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Microbial Responses to Microgravity and Other Low-Shear Environments

Microbial adaptation to environmental stimuli is essential for survival. While several of these stimuli have been studied in detail, recent studies have demonstrated an important role for a novel environmental parameter in which microgravity and the low fluid shear dynamics associated with microgravity globally regulate microbial gene expression, physiology, and pathogenesis. In addition to analyzing fundamental questions about microbial responses to spaceflight, these studies have demonstrated important applications for microbial responses to a ground-based, low-shear stress environment similar to that encountered during spaceflight. Moreover, the low-shear growth environment sensed by microbes during microgravity of spaceflight and during ground-based microgravity analogue culture is relevant to those encountered during their natural life cycles on Earth. While no mechanism has been clearly defined to explain how the mechanical force of fluid shear transmits intracellular signals to microbial cells at the molecular level, the fact that cross talk exists between microbial signal transduction systems holds intriguing possibilities that future studies might reveal common mechanotransduction themes between these systems and those used to sense and respond to low-shear stress and changes in gravitation forces. The study of microbial mechanotransduction may identify common conserved mechanisms used by cells to perceive changes in mechanical and/or physical forces, and it has the potential to provide valuable insight for understanding mechanosensing mechanisms in higher organisms. This review summarizes recent and future research trends aimed at understanding the dynamic effects of changes in the mechanical forces that occur in microgravity and other low-shear environments on a wide variety of important microbial parameters.

Review↗

Restraint of Liquid Jets by Surface Tension in Microgravity Modeled

Tension in Microgravity Modeled Microgravity poses many challenges to the designer of spacecraft tanks. Chief among these are the lack of phase separation and the need to supply vapor-free liquid or liquidfree vapor to the spacecraft processes that require fluid. One of the principal problems of phase separation is the creation of liquid jets. A jet can be created by liquid filling, settling of the fluid to one end of the tank, or even closing a valve to stop the liquid flow. Anyone who has seen a fountain knows that jets occur in normal gravity also. However, in normal gravity, the gravity controls and restricts the jet flow. In microgravity, with gravity largely absent, jets must be contained by surface tension forces. Recent NASA experiments in microgravity (Tank Pressure Control Experiment, TPCE, and Vented Tank Pressure Experiment, VTRE) resulted in a wealth of data about jet behavior in microgravity. VTRE was surprising in that, although it contained a complex geometry of baffles and vanes, the limit on liquid inflow was the emergence of a liquid jet from the top of the vane structure. Clearly understanding the restraint of liquid jets by surface tension is key to managing fluids in low gravity. To model this phenomenon, we need a numerical method that can track the fluid motion and the surface tension forces. The fluid motion is modeled with the Navier-Stokes equation formulated for low-speed incompressible flows. The quantities of velocity and pressure are placed on a staggered grid, with velocity being tracked at cell faces and pressure at cell centers. The free surface is tracked via the introduction of a color function that tracks liquid as 1/2 and gas as -1/2. A phase model developed by Jacqmin is used. This model converts the discrete surface tension force into a barrier function that peaks at the free surface and decays rapidly. Previous attempts at this formulation have been criticized for smearing the interface. However, by sharpening the phase function, double gridding the fluid function, and using a higher order solution for the fluid function, interface smearing is avoided. These equations can be rewritten as two coupled Poisson equations that also include the velocity. The method of solution is as follows: first, the phase equations are solved from this solution, a velocity field is generated, then a successive overrelaxation scheme is used to solve for a pressure field consistent with the velocity solution. After the code was implemented in axisymmetric form and verified by several test cases, the drop tower runs of Aydelott were modeled. The model handed the free-surface deformation quite nicely, even to the point of modeling geyser growth in the regime where the free surface was no longer restrained. A representative run is shown.

Chato, David J.↗

Ground Based Microgravity Emissions Testing Of Flight Hardware

To control microgravity environment on the International Space Station (ISS), NASA developed payloads have to meet the payload integration requirements of the Space Station Program, specifically a microgravity allocation plan. The Microgravity Emissions Laboratory (MEL) was developed at NASA Glenn Research Center (GRC) for verification of the payloads compliance with payload integration requirements. MEL is a 6 degree of freedom inertial measurement system capable of characterizing the microgravity emissions, generated by a disturber, down to a micro g. Microgravity Emissions tests provide a payload developer with a tool to assess payload's compliance with the requirements, i.e. forces and moments, generated by the payload at its center of gravity. Forces and moments are presented in time domain for both stationary and transient signals, and in frequency domain for the stationary signals. To date, MEL conducted over thirty tests of ISS hardware. The test results are being successfully used by the payload developers for design verification and improvement.

Samorezov, Sergey↗

The NASA Microgravity Fluid Physics Program: Research Plans for the ISS

Building on over four decades of research and technology development related to the behavior of fluids in low gravity environments, the current NASA Microgravity Fluid Physics Program continues the quest for knowledge to further understand and design better fluids systems for use on earth and in space. NASA's Biological and Physical Research Enterprise seeks to exploit the space environment to conduct research supporting human exploration of space (strategic research), research of intrinsic scientific importance and impact (fundamental research), and commercial research. The strategic research thrust will build the vital knowledge base needed to enable NASA's mission to explore the Universe and search for life. There are currently five major research areas in the Microgravity Fluid Physics Program: complex fluids, niultiphase flows and phase change, interfacial phenomena, biofluid mechanics, and dynamics and instabilities. Numerous investigations into these areas are being conducted in both ground-based laboratories and facilities and in the flight experiments program. Most of the future NASA- sponsored flight experiments in microgravity fluid physics and transport phenomena will be carried out on the International Space Station (ISS) in the Fluids Integrated Rack (FIR), in the Microgravity Science Glovebox (MSG), in EXPRESS racks, and in other facilities provided by international partners. This paper presents an overview of the near- and long-term visions for NASA's Microgravity Fluid Physics Research Program and brief descriptions of hardware systems planned to enable this research.

Kohl, Fred J.↗

Multiphase flow and phase change in microgravity: Fundamental research and strategic research for exploration of space

NASA is preparing to undertake science-driven exploration missions. The NASA Exploration Team's vision is a cascade of stepping stones. The stepping-stone will build the technical capabilities needed for each step with multi-use technologies and capabilities. An Agency-wide technology investment and development program is necessary to implement the vision. The NASA Exploration Team has identified a number of areas where significant advances are needed to overcome all engineering and medical barriers to the expansion of human space exploration beyond low-Earth orbit. Closed-loop life support systems and advanced propulsion and power technologies are among the areas requiring significant advances from the current state-of-the-art. Studies conducted by the National Academy of Science's National Research Council and Workshops organized by NASA have shown that multiphase flow and phase change play a crucial role in many of these advanced technology concepts. Lack of understanding of multiphase flow, phase change, and interfacial phenomena in the microgravity environment has been a major hurdle. An understanding of multiphase flow and phase change in microgravity is, therefore, critical to advancing many technologies needed. Recognizing this, the Office of Biological and Physical Research (OBPR) has initiated a strategic research thrust to augment the ongoing fundamental research in fluid physics and transport phenomena discipline with research especially aimed at understanding key multiphase flow related issues in propulsion, power, thermal control, and closed-loop advanced life support systems. A plan for integrated theoretical and experimental research that has the highest probability of providing data, predictive tools, and models needed by the systems developers to incorporate highly promising multiphase-based technologies is currently in preparation. This plan is being developed with inputs from scientific community, NASA mission planners and industry personnel. The fundamental research in multiphase flow and phase change in microgravity is aimed at developing better mechanistic understanding of pool boiling and ascertaining the effects of gravity on heat transfer and the critical heat flux. Space flight experiments conducted in space have shown that nucleate pool boiling can be sustained under certain conditions in the microgravity environment. New space flight experiments are being developed to provide more quantitative information on pool boiling in microgravity. Ground-based investigations are also being conducted to develop mechanistic models for flow and pool boiling. An overview of the research plan and roadmap for the strategic research in multiphase flow and phase change as well as research findings from the ongoing program will be presented.

Singh, Bhim S.↗

New Technologies Being Developed for the Thermophoretic Sampling of Smoke Particulates in Microgravity

The Characterization of Smoke Particulate for Spacecraft Fire Detection, or Smoke, microgravity experiment is planned to be performed in the Microgravity Science Glovebox Facility on the International Space Station (ISS). This investigation, which is being developed by the NASA Glenn Research Center, ZIN Technologies, and the National Institute of Standards and Technologies (NIST), is based on the results and experience gained from the successful Comparative Soot Diagnostics experiment, which was flown as part of the USMP-3 (United States Microgravity Payload 3) mission on space shuttle flight STS-75. The Smoke experiment is designed to determine the particle size distributions of the smokes generated from a variety of overheated spacecraft materials and from microgravity fires. The objective is to provide the data that spacecraft designers need to properly design and implement fire detection in spacecraft. This investigation will also evaluate the performance of the smoke detectors currently in use aboard the space shuttle and ISS for the test materials in a microgravity environment.

Sheredy, William A.↗

Modeled Microgravity Disrupts Collagen I/Integrin Signaling During Osteoblastic Differentiation of Human Mesenchymal Stem Cells

Spaceflight leads to reduced bone mineral density in weight bearing bones that is primarily attributed to a reduction in bone formation. We have previously demonstrated severely reduced osteoblastogenesis of human mesenchymal stem cells (hMSC) following seven days culture in modeled microgravity. One potential mechanism for reduced osteoblastic differentiation is disruption of type I collagen-integrin interactions and reduced integrin signaling. Integrins are heterodimeric transmembrane receptors that bind extracellular matrix proteins and produce signals essential for proper cellular function, survival, and differentiation. Therefore, we investigated the effects of modeled microgravity on integrin expression and function in hMSC. We demonstrate that seven days of culture in modeled microgravity leads to reduced expression of the extracellular matrix protein, type I collagen (Col I). Conversely, modeled microgravity consistently increases Col I-specific alpha2 and beta1 integrin protein expression. Despite this increase in integrin sub-unit expression, autophosphorylation of adhesion-dependent kinases, focal adhesion kinase (FAK) and proline-rich tyrosine kinase 2 (PYK2), is significantly reduced. Activation of Akt is unaffected by the reduction in FAK activation. However, reduced downstream signaling via the Ras-MAPK pathway is evidenced by a reduction in Ras and ERK activation. Taken together, our findings indicate that modeled microgravity decreases integrin/MAPK signaling, which likely contributes to the observed reduction in osteoblastogenesis.

Meyers, Valerie E.↗

Risk Assessment and Control through Countermeasure System Iplementation for Long-term Crew Exposure to Microgravity

Experience with the International Space Station (ISS) program demonstrates the degree to which engineering design and operational solutions must protect crewmembers from health risks due to long-term exposure to the microgravity environment. Risks to safety and health due to degradation in the microgravity environment include crew inability to complete emergency or nominal activities, increased risk of injury, and inability to complete safe return to the ground due to reduced strength or embrittled bones. These risks without controls slowly increase in probability for the length of the mission and become more significant for increasing mission durations. Countermeasures to microgravity include hardware systems that place a crewmember s body under elevated stress to produce an effect similar to daily exposure to gravity. The ISS countermeasure system is predominately composed of customized exercise machines. Historical treatment of microgravity countermeasure systems as medical research experiments unintentionally reduced the foreseen importance and therefore the capability of the systems to function in a long-term operational role. Long-term hazardous effects and steadily increasing operational risks due to non-functional countermeasure equipment require a more rigorous design approach and incorporation of redundancy into seemingly non- mission-critical hardware systems. Variations in the rate of health degradation and responsiveness to countermeasures among the crew population drastically increase the challenge for design requirements development and verification of the appropriate risk control strategy. The long-term nature of the hazards and severe limits on logistical re-supply mass, volume and frequency complicates assessment of hardware availability and verification of an adequate maintenance and sparing plan. Design achievement of medically defined performance requirements by microgravity countermeasure systems and incorporation of adequate failure tolerance significantly reduces these risks. Future implementation of on-site monitoring hardware for critical health parameters such as bone mineral density would allow greater responsiveness, efficiency, and optimized design of the countermeasures system.

Gernand, Jeremy M.↗

Motion of Air Bubbles in Water Subjected to Microgravity Accelerations

The International Space Station (ISS) serves as a platform for microgravity research for the foreseeable future. A microgravity environment is one in which the effects of gravity are drastically reduced which then allows physical experiments to be conducted without the over powering effects of gravity. During his 6-month stay on the ISS, astronaut Donald R. Pettit performed many informal/impromptu science experiments with available equipment. One such experiment focused on the motion of air bubbles in a rectangular container nearly filled with de-ionized water. Bubbles were introduced by shaking and then the container was secured in place for several hours while motion of the bubbles was recorded using time-lapse photography. This paper shows correlation between bubble motion and quasi-steady acceleration levels during one such experiment operation. The quasi-steady acceleration vectors were measured by the Microgravity Acceleration Measurement System (MAMS). Essentially linear motion was observed in the condition considered here. Dr. Pettit also created other conditions which produced linear and circulating motion, which are the subjects of further study. Initial observations of this bubble motion agree with calculations from many microgravity physical science experiments conducted on shuttle microgravity science missions. Many crystal-growth furnaces involve heavy metals and high temperatures in which undesired acceleration-driven convection during solidification can adversely affect the crystal. Presented in this paper will be results showing correlation between bubble motion and the quasi-steady acceleration vector.

DeLombard, Richard↗

The Effect of Acute Microgravity on Mechanically-Induced Membrane Damage and Membrane-Membrane Fusion Events

Although it is unclear how a living cell senses gravitational forces there is no doubt that perturbation of the gravitational environment results in profound alterations in cellular function. In the present study, we have focused our attention on how acute microgravity exposure during parabolic flight affects the skeletal muscle cell plasma membrane (i.e. sarcolemma), with specific reference to a mechanically-reactive signaling mechanism known as mechanically-induced membrane disruption or "wounding". This response is characterized by both membrane rupture and membrane resealing events mediated by membrane-membrane fusion. We here present experimental evidence that acute microgravity exposure can inhibit membrane-membrane fusion events essential for the resealing of sarcolemmal wounds in individual human myoblasts. Additional evidence to support this contention comes from experimental studies that demonstrate acute microgravity exposure also inhibits secretagogue-stimulated intracellular vesicle fusion with the plasma membrane in HL-60 cells. Based on our own observations and those of other investigators in a variety of ground-based models of membrane wounding and membrane-membrane fusion, we suggest that the disruption in the membrane resealing process observed during acute microgravity is consistent with a microgravity-induced decrease in membrane order.

Clarke, Mark, S. F.↗

In Vitro Disease Model of Microgravity Conditioning on Human Energy Metabolism

NASA and its partners are committed to introducing appropriate new technology to enable learning and living safely beyond the Earth for extended periods of time in a sustainable and possibly indefinite manner. In the responsible acquisition of that goal, life sciences is tasked to tune and advance current medical technology to prepare for human health and wellness in the space environment. The space environment affects the condition and function of biological systems from organ level function to shape of individual organelles. The objective of this paper is to study the effect of microgravity on kinetics of drug metabolism. This fundamental characterization is meaningful to (1) scientific understanding of the response of biology to microgravity and (2) clinical dosing requirements and pharmacological thresholds during long term manned space exploration. Metabolism kinetics of the anti-nausea drug promethazine (PMZ) were determined by an in vitro ground model of 3-dimensional aggregates of human hepatocytes conditioned to weightlessness using a rotating wall bioreactor. The authors observed up-regulated PMZ conversion in model microgravity conditions and attribute this to effect to model microgravity conditioning acting on metabolic mechanisms of the cells. Further work is necessary to determine which particular cellular mechanisms are governing the experimental observations, but the authors conclude kinetics of drug metabolism are responsive to gravitational fields and further study of this sensitivity would improve dosing of pharmaceuticals to persons exposed to a microgravity environment.

Snyder, Jessica↗

Microgravity-Enhanced Stem Cell Selection

Stem cells, both embryonic and adult, promise to revolutionize the practice of medicine in the future. In order to realize this potential, a number of hurdles must be overcome. Most importantly, the signaling mechanisms necessary to control the differentiation of stem cells into tissues of interest remain to be elucidated, and much of the present research on stem cells is focused on this goal. Nevertheless, it will also be essential to achieve large-scale expansion and, in many cases, assemble cells in 3D as transplantable tissues. To this end, microgravity analog bioreactors can play a significant role. Microgravity bioreactors were originally conceived as a tool to study the cellular responses to microgravity. However, the technology can address some of the shortcomings of conventional cell culture systems; namely, the deficiency of mass transport in static culture and high mechanical shear forces in stirred systems. Unexpectedly, the conditions created in the vessel were ideal for 3D cell culture. Recently, investigators have demonstrated the capability of the microgravity bioreactors to expand hematopoietic stem cells compared to static culture, and facilitate the differentiation of umbilical cord stem cells into 3D liver aggregates. Stem cells are capable of differentiating into functional cells. However, there are no reliable methods to induce the stem cells to form specific cells or to gain enough cells for transplantation, which limits their application in clinical therapy. The aim of this study is to select the best experimental setup to reach high proliferation levels by culturing these cells in a microgravity-based bioreactor. In typical cell culture, the cells sediment to the bottom surface of their container and propagate as a one-cell-layer sheet. Prevention of such sedimentation affords the freedom for self-assembly and the propagation of 3D tissue arrays. Suspension of cells is easily achievable using stirred technologies. Unfortunately, in conventional bioreactors, stirring invokes deleterious forces that disrupt cell aggregation and results in cell death. First-generation rotating bioreactors provided rotation on the horizontal axis, which resulted in the suspension of cells without stirring, thus providing a suitable environment to propagate cells without sedimentation to a surface. The rotating wall bioreactors did not provide a way to remove air bubbles that were causing shear and disrupting 3D cultures. Johnson Space Center successfully engineered the hydrofocusing bioreactor (HFB) that resolved the problem of removing the air bubbles from the fluid medium of NASA's rotating-wall space bioreactors. The HFB uses the principle of hydrodynamic focusing that simultaneously produces a low-shear fluid culture environment and a variable hydrofocusing force that can control the movement, location, and removal of suspended cells, tissues, and air bubbles from the bioreactor. The HFB is a rotating, domeshaped cell culture vessel with a centrally located sampling port and an internal viscous spinner. The vessel and spinner can rotate at different speeds either in the same or opposite directions. Rotation of the vessel and viscous interaction at the spinner generate a hydrofocusing force. Adjusting the differential rotation rate between vessel and spinner controls the magnitude of the force.

Claudio, Pier Paolo↗

Straight Ahead in Microgravity

INTRODUCTION The subjective straight-ahead direction is a very basic perceptual reference for spatial orientation and locomotion. The perceived straight-ahead along the horizontal and vertical meridian is largely determined by both otolith and somatosensory inputs which are altered in microgravity. The Straight Ahead in Microgravity (SAM) experiment will be conducted on the International Space Station (ISS) to examine how this spatial processing changes as a function of spaceflight. METHODS Data will be collected before the flight, at one-month intervals during long-duration stay (180 days) on board ISS, and after return to Earth. Control studies will also be performed during parabolic flights. Three different protocols will be used in each test session: (1) Fixation: The subject will be asked to look at actual targets (normal vision) and then to imagine these same targets (occluded vision) in the straight-ahead direction. Targets will be located at near distance (arm s length, ~0.5m), medium distance (~1 m), and far distance (beyond 2 m). This task will be successively performed with subject s body aligned with the spacecraft interior, and with subject s body tilted forward and backward by an operator. (2) Saccades: The subject will be asked to make horizontal and vertical saccades, first relative to the spacecraft interior reference system, and then relative to the subject s head reference system. This task will be successively performed with subject s body aligned with the spacecraft interior, and with subject s body tilted in roll or in pitch by an operator. (3) Linear Vestibulo-Ocular Reflex (VOR): The subject will be asked to stare at actual visual targets (normal vision) at various distances (near, medium, far) in the straight-ahead direction. Vision will then be occluded, and the subject will be asked to continue staring at the same imagined targets while he/she is passively translated forward-backward, up-down, or side-to-side. The subject's body motion will be performed by the restrained operator while the subject is free-floating. EXPECTED RESULTS The coupling of downward gaze with vergence eye movements observed on Earth is expected to increase in microgravity. Saccadic eye movements made in darkness along perceived axes are expected to be more closely aligned with the body s longitudinal axis in 0g compared to 1g, as the reference system for spatial orientation moves from an allocentric (gravitational) to an egocentric (idiotropic) vector. Changes in the linear VOR will reflect adaptive changes in otolith-ocular reflex contributions to the perceived straight-ahead. DISCUSSION A change in an individual's egocentric reference might have negative consequences on evaluating the direction of an approaching object or on the accuracy of reaching movements or locomotion. Consequently, investigating how microgravity affects the egocenter is important for understanding the problems associated with long-term effects of microgravity on astronauts' and how they re-adapt to the return of gravitational forces on Earth or other planetary surfaces. This project therefore has theoretical, practical and even clinical implications for the sensorimotor research gap "What are the changes in sensorimotor function over the course of a mission?"

Clement, G.↗

The Effects of Microgravity on Seated Height (Spinal Elongation)

ABSTRACT Many physiological factors, such as spinal elongation, fluid shifts, bone atrophy, and muscle loss, occur during an exposure to a microgravity environment. Spinal elongation is just one of the factors that can also affect the safety and performance of a crewmember while in space. Spinal elongation occurs due to the lack of gravity/compression on the spinal column. This allows for the straightening of the natural spinal curve. There is a possible fluid shift in the inter-vertebral disks that may also result in changes in height. This study aims at collecting the overall change in seated height for crewmembers exposed to a microgravity environment. During previous Programs, Apollo-Soyuz Test Project (ASTP) and Skylab, spinal elongation data was collected from a small number of subjects in a standing posture but were limited in scope. Data from these studies indicated a quick increase in stature during the first few days of weightlessness, after which stature growth reached a plateau resulting in up to a 3% increase of the original measurement [1-5]. However, this data was collected only for crewmembers in standing posture and not in a seated posture. Seated height may have a different effect than standing height due to a change in posture as well as due to a compounded effect of wearing restraints and a potential compression of the gluteal area. Seated height was deemed as a critical measurement in the design of the Constellation Program s (CxP) Crew Exploration Vehicle (CEV), called Orion which is now the point-of-departure vehicle for the Multi-Purpose Crew Vehicle (MPCV) Program; therefore a better understanding of the effects of microgravity on seated height is necessary. Potential changes in seated height that may not have impacted crew accommodation in previous Programs will have significant effects on crew accommodation due to the layout of seats in the Orion.. The current and existing configuration is such that the four crewmembers are stacked two by two with the commander and pilot seats on the top and the two remaining seats underneath, thereby limiting the amount of clearance for the crewmembers seated in the bottom seat. The inner mold line of these types of vehicles are fixed due to other design constraints; therefore, it is essential that all seats incorporate additional clearance to account for adequate spinal growth thereby ensuring that the crew can safely ingress the seat and be strapped in prior to its return to earth. If there is not enough clearance to account for spinal growth deltas between seats then there is the potential that crewmembers will not be able to comfortably and safely fit into their seats. The crewmember in the bottom stacked seat may even have negative clearance with the seat above him or her which could lead to potential ingress/egress issues or potentially injury of the crewmember during landing. These impacts are specific to these types of vehicles with stacked seat configuration. Without proper knowledge of the amount of spinal elongation, or growth, which occurs due to microgravity and space flight, the design of future vehicle(s) or suits may cause injury, discomfort, and limit crew accommodation and crew complements. The experiment primarily aimed to collect seated height data for subjects exposed to microgravity environments, and feed new information regarding the effect of elongation of the spine forward into the design of the Orion. The data collected during the experiment included, two seated height measurement and two digital pictures of seated height pre-, in-, and post-flight. In addition to seated height, crewmembers had an optional task of collecting stature , standing height. Seated height data was obtained from 29 crewmembers that included 8 ISS increment crew (2 females and 6 males) and 21 Shuttle crew (1 female, 20 males), and whose mean age was 48 years ( 4 years). This study utilized the last six Shuttle flights, STS-128 to STS-134. The results show that partipating crewmembers experienced growth up to 6% in seated height and up to 3% in stature. Based on the worst case statistical analysis of the subject data, the recommended seated height growth of 6% will be provided to the designers as the necessary seated height adjustment.

Young, K. S.↗

A Geology Sampling System for Microgravity Bodies

Human exploration of microgravity bodies is being investigated as a precursor to a Mars surface mission. Asteroids, comets, dwarf planets, and the moons of Mars all fall into this microgravity category and some are been discussed as potential mission targets. Obtaining geological samples for return to Earth will be a major objective for any mission to a microgravity body. Currently the knowledge base for geology sampling in microgravity is in its infancy. Humans interacting with non-engineered surfaces in microgravity environment pose unique challenges. In preparation for such missions a team at the NASA Johnson Space Center has been working to gain experience on how to safely obtain numerous sample types in such an environment. This paper describes the type of samples the science community is interested in, highlights notable prototype work, and discusses an integrated geology sampling solution.

Hood, Anthony↗

Systemic Response to Microgravity: Utilizing GeneLab Datasets to Identify Molecular Targets for Future Hypotheses-Driven Spaceflight Studies

Biological risks associated with microgravity are a major concern for long-term space travel. Although determination of risk has been a focus for NASA research, data examining systemic (i.e., multi- or pan-tissue) responses to space flight are sparse. To perform our analysis, we utilized the NASA GeneLab database which is a publicly available repository containing a wide array of omics results from experiments conducted with: i) with different flight conditions (space shuttle (STS) missions vs. International Space Station (ISS); ii) a variety of tissues; and 3) assays that measure epigenetic, transcriptional, and protein expression changes. Meta-analysis of the transcriptomic data from 7 different murine and rat data sets, examining tissues such as liver, kidney, adrenal gland, thymus, mammary gland, skin, and skeletal muscle (soleus, extensor digitorum longus, tibialis anterior, quadriceps, and gastrocnemius) revealed for the first time, the existence of potential master regulators coordinating systemic responses to microgravity in rodents. We identified p53, TGF(beta)1 and immune related pathways as the highly prevalent pan-tissue signaling pathways that are affected by microgravity. Some variability in the degree of change in their expression across species, strain and time of flight was also observed. Interestingly, while certain skeletal muscle (gastrocnemius and soleus) exhibited an overall down-regulation of these genes, some other muscle types such as the extensor digitorum longus, tibialis anterior and quadriceps, showed an up-regulated expression, indicative of potential compensatory mechanisms to prevent microgravity-induced atrophy. Key genes isolated by unbiased systems analyses displayed a major overlap between tissue types and flight conditions and established TGF(beta)1 to be the most connected gene across all data sets. Finally, a set of microgravity responsive miRNA signature was identified and based on their predicted functional state and subsequent impact on health, a theoretical health risk score was calculated. The genes and miRNAs identified from our analyses can be targeted for future research involving efficient countermeasure design. Our study thus exemplifies the utility of GeneLab data repository to aid in the process of performing novel hypothesis based spaceflight research aimed at elucidating the global impact of environmental stressors at multiple biological scales.

GeneLab↗

Systemic Microgravity Response: Utilizing GeneLab to Develop Hypotheses for Spaceflight Risks

Biological risks associated with microgravity are a major concern for long-term space travel. Although determination of risk has been a focus for NASA research, data examining systemic (i.e., multi- or pan-tissue) responses to space flight are sparse. To perform our analysis, we utilized the NASA GeneLab database which is a publicly available repository containing a wide array of omics results from experiments conducted with: i) with different flight conditions (space shuttle (STS) missions vs. International Space Station (ISS); ii) a variety of tissues; and 3) assays that measure epigenetic, transcriptional, and protein expression changes. Meta-analysis of the transcriptomic data from 7 different murine and rat data sets, examining tissues such as liver, kidney, adrenal gland, thymus, mammary gland, skin, and skeletal muscle (soleus, extensor digitorum longus, tibialis anterior, quadriceps, and gastrocnemius) revealed for the first time, the existence of potential master regulators coordinating systemic responses to microgravity in rodents. We identified p53, TGF1 and immune related pathways as the highly prevalent pan-tissue signaling pathways that are affected by microgravity. Some variability in the degree of change in their expression across species, strain and time of flight was also observed. Interestingly, while certain skeletal muscle (gastrocnemius and soleus) exhibited an overall down-regulation of these genes, some other muscle types such as the extensor digitorum longus, tibialis anterior and quadriceps, showed an up-regulated expression, indicative of potential compensatory mechanisms to prevent microgravity-induced atrophy. Key genes isolated by unbiased systems analyses displayed a major overlap between tissue types and flight conditions and established TGF1 to be the most connected gene across all data sets. Finally, a set of microgravity responsive miRNA signature was identified and based on their predicted functional state and subsequent impact on health, a theoretical health risk score was calculated. The genes and miRNAs identified from our analyses can be targeted for future research involving efficient countermeasure design. Our study thus exemplifies the utility of GeneLab data repository to aid in the process of performing novel hypothesis based spaceflight research aimed at elucidating the global impact of environmental stressors at multiple biological scales.

GeneLab↗

Gene Expression of Pathogens in Simulated Microgravity

Extended exposure to radiation and microgravity in space has been linked to astronauts developing chronic diseases upon returning to Earth. The Gram-negative pathogen Serratia marcescens has been shown to potentially cause significant infections in humans and in insect models on Earth. Our recent findings also showed that S. marcescens shows an increase in virulence after a short period of growth in the spaceflight environment, which raises initiatives to find the correlation between space environment and the increased virulence. Because we know that the health of astronauts is immunocompromised in space, it is possible that the combination of increased bacterial virulence and the weakened immune system will cause astronauts to be more susceptible to chronic diseases in extended spaceflight. With 75% of human disease genes being conserved in the fruit fly Drosophila melanogaster, these insects act as an ideal model organism to study the human immune system. The high accessibility, low cost, high rate of reproductivity, and short lifespans of D. melanogaster facilitate efficient, high-quality research that seeks to understand altered virulence of this opportunistic pathogen. In this ground-based study, we will use a rotating wall vessel apparatus to simulate microgravity and determine how pathogenicity changes by evaluating differences in gene expression for S. marcescens between bacteria grown in simulated microgravity conditions and controls. We will compare the results of our findings to gene expression patterns in actual spaceflight samples of S. marcescens grown on the ISS (International Space Station) during a recent validation mission, to see if there are common mechanisms across our simulated microgravity and actual spaceflight microgravity samples that both show increased virulence in the fruit fly. With extended space travel in the foreseeable future, understanding how human physiology will be affected by these different factors will help mitigate risks and deaths.

spaceflight analogue↗