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Biospecimen Culling: Temporal RNA Integrity Analysis Across Spaceflight Missions Dating from 1985 to 2011

The Ames Life Science Data Archive (ALSDA) at NASA Ames Research Center is managed by the Space Biosciences Division and has been operational since 1993. The ALSDA is responsible for archiving information and biospecimens collected from life science spaceflight experiments and matching ground control experiments. They are stored in the Ames biobank, which is located in the Biospecimen Storage Facility (BSF). The ALSDA also manages a Biospecimen Sharing Program, performs curation and long-term storage operations, and makes biospecimens available to the scientific community for research purposes via the Life Science Data Archive public website (https:lsda.jsc.nasa.gov). The BSF maintains both fixed and frozen spaceflight and ground tissues, collected from recent and past spaceflight missions. Due to the ever increasing demand for space to preserve current and future flight biospecimens, the ALSDA has initiated the development of a culling plan for biospecimens currently stored in the BSF. Culling enables the ALSDA to assess the quality of archived samples, and supports the development of standardized culling procedures that improve the operational efficiency of the BSF. The culling plan focuses on generating disposition recommendations for samples in the BSF, and currently is based on measuring ribonucleic acid (RNA) integrity number (RIN). The culling process includes (1) sorting and identification of candidate samples for RIN analysis, (2) completion of RIN analysis on select samples, and (3) development of disposition recommendations for specimens based on the RIN values. Furthermore, our approach allows for unique scientific opportunities, including development of a RIN-based methodology for culling, and temporal assessment of the quality of the tissues that have been stored in BSF since the 1980s. Results of this work will also support NASA open science initiatives.

biospecimen

Ground-Laboratory to In-Space Atomic Oxygen Correlation for the Polymer Erosion and Contamination Experiment (PEACE) Polymers

The Materials International Space Station Experiment 2 (MISSE 2) Polymer Erosion and Contamination Experiment (PEACE) polymers were exposed to the environment of low Earth orbit (LEO) for 3.95 years from 2001 to 2005. There were 41 different PEACE polymers, which were flown on the exterior of the International Space Station (ISS) in order to determine their atomic oxygen erosion yields. In LEO, atomic oxygen is an environmental durability threat, particularly for long duration mission exposures. Although spaceflight experiments, such as the MISSE 2 PEACE experiment, are ideal for determining LEO environmental durability of spacecraft materials, ground-laboratory testing is often relied upon for durability evaluation and prediction. Unfortunately, significant differences exist between LEO atomic oxygen exposure and atomic oxygen exposure in ground-laboratory facilities. These differences include variations in species, energies, thermal exposures and radiation exposures, all of which may result in different reactions and erosion rates. In an effort to improve the accuracy of ground-based durability testing, ground-laboratory to in-space atomic oxygen correlation experiments have been conducted. In these tests, the atomic oxygen erosion yields of the PEACE polymers were determined relative to Kapton H using a radio-frequency (RF) plasma asher (operated on air). The asher erosion yields were compared to the MISSE 2 PEACE erosion yields to determine the correlation between erosion rates in the two environments. This paper provides a summary of the MISSE 2 PEACE experiment; it reviews the specific polymers tested as well as the techniques used to determine erosion yield in the asher, and it provides a correlation between the space and ground laboratory erosion yield values. Using the PEACE polymers asher to in-space erosion yield ratios will allow more accurate in-space materials performance predictions to be made based on plasma asher durability evaluation.

Stambler, Arielle H.

Synthetic Biology to Support Human Exploration of Deep Space

The International Space Station (ISS) has enabled a continuous human presence in space since November 2000. The ISS is in low Earth orbit, facilitating regular resupply missions to deliver air, water, food, spare parts, and science experiments. NASA’s Moon to Mars campaign seeks to return humans to the Moon and prepare for crewed missions to Mars. Increased distance from Earth poses logistical challenges to provide all resources needed by humans for deep space missions. NASA Ames Research Center is conducting a series of synthetic biology projects to test the use of microbes for on-demand biosynthesis of human micronutrients. The BioNutrients spaceflight experiments test an implementation concept to produce fermented food products in which microbe growth enhances micronutrient content. On-demand production of carotenoids was engineered into two yeast (Saccharomyces cerevisiae) strains that have been tested in nearly 5 years of storage. One of the BioNutrients strains will be incorporated into the Lunar Explorer Instrument for Space Biology Applications (LEIA) project. LEIA is developing an instrument suite to be delivered to the lunar south pole region by the Commercial Lunar Payload Services (CLPS) program. The LEIA instrument suite will be used to measure multiple yeast strains for growth, metabolic activity, and synthetic biology-enabled production of carotenoids, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. LEIA data will be used to assess the impact of lunar surface radiation and reduced gravity on the production of engineered traits.

Synthetic Biology

Machine Learning Approaches to Increasing Value of Spaceflight Omics Databases

The number of spaceflight bioscience mission opportunities is too small to allow all relevant biological and environmental parameters to be experimentally identified. Simulated spaceflight experiments in ground-based facilities (GBFs), such as clinostats, are each suitable only for particular investigations -- a rotating-wall vessel may be 'simulated microgravity' for cell differentiation (hours), but not DNA repair (seconds) -- and introduce confounding stimuli, such as motor vibration and fluid shear effects. This uncertainty over which biological mechanisms respond to a given form of simulated space radiation or gravity, as well as its side effects, limits our ability to baseline spaceflight data and validate mission science. Machine learning techniques autonomously identify relevant and interdependent factors in a data set given the set of desired metrics to be evaluated: to automatically identify related studies, compare data from related studies, or determine linkages between types of data in the same study. System-of-systems (SoS) machine learning models have the ability to deal with both sparse and heterogeneous data, such as that provided by the small and diverse number of space biosciences flight missions; however, they require appropriate user-defined metrics for any given data set. Although machine learning in bioinformatics is rapidly expanding, the need to combine spaceflight/GBF mission parameters with omics data is unique. This work characterizes the basic requirements for implementing the SoS approach through the System Map (SM) technique, a composite of a dynamic Bayesian network and Gaussian mixture model, in real-world repositories such as the GeneLab Data System and Life Sciences Data Archive. The three primary steps are metadata management for experimental description using open-source ontologies, defining similarity and consistency metrics, and generating testing and validation data sets. Such approaches to spaceflight and GBF omics data may soon enable unique insight into which measured phenomena correlate to biological mechanisms that are truly affected by spaceflight conditions; which are most likely to be confounded by other variables; and which are insufficiently characterized, significantly increasing existing and future science return from ISS and spaceflight missions.

Gentry, Diana

Effects of microgravity on rat muscle

It is well known that humans exposed to long term spaceflight experience undesirable progressive muscle weakness and increased fatigability. This problem has prompted the implementation of inflight exercise programs because most investigators believe that the major cause of diminished muscle performance is a combination of disuse and decreased workload. Inflight exercise has improved muscle health, but deficits have persisted, indicating that either the regimens utilized were suboptimal or there existed additional debilitating factors which were not remedied by exercise. Clarification of this question requires an improved understanding of the cellular and molecular basis of spaceflight-induced muscle deterioration. To this end, multiple investigations have been performed on the muscles from rats orbited 5 to 22 days in Cosmos biosatellites and Spacelab-3 (2,4,5,8,10 to 14,16,18,19,21 to 23,25,27,28). The eight Cosmos 1887 investigations examined the structural and biochemical changes in skeletal and cardiac muscles of rats exposed to microgravity for 12.5 days and returned to terrestrial gravity 2.3 days before tissues were collected. Even though interpretation of these results was complicated by the combination of inflight and postflight induced alterations, the consensus is that there is marked heterogeneity in both degree and type of responses from the whole muscle level down to the molecular level. Collectively, the muscle investigations of Cosmos 1887 clearly illustrate the wide diversity of muscle tissue responses to spaceflight. Judging from the summary report of this mission, heterogeneity of responses is not unique to muscle tissue. Elucidating the mechanism underlying this heterogeneity holds the key to explaining adaptation of the organism to prolonged spaceflight.

Riley, D. A.

AMMPER: a user-friendly agent-based model that recapitulates simple metabolic responses of yeast to deep-space radiation

For humans venturing to deep space, radiation exposure poses a major health risk. Fundamental research into the biological effects of space radiation are essential for enabling exploration, and the first experimental organisms we send to deep space will be microbial. Yet there are many ways in which microorganisms are likely to experience the effects of high-energy particle radiation (such as Galactic Cosmic Rays) differently from multicellular animals, partly due to the simple fact that microbes are small and unicellular-- less likely to get hit in the first place, and less likely to communicate damage between cells. Computational modeling can aid in designing experiments and predicting the biological effects of radiation, but thus far particle radiation models have not focused on microbes. Here we present the latest developments in AMMPER, the Agent-based Model for Microbial Populations Exposed to Radiation. Originally written in 2021, AMMPER is a Python-based model that incorporates radiation track data from NASA's RITRACKS software and simulates the growth, damage, and death of yeast cells in 3D. It is now freely available as an open-source package on NASA's GitHub repository. Recent improvements include the ability to simulate the dynamics of alamarBlue, a color-changing redox dye commonly used to track metabolic activity in microbial spaceflight experiments. We demonstrate that a simple blue-pink-clear transition model is able to recapitulate key features observed in empirical data from ground studies. AMMPER also includes a new graphical user interface and introductory tutorial to facilitate ease of use by a wider audience. AMMPER can help us to understand how spatially heterogeneous particle radiation damage at the single-cell level can translate to growth differences at the population level, ultimately allowing us to better interpret experiments using microbes as model organisms and how well their results apply to humans.

yeast

Evolutionary Stability of Microbial Mutualism in Simulated Microgravity

Sustainable long-term space travel will require astronauts to live in built environments cohabited by microbes. Though much of space biology research has focused on microbial monocultures, microbes naturally live in multispecies communities in which they compete and cooperate. Understanding how communities experience spaceflight over generations can influence sanitation measures and the design of synthetic communities for applications in spaceflight. In this study, we investigated the evolutionary stability of mutualism in multispecies communities in microgravity using a model system of cooperating microbes. Our model consisted of bioengineered strains of mutually dependent E. coli and S. enterica. E. coli metabolized lactose and produced a carbon source for S. enterica, which secreted methionine for E. coli. By introducing a noncooperative ("cheater") strain of S. enterica, we tested the dynamic stability of mutualism. Using a rotating wall vessel, we simulated microgravity, specifically a low-shear, diffusion-limited environment where metabolite transfer is limited compared to 1g conditions. Fluorescent motile and non-motile strains were used to assess how motility affects the microbial experience of microgravity. We hypothesized that mutualism in non-motile cocultures would be favored in microgravity due to the diffusion-limited spatial structure, while motile strains would show less difference between simulated microgravity and other conditions. We cultured communities for approximately 20 generations and assessed community composition over time using flow cytometry. We compared simulated microgravity to cultures grown in agar plates as spatially structured controls, shaken flasks as well-mixed controls, and still flasks representing the convective mixing of 1g. A high, sustained cooperator-to-cheater ratio represented stable mutualism within the community. Our results show that microbial communities may better maintain mutualism in microgravity compared to their counterparts in 1g conditions and that the motility of a community’s constituents influences how the community experiences microgravity on generational timescales.

Nathan Moreno

Hollow cathode-based plasma contactor experiments for electrodynamic tether

The role plasma contactors play in effective electrodynamic tether operation is discussed. Hollow cathodes and hollow cathode-based plasma sources have been identified as leading candidates for the electrodynamic tether plasma contactor. Present experimental efforts to evaluate the suitability of these devices as plasma contactors are reviewed. This research includes the definition of preliminary plasma contactor designs, and the characterization of their operation as electron collectors from a simulated space plasma. The discovery of an 'ignited mode' regime of high contactor efficiency and low impedance is discussed, as well as is the application of recent models of the plasma coupling process to contactor operation. Results indicate that ampere-level electron currents can be exchanged between hollow cathode-based plasma contactors and a dilute plasma in this regime. A discussion of design considerations for plasma contactors is given which includes expressions defining the total mass flow rate and power requirements of plasma contactors operating in both the cathodic and anodic regimes, and correlation of this to the tether current. Finally, future ground and spaceflight experiments are proposed to resolve critical issues of plasma contactor operation.

Patterson, Michael J.

Electrodynamic Dust Shield Testing on the Materials on International Space Station Experiment 11

Dust is a major concern for lunar exploration. To combat the effects of dust, NASA, academia, and industry are developing solutions to the dust problem. One potential technology solution for this problem is the Electrodynamic Dust Shield (EDS). Many years of research and development have gone into this technology. The Materials on International Space Station Experiment – 11 (MISSE-11) provides a long term space exposure platform for this technology to verify compatibility of materials and manufacturing processes to the space environment. The MISSE-11 EDS experiment consists of 12 EDS panels. These panels are made of glass, polyimide, or prototype spacesuit fabric. Some panels are covered with a lotus leaf coating while others are covered with thermal paint. They are flown in the wake position of the ISS to simulate the lunar environment. Two panels are in an active configuration and are energized with a high voltage power supply, which generates high-voltage pulses to activate the dust shields. Current and voltage data are recovered from each of these trials to compare to baseline data. Also, each of the EDS panels are imaged on a monthly basis to track any changes with time that may occur with the EDS variants. In this paper, we report preliminary data and analysis from this spaceflight experiment.

Electrostatics

Electrodynamic Dust Shield Testing on the Materials on International Space Station Experiment 11

Dust is a major concern for lunar exploration. To combat the effects of dust, NASA, academia, and industry are developingsolutions to the dust problem. One potential technology solution for this problem is the Electrodynamic DustShield (EDS). Many years of research and development have gone into this technology. The Materials on InternationalSpace Station Experiment - 11 (MISSE-11) provides a long term space exposure platform for this technology to verifycompatibility of materials and manufacturing processes to the space environment. The MISSE-11 EDS experimentconsists of 12 EDS panels. These panels are made of glass, polyimide, or prototype spacesuit fabric. Some panels arecovered with a lotus leaf coating while others are covered with thermal paint. They are flown in the wake position ofthe ISS to simulate the lunar environment. Two panels are in an active configuration and are energized with a highvoltage power supply, which generates high-voltage pulses to activate the dust shields. Current and voltage data arerecovered from each of these trials to compare to baseline data. Also, each of the EDS panels are imaged on a monthlybasis to track any changes with time that may occur with the EDS variants. In this paper, we report preliminary dataand analysis from this spaceflight experiment.

Johansen, Michael R.

Germination and elongation of flax in microgravity

This experiment was conducted as part of a risk mitigation payload aboard the Space Shuttle Atlantis on STS-101. The objectives were to test a newly developed water delivery system, and to determine the optimal combination of water volume and substrate for the imbibition and germination of flax (Linum usitatissimum) seeds in space. Two different combinations of germination paper were tested for their ability to absorb, distribute, and retain water in microgravity. A single layer of thick germination paper was compared with one layer of thin germination paper under a layer of thick paper. Paper strips were cut to fit snugly into seed cassettes, and seeds were glued to them with the micropyle ends pointing outward. Water was delivered in small increments that traveled through the paper via capillary action. Three water delivery volumes were tested, with the largest (480 microliters) outperforming the 400 microliters and 320 microliters volumes for percent germination (90.6%) and root growth (mean=4.1 mm) during the 34-hour spaceflight experiment. The ground control experiment yielded similar results, but with lower rates of germination (84.4%) and shorter root lengths (mean=2.8 mm). It is not clear if the roots emerged more quickly in microgravity and/or grew faster than the ground controls. The single layer of thick germination paper generally exhibited better overall growth than the two layered option. Significant seed position effects were observed in both the flight and ground control experiments. Overall, the design of the water delivery system, seed cassettes and the germination paper strip concept was validated as an effective method for promoting seed germination and root growth under microgravity conditions. c2003 COSPAR. Published by Elsevier Ltd. All rights reserved.

Non-NASA Center

Effects of Gravity on Insect Circadian Rhythmicity

Circadian rhythms - endogenous daily rhythmic fluctuations in virtually all characteristics of life - are generated and coordinated by the circadian timing system (CTS). The CTS is synchronized to the external 24-hour day by time cues such as the light/dark cycle. In an environment without time cues, the length of an animal's day is determined by the period of its internal pacemaker (tau) and the animal is said to be free-running. All life on earth evolved under the solar day; the CTS exists as an adaptation that allows organisms to anticipate and to prepare for rhythmic environmental fluctuations. All life on earth also evolved under the force of earth's gravitational environment. While it is therefore not surprising that changes in the lighting environment affect the CTS, it is surprising that changes in the gravitational environment would do so. However, recent data from one of our laboratories using the brn-3.1 knockout mouse revealed that this model, which lacks the sensory receptor hair cells within the neurovestibular system, does not respond to exposure to a hyperdynamic environment in the same fashion as normal mice. The brn-3.1 mice did not show the expected suppression of circadian rhythmicity shown by control mice exposed to 2G. Exposure to altered ambient force environments affects the amplitude, mean and timing of circadian rhythms in species from unicellular organisms to man. In addition, there is a circadian influence on the homeostatic response to acute 2G acceleration and pulses of 2G can act as a time cue, synchronizing the CTS. This is of significance because maintenance of internal and external temporal coordination is critical for normal physiological and psychological function. Typically, during adaptation to an increased gravitational environment (+G), an initial acute reaction is followed by adaptation and, eventually, a new steady state (14-16), which can take weeks to months to establish. Until the development of space stations, exposure to microgravity was, of necessity, relatively short in duration. In early spaceflight experiments an organism's internal rhythms often expressed periods that were different from each other, even in the presence of a 24.0 hour light-dark cycle, suggesting that the organism was experiencing internal desynchronization (17, 18). In (micro)G, the body temperature rhythm was delayed with respect to other body rhythms and to the light-dark cycle in rhesus macaques (19) and man (20, 21). In the absence of a light-dark cycle, the circadian rhythm of spore formation persisted in Neurospora crassa, however, both the variability and average period of the rhythm increased (22). The beetle Trigonoscelis gigas, exhibited changes in period during and following 11-13 days in (micro)G (23, 24). Resynchronization of the urinary calcium rhythm following a 1800 phase shift of the LID cycle was retarded in rats exposed to (micro)G compared to 1G controls (25). With the development of the Russian Mir Space Station, long-term controlled microgravity exposure became possible. We recorded activity rhythms from black-bodied Tenebrionid beetles, Trigonoscelis gigas, in (micro)G (spaceflight). Each insect was housed individually within an activity monitor (26) and data (activity counts) were collected and stored in five-minute bins. Thirty-two individual activity monitors were housed within each of 2 experimental kits. The beetles within each kit were divided into two groups and the lighting was controlled separately for each group.

Hoban-Higgins, Tana M.

Operating a Crewed Spacecraft in the Age of Commercial Space Using Private/Government Partnership

Fifty years after humans completed a large-scale United States government-funded and government-engineered effort of landing humans on the moon, human spaceflight has entered a new paradigm. Private companies are now investing their own money and taking on an ever-increasing role in human spaceflight, in partnership with the U.S. government. This paper will describe the development of one of these partnerships through the lens of its mission operations team. As part of the Commercial Crew Program (CCP), NASA selected Boeing’s CST-100 Starliner as one of the next generation of crewed vehicles. Boeing opted to partner with the US Government for their Starliner operations by contracting with NASA’s Mission Control teams in the Flight Operations Directorate (FOD) at the Johnson Space Center to create its own Mission Operations (MO) flight controllers. Partnering with FOD provided benefits to both Boeing and NASA while also creating new challenges. MO brought over 60 years of crewed spaceflight experience and infrastructure to Boeing’s new program. Within certain legal constraints, MO was able to work closely and efficiently with their FOD counterparts who were performing both integration duties and, under the auspices of the CCP, insight of the contractors, in this case Boeing. Involvement of NASA as the Boeing operations agent did lead to what management deemed a ‘healthy tension’ within FOD, challenging old processes and often creating better, more robust teamwork. Successful development of the framework and boundaries of both the legal aspects and the oversight tensions has been one of the keys to developing a successful corporate/government partnership. Due to the highly automated nature of the Starliner, the MO organization was designed to be much smaller than previous NASA’s flight control teams for past programs. NASA has learned through the decades that spacecraft design and operations need to be as flexible and forgiving as possible. NASA’s Commercial Crew Program was established to sponsor corporate development of economical vehicles that could get humans to and from low Earth orbit. These companies, of course, need to meet contractual obligations in providing a safe means of transporting astronauts to the International Space Station (ISS), but also need to do so in a manner that leads to the venture resulting in a profit at the same time. Through MO’s involvement in the development of this spaceflight paradigm shift, there are ample lessons to be conveyed to future teams and programs working to develop similar missions.

Robert C Dempsey

Operating a Crewed Spacecraft in the Age of Commercial Space Using Private/Government Partnership

Fifty years after humans completed a large-scale United States government-funded and government-engineered effort of landing humans on the moon, human spaceflight has entered a new paradigm. Private companies are now investing their own money and taking on an ever-increasing role in human spaceflight, in partnership with the U.S. government. This paper will describe the development of one of these partnerships through the lens of its mission operations team. As part of the Commercial Crew Program (CCP), NASA selected Boeing’s CST-100 Starliner as one of the next generation of crewed vehicles. Boeing opted to partner with the US Government for their Starliner operations by contracting with NASA’s Mission Control teams in the Flight Operations Directorate (FOD) at the Johnson Space Center to create its own Mission Operations (MO) flight controllers. Partnering with FOD provided benefits to both Boeing and NASA while also creating new challenges. MO brought over 60 years of crewed spaceflight experience and infrastructure to Boeing’s new program. Within certain legal constraints, MO was able to work closely and efficiently with their FOD counterparts who were performing both integration duties and, under the auspices of the CCP, insight of the contractors, in this case Boeing. Involvement of NASA as the Boeing operations agent did lead to what management deemed a ‘healthy tension’ within FOD, challenging old processes and often creating better, more robust teamwork. Successful development of the framework and boundaries of both the legal aspects and the oversight tensions has been one of the keys to developing a successful corporate/government partnership. Due to the highly automated nature of the Starliner, the MO organization was designed to be much smaller than previous NASA’s flight control teams for past programs. NASA has learned through the decades that spacecraft design and operations need to be as flexible and forgiving as possible. NASA’s Commercial Crew Program was established to sponsor corporate development of economical vehicles that could get humans to and from low Earth orbit. These companies, of course, need to meet contractual obligations in providing a safe means of transporting astronauts to the International Space Station (ISS), but also need to do so in a manner that leads to the venture resulting in a profit at the same time. Through MO’s involvement in the development of this spaceflight paradigm shift, there are ample lessons to be conveyed to future teams and programs working to develop similar missions.

Robert C. Dempsey

Operating a Crewed Spacecraft in the Age of Commercial Space Using Private/Government Partnership

Fifty years after humans completed a large-scale United States government-funded and government-engineered effort of landing humans on the moon, human spaceflight has entered a new paradigm. Private companies are now investing their own money and taking on an ever-increasing role in human spaceflight, in partnership with the U.S. government. This paper will describe the development of one of these partnerships through the lens of its mission operations team. As part of the Commercial Crew Program (CCP), NASA selected Boeing’s CST-100 Starliner as one of the next generation of crewed vehicles. Boeing opted to partner with the US Government for their Starliner operations by contracting with NASA’s Mission Control teams in the Flight Operations Directorate (FOD) at the Johnson Space Center to create its own Mission Operations (MO) flight controllers. Partnering with FOD provided benefits to both Boeing and NASA while also creating new challenges. MO brought over 60 years of crewed spaceflight experience and infrastructure to Boeing’s new program. Within certain legal constraints, MO was able to work closely and efficiently with their FOD counterparts who were performing both integration duties and, under the auspices of the CCP, insight of the contractors, in this case Boeing. Involvement of NASA as the Boeing operations agent did lead to what management deemed a ‘healthy tension’ within FOD, challenging old processes and often creating better, more robust teamwork. Successful development of the framework and boundaries of both the legal aspects and the oversight tensions has been one of the keys to developing a successful corporate/government partnership. Due to the highly automated nature of the Starliner, the MO organization was designed to be much smaller than previous NASA’s flight control teams for past programs. NASA has learned through the decades that spacecraft design and operations need to be as flexible and forgiving as possible. NASA’s Commercial Crew Program was established to sponsor corporate development of economical vehicles that could get humans to and from low Earth orbit. These companies, of course, need to meet contractual obligations in providing a safe means of transporting astronauts to the International Space Station (ISS), but also need to do so in a manner that leads to the venture resulting in a profit at the same time. Through MO’s involvement in the development of this spaceflight paradigm shift, there are ample lessons to be conveyed to future teams and programs working to develop similar missions.

Robert C. Dempsey

A Multi-Omics Approach Demonstrates that Spaceflight Leads to Lipid Accumulation in Mouse Livers

Spaceflight has several detrimental effects on the physiology of astronauts, many of which are recapitulated in rodent models. We analyzed liver transcriptomic and proteomic data from three mouse spaceflight experiments flown aboard the International Space Station (Rodent Research-1 NASA (RR-1 NASA), Rodent Research-1 CASIS (RR-1 CASIS), Rodent Research-3 (RR-3)), and one mouse experiment flown on the Space Shuttle (Commercial Biomedical Testing Module-3 (CBTM-3) aboard STS-135). Despite the differences in genetic background and time of exposure to microgravity it was shown through Oil Red staining and histology that increased lipid accumulation was occurring in the liver of all mice flown in space compared to the ground controls. This led to further pursue the existing GeneLab datasets related to liver omics data from these mice. We were able to discover key conserved pathways across all the mice independent of the flight conditions that were related to increased lipid metabolism, fatty acid metabolism, both lipid and fatty acid processing, lipid catabolic processing, and lipid localization. In addition, key upstream regulators were predicted to be commonly regulated across all conditions which include ESR1, GCG, and NR1I2 being inhibited and INS being activated. Interestingly, estrogen receptor alpha (ESR1) expression has been known to be heavily involved with lipoprotein metabolism. In addition, insulin (INS) is the primary driver for fat metabolism and increased INS has been associated with increased fatty acids in the liver. Through additional proteomic analysis we were able to identify the majority of the key proteins related to lipids for both the RR-1 and RR-3 rodents were being up-regulated in the livers when comparing flight to ground controls. This additional confirmation of the lipid associated activity also showed that the lipid related proteins are heavily involved with lipid metabolism, cholesterol binding, and cholesterol metabolism. Lastly, the analysis also revealed that the circadian clock related pathways in the liver are commonly being increased across all space flight conditions which has also been reported in the literature to potentially cause increased liver damage. The combination of the very strong lipid uptake in the liver and the transcriptomic/proteomic signatures (including the circadian clock pathways) following spaceflight are consistent with early onset of liver disease. Taken together, these data indicate that, activation of lipotoxic pathways could persist during longer duration spaceflight which might result in the development of liver disease

Lipids

Functional Data Analysis of Spaceflight-Induced Changes in Coordination and Phase in Head Pitch Acceleration During Treadmill Walking

Astronauts returning from spaceflight experience neurovestibular disturbances during head movements and attempt to mitigate them by limiting head motion. Analyses to date of the head movements made during walking have concentrated on amplitude and variability measures extracted from ensemble averages of individual gait cycles. Phase shifts within each gait cycle can be determined by functional data analysis through the computation of time-warping functions. Large, localized variations in the timing of peaks in head kinematics may indicate changes in coordination. The purpose of this study was to determine timing changes in head pitch acceleration of astronauts during treadmill walking before and after flight. Six astronauts (5M/1F; age = 43.5+/-6.4yr) participated in the study. Subjects walked at 1.8 m/sec (4 mph) on a motorized treadmill while reading optotypes displayed on a computer screen 4 m in front of their eyes. Three-dimensional motion of the subject s head was recorded with an Inertial Measurement Unit (IMU) device. Data were recorded twice before flight and four times after landing. The head pitch acceleration was calculated by taking the time derivative of the pitch velocity data from the IMU. Data for each session with each subject were time-normalized into gait cycles, then registered to align significant features and create a mean curve. The mean curves of each postflight session for each subject were re-registered based on their preflight mean curve to create time-warping functions. The root mean squares (RMS) of these warping functions were calculated to assess the deviation of head pitch acceleration mean curves in each postflight session from the preflight mean curve. After landing, most crewmembers exhibited localized shifts within their head pitch acceleration regimes, with the greatest deviations in RMS occurring on landing day or 1 day after landing. These results show that the alteration of head pitch coordination due to spaceflight may be assessed using an analysis of time-warping functions.

Miller, Christopher