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Real-Time Culture-Independent Microbial Profiling Onboard the International Space Station using Nanopore Sequencing

For the past two decades, microbial monitoring of the International Space Station (ISS) has relied on culture-dependent methods that require return to Earth for analysis. This has a number of limitations, with the most significant being bias towards the detection of culturable organisms and the inherent delay between sample collection and ground-based analysis. In recent years, portable and easy-to-use molecular-based tools, such as Oxford Nanopore Technologies’ MinION™ sequencer and miniPCR bio’s miniPCR™ thermal cycler, have been validated onboard the ISS. Here, we report on the development, validation, and implementation of a swab-to-sequencer method that provides a culture-independent solution to real-time microbial profiling onboard the ISS. Method development focused on analysis of swabs collected in a low-biomass environment with limited facility resources and stringent controls on allowed processes and reagents. ISS-optimized procedures included enzymatic DNA extraction from a swab tip, bead-based purifications, altered buffers, and the use of miniPCR and the MinION. Validation was conducted through extensive ground-based assessments comparing current standard culture-dependent and newly developed culture-independent methods. Similar microbial distributions were observed between the two methods; however, as expected, the culture-independent data revealed microbial profiles with greater diversity. Protocol optimization and verification was established during NASA Extreme Environment Mission Operations (NEEMO) analog missions 21 and 22, respectively. Unique microbial profiles obtained from analog testing validated the swab-to-sequencer method in an extreme environment. Finally, four independent swab-to-sequencer experiments were conducted onboard the ISS by two crewmembers. Microorganisms identified from ISS swabs were consistent with historical culture-based data, and primarily consisted of commonly observed human-associated microbes. This simplified method has been streamlined for high ease-of-use for a non-trained crew to complete in an extreme environment, thereby enabling environmental and human health diagnostics in real-time as future missions take us beyond low-Earth orbit.

microbiology

An Object-Oriented Interface to the CCSDS Ground Telecommand Services

The Telecommand Data Routing and Channel Services defined by the Consultative Committee for Space Data Systems (CCSDS) are flexible enough to support a myriad of commanding models. Because the standard is so broad, the traditional approach has been to implement only the portion of the standard needed by the particular spacecraft being tested/operated. Tasked with providing Telecommand Services for an entire class of spacecraft, where each spacecraft may choose any valid CCSDS commanding model, NASA Code 584 designed a common architecture capable of handling the full CCSDS protocol. The solution uses another CCSDS standard - the Standard Formatted Data Unit (SFDU) as the interface to the Telecommand Services. SFDUs provide a consistent way of labelling data objects, as well as allowing data objects to encapsulate other data objects. The resulting interface is: - Flexible: The full Data Routing and Channel Services are available via a single interface. The client (i.e. the command source) may enter commands at any layer within the protocol stack, specify any of the data aggregation or segmentation methods, and dynamically set any configuration parameter defined in the standard. - Object-oriented: Each object specifies both the data and the actions to be performed with the data. An object may contain other objects. - Expandable: New capabilities are added by defining new objects. Objects pass thru the protocol layers until they reach the applicable layer. The resulting design is: - Modular: The logic for each protocol layer is contained in a separate Application Program Interface (API). The objects used for the external interface are also used for communication between layers. - Distributable: The design can be split along any layer boundary for distribution across multiple machines. The objects ensure data consistency across platforms. This paper describes the SFDU-based interface and the resulting protocol implementation. The implementation is currently used by NASA (National Aeronautics and Space Administration) for integration & test of the microwave Anisotropy Probe (MAP) and Earth observer-I (EO-l) spacecraft. It will be used for post-launch operations of these spacecraft as well as the Imager for Magnetopause to Aurora Global Exploration (IMAGE) spacecraft.

Ray, Timothy Joseph

Performance Modeling of Network-Attached Storage Device Based Hierarchical Mass Storage Systems

Network attached storage devices improve I/O performance by separating control and data paths and eliminating host intervention during the data transfer phase. Devices are attached to both a high speed network for data transfer and to a slower network for control messages. Hierarchical mass storage systems use disks to cache the most recently used files and a combination of robotic and manually mounted tapes to store the bulk of the files in the file system. This paper shows how queuing network models can be used to assess the performance of hierarchical mass storage systems that use network attached storage devices as opposed to host attached storage devices. Simulation was used to validate the model. The analytic model presented here can be used, among other things, to evaluate the protocols involved in 1/0 over network attached devices.

Menasce, Daniel A.

Performance, Capacity and Limitations of AMSS Communications

This paper reports on the performance of Aeronautical Mobile Satellite Service (AMSS), based on simulation results, when incorporated into the Aeronautical Telecommunications Network (ATN). A comparison between its performance under current data traffic load and under future ATN traffic load was used to determine the reliability of AMSS in providing continuous communications between the ground and the aircraft over remote regions of the globe. The simulation involved modeling AMSS through the use of the published standards to ensure the accuracy of the results. Although the results indicated that there is indeed a reduced capacity (to about one-fourth) and efficiency of the system at the increased traffic load, the reduction in performance does not lead to the conclusion that an alternative to AMSS must be found. On the contrary, some modifications to the protocols controlling the operation of AMSS and updating the system with new technologies, would make it as valid a solution to the problem as it is today.

Ripamonti, Claudio

Advanced Extravehicular Activity Pressure Garment Requirements Development

The NASA Johnson Space Center advanced pressure garment technology development team is addressing requirements development for exploration missions. Lessons learned from the Z‐2 high fidelity prototype development have reiterated that clear low‐level requirements and verification methods reduce risk to the government, improve efficiency in pressure garment design efforts, and enable the government to be a smart buyer. The expectation is to provide requirements at the specification level that are validated so that their impact on pressure garment design is understood. Additionally, the team will provide defined verification protocols for the requirements. However, in reviewing exploration space suit high level requirements there are several gaps in the team's ability to define and verify related lower level requirements. This paper addresses the efforts in requirement areas such as mobility/fit/comfort and environmental protection (dust, radiation, plasma, secondary impacts) to determine the by what method the requirements can be defined and use of those methods for verification. Gaps exist at various stages. In some cases component level work is underway, but no system level effort has begun, in other cases no effort has been initiated to close the gap. Status of ongoing efforts and potential approaches to open gaps are discussed.

Ross, Amy

Advanced Extra-Vehicular Activity Pressure Garment Requirements Development

The NASA Johnson Space Center advanced pressure garment technology development team is addressing requirements development for exploration missions. Lessons learned from the Z-2 high fidelity prototype development have reiterated that clear low-level requirements and verification methods reduce risk to the government, improve efficiency in pressure garment design efforts, and enable the government to be a smart buyer. The expectation is to provide requirements at the specification level that are validated so that their impact on pressure garment design is understood. Additionally, the team will provide defined verification protocols for the requirements. However, in reviewing exploration space suit high level requirements there are several gaps in the team's ability to define and verify related lower level requirements. This paper addresses the efforts in requirement areas such as mobility/fit/comfort and environmental protection (dust, radiation, plasma, secondary impacts) to determine the method by which the requirements can be defined and use of those methods for verification. Gaps exist at various stages. In some cases component level work is underway, but no system level effort has begun; in other cases no effort has been initiated to close the gap. Status of on-going efforts and potential approaches to open gaps are discussed.

Ross, Amy

The Jade File System

File systems have long been the most important and most widely used form of shared permanent storage. File systems in traditional time-sharing systems, such as Unix, support a coherent sharing model for multiple users. Distributed file systems implement this sharing model in local area networks. However, most distributed file systems fail to scale from local area networks to an internet. Four characteristics of scalability were recognized: size, wide area, autonomy, and heterogeneity. Owing to size and wide area, techniques such as broadcasting, central control, and central resources, which are widely adopted by local area network file systems, are not adequate for an internet file system. An internet file system must also support the notion of autonomy because an internet is made up by a collection of independent organizations. Finally, heterogeneity is the nature of an internet file system, not only because of its size, but also because of the autonomy of the organizations in an internet. The Jade File System, which provides a uniform way to name and access files in the internet environment, is presented. Jade is a logical system that integrates a heterogeneous collection of existing file systems, where heterogeneous means that the underlying file systems support different file access protocols. Because of autonomy, Jade is designed under the restriction that the underlying file systems may not be modified. In order to avoid the complexity of maintaining an internet-wide, global name space, Jade permits each user to define a private name space. In Jade's design, we pay careful attention to avoiding unnecessary network messages between clients and file servers in order to achieve acceptable performance. Jade's name space supports two novel features: (1) it allows multiple file systems to be mounted under one direction; and (2) it permits one logical name space to mount other logical name spaces. A prototype of Jade was implemented to examine and validate its design. The prototype consists of interfaces to the Unix File System, the Sun Network File System, and the File Transfer Protocol.

Rao, Herman Chung-Hwa

Marine Optical Characterizations

The team's major emphasis during this reporting period has been focused on the completion of the operational versions of the Marine Optical Buoys (MOBY's). Other work areas consisted of designing and testing bio-optical instrumentation, evaluating several of the SeaWiFS bio-optical protocols, processing data collected during field experiments, and reprocessing several of the Marine Optical Characteristics Experiment (MOCE) 2 and 3 bio-optical data sets. The team conducted one trip to the operations site in Honolulu, Hawaii, making necessary preparations for future field experiments. Part of the team also traveled to Moss Landing Marine Laboratories, Salinas, CA, and to American Holographic Co. Fitchburg MA, to assist with the fabrication of the next generation Marine Optical Buoys. Technical memoranda are being written to address the remote sensing reflectance, and instrument self-shading protocols. During the Ocean Color 96 meeting discussions with the Spanish on acquiring research vessel support during the MODIS validation period were conducted. A proposal will be generated towards this purpose for an experiment to be conducted off the North African coast during the summer of 1999.

Clark, Dennis K.

Effects of the 8 psia / 32% O2 Atmosphere on the Human in the Spaceflight Environment

Extravehicular activity (EVA) is at the core of a manned space exploration program. There are elements of exploration that may be safely and effectively performed by robots, but there are critical elements of exploration that will require the trained, assertive, and reasoning mind of a human crewmember. To effectively use these skills, NASA needs a safe, effective, and efficient EVA component integrated into the human exploration program. The EVA preparation time should be minimized and the suit pressure should be low to accommodate EVA tasks without undue fatigue, physical discomfort, or suit-related trauma. Commissioned in 2005, the Exploration Atmospheres Working Group (EAWG) had the primary goal of recommending to NASA an internal environment that allowed efficient and repetitive EVAs for missions that were to be enabled by the former Constellation Program. At the conclusion of the EAWG meeting, the 8.0 psia and 32% oxygen (O2) environment were recommended for EVA intensive phases of missions. As a result of selecting this internal environment, NASA gains the capability for efficient EVA with low risk of decompression sickness (DCS), but not without incurring additional negative stimulus of hypobaric hypoxia to the already physiologically challenging spaceflight environment. This paper provides a literature review of the human health and performance risks associated with the 8 psia/32% O2 environment. Of most concern are the potential effects on the central nervous system including increased intracranial pressure, visual impairment, sensorimotor dysfunction, and oxidative damage. Other areas of focus include validation of the DCS mitigation strategy, incidence and treatment of acute mountain sickness (AMS), development of new exercise countermeasures protocols, effective food preparation at 8 psia, assurance of quality sleep, and prevention of suit-induced injury. As a first effort, the trade space originally considered in the EAWG was re-evaluated looking for ways to decrease the hypoxic dose by further enriching the O2% or increasing the pressure. After discussion with the NASA engineering and materials community, it was determined that the O2 could be enriched from 32% to 34% and the pressure increased from 8.0 to 8.2 psia without significant penalty. These two small changes increase alveolar O2 pressure by 11 mmHg, which is expected to significantly benefit crewmembers. The 8.2/34 environment (inspired O2 pressure = 128 mmHg) is also physiologically equivalent to the staged decompression atmosphere of 10.2 psia / 26.5% O2 (inspired O2 pressure = 127 mmHg) used on 34 different shuttle missions for approximately a week each flight. Once decided, the proposed internal environment, if different than current experience, should be evaluated through appropriately simulated research studies. In many cases, the human physiologic concerns can be investigated effectively through integrated multi-discipline ground-based studies. Although missions proposing to use an 8.2/34 environment are still years away, it is recommended that these studies begin early enough to ensure that the correct decisions pertaining to vehicle design, mission operational concepts, and human health countermeasures are appropriately informed.

Environmental control

Railroad Valley Radiometric Calibration Test Site (RadCaTS) as Part of a Global Radiometric Calibration Network (RadCalNet)

The Radiometric Calibration Network (RadCalNet) is a coordinated multinational effort to provide in situ data that are suitable for the radiometric calibration and validation of Earth observation sensors that operate in the visible to shortwave infrared solar reflective spectral region (400 nm to 1000 nm). The main goals of RadCalNet are to provide top-of-atmosphere reflectance data to the scientific community, standardize data collection protocols for automated test sites, and to document the SI-traceable uncertainty budgets for each automated test site, of which there are currently four. The data available from RadCalNet are suitable for the calibration and validation of spaceborne imaging spectrometers. The work presented here provides a description of RadCalNet as well as a sample of the current results from the Radiometric Calibration Test Site (RadCaTS), which is located at Railroad Valley, Nevada, USA. Selected sensors for comparison include Terra and Aqua MODIS, SNPP and NOAA-20 VIIRS, and Sentinel-3A and -3B OLCI.

RadCalNet

Evidence Report: Risk of Hypobaric Hypoxia from the Exploration Atmosphere

Extravehicular activity (EVA) is at the core of a manned space exploration program. Some elements of exploration may be safely and effectively performed by robots, but certain critical elements will require the trained, assertive, and reasoning mind of a human crewmember. To effectively use these skills, NASA needs a safe, effective, and efficient EVA component integrated into the human exploration program. The EVA preparation time should be minimized and the suit pressure should be low to accommodate EVA tasks without causing undue fatigue, physical discomfort, or suit-related trauma. Commissioned in 2005, the Exploration Atmospheres Working Group (EAWG) had the primary goal of recommending to NASA an internal environment that allowed efficient and repetitive EVAs for missions that were to be enabled by the former Constellation Program. At the conclusion of the EAWG meeting, the 8.0 psia and 32% oxygen (O2) environment were recommended for EVA-intensive phases of missions. After re-evaluation in 2012, the 8/32 environment was altered to 8.2 psia and 34% O2 to reduce the hypoxic stress to a crewmember. These two small changes increase alveolar O2 pressure by 11 mmHg, which is expected to significantly benefit crewmembers. The 8.2/34 environment (inspired O2 pressure = 128 mmHg) is also physiologically equivalent to the staged decompression atmosphere of 10.2 psia / 26.5% O2 (inspired O2 pressure = 127 mmHg) used on 34 different shuttle missions for approximately a week each flight. As a result of selecting this internal environment, NASA gains the capability for efficient EVA with low risk of decompression sickness (DCS), but not without incurring the additional negative stimulus of hypobaric hypoxia to the already physiologically challenging spaceflight environment. This report provides a review of the human health and performance risks associated with the use of the 8.2 psia / 34% O2 environment during spaceflight. Of most concern are the potential effects on the central nervous system (CNS), including increased intracranial pressure, visual impairment, sensorimotor dysfunction, and oxidative damage. Other areas of focus include validation of the DCS mitigation strategy, incidence and treatment of transient acute mountain sickness (AMS), development of new exercise countermeasure protocols, effective food preparation at 8.2 psia, assurance of quality sleep, and prevention of suit-induced injury. Although missions proposing to use an 8.2/34 environment are still years away, it is recommended that these studies begin early enough to ensure that the correct decisions pertaining to vehicle design, mission operational concepts, and human health countermeasures are appropriately informed.

Norcross, Jason R.

In Flight MiRNA Isolation and Recovery on the ISS Using the Wetlab-2 System

Due to advancements in RNA research, mi (micro) RNAs and other small nucleotide RNAs have become a major research field in biology including spaceflight research. The regulation of RNA transcription and processing by miRNAs makes miRNAs an appealing topic for genetics and molecular research. It has been estimated that over 60% of human gene transcripts are targets of miRNA regulation. In fact, this is true for all organisms, including plants and insects. Small nucleotide RNAs can also play a role in regulating gene expression, meaning that gene expression alone is not a complete picture of the potential genetic changes that occur in an organism during spaceflight. The goal of the WetLab-2 project is to isolate and recover miRNAs from various tissue sources on the International Space Station (ISS). No system currently exists that can isolate and recover small nucleotide RNA in space. However, the WetLab-2 system that was validated on the ISS in 2016 can be adapted to fit this purpose. We are currently testing the new modified protocols by running plant and mouse blood experiments in parallel, allowing us to demonstrate the effectiveness of the procedure on different sample types. We expect to be able to optimize and implement the modified miRNA protocols for use on future ISS flights.

miRNA

In Flight miRNA Isolation and Recovery on the ISS

Due to advancements in RNA research, mi (micro) RNAs and other small nucleotide RNAs have become a major research field in biology including spaceflight research. The regulation of RNA transcription and processing by miRNAs makes miRNAs an appealing topic for genetics and molecular research. It has been estimated that over 60% of human gene transcripts are targets of miRNA regulation. In fact, this is true for all organisms, including plants and insects. Small nucleotide RNAs can also play a role in regulating gene expression, meaning that gene expression alone is not a complete picture of the potential genetic changes that occur in an organism during spaceflight. The goal of the WetLab-2 project is to isolate and recover miRNAs from various tissue sources on the International Space Station (ISS). No system currently exists that can isolate and recover small nucleotide RNA in space. However, the WetLab-2 system that was validated on the ISS in 2016 can be adapted to fit this purpose. We are currently testing the new modified protocols by running plant and mouse blood experiments in parallel, allowing us to demonstrate the effectiveness of the procedure on different sample types. We expect to be able to optimize and implement the modified miRNA protocols for use on future ISS flights.

miRNA

Snowex 2017 Community Snow Depth Measurements: A Quality-Controlled, Georeferenced Product

Snow depth was one of the core ground measurements required to validate remotely-sensed data collected during SnowEx Year 1, which occurred in Colorado. The use of a single, common protocol was fundamental to produce a community reference dataset of high quality. Most of the nearly 100 Grand Mesa and Senator Beck Basin SnowEx ground crew participants contributed to this crucial dataset during 6-25 February 2017. Snow depths were measured along ~300 m transects, whose locations were determined according to a random-stratified approach using snowfall and tree-density gradients. Two-person teams used snowmobiles, skis, or snowshoes to travel to staked transect locations and to conduct measurements. Depths were measured with a 1-cm incremented probe every 3 meters along transects. In shallow areas of Grand Mesa, depth measurements were also collected with GPS snow-depth probes (a.k.a. MagnaProbes) at ~1-m intervals. During summer 2017, all reference stake positions were surveyed with <10 cm accuracy to improve overall snow depth location accuracy. During the campaign, 193 transects were measured over three weeks at Grand Mesa and 40 were collected over two weeks in Senator Beck Basin, representing more than 27,000 depth values. Each day of the campaign depth measurements were written in waterproof field books and photographed by National Snow and Ice Data Center (NSIDC) participants. The data were later transcribed and prepared for extensive quality assessment and control. Common issues such as protocol errors (e.g., survey in reverse direction), notebook image issues (e.g., halo in the center of digitized picture), and data-entry errors (sloppy writing and transcription errors) were identified and fixed on a point-by-point basis. In addition, we strove to produce a georeferenced product of fine quality, so we calculated and interpolated coordinates for every depth measurement based on surveyed stakes and the number of measurements made per transect. The product has been submitted to NSIDC in csv format. To educate data users, we present the study design and processing steps that have improved the quality and usability of this product. Also, we will address measurement and design uncertainties, which are different in open vs. forest areas.

Brucker, L.

Measuring sleep habits without using a diary: the sleep timing questionnaire

STUDY OBJECTIVES: To develop a single-administration instrument yielding equivalent measures of sleep to those obtained from a formal (2-week) sleep diary. DESIGN & SETTING: A single-administration Sleep riming Questionnaire (STQ) is described (and reproduced in the Appendix). Test-retest reliability was examined in 40 subjects who were given the STQ on two occasions separated by less than 1 year. Convergent validity was measured both by comparing STO-derived measures with objective measures derived from wrist actigraphy (n=23) and by comparing STQ-derived measures with other subjective measures derived from a detailed 2-week sleep diary in two nonoverlapping samples (n=101, 93). Correlations of STQ measures with age and momingness-eveningness (chronotype) were also examined. SUBJECTS: The analyses used sample sizes of 40, 23, 101, and 93 (both genders, overall age range 20y-89y). Most subjects were healthy volunteers; some Study 4 subjects were patients (enrolled in research protocols). RESULTS: Test-retest reliability for the STQ was demonstrated for estimates of bedtime (r = 0.705, p < 0.001) and waketime (r = 0.826, p < 0.001). Convergent validity using wrist actigraphy was demonstrated by correlations of 0.592 (p < 0.005) for bedtime, and of 0.769 (p < 0.001) for waketime. Diary studies indicated STQ bedtime and waketime data to be highly correlated (at about 0.8) with those obtained from a formal 2-week sleep diary. The STQ also provided data on estimated sleep latency and wake after sleep onset (WASO), which correlated reliably (at about 0.7) with average nightly ratings of these variables from a 2-week sleep diary. Mean estimated values of sleep latency and WASO from the two instruments were within 1 minute of each other. ST-derived bedtimes and waketimes correlated with both age and chronotype in the expected direction (older subjects earlier, morning types earlier). CONCLUSION: The STQ may be a reliable valid measure of sleep timing that could provide a time-efficient alternative to traditional sleep diaries.

Validation Studies

A Pilot Study for Applying an Extravehicular Activity Exercise Prebreathe Protocol to the International Space Station

Decompression sickness (DCS) is a serious risk to astronauts performing extravehicular activity (EVA). To reduce this risk, the addition of ten minutes of moderate exercise (75% VO2pk) during prebreathe has been shown to decrease the total prebreathe time from 4 to 2 hours and to decrease the incidence of DCS. The overall purpose of this pilot study was to develop an exercise protocol using flight hardware and an in-flight physical fitness cycle test to perform prebreathe exercise before an EVA. Eleven subjects volunteered to participate in this study. The first objective of this study was to compare the steady-state heart rate (HR) and oxygen consumption (VO2) from a submaximal arm and leg exercise (ALE) session with those predicted from a maximal ALE test. The second objective was to compare the steady-state HR and V02 from a submaximal elastic tube and leg exercise (TLE) session with those predicted from the maximal ALE test. The third objective involved a comparison of the maximal ALE test with a maximal leg-only (LE) test to conform to the in- flight fitness assessment test. The 75% VO2pk target HR from the LE test was significantly less than the target HR from the ALE test. Prescribing exercise using data from the maximal ALE test resulted in the measured submaximal values being higher than predicted VO2 and HR. The results of this pilot study suggest that elastic tubing is valid during EVA prebreathe as a method of arm exercise with the flight leg ergometer and it is recommended that prebreathe countermeasure exercise protocol incorporate this method.

Woodruff, Kristin K.

Data management

The following tasks were prioritized: software acquisition management plan; space station flight data system architectural study; space station user data system interface; automation of software development process; automation of software testing; distributed data base management; ADA (automated data acquisition) evaluation and transition and planning; network operating system software; fault tolerant computer validation methodology for onboard data management system; systems integration; artificial intelligence/expert systems; space station data network concept; space station standard interface protocols; space station data networks systems; integrated software development facility; and language trade studies.

Love, G.

Medical Lessons Learned from the Exploration Atmospheres Study

Background: The National Aeronautics and Space Administration’s (NASA) Exploration Atmospheres study (EA) was done to evaluate alternative cabin atmospheres for future spacecraft designs and planetary surface exploration of the Moon, Mars and beyond. Deep space exploration involves creating habitats and environments safe for human occupancy and means to explore the outside environment (extravehicular activities, EVA). In order to validate alternative atmospheres and pre-spacewalk procedures, the EA study was conducted to evaluate factors such as hypoxia risks, denitrogenation protocols, food limitations, medications, and the impact of other factors on human performance. Overview: Space travel is constrained by mass, volume, power and the cost of vehicle development, which creates tradeoffs in various capabilities, including breathing gasses. Higher atmospheric pressure in a vehicle means more gas, and a stronger containment vessel/habitat; while lower pressure requires higher oxygen partial pressure, which may increase fire risk. NASA’s EA study evaluated a proposed alternative cabin environment (8.2 psia, 34% Oxygen), for future spacecraft habitat, and planetary EVAs. EA included both a 3-day and a 11-day trial. These trials included a depressurization and saturation to 8.2psi at 34% O2 with additional depresses to 4.3 psia at 85% O2 for simulated EVAs, (1 EVA during the 3-day and 5 during the 11-day trials). Discussion: Planning for and executing the medical monitoring and response plan for a trial of this scope was a huge undertaking with no prior practice to fall back on. Food obstacles, sleeping issues, medications, joint injury, equipment limitations, medical privacy, multiple cases of decompression sickness, and even a COVID outbreak among the support team proved challenging. Conclusion: Testing of this nature is an essential part NASA’s preparation for the upcoming Lunar Artemis missions. As spaceflight transitions beyond low earth orbit, to planetary, even more trials of this nature will be required to learn what are the optimal atmospheric and associated operational constraints to maintain the optimal health of the crew and achieve mission objectives.

R Sanders