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Planetary Protection at Marshall Space Flight Center

Introduction: NASA Marshall Space Flight Center (MSFC) is historically known for its role in propulsion. While this is still the mainstay of MSFC’s expertise, many unique capabilities exist at MSFC which pertain to Planetary Protection (PP), including 1) identifying PP threats, and 2) developing novel methods to neutralize those threats. Furthermore, because these capabilities exist among diverse groups at MSFC, this work promotes collaboration both within and outside MSFC to expand and develop PP studies related to a full spectrum of NASA research, design, manufacture, and test interests. This abstract describes the PP research ongoing at MSFC and describes how it contributes to NASA’s overall PP objectives. Microbial Identification in Cleanrooms: One of the greatest threats to successful implementation of PP requirements is recontamination post bioburden reduction. One method to prevent recontamination is to keep the spacecraft in clean environments (i.e. cleanrooms) as much as possible during assembly and integration. However, cleanrooms are not without their own sources of contamination, which is why NASA is interested in monitoring the cleanliness of cleanrooms and characterizing the microbial species present. Such information allows a greater understanding of the resistance of these microbes to cleaning methods, as well as the risk of their contaminating the targeted planetary body of a given mission. MSFC has multiple cleanrooms of various ISO cleanliness levels onsite. We sampled the air and surfaces of three of these rooms, isolated microbes, and then sequenced the 16S rRNA gene or ITS region of the 18S rRNA gene for bacterial and fungal isolates, respectively. This has resulted in a microbial library which currently includes nearly 100 isolates. Microbial Enumeration of Spacecraft Materials: Currently, there are only a couple bioburden reduction methods approved by NASA, and often the harshness of these methods presents additional concerns or risks related to material properties. The goal of this research is to assess the microbial content of solid rocket motor (SRM) materials potentially used for lander missions. This work aims to more accurately define the risk of planetary contamination by providing empirical data associated with commonly used SRM raw materials. In this study, we pulverized nonmetallic SRM materials using a cryogenic grinder, then analyzed the resulting substrate for microbial colony forming units (CFU). We found that many SRM nonmetallic materials do not harbor detectable bioburden, though a range existed depending on the material. The results from this work provide quantitative data to potentially reduce concerns of contamination, while also providing a foundation for follow up studies into additional sterilization methods and molecular identification of contaminating microbes. Space Environmental Effects on Microbial Survival: One potential area of microbial reduction is the space environment. Understanding the survivability of hardy microbes in space-like conditions is a crucial first step in answering how space may reduce bioburden and if it can be relied upon for adherence to PP requirements. This work studied the effects of ultraviolet (UV) and ionizing radiation on survival of Bacillus atrophaeus spores. Microbes were dried on relevant polymeric materials then exposed to space environmental stressors. Coupons were submerged in water, diluted, and plated to determine survival compared with controls. We found that both UV and ionizing radiation were capable of reducing viability by nearly 99%, but there were still survivors, some with changed morphology indicating resistance mechanisms within certain cells. Manufacturing credit: Finally, given the above-mentioned limitations of the NASA-approved bioburden reduction methods, there is interest in understanding if manufacturing processes may provide enough bioburden reduction without additional PP-specific bakeouts. For instance, some material additives may be antimicrobial. Given this, we investigated the effects of several commonly used rubber additives on the growth of B. atrophaeus spores. We found that some of the materials inhibited growth of the spores, possibly supporting the use of these additives on missions with PP constraints. Future work into manufacturing credit for bioburden reduction includes inoculation of green insulation with B. atrophaeus spores, followed by a typical cure. Thermal profiles will be verified for appropriate temperature and durations to meet PP requirements, and cured samples will be analyzed using a cryogenic grinder to determine survivability of spores.

Chelsi D. Cassilly↗

Transcriptomics-based Machine Learning (ML) Analysis Predicts Space-Exposed Murine Livers

Limited sample sizes, high data dimensionality, and sensitivity to technical and biological variability of next generation sequencing (NGS), typically limits machine learning (ML) approaches in spaceflight studies that include radiation effects. However, pooling smaller studies while addressing intra- and inter-study variabilities allows for ML predictive modeling. Here, integration methods were applied to whole transcriptome shotgun sequencing (RNA-seq) data from six mouse liver GeneLab datasets (GLDS) (n ranging from 6 to 39 samples) from with a total of 81 spaceflight and ground-control samples to determine top features (i.e. genes) relevant to spaceflight including the effect of radiation exposure. RNASeq counts were normalized for each study, then merged and scaled across all datasets. Data dimensionality was reduced using a minimum redundancy maximum relevance (MRMR) methodology. Redundancy and relevance were computed using the Pearson correlation and F-statistic, respectively. The top 100 MRMR features were used to predict spaceflight vs. ground-control samples using Random Forest (RF), Support Vector Machine (SVM), and Linear Discriminant Analysis (LDA) classifiers with 5-fold cross validation (CV). Principal component analysis (PCA) on the complete feature set versus the MRMR features shows separation between spaceflight samples and ground controls (Figure 1A). The ML-based gene sets were compared against differential gene expression results obtained with DESeq2 from individual GLDS. Using all features or randomly sampled subsets at matching set sizes with MRMR, a maximum classifier accuracy of 69% on the test set over 5 folds. For all classifiers, CV training using at least the top 30 MRMR genes show minimum 89% accuracy and 0.95 AUC value on the test set over 5 folds (Figure 1B). Baseline set analysis on differentially expressed genes (DEGs) identified using padj ≤ 0.05 show 295 DEGs that overlap at least two studies and 13 DEGs that overlap three studies (Figure 1C). Set analysis between the top 100 MRMR features and the DEGs showed 47 genes that overlap at least one study and 24 genes that overlap two studies. Over-representation analysis showed overlapping biological processes related to fatty acid and lipid metabolism which may indicate these processes in the response to spaceflight stressors. MRMR feature selection for the selected ML methods improve performance relative to a classifier built on all features or randomly sampled subsets. Permutation feature importance within the decorrelated MRMR features showed concordance in feature ranking between ML methods. A challenge of applying ML methods across heterogeneous NGS data is accounting for signal:noise. Here, signal validation across studies was shown by intersecting sets between top MRMR genes and DEGs from DESeq2 analysis. Non-intersecting sets introduce opportunity to explore genes relevant to differentiating space flight exposed groups and implementing ML methods across existing NGS datasets may overcome sample size limitations.

Machine Learning↗

Validation of Fitness for Duty Standards Using Pre- and Post-Flight Capsule Egress and Suited Functional Performance Tasks in Simulated Reduced Gravity

The transition between gravity environments will involve one of the most complex, high-risk phases of any mission. For example, the reduced functional capacity caused by physiological deconditioning adaptations in microgravity coupled with the stressors of re-entry into partial gravity environments will increase risks to crew, even with rigorous adherence to inflight countermeasures. Quantification of the astronauts’ post-landing functional performance is necessary to design ConOps for exploration missions. Specifically, these two high-risk scenarios may be required to be performed soon after gravity transitions: • Nominal and/or emergency unassisted capsule egress task after return to Earth • Planetary extravehicular activity (EVA) soon after landing on Mars or the Moon This study is broken down into two phases. Phase 1: A feasibility study that will assess the overall feasibility and demonstrate the capability to do these tasks shortly after landing. Phase 2: the full Egress Fitness study, which is part of the CIPHER complement. This study uses a task-based approach to characterize functional performance of these high-risk scenarios in long-duration ISS crewmembers before flight and shortly after return to Earth. Prior to any testing, each astronaut subject completes a suit fit check to ensure adequate sizing and a mobility assessment to confirm completion of the EVA tasks. Pilot Egress Fitness pre-flight and post-flight testing includes an Earth-based emergency egress out of a functional capsule mockup and a short Mars gravity EVA simulation including suit donning, hatch egress, ladder descent, task board cable operations, baggage transfer over sand/rocky regolith, alignment with a rear entry port, and suit egress. Pre-flight testing will occur at any time point prior to flight. Post-flight assessment is much more critical with the capsule egress test that will occur 1–4 hours after landing and the planetary EVA approximately 18–36 hours after landing. The CIPHER study will incorporate additional pre-flight sessions, longer EVA tasks that include traverse and geology sampling, and post-flight sessions on R+1, 4, and 8 to characterize the timeframe of recovery. Data collected for both tasks include task completion time, photo, and video. The EVA portion also includes collection of metabolic and heart rate. Pilot Egress Fitness study has completed both baseline and post-flight testing on four astronaut subjects, with all four able to complete post-flight EVA testing and three able to complete postflight capsule egress testing. Preliminary results observe individual physiological variations, which were to be expected but also suggest the need to carefully track the timeline from undock to landing to testing. Furthermore, some task performance instructions and equipment may need to be adjusted to ensure results are primarily physiological. These and other lessons learned will be addressed with minor protocol changes in the full CIPHER Egress Fitness study.

J R Norcross↗

Method Development for In Situ Microbiome Profiling of the Water Recovery System’s Wastewater Tank Onboard the International Space Station

A distinctive microbial community has inhabited the International Space Station (ISS) Water Recovery System (WRS) for over 14 years and has experienced the stressors associated with the microgravity environment. The WRS generates potable water for the crew from urine distillate, humidity condensate, Sabatier product water, and the occasional off-loading of ground-supplied water (1). The reservoir for these products, the wastewater tank, does not have a means of microbial control. Current in situ microbial monitoring of the WRS is limited to quarterly culture-based assessments of the potable water product using a microbial capture device and coliform detection bag. Additional analysis of the wastewater and condensate sources are collected into Teflon bags for analysis following return to the ground. The time between sample collection and the return to Earth, as well as the lack of preservation, results in a skewed depiction of the microbiome. Routinely observed from these returned wastewater samples are high counts (105 – 106 colony forming units per mL) and two prevailing genera, Ralstonia and Cupriavidus, as well as a high abundance of unidentified organisms (Table 1). The wastewater tank likely contains a more diverse microbiome, as a higher diversity of bacteria and fungi has been noted upstream and downstream of the tank.

Sarah Stahl-Rommel↗

Lessons Learned From Plant Research in Low Earth Orbit

This presentation is part of the pre-ASGSR workshop that will be held in Houston, Texas on November 9, 2022. The overall objective of the workshop is to discuss conducting biology experiments beyond low Earth orbit (LEO) and present the challenges and the paths forward. In part, the workshop will highlight lessons learned in plant space biology, some of the plant spaceflight stressors that are anticipated to cause biological changes beyond LEO and discuss what is currently known about these changes, and how results can be leveraged for future missions to the Moon and Mars. Platforms that can be utilized to conduct plant science experiments beyond LEO will also be discussed.

Plant Space Biology↗

Reanalysis of Rodent Data from Spacelab Life Science-1

The space bioscience field has long been plagued by the challenge of spaceflight with effects of radiation and microgravity. Having multiple and repeated spaceflight experiments for model organisms to solve these space stressors is costly and time consuming. Therefore, reusing and reanalyzing legacy experiments is one way that scientists can draw new conclusions in a timely manner and without using too many resources. Moreover, advances in general biological knowledge allows legacy experiments to be placed into more complete context.Here we aim to analyze all data and metadata taken from rats flown on the SLS-1 mission to create a comprehensive biological model that can be supplemented with current data to allow new discoveries in how space flown organisms adapt to the space environment. Our approach begins with the identification of all the data and metadata, including graphs and tables, for SLS-1 in NASA archives and other sources. Then, each piece of data and metadata will be digitized, reformatted and analyzed. Lastly, a previously developed astronaut model will be used to create the data framework and a comprehensive biological rodent model. The datasets we are using is from the 1991 SpaceLab Life Science 1 (SLS-1) NASA Mission. This was the first designated spacelab mission flown. All 29 rodents were tested for nine days in two different habitats: Research Animal Holding Facility (RAHF) and Animal Enclosure Module (AEM). The rodents were prepared for a live return and compared to a ground control. A total of 30 rodent experiments were accepted as flight studies on the mission. By digitization and reorganizing SLS-1 rat data we will both directly generate new insights and indirectly enable other scientists to by providing the data and metadata in a digitized form.

Space Biology↗

Interactive Biogenic Emissions and Drought Stress Effects on Atmospheric Composition in NASA GISS ModelE

Drought is a hydroclimatic extreme that causes perturbations to the terrestrial biosphere and acts as a stressor on vegetation, affecting emissions patterns. During severe drought, isoprene emissions are reduced. In this paper, we focus on capturing this reduction signal by implementing a new percentile isoprene drought stress (y d ) algorithm in NASA GISS ModelE based on the MEGAN3 (Model of Emissions of Gases and Aerosols from Nature Version 3) approach as a function of a photosynthetic parameter (V c,max ) and water stress (β). Four global transient simulations from 2003–2013 are used to demonstrate the effect without y d (Default_ModelE) and with online y d (DroughtStress_ModelE). DroughtStress_ModelE is evaluated against the observed isoprene measurements at the Missouri Ozarks AmeriFlux (MOFLUX) site during the 2012 severe drought where improvements in the correlation coefficient indicate it is a suitable drought stress parameterization to capture the reduction signal during severe drought. The application of y d globally leads to a decadal average reduction of ∼ 2.7 %, which is equivalent to ∼14.6 Tg yr -1 of isoprene. The changes have larger impacts in regions such as the southeastern US. DroughtStress_ModelE is validated using the satellite ΩHCHO column from the Ozone Monitoring Instrument (OMI) and surface O 3 observations across regions of the US to examine the effect of drought on atmospheric composition. It was found that the inclusion of isoprene drought stress reduced the overestimation of ΩHCHO in Default_ModelE during the 2007 and 2011 southeastern US droughts and led to improvements in simulated O 3 during drought periods. We conclude that isoprene drought stress should be tuned on a model-by-model basis because the variables used in the parameterization responses are relative to the land surface model hydrology scheme (LSM) and the effects of y d application could be larger than seen here due to ModelE not having large biases of isoprene during severe drought.

drought stress effects↗

Assessing Environmental and Socioeconomic Factors of Urban Flood Vulnerability in Kansas City, Kansas

Pluvial flooding, over-saturated ground, and drainage systems disproportionately impact historically marginalized urban neighborhoods during extreme rainfall events. These communities are impacted by physical and socioeconomic factors that make them vulnerable to flooding events, such as high concentrations of impervious landcover, high precipitation rates, and a combined sewer system framework. Despite known vulnerability to environmental hazards, understanding potential pluvial street-level flooding events are largely unknown. Using the open-source National Capital Project’s Integrated Valuation of Ecosystem Services and Tradeoffs (InVEST) Urban Flood Risk Mitigation model, NASA DEVELOP examined neighborhood scale runoff retention and potential economic damages for risk mapping throughout Kansas City, Kansas. The generated outputs aid in identifying vulnerable neighborhoods susceptible to flooding and in need of future intervention. Previous studies have applied this model framework to understand urban flood vulnerability regarding ecosystem services, urban planning, and flood mitigation strategies. We utilized the InVEST outputs to develop indices pertaining to environmental justice factors of race, socioeconomic status, social vulnerability, and health. The findings indicate that historically redlined neighborhoods in Kansas City, Kansas face disproportional impacts from flood events and are subject to greater environmental stressors. This research provides an approach to utilizing an open-source flood vulnerability model to empower neighborhood-scale environmental justice analysis, enhancing the local communities' understanding of present-day impacts of historical environmental injustices.

Hadwynne Gross↗

Analog Environments for Venus Aerosol Instrument and Mission Concept Testing

The most common hypothesis discussed for life on Venus is a habitat in the dense, persistent clouds, which are primarily sulfuric acid with a secondary water component. Although Earth lacks a complete Venus analog environment, several partial analogs exist which can inform our understanding of the requirements for cloud habitability as well as serving as potential testbeds for future Venus instruments or missions. Earth’s tropospheric clouds have larger particles, more water, and shorter lifetimes than Venus’s; however, they have a partial overlap in pressure and temperature, and are our only example of an airborne habitat.They can inform how particle residence time and microbial generation time constrain habitable atmospheric regions. On the biological side, this includes available nutrients and energy, environmental stressors such as radiation, osmotic potential and acidity, and time spent in protective but inactive forms (endospores, cysts, akinetes); on the physical side, gravity, air density and viscosity, thermal lofting, gravity waves, scavenging due to precipitation, and other airflow dynamics, as well as effective particle radius and electrical charge. They can also enable sampling strategy tests for airborne life detection, ranging from10 2 to10 8 viable cells/mL (10 -3 to 10 -9 cells/particle). Conversely, Earth's stratospheric sulfate layer lacks an active microbial presence, and is at a colder and less dense altitude but has partially analogous values in terms of H2SO4 and water concentration, particle size, and number density. These aerosols are accessible to aircraft and high-altitude balloons. Even more accessible potential analogs include sea spray, which overlaps with the smaller (haze) particle sizes and densities at Venus and contains trace organic constituents; marine fog, which at higher densities can match Venus’s larger (Mode 2) particle properties and contains a range of trace compounds including ammonia, phosphates, sulfates, and nitrates; and spray from acidic hydrothermal systems, which can cover a wide variety of sizes, densities, and chemical compositions. These analogs could improve the development of Venus cloud sampling instruments or sondes, including field tests of material compatibility, particle capture efficiency, and analysis sensitivity.

Analog↗

Climate Anomalies Correlation with Population Trends of the Eastern Towhee in the Mid-Eastern U.S.

The species of sparrow identified by foraging under dense thickets, the Eastern Towhee (Pipilo erythrophthalmus) resides in the Eastern United States, some populations either permanently living in the southeastern US or migratory populations living in the North. The mostly migratory Eastern Towhee has been experiencing population changes since the 1980s in the Eastern Seaboard. This species is increasing during the winter and decreasing during the summer, which is their breeding season. The Christmas Bird Count shows that in the winter, the Eastern Towhee populations have been increasing in their northern wintering range by 0.2625% in Maryland, 0.056% in West Virginia, 0.133% in New Jersey, 0.049% in Virginia, and 0.224% in Delaware from 1984-2019. The Breeding Bird Survey Data shows that in the summer, the Eastern Towhee populations have been declining in the Eastern United States by -4.0635% in Maryland, -3.073% in West Virginia, -15.0722% in New Jersey, -3.9375% in Virginia, and -6.3735% in Delaware from 1984-2015. The average climate anomalies in June range from -0.108 to 0.3705 degrees Celsius while the average climate anomalies in January range from 0.1395 to 0.5014 degrees Celsius. The Eastern Towhee are moving northward towards their northern wintering range during the winter and we explore the hypothesis of northward range shift in association with climate change. Our research displays how the behavior patterns of species are varying due to this stressor and how to better conserve species like the Eastern Towhee.

Mahlet Mesfin↗

Risk of Impaired Performance Due to Reduced Muscle Mass, Strength &, Endurance (Short Title: Muscle) and Risk of Reduced Physical Performance Capabilities Due to Reduced Aerobic Capacity (Short Title: Aerobic)

This report reviews the scientific literature regarding the human system risks to the microgravity environment of space flight in relation to human performance. The primary human performance-related risks involve deconditioning of the cardiovascular and skeletal muscles systems due to prolonged exposure to the reduced gravitational input. The chronological history of U.S. space flight is reviewed as a starting point to inform and understand the gaps in the knowledge to these risks. Maintenance of physical performance capabilities involves understanding the health of many organ systems (peripheral [vascular, heart, blood volume, skeletal muscle] and central [brain]) that ultimately contribute to the submaximal and maximal capacity of the aerobic (VO2peak), skeletal muscle (strength and endurance) systems. Maintaining astronaut VO2peak, muscle mass, strength, and endurance before, during, and after space flight is a significant priority to NASA for the current International Space Station (ISS) era, as well as for future exploration missions. A growing research database from both space flight and ground-based analog studies finds that the cardiorespiratory system is compromised and skeletal muscles (predominantly postural muscles of the lower limbs) undergo atrophy. These structural and metabolic responses to living in microgravity conditions contribute to physiological deconditioning during space flight that potentially increase the risks to astronauts returning to surface operations (i.e., Moon, Mars, or Earth). The time course changes from short to long-duration space flight and the relationships between in-flight performance deconditioning levels are not well characterized. Moreover, there are large interindividual variabilities that may be dependent on genetics, age, sex, preflight fitness levels, and individual exercise prescriptions that need further careful evaluations. Efforts should be made to understand the current status of preflight, in-flight, and postflight exercise performance capability and to define the operational goals and target areas for protection with the in-flight exercise program. There is a bi-directional relationship between exercise prescription and hardware countermeasures that need further understanding in-flight. For example, hardware with limited capabilities/modalities may be counterbalanced by changes in exercise prescription (i.e., frequency, time, intensity, volume) for providing effective responses to maintain fitness. Importantly, the minimal requirements for exercise prescription on ISS hardware may not translate to lower capability hardware on exploration missions. Due to limited volume on exploration vehicles, future Artemis missions to the Lunar surface will not have similar exercise hardware capabilities as ISS. This may alter the effectiveness of hardware to provide adequate physiological stress on bodily systems allowing for adaptations to maintain aerobic capacity, strength, and bone density. Thus, it will be important to understand the exercise responses of current ISS countermeasures to develop individualized exercise prescriptions that minimize aerobic and muscular risks, accounting for the large variability of responses among crewmembers. Newer exploration exercise hardware is currently being evaluated that is more compact (i.e., E4D and Orion Flywheel) and will require careful evaluation of the hardware on the stressor (i.e., metabolic rate, oxygen uptake, and heart rate work relationships, and force plate load profiles) needed the human body to protect and maintain crew health and performance. Moreover, exercise responses on the hardware need careful evaluation on the chronic adaptations. Lastly, in-flight evaluation of hardware exercise response may differ in 0-g or partial-g compared to 1-g. Therefore, it cannot be assumed that the stress on the body will be the same in each environment. Understanding this has a direct impact on exercise prescriptions. This document provides an overview of key scientific investigations that have been conducted before, during, and after human space flight missions, as well as from human ground-based analog studies that contribute to the evidence base on changes in aerobic capacity and muscle mass, strength, and endurance. Additional data from rodent and nonhuman primate experiments of skeletal muscle unloading completed during space flight or ground-based flight-simulations provide supportive information about this risk topic. Most importantly, a recent, large dataset from long-duration ISS crew has been added to give improved insight into the variability of exercise response of crew, demonstrating that a large portion of the crew population return to Earth with greater than 10-20% loss of aerobic capacity and muscle strength and endurance. Data from human space flight and ground-based studies are narrowing in on the required exercise paradigms but thus far still provide an incomplete answer to an effective approach for maintaining skeletal muscle function and aerobic fitness of all human space travelers. Finally, the relationship of this risk topic to various space flight operational scenarios is examined and discussed.

Eric Rivas↗

Transcriptomics-based Machine Learning Analysis Predicts Space-Exposed Murine Livers

Limited sample sizes, high data dimensionality, and sensitivity to technical and biological variability of next generation sequencing (NGS), typically limits machine learning (ML) approaches in spaceflight studies that include radiation effects. However, pooling smaller studies while addressing intra- and inter-study variabilities allows for ML predictive modeling. Here, integration methods were applied to whole transcriptome shotgun sequencing (RNA-seq) data from six mouse liver GeneLab datasets (GLDS) (n ranging from 6 to 39 samples) from with a total of 81 spaceflight and ground-control samples to determine top features (i.e. genes) relevant to spaceflight including the effect of radiation exposure. RNASeq counts were normalized for each study, then merged and scaled across all datasets. Data dimensionality was reduced using a minimum redundancy maximum relevance (MRMR) methodology. Redundancy and relevance were computed using the Pearson correlation and F-statistic, respectively. The top 100 MRMR features were used to predict spaceflight vs. ground-control samples using Random Forest (RF), Support Vector Machine (SVM), and Linear Discriminant Analysis (LDA) classifiers with 5-fold cross validation (CV). Principal component analysis (PCA) on the complete feature set versus the MRMR features shows separation between spaceflight samples and ground controls (Figure 1A). The ML-based gene sets were compared against differential gene expression results obtained with DESeq2 from individual GLDS. Using all features or randomly sampled subsets at matching set sizes with MRMR, a maximum classifier accuracy of 69% was shown on the test set over 5 folds. For all classifiers, CV training using at least the top 30 MRMR genes show minimum 89% accuracy and 0.95 AUC value on the test set over 5 folds (Figure 1B). Baseline set analysis on differentially expressed genes (DEGs) identified using padj ≤ 0.05 show 295 DEGs that overlap at least two studies and 13 DEGs that overlap three studies (Figure 1C). Set analysis between the top 100 MRMR features and the DEGs showed 47 genes that overlap at least one study and 24 genes that overlap two studies. Over-representation analysis showed overlapping biological processes related to fatty acid and lipid metabolism which may indicate these processes in the response to spaceflight stressors. MRMR feature selection for the selected ML methods improve performance relative to a classifier built on all features or randomly sampled subsets. Permutation feature importance within the decorrelated MRMR features showed concordance in feature ranking between ML methods. A challenge of applying ML methods across heterogeneous NGS data is accounting for signal:noise. Here, signal validation across studies was shown by intersecting sets between top MRMR genes and DEGs from DESeq2 analysis. Non-intersecting sets introduce opportunity to explore genes relevant to differentiating space flight exposed groups and implementing ML methods across existing NGS datasets may overcome sample size limitations.

Machine Learning↗

A Review of the Safety of Selective Serotonin Reuptake Inhibitors for Long Duration Spaceflight

Introduction: As humankind ventures further into the depths of space, planning is already underway for long-duration exploration missions that will test the bounds of human performance. Deep space travel will include added risk related to stressors from the isolated, confined, and extreme (ICE) environment that lies outside the boundaries of low-earth orbit. Currently, selective serotonin reuptake inhibitors (SSRIs) are considered the gold standard treatment for many mental health diagnoses including anxiety and depression; however, SSRIs are also associated with several undesired side effects. The utility of non-pharmacologic therapies for the management of behavioral health conditions has not yet been fully explored. Methods: A comprehensive literature search was performed using PubMed and relevant articles pertaining to the psychological impacts of ICE environments, use of SSRIs in spaceflight, side effects associated with SSRIs, and non-pharmacologic treatments for anxiety and depression were reviewed. Over 100 studies were reviewed in total. Results: Reduced bone mineral density, impaired hemostatic function, significant individual variability resulting from gene polymorphisms, and drug-drug interactions are well described adverse effects of SSRIs that may complicate their operational use in the deep space environment. Many non-pharmacologic therapies have been utilized with success for the management of behavioral health conditions in the terrestrial environment that may show promise for long duration missions. Discussion: Although SSRIs have long been considered standard of care treatment for many behavioral health conditions, we cannot trivialize the risk that prolonged pharmacologic therapy may pose. The need to mitigate these risks by exploring alternative non-pharmacologic therapies has never been more relevant.

Charles H Dukes↗

Differential Saccade-Pursuit Coordination Under Sleep Loss and Low-Dose Alcohol

Introduction: Ocular tracking of a moving object requires tight coordination between smooth pursuit and saccadic eye movements. Normally, pursuit drives gaze velocity to closely match target velocity, with residual position offsets corrected by catch-up saccades. However, how/if common stressors affect this coordination is largely unknown. This study seeks to elucidate the effects of acute and chronic sleep loss, and low-dose alcohol, on saccade-pursuit coordination, as well as that of caffeine. Methods: We used an ocular tracking paradigm to assess three metrics of tracking (pursuit gain, saccade rate, saccade amplitude) and to compute “ground lost” (from reductions in steady-state pursuit gain) and “ground recouped” (from increases in steady-state saccade rate and/or amplitude). We emphasize that these are measures of relative changes in positional offsets, and not absolute offset from the fovea. Results: Under low-dose alcohol and acute sleep loss, ground lost was similarly large. However, under the former, it was nearly completely recouped by saccades, whereas under the latter, compensation was at best partial. Under chronic sleep restriction and acute sleep loss with a caffeine countermeasure, the pursuit deficit was dramatically smaller, yet saccadic behavior remained altered from baseline. In particular, saccadic rate remained significantly elevated, despite the fact that ground lost was minimal. Discussion: This constellation of findings demonstrates differential impacts on saccade-pursuit coordination with low-dose alcohol impacting only pursuit, likely through extrastriate cortical pathways, while acute sleep loss not only disrupts pursuit but also undermines saccadic compensation, likely through midbrain/brainstem pathways. Furthermore, while chronic sleep loss and caffeine-mitigated acute sleep loss show little residual pursuit deficit, consistent with uncompromised cortical visual processing, they nonetheless show an elevated saccade rate, suggesting residual midbrain and/or brainstem impacts.

smooth pursuit↗

Space Algae-2: Preflight Testing for A Long-Duration, Multi-Omics Analysis of Arthrospira Platensis

The cyanobacteria Arthrospira platensis NIES-39, commonly known as spirulina, could provide a fresh supply of nutrients for crew on long-duration spaceflight missions. Spirulina is a readily digestible food that is high in protein with all essential amino acids as well as significant levels of B vitamins, antioxidants, and anti-inflammatory metabolites. Spaceflight has multiple abiotic stressors such as increased ionizing radiation and microgravity, which causes a lack of convective mixing. These environmental conditions may impact productivity, nutritional composition, and in long-duration propagation, spaceflight stress may impact the genetic stability of spirulina cultures. We are developing an International Space Station experiment to continuously culture A. platensis for six months. Multi-omics profiling will be used to monitor for changes in the genome, transcriptome, proteome, and metabolome to determine if A. platensis is a suitable nutritional supplement on long-duration missions. During preflight testing we developed a protocol for inoculated liquid cultures to survive a 10-week storage period prior to photo-incubation. The bioreactor bag, temperature, and lighting conditions that support a 14-day growth cycle between passages were also determined. Media testing identified minimal salts supporting robust growth that can be stored in liquid or dry form. A simple filtration method was developed to dewater cultures and harvest biomass for frozen sample return. We optimized a cryopreservation method to enable return of live cells for isolation of individual A. platensis clones. The concept of operations for Space Algae-2 developed from these test results as well as progress on multi-omics analysis methods will be presented.

Algae↗

Results of First Long Duration Space Flight of Hybrid Perovskite Thin Film

In support of NASA's Artemis program with the goal of a sustained human-lunar presence, there is a need for very large (>100kW) and high-voltage-capable solar arrays, estimated to cost over $150M. Perovskite-based thin-film photovoltaics offer substantial advantages over state-of-the-art solar arrays from the perspective of manufacturing large arrays. Perovskites have also demonstrated some of the lowest temperature coefficients and highest defect tolerance, which make them excellent candidates for aerospace applications. However, MHPs must demonstrate durability in space which presents different challenges than terrestrial operating environments. To decisively test the viability of perovskites being used in space, a perovskite thin film is positioned in low earth orbit for 10 months on the International Space Station, which was the first long-duration study of an MHP in space. Postflight high-resolution ultrafast spectroscopic characterization and comparison with control samples reveal that the flight sample exhibits superior photo-stability, no irreversible radiation damage, and a suppressed structural phase transition temperature by nearly 65 K, broadening the photovoltaic operational range. Further, significant photo-annealing of surface defects is shown following prolonged light-soaking postflight. These results emphasize that methylammonium lead iodide can be packaged adequately for space missions, affirming that space stressors can be managed as theorized.

William Delmas↗

Open Science for Plants in Space: Data Sharing, Standards, and Informatics for Reuse and Knowledge Discovery

Upcoming deep space missions will rely on plants for crew and ecosystem health. Open access space biology data enables scientists to examine the biological responses of plants to ionizing radiation, altered gravity, low atmospheric pressure, elevated CO2, altered photoperiods and many other abiotic stressors. Open Science is the practice of making research available to all, while respecting diverse cultures, and fostering collaborations with equity. 2023 is the ‘Year of Open Science’, and NASA has a 5-year Transform to Open Science (TOPS) initiative designed to rapidly transform the agency toward an inclusive culture of open science. NASA’s Open Science Data Repository (OSDR) within NASA’s Biological and Physical Sciences Division provides access to data from space-relevant biological experiments. OSDR combines two databases, GeneLab and Ames Life Sciences Data Archive (ALSDA) to maximize access to standardized ‘omics (e.g., transcriptomics, proteomics) and phenotypic data (e.g., microscopy, biomass), respectively. GeneLab started in 2014 with the creation of the first space-relevant FAIR (Findable, Accessible, Interoperable, Reusable) biological ‘omics repository, providing detailed metadata on investigation, sample, and assay levels. The addition of ALSDA to OSDR expands plant data analysis capabilities across both phenotypic and ‘omics data. Today, OSDR hosts 62+ plant datasets and has enabled 58 peer-reviewed publications. Most of these publications were collaboration efforts under the OSDR Analysis Working Groups (AWGs). AWGs provide great opportunities for investigators to collaborate and set new standards for space-relevant data and metadata. The AWGs welcome any ASPB members interested in contributing plant expertise for space biology, and to serve as subject matter experts as we establish the framework for modern plant data archiving. Investigators are invited to submit their space-relevant plant datasets to OSDR and visit the site to learn about the tools OSDR has to offer (osdr.nasa.gov/bio).

FAIR↗

Testing A Concept of Operations for the Space Algae-2 Spaceflight Experiment

Algae has abundant potential spaceflight applications to enable future crewed missions and habitation beyond low-Earth orbit, including production of essential nutrients, oxygen, and biofuels. Implementing algae production in space requires understanding how different algae species respond to spaceflight stressors such as microgravity and radiation over a realistic production time course. Space Algae-2 is focusing on Arthrospira platensis, a filamentous cyanobacteria commonly known as Spirulina. A. platensis is a human nutritional supplement on Earth that is a good source of essential amino acids, β-carotene, thiamin, riboflavin, and antioxidants. The Space Algae-2 experiment plans to grow and passage A. platensis continuously aboard the International Space Station for six months. The algae produced will be analyzed with multi-omics profiling to monitor for genetic and phenotype stability in the spaceflight environment. This presentation will report the results from pre-flight science verification tests of the concept of operations for Space Algae-2. The purpose of these tests was to determine if the proposed crew operations for serial passages, sample collection, and sample preservation are feasible to meet science requirements for axenic culturing and preservation of DNA, RNA, protein, and nutritional metabolites.

Microalgae↗