schedule. It did not come without its exciting moments and challenges. First contact with the spacecraft uncovered that the vehicle was unexpectedly tumbling after deployment, a situation that needed to be corrected urgently. The mission operations team executed a contingency plan to stabilize the spacecraft, with just moments to spare before the battery ran out of power. The BioSensor payload onboard the spacecraft is a complex instrument that includes microfluidics, fluid systems, sensor control electronics, as well at the living yeast cells. BioSentinel also included a TimePix radiation sensor implemented by JSC’s RadWorks group. Dose and Linear Energy Transfer (LET) data is compared directly to the rate of DSB-and repair events measured by the S. cerevisiae cells. BioSentinel mature nanosatellite technologies included: deep space communications and navigation, autonomous attitude control and momentum management, and micro-propulsion systems, to provide an adaptable nanosatellite platform for deep space uses. This paper discusses the performance of the BioSentinel spacecraft through the mission phase, and includes lessons learned from challenges and anomalies. BioSentinel had many successes and will be a pathfinder for future deep space CubeSats and biology missions.

BioSentinel↗

NASA NTRSDate not supplied

NASA SmallSat LEARN Forum Briefing: BioSentinel

We will focus on the technologies developed for BioSentinel, primarily the science payloads. One of them, the BioSensor, is a microfluidics-based instrument that houses a series of fluidic cards and manifolds. The fluidic cards can be loaded with microbial organisms to study the effect of the space environment on biology. For BioSentinel, we used the budding yeast as our model organism to study the response to the radiation environment in both low Earth orbit (ISS) and in deep space. The second instrument is a radiation spectrometer. This sensor allows the quantification of ionizing radiation as well as the characterization of the different particles based on the linear energy transfer or LET. Given that BioSentinel launched this past November (Artemis I), part of the presentation will include lessons learned and how we will apply them to future missions, including LEIA. LEIA was recently selected for a mission to the lunar surface on a CLPS lander vehicle, and is primarily based at NASA Ames.

BioSentinel↗

NASA NTRS2023

Development of an Efficient Alternative to Recovery O2 From Metabolic CO2 Via Electrolysis Operated at Ambient Temperature and Driven By A Highly Selective Catalysis

The current State of Art (SOA) on oxygen recovery onboard the Environmental Control and Life Support System (ECLSS) at the International Space Station (ISS) is complex, heavy, and power consuming system that recovers approximately 50% of the oxygen (O 2 ) from metabolic carbon dioxide (CO 2 ). For future long duration beyond low earth orbit missions (LEO), O 2 recovery systems will need to be highly reliable, efficient, and recover a minimum of 75% O 2 from metabolic CO 2 . An alternative technology development effort currently underway at NASA Marshall Space Flight Center (MSFC) has the potential to significantly increase O 2 recovery currently limited to 50% (Sabatier) and reduce the complexity of ECLSS O 2 recovery. MSFC and University of Texas in Arlington (UTA) have jointly designed and fabricated a microfluid electrochemical reactor (MFECR) that operates at ambient conditions and utilizes a proprietary catalysis highly selective on reducing CO 2 to ethylene (C 2 H 4 ) at the cathode while O 2 is generated at the anode. The MFECR would replace three pieces of hardware for future ECLSS architectures: the current CO 2 Reduction Assembly (CRA) (Sabatier reactor), the Plasma Pyrolysis Assembly (PPA), and the Oxygen Generation Assembly (OGA). It is designed to interface directly with the CO 2 Removal Assembly (CDRA) and the Water Processing Assembly (WPA) to supply CO 2 reactant and water replenish respectively. This is expected to substantially improve sustainability of the ISS ECLSS and reduce requirement on power and weight. Here, we discuss the current development and evaluation efforts on different alternatives on not only the configuration and setup of the MFECR at an Engineering Design Unit (EDU) scale but also the selection of component materials.

Jesus A Dominguez↗

NASA NTRS2023

Automated Fluidics Device for Extraction and Quantification of miRNA Biomarkers From Blood

Radiation Assessment DuRing Exposure And long-Duration Spaceflight (RADREADS) demonstrates space-compatible point-of-care technology for quantitative biological monitoring of blood miRNA biomarkers in response to long-term low dose radiation exposure. This individualized monitoring approach will inform targeted treatment strategies to maximize medical resource utilization by accounting for individual susceptibility to radiation-related illnesses. As human spaceflight progresses beyond Earth’s magnetic shielding, radiation exposure poses a significant risk to astronaut health and safety. Extended operation in this environment comes with an increased risk of radiation exposure, leading to higher risks of radiation sickness, cancer, central nervous system effects, and degenerative diseases. While conventional physical dosimetry techniques capture radiation dose, individualistic susceptibility to radiation damage is varied. Multiple characteristics, including age, body weight, sex, genetics, and immune status, have been found to influence radiosensitivity (Liu et al. 2011, and Bouffler 2016). This differential response necessitates individualized monitoring and targeted treatment strategies to maximize medical resource utilization; however, a practical diagnostic platform for quantifying long-term, low dose radiation-induced tissue damage does not currently exist. MicroRNAs (miRNAs) are a class of small, non-coding RNAs that regulate gene expression by mediating the degradation of messenger RNA. The levels of particular miRNAs are influenced by biological processes such as inflammation and serve as biomarkers for a variety of conditions including cancer (Singh et al. 2017). MicroRNAs are found in various bodily fluids and are amenable to collection via liquid biopsies, providing a minimally invasive and easily quantifiable readout for a variety of radiosensitive reporters. A preliminary signature of 15 spaceflight sensitive miRNA has been identified in rodent and human studies, including miR-21-5p, miR-24-3p, miR-92a-3p, miR-17-5p, miR-16a-3p, miR-34a-3p, and miR-223-3p. These targets generally increased expression with radiation dose and linear energy transfer, though variation between individuals is not yet described. Current gaps in the field include a lack of understanding of longitudinal biological responses to long-term, low dose radiation exposure and the absence of space-compatible point-of-care technology for quantitative biological monitoring. In this body of work, we aim to develop an automated bleed-to-read system to process whole blood for the detection of miRNA biomarkers in order to monitor individualistic responses to radiation exposure. This will be achieved via separating serum (or plasma) from whole blood, followed by extraction, amplification, and quantification of the miRNA using a RT-qPCR reaction. Previously, the WetLab-2 hardware enabled execution of a RT-qPCR reaction aboard ISS; however, it is a manual system that requires crew manipulation and bulky components (Parra et al. 2017). To address these issues, automated fluid handling hardware was developed for each stage of sample preparation. Extraction of total RNA is achieved by sequentially pumping reagents through an off-the-shelf nucleic acid binding column (miRNeasy Serum/Plasma Advanced Kit, Qiagen). This approach eliminates several manual pipetting and centrifuging steps and limits the use of toxic chemicals commonly found in other sample processing techniques. The resulting elution will then be automatically dispensed for RT-qPCR analysis using a compact rotary qPCR (Mic qPCR Cycler, Bio Molecular Systems) that will improve spaceflight compatibility by removing bubbles from the detection region, another challenge highlighted by WetLab-2 (Parra et al. 2017). Efforts are also being made to simplify the RT-qPCR reaction to a 1-step air-dryable mix to improve long-term reagent stability at room temperature and reduce system complexity. By automating the RT-qPCR processes via microfluidic manipulation, RADREADS will reduce crewmember hands-on time and enable the personalized detection of radiation-induced tissue damage during long duration missions. Minimally invasive, longitudinal monitoring of individual’s response to radiation exposure will inform how the physiological system responds to long-term low dose space radiation and enables development of targeted countermeasures by the medical team. Ultimately, this portable technology will require minimal technical expertise and can also be used to monitor miRNA biomarkers associated with other diseases.

Tristen Head↗

NASA NTRSDate not supplied

Biosentinel: Mission Summary and Lessons Learned From the First Deep Space Biology CubeSat Mission

BioSentinel↗