ALBERT

Description: Space Biology current Rodent Research hardware and capabilities as of October 2017. These include the Life Sciences Glovebox, Rapid Freeze, sample cartridges and simulated carcass freezing, mass measurement device, habitat configuration with enrichment, and the non-Rodent capabilities they feature.

Description: No abstract available

Description: No abstract available

Description: For over 100 years, neurologists have used eye movements to identify neural impairment, disease, or injury. Prior to the age of modern imaging, qualitative assessment of eye movements was a critical, routine component of diagnosis and remains today a routine law-enforcement tool for detecting impaired driving due to drugs or alcohol. We will describe the application of a simple 5-minute oculomotor tracking task coupled with a broad range of quantitative analyses of high-resolution oculomotor measurements for the sensitive detection of sub-clinical neural impairment and for the potential differentiation of various causes. Specifically, we will show that there are distinct patterns of impairment across our set of oculometric parameters observed with brain trauma, sleep and circadian disruption, and alcohol consumption. Such differences could form the basis of a self-administered medical monitoring or diagnostic support tool.

Description: Behavioral characteristics of D.melanogaster are strongly influenced by intrinsic and extrinsic factors, allowing scientists to assess how changes in physiology or environment manifest into behavior. Conversely, assessing changes in behavior of specimens provides valuable information about how the physiology of that organism responds to external changes. In this project, we developed a computer program to automate behavioral analyses of larvae and adult D. melanogaster aboard the International Space Station using on-board video recordings. Utilizing freely available libraries for Python, we set parameters to compute the number of animals, amount of locomotion as distance or movement, and the change in the perimeter of the larvae's outer shape to quantify behaviors such as curling or peristaltic full body wall contractions. Results show that our program is an efficient tool for analysis of larvae and adult locomotive behavior, thus providing scientists with a low-cost, efficient, and reliable method of quantifying behavioral data.

Description: Tardigrades are microscopic invertebrates that are uniquely radio tolerant among animals, and while the mechanisms of radiotolerance in some species is becoming understood, such mechanisms in Hypsibius dujardini, the most radio tolerant fully aquatic tardigrade, are unknown. We asked 1) Is H. dujardini resistant to direct or indirect DNA damage due to ionizing radiation? and 2) Is this resistance through initial DNA protection or efficient repair once damage has occurred? We confirmed H. dujardinis extraordinary radiotolerance but encountered challenges in performing molecular techniques, thus identifying a need for standardization of tardigrade experimental protocols.

Description: No abstract available

Description: One potentially important bone quality characteristic is the response of bone to cyclic (repetitive) mechanical loading. In small animals, such as in rats and mice, cyclic loading experiments are particularly challenging to perform in a precise manner due to the small size of the bones and difficult-to-eliminate machine compliance. Addressing this issue, we developed a precise method for ex vivo cyclic compressive loading of isolated mouse vertebral bodies. The method has three key characteristics: 3D-printed support jigs for machining plano-parallel surfaces of the tiny vertebrae; pivotable loading platens to ensure uniform contact and loading of specimen surfaces; and specimen-specific micro-CT-based finite element analysis to measure stiffness to prescribe force levels that produce the same specified level of strain for all test specimens. To demonstrate utility, we measured fatigue life for three groups (n = 5-6 per group) of L5 vertebrae of C57BL/6J male mice, comparing our new method against two methods commonly used in the literature. We found reduced scatter of the mechanical behavior for this new method compared to the literature methods. In particular, for a controlled level of strain, the standard deviation of the measured fatigue life was up to 5-fold lower for the new method (F-ratio = 4.9; p 〈 0.01). The improved precision for this new method for biomechanical testing of small-animal vertebrae may help elucidate aspects of bone quality.

Description: NASA in its plans to send humans to distant destination such as Mars faces the health and physiological performance problems caused by microgravity and space radiation. While most of the environmental conditions in spacecraft during flight can be made to mimic terrestrial conditions, microgravity cannot yet be managed. This space environmental factor has a major impact on the bodys biological system forcing alterations, in order to adapt to this new environment. Most space flight and ground-based studies suggest that prolonged exposure to microgravity leads to significant skeletal muscle atrophy, bone loss, and results in suppression of total metabolism. Due to microgravity, unloaded crewmembers lose up to 1.5% of their skeletal mass and 1.8% of bone strength each month during ISS missions. Remarkably many animals, including human-size bears, which are largely inactive during the 6 to 8 months of hibernation, show no loss in bone mass and much less muscle atrophy than would be anticipated over such a prolonged period of physical inactivity. This suggests that while in a suppressed metabolic state animals have unique natural mechanisms to prevent muscle disuse and bone atrophy. The molecular mechanisms underlying these important adaptations are not yet known. Radiation exposure is the second health hazard encountered during spaceflight that can cause radiation sickness, cancer or death. This study provides new evidence that metabolic activity levels play a critical role in radioprotection. Metabolic suppression, as an adaptive response of cells to minimize damage caused by radiation, enables cells to reduce cellular dysfunction and damage, and prolong their survival despite persistent oxidative stress. Thus mechanistic understanding of metabolism offers a means for sustaining astronauts in long-duration missions. The ultimate goals of this study are to demonstrate that induced metabolic suppression in animals and humans will profoundly reduce their sensitivity to the damaging effects of radiation and microgravity as well as other kinds of stresses caused by spaceflight. The beneficial effects of suppressed metabolism induced by different factors such as temperature, nutrition, and medications, will not only mitigate the most detrimental hazards of spaceflight but also radically reduce mission life support requirements and spaceflight logistics.

Description: NASA Ames Research Centers WetLab-2 system brings new capabilities to the ISS for researchers. The system can lyse cells and extract RNA on-orbit from different sample types ranging from microbial cultures to animal tissues. Our purification method has the advantage of using non-toxic chemicals and does not require alcohols or other organics. The purified RNA can then either be stabilized for return to Earth or can be used to conduct on-orbit quantitative Reverse Transcriptase PCR (qRT-PCR) analysis without the need for sample return. qRT-PCR reactions are performed by dispensing the RNA into reaction tubes that contain all lyophilized reagents needed to perform the analysis. The system uses a Cepheid SmartCycler that allows for multiplexing of assays, this can be used to normalize for RNA concentration and integrity and to study multiple genes of interest in each tube. There are a total of 16 independent PCR modules each capable of detecting up to four fluorescent channels. The WetLab- 2 system can downlink data from the ISS to the ground after a completed run and uplink new thermal cycling programs. The ability to purify and stabilize RNA on-orbit can eliminate the confounding effects of reentry stresses and shock acting on live cells and organisms or the concern of RNA degradation of some samples. It also has the benefit of minimizing the needed downmass. Conducting qRT-PCR and generating results on-orbit is also an important step towards utilizing the ISS as a National Laboratory facility. Specifically, the ability to get on-orbit data will provide investigators with the opportunity to adjust experimental parameters in real time without the need for sample return and re-flight. On orbit gene expression analysis can also provide benchmarking prior to sample return. The system can also be used for analysis of air, surface, water, and clinical samples to monitor environmental pathogens and crew health. The validation flight of the WetLab-2 system using E. coli bacteria and mouse liver is scheduled to launch on SpaceX-8 this spring. Pending operations, the preliminary results from the validation flight will be presented. To support the needs of future researchers, we are adapting our system to purify RNA from two additional sample types: fibrous tissue such as muscle and mammalian adherent cells grown on alginate beads. Progress of this work will also be presented. The WetLab-2 Project is supported by the Research Integration Office in the ISS Program.

Description: Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small "production pack" system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.

Description: The Micro-12 flight experiment was launched on SpaceX-15 and completed during berthing on the International Space Station. The goal of this experiment was to understand the effects of spaceflight and microgravity on the physiology of the model exoelectrogen Shewanella oneidensis MR-1. BioServe Fluid Processing Apparatus (FPA) and Group Activation Pack (GAP) hardware systems were used for both flight and ground control tests. Under spaceflight conditions, extracellular electron transfer (EET) rates were found to be significantly increased on insoluble substrates, while biofilm development appeared to be unchanged under the conditions tested; these processes are critical for microbial-assisted bioelectrochemical systems. Additionally, RNAseq analysis, proteomic profiling, and competitive mutant fitness profiling were performed to gain further understanding of microbial physiology under EET-respiring conditions during spaceflight. Overall, the results of the Micro-12 project support the idea that Shewanella oneidensis MR-1, in particular, and exoelectrogens in general could be useful chassis organisms for synthetic biology applications using microbial bioelectrochemical systems. These findings will assist bioengineering and synthetic biology development efforts harnessing the unique capabilities of exoelectrogens for life support and in situ resource utilization.

Description: No abstract available

Description: Since Apollo 17 in 1972, NASA has sent no humans or other biological organisms outside of Earth's protective magnetosphere. Recently, NASA has set its sights on human exploration in deep space, with an ambitous plan to put astronauts back on the Moon by 2024 and to eventually land human missions on Mars. Such missions will require significant countermeasures, likely both technological and biomedical, to protect biology from chronic radiation exposure. CubeSats can inform these countermeasures by querying relevant space environments with model organisms.NASA has launched five biological CubeSat missions into low-Earth orbit (LEO). GeneSat-1 was launched in 2006 to study gene expression and increase our knowledge of how spaceflight affects microbes. Similar life-support technologies were then used in PharmaSat and O/OREOS, which launched in 2009 and 2010, respectively. PharmaSat contained optical systems to examine how yeast cells responded to an antifungal treatment. One of O/OREOS payloads, SESLO (Space Environment Survivability of Living Organisms), housed dormant microorganisms, which were rehydrated on orbit to track alterations to growth and metabolism induced by microgravity and radiation. In 2014, NASA launched SporeSat to study the mechanisms of plant cell gravity sensing using lab-on-a-chip devices. Most recently, in 2017, NASA launched EcAMSat (E. coli AntiMicrobial Satellite), which investigated the effects of microgravity on antibiotic resistance of a pathogenic bacterium. Each one of these missions increased our understanding of the biological effects of spaceflight in LEO, while refining technologies and imparting valuable lessons to the next generation of CubeSats.CubeSats housing translational biological models are therefore ideal for defining the hazards of deep space travel, as they can provide critical data over relevant durations. BioSentinel, a next-generation deep-space CubeSat, is planned to launch as a secondary payload on Artemis 1 in 2020. BioSentinel will study the DNA damage response to deep space radiation in yeast.

Description: Space crop production will be important in future long duration exploration missions to supplement the packaged diet with fresh bioactive nutrients. Plant care and the addition of fresh veggies to the diet may also have a role in astronaut well-being. Pick-and-eat salad crops are the best candidates for this near-term supplementation since they require minimal processing or preparation to add to meals. While light quality can strongly influence plant responses on Earth, the impacts of light quality on plant growth and composition in spaceflight remain unclear. The VEG-04 experiment uses two Veggie plant growth chambers on the International Space Station to simultaneously test different red: blue light ratios on the growth of Mizuna mustard, a leafy green salad crop. In addition to plant health and yield, the composition of key nutrients is assessed. Astronauts conduct on-board organoleptic evaluation of the fresh produce. Microbial food safety of returned produce is examined, and a Hazard Analysis Critical Control Point (HACCP) plan has been developed for this crop. VEG-04 consists of two experiments, one lasting 28 days with a single harvest, and the second lasting 56 days, with three cut-and-come-again harvests. These different scenarios provide an opportunity to test two production concepts, examine different fertilizers, monitor microbial changes over time for this crop, and assess potential impacts of interacting with plants on crew behavioral health and performance in spaceflight operations. In ground testing, plant growth was not significantly different across the different light treatments, however nutrient composition did differ significantly. Flight test results will be compared with ground data. This research was co-funded by NASA's Human Research Program and Space Biology in the ILSRA 2015 NRA call.

Description: The impact of spaceflight on immune function is undoubtedly a critical focus in the area of space biology and human health research. Heat shock proteins (Hsp) are an evolutionarily conserved family of proteins that are expressed in response to cellular and physiological stressors, experienced during radiation exposure, confinement, circadian rhythm disruption, and altered gravity (hypergravity experienced at launch/landing and microgravity experienced in-flight). In particular, Hsp70 aids in the folding of proteins, facilitates the movement of proteins across the membranes during signal transductions and can stimulate innate immunity. Since Hsp70 is induced during cellular stress, and can act as a stimulator for innate immunity, we sought to address how a loss of Hsp70 affects immunity, under the stress-inducing model of acute and chronic hypergravity. Moreover, the effects of gravity as a continuum on the induction of Hsps and key immune genes were also assessed to determine if increased cellular stress, via increased gravity (g)-force, contributes to immune dysfunctions. For this, wildtype (W1118) and Hsp70 deficient (Hsp70null) Drosophila melanogaster were subjected to simulated hypergravity at increasing levels of g-force (1.2g, 3g, and 5g) for acute (1hr) and chronic (7-day) timepoints and were compared to 0g 'non-hypergravity' controls. Following simulation, whole bodies were sex-segregated, RNA was isolated and quantitative (q)PCR was performed to determine differential immune gene expression profiles. Further, functional output of hemocytes were assessed by a phagocytosis assay. Collectively, these studies evaluated the effects of Hsp70 in the context of immunity during acute and chronic hypergravity. Indeed, relevance for this work can directly translate to acute effects of launch/landing gravitational forces upon liftoff (~1.7g) and entry (~3.4g) that astronauts experience. In addition, the effects of chronic cellular stress is directly relevant to the immune health of astronauts on long duration missions, as well. Thus, as we approach the goal of returning to the Moon and landing the first humans on Mars, an evaluation of gravity as a continuum and the stress-inducing effects of altered gravity experienced during spaceflight on astronaut immunity and health are necessary.

Description: Extra-terrestrial colonization is of growing interest to space agencies and private entities, emphasizing the importance of research on reproduction and development in the absence of Earth's 1G. Maternal stressors can modify offspring development, exerting significant lifespan and crossgenerational changes through prenatal programming. The space environment is stressful, therefore exposure to altered gravity during pregnancy may impact later life outcomes in offspring. In ground-based studies, we exposed pregnant rats to continuous +G (above Earth gravity), and observed overweight and elevated anxiety in adult male (but not female) offspring, common phenotypes associated with prenatal maternal stress. Here we hypothesize that exposure to increased gravity during pregnancy elicits changes in the expression of stress-related genes in placenta that may mediate emergence of later life outcomes. While the placenta transports maternal factors to the fetus and produces endogenous fetal hormones, stress-induced changes at the placental-uterine interface may also alter communication between mother and fetus, facilitating prenatal transmission of unfavorable later life outcomes and cross-generational epigenetic alterations. Maternal stress elevates maternal glucocorticoids however placental 11b-hydroxysteroid dehydrogenase type 2 (HSD11B2) buffers fetal exposure by converting cortisol/corticosterone into inactive metabolites. Maternal stress during pregnancy down-regulates this enzyme and can induce epigenetic changes in placental and fetal tissues accounting for heightened adult HPA reactivity. Past studies have shown a placenta-specific increase in DNA methyltransferase (DNMT3a) mRNA in stressed mothers, an effect with implications for genome-wide epigenetic changes that may account for diverse phenotypic outcomes following maternal stress. Here we exposed groups of pregnant rats to one of five gravity loads (1, 1.5, 1.75 and 2G) and analyzed placental samples during late gestation. We predicted a systematic dose-response relationship between gravity load and the expression of the HSD11B2 and DNMT3 genes, thereby linking maternal exposure to altered gravity during pregancy with maternal stress.

Description: 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

Description: Research on human acclimation to spaceflight, including the recent NASA's Twin Study, reports complex effects of the spaceflight environment on health, with both acute and prolonged changes in multiple tissues. Spaceflight includes multiple factors such as microgravity, ionizing radiation, physiological stress, and disrupted circadian rhythms, that have been shown to contribute to pathophysiological responses that target immunity, bone and muscle integrity, cardiovascular and nervous systems. In this study, we used a well-established spaceflight model organism, Drosophila melanogaster, to assess spaceflight-associated changes on the nervous system. With 75% disease gene orthology to humans, short generation time, large sample size and ease of genetic, neuronal and behavioral studies, Drosophila is an excellent model to study nervous system dysfunction. Here, we present results from MVP-Fly-01 spaceflight mission that was launched on SpaceX CRS-14. The MVP hardware (developed by Techshot) used in this mission enabled us to have an in-flight 1g centrifuge, to distinguish the changes resulting from gravity versus those induced by other environmental factors associated with spaceflight. We observe behavioral impairments (p〈0.001) and synaptic deficits, including decreased synaptic connections (p〈0.05), in 3rd instar larvae which were developed in space. Furthermore, space-grown microgravity adults show a decrease in neuronal (p〈0.05) and dendritic field (p〈0.01) in adult brains coupled with an increased number of apoptotic cells (p〈0.001) compared to in-flight 1g controls, suggesting increased neuronal loss under spaceflight conditions. In summary, we observe that altered gravity leads to gross neurological deficits. To better understand the long-term effects of spaceflight on the nervous system, longitudinal and multigenerational changes were also identified. This study will help elucidate the different approaches to prevent nervous system dysfunction in astronauts during spaceflight, while also contributing to a better understanding of the pathways that are related to some CNS disorders on Earth.

Description: The Veggie facility on ISS was used to demonstrate the suitability of the leafy greens, red romaine 'Outredgeous' lettuce, Waldman's green lettuce, and mizuna in a mixed crop configuration.

Description: In-situ food production is a necessary step for human exploration of the solar system and requires a deep understanding of plant growth in reduced gravity environments. In particular, the lack of buoyancy-driven convection changes the gas exchange at the leaf surface, which decreases photosynthesis and transpiration rates, and ultimately biomass production. To understand the intricate relations between physical, chemical, and biochemical processes, the following methodology combines the development of a mechanistic model of plant growth in reduced gravity environments, computational fluid dynamics (CFD) simulations, and experiments in different time frames.The model presented here is a coupled mass and energy balance using the single round leaf assumption, including gravity as an entry parameter, and the leaf surface temperature as an output variable. Measures of the leaf surface temperature using infra-red cameras allow for a computation of the transpiration rate. This approach was followed to design a parabolic flight experiment, which performed 7 flights, and enabled data collection for model validation in different gravity and ventilation settings on a short time frame. Current measures of carbon assimilation and transpiration rate at the leaf and canopy level using an infra-red gas analyzer (Li-6800) in 1g lab conditions on several species will enable a validation on longer time frames and further calibration of the model. CFD studies both on the parabolic flight and on the lab experimental set-up allow the precise assessment of ventilation above the canopy and plants' leaves.Ultimately, this work will provide recommendations for the design of future plant growth hardware, especially on the lowest adequate ventilation for optimal plant growth in reduced gravity environments, as well as assessing biomass and oxygen production rates on planetary surfaces and space stations. This work was funded by CNES, CNRS, Clermont Auvergne Metropole, and NASA Space Biology through NASA postdoctoral program / USRA.

Description: Spaceflight poses many challenges for humans. Ground-based analogs typically focus on single parameters of spaceflight and their associated acute effects. This study assesses the long-term transcriptional effects following single and combination spaceflight analog conditions using the mouse model, simulated microgravity via hindlimb unloading (HLU) and/or low-dose irradiation (LDR) for 21-days, followed by 4 months of readaptation. Changes in gene expression and epigenetic modifications in whole brain samples during readaptation were analyzed by DESeq2 and reduced representation bisulfite sequencing (RRBS). The results showed minimal gene expression alterations at 4-months within single treatment conditions of HLU and LDR. Following combined HLU+LDR, gene ontology and methylation analyses showed multiple altered pathways involved in neurogenesis and neuroplasticity, regulation of neuropeptides and cellular signaling. In brief, neurological readaptation following combined chronic LDR and HLU is a dynamic process that impacts brain structure and function and may lead to late onset neurological sequelae

Description: The Advanced Plant Habitat (APH) was installed on the International Space Station (ISS) in October 2017. Following a successful EVT (Experiment Verification Test) study at Kennedy Space Center (KSC), using Arabidopsis lines with varying levels of lignin, two inaugural studies were carried out on ISS in 2018 under the same experimental design, with the corresponding ground controls at KSC. The APH for this study deploys a substrate-based root module designed for plant growth in microgravity. Upon experiment initiation (such as for the EVT), the root module is primed (liquid imbibition) by flooding the root zone to initiate seed germination and to remove air from the porous tubing and particulate media. In the APH ISS inaugural study, the speed of supplying water to initially dry media was found to adversely affect the overall moisture distribution within the root module in microgravity (but not at 1g). Non-destructive estimations of Arabidopsis plant growth were carried out by monitoring changes in rosette leaf area on a daily basis. These data indicated that the original priming procedure caused patchy moisture distribution that affected plant growth and survival. An improved methodology for priming the second root module of PH-01 was devised and implemented in the second experiment. Leaf area and color estimates suggested that the modified priming scheme improved moisture distribution and plant growth. These data, when compared with the EVT study, suggest that nondestructive measurements of plant growth can aid towards optimization of plant growth conditions in microgravity.

Description: The rodent hindlimb unloading (HU) model was initially developed to simulate the cephalad fluid shift and musculoskeletal disuse in astronauts. Since then, the HU model has been applied to explore how other systems (e.g. immune, cardiovascular and CNS) respond to weightlessness. Most HU studies are performed with singly-housed animals, although social isolation also can substantially impact behavior and physiology, and therefore may confound HU experimental results. We hypothesized that relative to social housing, single housing exacerbates HU-induced dysfunction in select organ systems. We refined the standard NASA-Ames HU model to accommodate social housing in HU pairs, retaining advantageous features of traditional housing but using commercial off-the-shelf components to facilitate adoption by others. We conducted a 30 day HU experiment with adult, female C57Bl6/NJ mice that were either singly or socially housed. HU animals in both single and social HU housing displayed expected musculoskeletal deficits compared to housing matched, normally loaded (NL) controls. However, select immune, HPA axis, and CNS responses were differentially impacted by the HU social environment relative to NL controls. HU reduced % CD4+ T cells in singly-housed, but not socially-housed mice. Surprisingly, HU increased adrenal gland mass in socially-housed but not singly-housed mice, while social isolation increased adrenal gland mass in NL controls. HU also increased plasma corticosterone levels (day 30) in both singly and socially-housed mice. Thus, the social environment altered select adrenal and immune, but not musculoskeletal, responses to simulated weightlessness. We refine our original hypothesis since our results show combined stressors can mask, not only exacerbate, tissue responses to HU. These findings further expand the utility of the HU model for studying possible combined effects of the various spaceflight stressors.

Description: The effects of microgravity, and social isolation on the CNS are poorly understood. We hypothesize that mitochondrial reactive oxygen species (ROS) play an important role in this process. Since mice are social animals, our lab developed a novel social model of hindlimb unloading (HU), enabling us to determine the effects of both social isolation and simulated microgravity. Responses to 30d of HU were compared in wildtype or transgenic MCAT mice who over-express human catalase in mitochondria. Abundance of 4-Hydroxynonenal, Park7 (a redox-sensitive chaperone and sensor of oxidative stress) and corticosterone were measured by ELISA. Cytokines related to inflammation in the hippocampus and in plasma were analyzed by a protein array. Behavioral data was collected over a 24-hour period.Socially housed HU mice were more active and conducted at least two times more exploratory activities, compared to normally loaded mice. Correlation analysis revealed that specific brain and plasma cytokines correspond with specific behaviors. Simulated microgravity and/or social isolation caused changes in cytokine patterns in the hippocampus and in plasma, with significant interaction effects of HU and genotype in expression levels of five cytokines (out of 35). Interestingly, elevation of these generally pro-inflammatory cytokines by HU in WT mice was mitigated in MCAT mice, suggesting a role for mitochondrial ROS signaling in inflammatory CNS responses to microgravity. Interestingly, socially housed mice had also lower level of 4HNE and higher level of Park7 in the hippocampus compared to singly housed animals. The cytokine responses to social isolation were more extensive in brain vs plasma. Further, there was no overlap in the cytokine repertoire regulated in response to microgravity versus, isolation suggesting divergent mechanisms or downstream signaling. These findings implicate a potentially important role for mitochondrial ROS in CNS responses to the challenges posed both by prolonged missions in space and bedrest on Earth

Description: Spaceflight and the ensuing fluid shifts, together with an overall reduction in physical activity, lead to acute and latent effects on the cardiovascular system. This current study makes use of the rodent hindlimb unloading (HU) model to determine how factors such as sex, age, and duration of exposure impact cardiac responses to weightlessness. We hypothesize that extended exposure to simulated weightlessness and the ensuing recovery alters cardiac structure and expression of select genes, including those involved in redox signaling which together, negatively impact long-term cardiac tissue health. To begin to test this hypothesis, male and female rats underwent HU at various durations up to 90 days, with a subset reambulated after 90 days of HU. Physiological stress or contractility changes lead to alterations in ventricular cardiomyocyte size and ventricular wall thickness to adapt to greater functional demand and mitigate mechanical stress to ventricular tissue; under certain conditions, these changes also may mark progression to cardiac failure. Hence, left ventricular cardiomyocyte size (cardiomyocyte cross sectional area, CSA) was quantified to determine if HU leads to structural adaptation responses in cardiac tissue and if age and sex had any impact on this outcome. Cardiomyocyte CSA of older males (9 months) were altered by HU in a time-dependent manner, where HU led to decreases in CSA at 14 days and increases at 90 days. In contrast, younger males (3 months) did not show any changes at day 14 of HU. CSA of females (3 months) was increased in response to short-term HU (14 days) suggesting sex-dependence of structural changes. In older HU males, cardiomyocyte CSA was comparable to controls after 90 days of re-ambulation. Levels of the DNA oxidative damage marker, 8-hydroxydeoxyguanosine (8-OHdG) were greater in left ventricular tissue of females that underwent HU compared to sex-matched controls, while there were no such differences in older or younger males. To gain insight into the signals that drive cardiac adaptations to HU, global transcriptomic analysis (RNAseq) was performed on left ventricular tissue of older males that underwent 14 days of HU. Short-term simulated weightlessness led to differential expression of genes involved in immune and pro-inflammatory signaling. A subset of these genes play a role in autoimmune and cardiovascular disease and are targets of current drugs used to treat bradycardia, hypertension, atherosclerosis and rheumatoid arthritis, amongst others. Oxidative damage/redox signaling pathways were not enriched at the timepoint tested in older males. Since young females displayed greater oxidative damage to DNA, activation of oxidative stress responses at earlier or later time points cannot be ruled out. In summary, simulated weightlessness in adult rats caused changes in cardiomyocyte structure in a sex and age-dependent manner, and the transcriptional regulation of key mediators of immunity and cardiovascular disease, meriting further study to define cardiac risks for interplanetary travel of human crew. Our findings also confirm the value of the rat HU model for cardiac health and countermeasure research.

Description: Future long-duration missions face significant challenges maintaining crew health. A critical area is supplying adequate nutrition, as certain vitamins and nutrients in supplied foods and supplements demonstrate substantial degradation during extended storage. To address this issue, we are developing and flight-testing a platform technology that demonstrates in situ microbial production of targeted nutrients over extended mission durations. This 5-year experiment, known as BioNutrients-1, was started on the International Space Station in May 2019. It involves two components: an on-orbit hydration and production experiment; and the development of space-compatible, key bio-manufacturing microorganisms. On-orbit testing utilizes a small production pack system that encloses sterile edible growth substrate and desiccated Saccharomyces cerevisiae strains genetically engineered to produce the nutrients beta-carotene or zeaxanthin. On hydration and mixing of the production pack, the organisms revive and grow until limited by the depletion of growth media, hypothetically leading to consistent amounts of biomass and nutrients. In eventual mission applications, the packet contents would be heat treated to inactivate the microorganisms prior to consumption. For these flight experiments, the packet will not be heat treated, but will instead be frozen for return to Earth for analyses. In addition to the production pack trials, 14 different microorganisms/treatments were also delivered to ISS for long-duration storage. These samples will be intermittently returned to Earth and analyzed to determine survival rates and genomics. For this presentation, initial data from returned samples and ground controls will be discussed.

Description: Plant associated microbiomes, the rhizosphere and phyllosphere, are composed of communities of bacteria and fungi that may be mutualistic or pathogenic. These communities have the potential to influence plant health and development and can affect plant growth. Crop plants are being investigated as a fresh and safe supplement to astronauts diet and it is critical to understand and characterize these microbial communities. Multi-species crops, Mizuna mustard (Brassica rapa var japonica), Outredgeous red romaine lettuce (Lactuca sativa), and Waldmans Green lettuce (Lactuca sativa) were grown in two Veggie units on the International Space Station (ISS) for three grow outs in various combinations of plant types. Upon harvest, plant and pillow samples were frozen and returned to Earth for analysis. Bacterial and fungal community analyses for plant leaf and root, as well as pillow components, wick and media, were completed using next generation sequencing with the goal of surveying the composition of the entire community and identifying any potential pathogens. Bacteria were identified using the 16S rRNA gene whereas, fungi were identified with the internal transcribed spacer (ITS). The community composition for these three crops was compared between crop types and between plant tissue types. It is vital to mission success for the short term and long term to add nutritious, safe to eat vegetables providing a supplement to the crew members dietary requirements as well as to develop planning for deep space missions as we reach for the moon and on to Mars. Veggie technology validation tests were supported by NASAs Space Biology Program.

Description: It is important to determine the health risks and potential survival for astronauts associated with long-term space missions. This entails not only understanding the impact the space environment will have on humans, but also how it will affect other organisms needed for humans to survive in space such as plants. In addition, it has been reported in the literature that hundreds of genes seem to be conserved and/or transferred between different organisms from bacteria, archaea, fungi, microorganisms, and plants to animals. Since space travel involves humans in a closed environment over a long period of time, we hypothesize that potential conserved biological factors will occur between the different organisms in that environment possibly due to transfer of genes. Determining the conserved factors that are commonly being regulated in space can shed insight into possible universal master regulators and also determine the symbiotic relationship between the organisms in space. Utilizing NASA's GeneLab Data Repository (a rapidly expanding, curated clustering of spaceflight-related omics-level datasets for all organisms), we were able to uncover a novel pathway and factors that were commonly shared between humans, mice, plants, C. Elegans, and drosophilas. Through ChIP-Seq enrichment analysis techniques utilizing various GeneLab datasets from each species that were flown in space, we found the following factors to be conserved across all species: oxidative stress, DNA damage (through GABPA/NRFs and NFY), SIX5, GTF2B and glutamine synthetase. Such commonalities would likely reflect the effects of factors such as microgravity and the increased radiation exposure inherent in spaceflight on basic physical processes shared by all biological systems at the cellular level. Differences between organismal responses revealed by GeneLab's data should also help understand the unique reactions to life in space that arise from the very different lifestyles of microbes, animals and plants.

Description: Prokaryotic lifeforms can be observed to demonstrate many keen adaptive advantages, perhaps facilitated by a nature simplistic relative to divergent domains of life. In particular, decompartmentalized gene expression facilitates adaptation by allowing free exchange of genetic material, albeit at the cost of increased susceptibility to genetic damage. Thus, these lifeforms must compensate by embracing diverse investment strategies in an attempt to brute force the evolvability equation through precipitous genesis, lean metabolic efficiency, and sheer population. This prokaryotic archetype also enables symbiotic relationships with secondary mobile genetic elements known as plasmids, which have been shown to drive evolution on rapid temporal scales through processes such as conjugation and transformation. This study attempts to decipher whether these mechanisms of horizontal gene transfer (HGT) are major factors in determining prokaryote fitness within a unique isolated environment, the International Space Station (ISS). The ISS Microbial Tracking (MT) project has generated a wealth of data concerning the successive reigns of microbial genera that appear to thrive amidst harsh conditions for life. Despite relatively higher doses of ionizing radiation as compared to Earth, complications associated with microgravity, and the anti-microbial mlange deployed, microbial life still persists in this environment. The NASA GeneLab serves as a data repository and analysis platform to enable researchers to access space flight factor related data. With the use of GeneLabs modern computational suites (computomics), phylogenetic and functional genomic investigations of HGT events were conducted on the data generated from the MT-1 project. The putative data concerning the plasmid population (plasmidome) of the ISS was algorithmically derived and compared to those of habitats with similar environmental dynamics- such as living quarters and hospitals- to investigate whether these HGT elements may play crucial role(s) in shaping the microbiome of this closed habitat that serves as the only inhabited structure in space.

Description: The ISS provides a platform for conducting Rodent Research (RR) in microgravity and 9 missions have been successfully conducted. The results from these experiments have begun to provide new insights into the effects of spaceflight on mammalian physiological systems. After RR-1-4, the Flight IACUC required inclusion of additional cage enrichment into the Rodent Habitats (RH) to "enhance animal well-being by providing animals with sensory and motor stimulation, through structures and resources that facilitate the expression of species typical behaviors". A Hut, in the form of a rigid, mesh igloo-like shelter was implemented beginning with RR-5. The potential influence of the Hut in the novel cage environment of RH on various spaceflight-sensitive physiological systems has not been fully explored. To understand the effects of the Hut, mice (female C57Bl/6J, 15wks) were housed in Vivarium cage (n=5), RH with Hut (n=5), No Hut (n=5), Nestlet (n=10), and Cocoon (n=10) for 7 weeks. There were no differences in weekly body mass or food consumption. Tail blood draw indicated no differences in plasma corticosterone levels, immune cell types, or IgA levels. 24hrs prior to euthanasia, Open Field (OF) and Novel Object (NO) tests were performed. There were no differences across groups, all mice engaged in thigmotaxis (arena wall proximity) in the OF over 50% of the recorded time, and thigmotaxis declined when a NO was introduced. Additional behavioral analysis from daily videos are in progress to quantify activity levels. Post-euthanasia, there were no differences in soleus muscle or adrenal gland mass. Analysis of distal femur cancellous revealed some differences in microarchitecture. These results show that introduction of the Hut may diminish differences observed between spaceflight and ground controls, warranting improved validation of Hut effects in space, and also underscore the value of thorough preflight, ground based testing.

Description: A comprehensive understanding of the effects of spaceflight and altered gravity on human physiology is necessary for continued human space exploration and long-term space habitation. The oxidative stress response has been identified in astronauts exposed to short- and long-term space missions that are exposed to the multitude of stress factors of spaceflight, including altered gravity and radiation exposure. Reactive oxygen species (ROS) are byproducts of homeostatic cellular metabolism, yet when overproduced the oxidative stress response ensues, rendering molecules destructive causing cell death and inflammation. Controlling aberrant ROS production is necessary to prevent pathological consequences, in particular within the nervous system, since neurons are extremely sensitive overexpressed ROS insults. We hypothesize that exposure to altered gravity triggers the oxidative stress response, leading to impairments in the nervous system. In this study, we used a well-established spaceflight model organism, Drosophila melanogaster, to assess altered gravity associated changes in the nervous system using a ground-based hypergravity model. Acute hypergravity resulted in an induction of oxidative stress-related genes with an increase in reactive oxygen species (ROS) in fly brains (p〈0.001). Also, qPCR analysis shows that parkin gene expression is significantly reduced in these fly brains(p〈0.05). Additionally, chronic hypergravity resulted in depressed locomotor phenotype in these flies (p〈0.05) in conjunction to decreased dopaminergic neuron counts (p〈0.0001) and increased apoptosis in these fly brains (p〈0.0001). Further, assessment of neurological changes, including the neuronal architecture, synaptic integrity and genetic regulation caused by hypergravity conditions were noted. Overall, our results validate chronic hypergravity simulation as a behavioral model to study spaceflight effects, and oxidative stress pathway as a potential avenue for countermeasure development for astronauts undergoing short- and long-term missions and for neurodegenerative research on Earth.

Description: With humans pushing to live further off Earth for longer periods of time, it is increasingly important to understand the changes that occur in biological systems during spaceflight whether these be astronauts, their microbial commensals, or their plant-based life support systems. In a three-part presentation, we discuss GeneLab and recent discoveries regarding the microbiota of spacecrafts and space-flown animals. Part 1: GeneLab: Open Science for Life in Space, Jonathan Galazka, NASA Ames Research Center To accelerate the pace of discovery from precious spaceflight biological experiments, NASA as develop the GeneLab data system (genelab.nasa.gov), which allows unfettered access to omics data from spaceflight and spaceflight relevant experiments. GeneLab houses metagenomic datasets from spacecraft and relevant spacecraft models. Users can download this data and associated metadata to make new discoveries about how microbial communities may change and adapt to spaceflight.

Description: High-LET ionizing radiation is a major occupational health hazard for astronauts, but risk assessment remains elusive due to limited epidemiological data. Identifying genetic factors modulating the individual radiation response may be the most effective strategy to provide individualized risk management for long-duration high-radiation missions. We have started tackling the challenge of predicting individual risks by identifying human genetic loci associated with various radiation sensitivity phenotypes in primary blood mononuclear cells from a relatively large healthy human cohort. To date, we have performed the isolation of PBMCs from 768 subjects of the same ethnicity, and irradiated PBMCs from 576 subjects with 1 and 3 particles/100m2 of 600 MeV/n 56Fe, 350 MeV/n 40Ar and 350 MeV/n 28Si ions. The phenotypes of interest were: number of radiation-induced foci (or RIFs), CellROX oxidative stress responses and cell death, at 4h and 24h following irradiation. We have observed a significant inter-individual variability at 0 Gy between the 576 studied subjects, with a mean fold difference between the 10% lowest and highest responders of 5.6 of RIFs/cell, 7.9 in mean CellRox intensity, and 9.3 in percentage of dead cells. In order to better assess genetic factors influencing DNA repair, we used a metric previously introduced by our group to sort out radiation sensitivity phenotypes in mice: i.e. the ratio of the first to the second slope of RIFs/cell (between 0 and 1, and between 1 and 3 particle/100m2). Preliminary data on 192 individuals showed a distribution of low-dose responders (ratio 〉 1) to high-dose responders (ratio 〈 1) at 4h of 12%, 55% and 52% respectively for Fe, Ar and Si. The average value for the first and the second slopes was very similar for the two lowest LET (0.10 [-0.26;0.58] and 0.09 [-0.45;0.41] for Ar, 0.07 [-0.27;0.38] and 0.08 [-0.19;0.42] for Si), indicating a linear dose response across both fluence. Fe showed clear saturation for the highest dose with a slope of -0.09 [-0.86;1.51] against 0.68 [-2.21;2.20] for the low dose range, which probably reflects that many PBMCs are beyond repair at the high dose. Note that other significances were found for additional factors such as BMI and age whereas none were found for sex. GWAS will be performed on all phenotypes upon completion of measurements.

Description: The Veggie system on the International Space Station (ISS) intermittently supplements the crew diet with fresh, leafy green crops. For 120 days, Sustained Veggie assessed the potential of continuous on-orbit crop production. Crops grown in Veggie have been grown concurrently, but Sustained Veggie staggered plant initiation and harvest to provide more constantly available produce. The objective of this preliminary study was to compare two growth schemes to determine the methodology for required inputs, optimal yield, food safety, and crew considerations.

Description: In a microgravity setting, such as the environment aboard the International Space Station (ISS), an ideal plant water delivery system is one that can grow edible crops with minimal resource consumption and minimal risk to crew members. There are also concerns associated with the ability to control fluid escape and biofilm formation resulting in potential dangers to systems, crops, or crewmembers. To identify an appropriate system, candidate systems were assembled and operated under simulated ISS environmental conditions (T,CO2,and RH) with red romaine lettuce (Lactuca sativa cultivar 'Outredgeous') as a model crop. Fluid reservoirs and randomly selected planting sites were sampled every seven days until maturity at which point edible plant biomass and root samples were also taken. Heterotrophic bacteria and fungi growth patterns throughout each planting cycle were determined by plate counts on appropriate agar media. The candidate systems were compared to a classic hydroponics system as a control and harvested crops were compared to controls as well as Veggie-grown and market produce. Plants harvested from candidate systems yielded lower average heterotrophic bacteria and fungi per gram of plant mass levels when compared to market and Veggie samples as well as those from the control system. Additional studies to evaluate the system sanitation regimen as well as testing additional crops should be considered to aid in the selection of an ideal system.

Description: Spaceflight can dysregulate immunity, by way of increasing granulocytes numbers with impaired function. Polymorphonuclear neutrophils (PMN) are granulocytes that are first responders to infection or injury, and consist of the largest pool of immune cells in humans. PMNs function during innate immunity, through phagocytosis and promotion of inflammation, via the release of reactive oxygen species (ROS) mediators and granule-containing enzymes, such as myeloperoxidase (MPO) and NADPH oxidase-2 (NOX-2). In addition, neutrophil extracellular trap (NET) formation is another mechanism of PMN surveillance that works independently of engulfment phagocytosis, and is a last resort function that can induce NETosis or PMN-specific cell death. Previous studies in our lab have identified increased mature neutrophils, ROS and MPO production, and reduced phagocytosis in granulocytes in simulated microgravity (sug) models of hindlimb unloading (HU) in adult mice and leukocytes cultured in high-aspect rotating wall vessels (HARV-RWV). Since sug impaired phagocytosis, but improved enzymatic mediator production of MPO and redox molecules, we sought to address the third known function of PMNs, NETosis. For this, PMNs were culture in the presence or absence of the anti-oxidant N-acetyl cysteine (NAC), which rescued impaired phagocytosis that was present in sug without NAC treatment. Further, NETosis was induced in sug that was no different in the presence of NAC, suggesting NAC targets independent functions of PMNs under sug. Collectively, these results suggest modeled microgravity induced NETosis, which opens a new avenue for spaceflight studies in immune dysfunction.

Description: A combination of spaceflight-relevant factors (fluid-shift and radiation) created a different gene expression profile than either factor individually. Some gene pathways including reduced transcriptional machinery, increased neurogenesis and neuropeptide production, and dysregulated cell structure and cell signaling. Gene expression differences can persist for at least 4 months after a 21- day exposure to a combination of fluid-shift and radiation in the brain tissue of mice. Brain-related transcriptional changes are dynamic during readaptation phase from exposure to spaceflight-like conditions, which may lead to long-term neurological consequences.

Description: One of the main health risks in human space exploration is central nervous system (CNS) damage by ionizing radiation. Irradiation with simulated GCRs or their components, or high doses of low-LET radiation such as gamma rays, in animal models has been shown to cause neuronal damage together with glial cell activation and neuroinflammation and has been associated with prolonged cognitive and behavioral dysfunction. The extent of CNS damage in response to any insult, including ionizing radiation, is partially regulated by the blood-brain barrier (BBB), which enables immune cells to enter the CNS. The main cellular regulators of BBB permeability are astrocytes, which also modulate neuronal death, immune responses and oxidative stress, and thus could serve as a robust CNS-specific target for countermeasure development. However, studies on BBB permeability and astrocyte functions in regulating CNS responses to ionizing radiation have been limited, especially in human tissue/organ analogs. Therefore, we have established a high throughput 3D organ-on-a-chip system to study human CNS functions in response to ionizing radiation, with the eventual goal of adapting it to spaceflight missions. We utilized commercially available OrganoPlate system (Mimetas, Inc.) seeded with primary or induced pluripotent stem cell-derived human cells for developing 3D neuronal-astrocytic and BBB models. We investigated both immediate and delayed CNS dose responses to 0.5-1 Gy X-rays by measuring BBB permeability and morphology, and astrocyte activation. We have also quantified secreted markers of oxidative stress and cell viability. In the future, we are planning to monitor dendritic, axonal and synaptic changes in neurons, evaluate the combined exposures to simulated microgravity and ionizing radiation, and compare the responses to low and high-LET ionizing radiation. We anticipate these studies could indicate novel cellular and mechanistic targets for countermeasure developments to improve CNS functions in astronauts.

Description: High-LET ionizing radiation is a major occupational health hazard for astronauts, but risk assessment remains elusive due to limited epidemiological data. Identifying genetic factors modulating the individual radiation response may be the most effective strategy to provide individualized risk management for long-duration high-radiation missions. We have started tackling the challenge of predicting individual risks by identifying human genetic loci associated with various radiation sensitivity phenotypes in primary blood mononuclear cells from a relatively large healthy human cohort. To date, we have performed the isolation of PBMCs from 768 subjects of the same ethnicity, and irradiated PBMCs from 576 subjects with 1 and 3 particles/100m2 of 600 MeV/n 56Fe, 350 MeV/n 40Ar and 350 MeV/n 28Si ions. The phenotypes of interest were: number of radiation-induced foci (or RIFs), CellROX oxidative stress responses and cell death, at 4h and 24h following irradiation. We have observed a significant inter-individual variability at 0 Gy between the 576 studied subjects, with a mean fold difference between the 10% lowest and highest responders of 5.6 of RIFs/cell, 7.9 in mean CellRox intensity, and 9.3 in percentage of dead cells. In order to better assess genetic factors influencing DNA repair, we used a metric previously introduced by our group to sort out radiation sensitivity phenotypes in mice: i.e. the ratio of the first to the second slope of RIFs/cell (between 0 and 1, and between 1 and 3 particle/100m2). Preliminary data on 192 individuals showed a distribution of ?low-dose responders? (ratio 〉 1) to ?high-dose responders? (ratio 〈 1) at 4h of 12%, 55% and 52% respectively for Fe, Ar and Si. The average value for the first and the second slopes was very similar for the two lowest LET (0.10 [-0.26;0.58] and 0.09 [-0.45;0.41] for Ar, 0.07 [-0.27;0.38] and 0.08 [-0.19;0.42] for Si), indicating a linear dose response across both fluence. Fe showed clear saturation for the highest dose with a slope of -0.09 [-0.86;1.51] against 0.68 [-2.21;2.20] for the low dose range, which probably reflects that many PBMCs are beyond repair at the high dose. Note that other significances were found for additional factors ? such as BMI and age ? whereas none were found for sex. GWAS will be performed on all phenotypes upon completion of measurements.

Description: GeneLab must establish data processing pipelines for common data types including microarray, RNA-sequencing, and metagenomic profiling. Here we give an overview of current microarray and RNA-seq pipelines and discuss future pipelines including metagenomic profiling pipelines

Description: The NASA GeneLab project capitalizes on multi-omic technologies to maximize the return on spaceflight experiments. To do this, GeneLab maintains a publicly accessible database (GLDS) that houses spaceflight and spaceflight relevant multi-omics data, and collaborates with NASA principal investigators and projects to generate additional omics data. GeneLab houses more than 200 transcriptomic, proteomic, metabolomic and epigenomic datasets from plant, animal and microbial experiments, with a growing number of these having been produced by the GeneLab sample processing lab. The GLDS contains rich metadata about each experiment and has recently integrated radiation dosimetery data from experiments flown on the Space Shuttle. GeneLab has also recently implemented an effort to present processed data in the GLDS in addition to the raw omics data. The processed data will enable interpretation of the data by a larger group of students, scientists and the general public. Standard pipelines for the transformation of raw data into visualizations were developed by four GeneLab Analysis Working Groups (animals, plants, microbes, multi-omics) comprised of over 100 scientists from NASA and academia. These pipelines are now being used by a group of bioinformatics interns to provide standard basic analysis of the data for incorporation into GLDS.

Description: Spaceflight can cause immune system dysfunction, such as elevated white blood cells (WBC) and polymorphonuclear neutrophils (PMN), along with unchanged or reduced lymphocyte counts. A high PMN to lymphocyte ratio (NLR) can acts as a poor prognosis in cancer and a biomarker for subclinical inflammation however, the NLR has not been identified as a predictor of astronaut health during spaceflight. CBC data collected on board the International Space Station (ISS) was repurposed to determine the granulocyte to lymphocyte ratio (GLR) in humans and the NLR in rodents. The results displayed a progressive increase in GLR and NLR during spaceflight and at landing. The mechanism for increased NLR was assessed in vitro using the microgravity-analog, rotating wall vessel (RWV), with human WBCs. The results indicated that simulated microgravity led to increased GLR and NLR profiles, and production of reactive oxygen species (ROS) and myeloperoxidase (MPO). Interestingly, simulated microgravity increased the number of matured PMNs that showed impaired phagocytic function, while treatment with tert-Butyl hydroperoxide (TBHP), also reduced PMN phagocytosis. In addition, 30-days of simulated microgravity (hindlimb unloading) in mice, indicated an increased NLR and MPO gene expression, which were mitigated in mitochondrial catalase overexpressing transgenic mice, suggesting ROS scavenging is essential for maintaining homeostatic immunity. Collectively, we propose that the health status of astronauts during future short- and long-term space missions can be monitored by their NLR profile, in addition to utilizing this measurement as a tool for oxidative stress response countermeasure development to restore homeostatic immunity.

Description: Almost all modern proteins possess well-defined, relatively rigid scaffolds that provide structural preorganization for desired functions. Such scaffolds require the sufficient length of a polypeptide chain and extensive evolutionary optimization. How ancestral proteins attained functionality, even though they were most likely markedly smaller than their contemporary descendants, remains a major, unresolved question in the origin of life. On the basis of evidence from experiments and computer simulations, we argue that at least some of the earliest water-soluble and membrane proteins were markedly more flexible than their modern counterparts. As an example, we consider a small, evolved in vitro ligase, based on a novel architecture that may be the archetype of primordial enzymes. The protein does not contain a hydrophobic core or conventional elements of the secondary structure characteristic of modern water-soluble proteins, but instead is built of a flexible, catalytic loop supported by a small hydrophilic core containing zinc atoms. It appears that disorder in the polypeptide chain imparts robustness to mutations in the protein core. Simple ion channels, likely the earliest membrane protein assemblies, could also be quite flexible, but still retain their functionality, again in contrast to their modern descendants. This is demonstrated in the example of antiamoebin, which can serve as a useful model of small peptides forming ancestral ion channels. Common features of the earliest, functional protein architectures discussed here include not only their flexibility, but also a low level of evolutionary optimization and heterogeneity in amino acid composition and, possibly, the type of peptide bonds in the protein backbone.

Description: The National Aeronautics and Space Administration Animal Enclosure Module (AEM) was developed as a self-contained rodent habitat for shuttle flight missions that provides inhabitants with living space, food, water, ventilation, and lighting for shuttle flight missions, and this study reports whether, after minimal hardware modification, the AEM could support an extended term up to 35 days for Sprague-Dawley rats and C57BL/6 female mice for use on the International Space Station. Success was evaluated based on comparison of AEM housed animals to that of vivarium housed and to normal biological ranges through various measures of animal health and well-being, including animal health evaluations, animal growth and body masses, organ masses, rodent food bar consumption, water consumption, and analysis of blood contents. The results of this study confirmed that the AEMs could support 12 adult female C57BL/6 mice for up to 35 days with self-contained RFB and water, and the AEMs could also support 5 adult male Sprague-Dawley rats for 35 days with external replenishment of diet and water. This study has demonstrated the capability and flexibility of the AEM to operate for up to 35 days with minor hardware modification. Therefore, with modifications, it is possible to utilize this hardware on the International Space Station or other operational platforms to extend the space life science research use of mice and rats.

Description: Neurolab, the final Spacelab mission, launched on STS-90 on April 17, 1998, was dedicated to studying the nervous system. NASA cooperated with domestic and international partners to conduct the mission. ARC's (Ames Research Center's) Payload included 15 experiments designed to study the adaptation and development of the nervous system in microgravity. The payload had the largest number of Principal and Co-Investigators, largest complement of habitats and experiment unique equipment flown to date, and most diverse distribution of live specimens ever undertaken by ARC, including rodents, toadfish, swordtail fish, water snails, hornweed and crickets To facilitate tissue sharing and optimization of science objectives, investigators were grouped into four science discipline teams: Neuronal Plasticity, Mammalian Development, Aquatic, and Neurobiology. Several payload development challenges were experienced and required an extraordinary effort, by all involved, to meet the launch schedule. With respect to hardware and the total amount of recovered science, Neurolab was regarded as an overall success. However, a high mortality rate in one rodent group and several hardware anomalies occurred inflight that warranted postflight investigations. Hardware, science, and operations lessons were learned that should be taken into consideration by payload teams developing payloads for future Shuttle missions and the International Space Station.

Description: No abstract available

Description: In this innovation project, we investigated the usage of narrow band violet light (408 nm) to attenuate the growth of bacteria typically found on ISS. Violet light is less hazardous than UV light and it can transmit through plastics, such as clear acrylics, making it possible to incorporate into large surface lamps and acrylic or polycarbonate based optical light guides. This study built a custom LED surface panel that was edge lit by an array of 408 nm violet LEDS. The optical light guide technology used in the lamp and the 408 nm violet LEDs are available on the market from multiple vendors. The application of the concept of using violet light driven LED panels and optical light guides is to integrate the paneling into spacecraft architectural surfaces for the automation of a light based microbial countermeasure that could enhance current cleaning methods used in areas on spacecraft prone to microbial growth.

Description: No abstract available

Description: The skeleton interacts with its environment in a way that resembles a mechanostat - through a controlled process of bone remodeling, namely local formation and resorption, to maintain a healthy structure. During weightlessness, astronauts lose structure in weight-bearing bones due to decreased formation by osteoblasts and increased resorption by osteoclasts. In contrast, increased mechanical loading through exercise targets bone remodeling to remove and repair microdamage, improving structural integrity. In fact, recent advances in astronaut exercise regimens have prevented the deleterious changes in skeletal structure during spaceflight. However, knowledge of the molecular underpinnings of the skeletal response to spaceflight and to mechanical stimulation is limited. We propose that epigenetic modification, specifically DNA methylation, may influence osteoblast differentiation and activity during spaceflight and exercise. We hypothesize that simulated weightlessness hypermethylates pro-osteoblastogenic gene promoters and decreases expression of osteoblastogenic genes. Oppositely, we hypothesize that mechanical loading hypomethylates pro-osteoblastogenic gene promoters and increases expression of osteoblastogenic genes.

Description: ISS crew health research indicate risk for immune system degradation for long duration missions, necessitating the need to maximize countermeasures on microbial growth (1). This project will investigate applications using violet (400nm) LEDs to mitigate growth of bacteria on architectural surfaces. The application is primarily for architectures and surfaces where a higher amount of bacterial growth is expected. The project will build two different types of violet light sources that represent lamp types that could be installed in spacecraft applications. Under a controlled setting, using violet LEDs, the evaluation will test the effectiveness of a glowing work surface using LED Panel verses an LED array overhead lamp. The effectiveness of the lighting systems to attenuate bacterial growth will be compared to a control. The results of this study can be used to inform spacecraft system architects on novel ways to improve habitat health, and reduce reliance on manual cleaning methods.

Description: No abstract available

Description: No abstract available

Description: W and c Any air borne vehicle needs incorporating safety as key parameter of measure, and inclusion of autonomy raises the critical need for safety under autonomous operations. Management of faults and component degradation is key as complexity in autonomous operations grow over the period of time. Therefore, in addition to basic operational requirements, an autonomous electric vehicle should be able to make accurate estimates of its current system health and take the correct decisions to complete its mission successfully. Real-time safety and state-awareness tools are therefore essential for the vehicle to be able to reach its destination in a safe and successful manner. The need for safety assurance and health management capabilities is particularly relevant for aircraft electric propulsion systems, which are relatively new and with limited historical to learn. They are critical systems requiring high power density along with reliability, resilience, efficient management of weight, and operational costs. A model- based fault diagnosis and prognostics approach of complex critical systems can successfully accomplish the safety and state awareness goal for such electric propulsion systems, enabling autonomous decision making capability for safe and efficient operation. To identify critical components in the system a Qualitative Bayesian approach using FMECA is implemented. This requires the assessment of some quantities representing the state of the electric unmanned aerial systems (e-UAS), as well as look-ahead forecasts of such states during the entire flight, presented in form of safety metrics (SM). In-service data and performance data gathered from degraded components sup- ports diagnostic and prognostic methods for these systems, but this data can be difficult to obtain as weight and packaging restrictions reduce redundancy and instrumentation on-board the vehicle. Therefore, an model-based framework should be capable or operating with limited data. In addition to data scarcity, the variability of such complex critical systems re- quires the model-based framework to reason in the presence of uncertainty, such as sensor noise, and modeling imperfections. Quantification of errors and uncertainties in the measured states and quantities is therefore a fundamental step for a precise estimation of such SMs; un-modeled uncertainty may result in erroneous state assessment and un- reliable predictions of future states of e-UAVs. Typical, centralized model-based schemes suffer from inherent disadvantages such as computational complexity, single point of failure, and scalability issues, and therefore may fail in such a complex scenario. This paper presents a methodology for developing a system level diagnostics and prognostics approach using a Qualitative Bayesian FMECA approach along with a formal uncertainty management framework for an e-UAS. In this work we demonstrate the efficacy of the framework to predict effects of sub-system level degradation on vehicle operation incorporating uncertainty management to predict future behavior under different operating conditions.

Description: Terrestrial ecosystems contribute most of the interannual variability (IAV) in atmospheric carbon dioxide (CO2) concentrations, but processes driving the IAV of net ecosystem CO2 exchange (NEE) remain elusive. For a predictive understanding of the global C cycle, it is imperative to identify indicators associated with ecological processes that determine the IAV of NEE. Here, we decompose the annual NEE of global terrestrial ecosystems into their phenological and physiological components, namely maximum carbon uptake (MCU) and release (MCR), the carbon uptake period (CUP), and two parameters, and , that describe the ratio between actual versus hypothetical maximum C sink and source, respectively. Using longterm observed NEE from 66 eddy covariance sites and global products derived from FLUXNET observations, we found that the IAV of NEE is determined predominately by MCU at the global scale, which explains 48% of the IAV of NEE on average while , CUP, , and MCR explain 14%, 25%, 2%, and 8%, respectively. These patterns differ in waterlimited ecosystems versus temperature and radiationlimited ecosystems; 31% of the IAV of NEE is determined by the IAV of CUP in waterlimited ecosystems, and 60% of the IAV of NEE is determined by the IAV of MCU in temperature and radiationlimited ecosystems. The LundPotsdamJena (LPJ) model and the Multiscale Synthesis and Terrestrial Model Intercomparison Project (MsTMIP) models underestimate the contribution of MCU to the IAV of NEE by about 18% on average, and overestimate the contribution of CUP by about 25%. This study provides a new perspective on the proximate causes of the IAV of NEE, which suggest that capturing the variability of MCU is critical for modeling the IAV of NEE across most of the global land surface.

Description: Molecular biosignatures are key targets for current, proposed, and future life detection missions. With the high accuracy and low limit of detection (LOD) that new and future instruments will require, decontamination of life detection hardware is necessary to prevent false positives. Lipids are a molecular biosignature of interest, as they are ubiquitous to all life as we know it, can survive unaltered in the geologic record for longer than any other biomolecule (i.e. billions of years), and form through both biotic and abiotic processes. Lipids display origin-diagnostic molecular patterns that can reveal biotic or abiotic synthesis, so finding them and ascertaining their molecular features is important for potentially detecting evidence of life elsewhere. Traditional methods of decontamination, or contamination control (CC), primarily clean hardware through fabrication in sterile (cleanroom) environments, killing microbes, and removing/flushing contaminants off instrument and spacecraft components. However, research suggests that some standard cleaning methods are either unlikely to remove lipid contaminants or are incompatible with life detection instrument materials. To solve this problem, I propose to find, test, and verify a decontamination method that thoroughly cleans instruments by destroying lipid molecules, but is simultaneously compatible with major materials used in these instruments. I will study the effects of traditional CC methods (including Dry Heat Microbial Reduction and Vapor phase Hydrogen Peroxide) and experimental CC methods (Electron Beam Irradiation) on lipid molecules for application to life detection instrumentation. I will then develop a CC plan for a novel lipid detector (ExCALiBR, Extractor for Chemical Analysis of Lipid Biomarkers in Regolith) searching for lipids in either soil or icy world scenarios. This plan will uphold planetary protection regulation requirements and validate experimental analyses of in-situ life detection tests.

Description: No abstract available

Description: Traces of life are nearly ubiquitous on Earth. However, a central unresolved question is whether these traces always indicate an active microbial community or whether, in extreme environments, such as hyperarid deserts, they instead reflect just dormant or dead cells. Although microbial biomass and diversity decrease with increasing aridity in the Atacama Desert, we provide multiple lines of evidence for the presence of an at times metabolically active, microbial community in one of the driest places on Earth. We base this observation on four major lines of evidence: (i) a physico-chemical characterization of the soil habitability after an exceptional rain event, (ii) identified biomolecules indicative of potentially active cells [e.g., presence of ATP, phospholipid fatty acids (PLFAs), metabolites, and enzymatic activity], (iii) measurements of in situ replication rates of genomes of uncultivated bacteria reconstructed from selected samples, and (iv) microbial community patterns specific to soil parameters and depths. We infer that the microbial populations have undergone selection and adaptation in response to their specific soil microenvironment and in particular to the degree of aridity. Collectively, our results highlight that even the hyperarid Atacama Desert can provide a habitable environment for microorganisms that allows them to become metabolically active following an episodic increase in moisture and that once it decreases, so does the activity of the microbiota. These results have implications for the prospect of life on other planets such as Mars, which has transitioned from an earlier wetter environment to todays extreme hyperaridity.

Description: This presentation will cover the the basic pipelines for transcriptomics and proteomics that the GeneLab Analysis Working Groups (AWGs) have so far determined to be optimal. Basic transcriptomic pipelines will first be presented from primary analysis to higher-order systems analysis. Examples of how the data has been analyzed will be presented. Proteomics pipelines will also be presented compiled from various AWG members. Discussion will be generated from the AWG members to reach a consensus for each omic type.

Description: This presentation is a summary of the continuing effort to determine options for studying artificial gravity with rodents. Results of an engineering trade study are presented and an overview of past and planned short radius centrifugation studies are presented. A leading proposal for a future flight centrifuge capable of housing rodents, the Techshot RCF, is presented in only enough detail as is approved by Techshot for public domain use.

Description: No abstract available

Description: The First Biennial Space Biomedical Investigators' Workshop, held January 11-13, 1999, was unique in that it assembled, for the first time, a broad cross section of NASA-funded biomedical researchers to present the current status of their projects and their plans for future investigations. All principal investigators with active, or recently-completed ground-based projects in NASA's Biomedical Research and Countermeasures Program that were funded through NASA's Office of Life and Microgravity Sciences and Applications were invited. Included were individual investigators funded through NASA Research Announcements, investigators with NASA Specialized Centers of Research and Training, investigators with the recently established National Space Biomedical Research Institute (NSBRI), and NASA civil servant investigators. Seventy-seven percent of all eligible projects were presented at the workshop. Thus, these Proceedings should provide a useful snapshot of the status of NASA-funded space biomedical research as of January 1999. An important workshop objective was to achieve free and open communication among the presenting investigators. Therefore, presentation of new and incomplete results, as well as hypotheses and ideas for future research, was encouraged. Comments and constructive criticisms from the presenters' colleagues were also encouraged. These ground rules resulted in many lively and useful discussions, during both the presentation sessions and informal evening gatherings and breaks.

Description: As NASA's effort to establish a permanent residence in space continues, research on the effects of microgravity onbiological microorganisms is vital to protect or promote the health of plants and their astronaut counterparts. The purpose of this study is to determine the effects of microgravity on Pantoea agglomerans (P. agglomerans), using an analog microgravity simulator; the Rotary Cell Culture System (RCCS) developed at Johnson Space Center (JSC) in Houston, TX. P. agglomerans has been shown to be a plant growth promoter (PGPR) in ground based studies, but has also been shown to be a pathogen in both plants and immunocompromised patients. In this study, we will determine changes in the growth rate and antibiotic susceptibility of P. agglomerans when exposed to simulated microgravity.

Description: A major stressor in the space environment is microgravity. Microgravity has profound effects on biological processes that are vital to normal functioning. This is most prevalently seen in microorganisms, which have altered growth rates and increased antibiotic susceptibility in microgravity. This is a concern for both astronauts and plants onboard spacecraft. Pantoea agglomerans is a soil bacterium that has been shown to be a plant growth promoter, plant pathogen, and an opportunistic pathogen to immunocompromised patients. Using the ground based microgravity analog, the Rotary Cell Culture System along with the High Aspect-Ratio Vessel, we analyzed the growth and the antibiotic susceptibility of Pantoea agglomerans grown in simulated microgravity. In certain parameters, we discovered an increased growth rate and no change in the antibiotic susceptibility. We found that there were differences in results when certain aspects of the protocol were altered. Further work will need to be conducted to get a better understanding of the changes in the microorganisms exposed to microgravity.

Description: NASA GeneLab is an open-access repository for omics datasets generated by biological experiments conducted in space or ground experiments relevant to spaceflight (e.g. simulated cosmic radiation, simulated microgravity, bed rest studies). The GeneLab Data Systems (GLDS) version 4.0 will be available on October 1st 2019, and will provide a state-of-the-art bioinformatics platform for the space biology and radiation communities to upload their data into an omics data commons, to process their data with vetted standard workflows and to compare with existing analyses. Started in 2015 as a repository designed to archive omics data from space experiments, GeneLab has expanded its scope to all ionizing radiation omics experiments conducted on the ground and has put considerable effort in providing carefully characterized radiation metadata on all datasets. GeneLab is also providing processed data derived from the raw data covering a large spectrum of omics (genome, epigenome, transcriptome, epitranscriptome, proteome, metabolome) to help users explore important questions: 1) Which genes or proteins are expressed differently in space for various living organisms? 2) What specific DNA mutations or epigenetic changes happen in space or after exposure to ionizing radiation? and 3) How does genetics affect these responses? Processed data available on GeneLab are derived by standard data analysis workflows vetted by hundreds of scientists who volunteered to join one of the four GeneLab Analysis Working Groups (Animal AWG, Plant AWG, Microbe AWG, Multi-Omics AWG). In this presentation, we will discuss how to bridge the gap between irradiation studies performed on earth and biological experiments conducted in space since the early 1990's. We will discuss how radiation dosimetry was estimated for datasets derived from samples collected during the Space Shuttle era on the International Space Station and on other orbiting platforms. Finally, we will address future strategies regarding dose monitoring in future missions into space, inter-agency efforts to unify data under one umbrella, and knowledge dissemination across the radiation research community and the space biology community.

Description: The many known health risks currently associated with space travel include increased risk of cardiovascular disease, cancer, central nervous system related diseases, muscle degeneration, and changes with host-gut microbiome interactions that can have profound impact with these and other health risks. The majority of the risk from space travel stem of the two components of the space environment which are microgravity and radiation. Two specific systemic effects have been uncovered by us to impact the body as a whole due to the space environment. One factor is related from our earlier work (Beheshti et al, PLOS One, 2018), we predicted that there is a systemic component of the host that causes general increased health risks due to spaceflight driven by a circulating microRNA (miRNA) signature consisting of 13 miRNAs that directly regulates both p53 and TGF1. MiRNAs are small non-coding RNA molecules with a negative and post-transcriptional regulation on gene expression) are increasingly recognized as major systemic regulators of responses to stressors, including microgravity, oxidative stress, and DNA damage. In addition, due to the size and stability of miRNAs, it is known that miRNAs can circulate throughout the body and have been found in the majority of the bodily fluids including blood, urine, saliva, and tears. Here, we start to dissect the actual impact of this miRNA signature on both the radiation and microgravity components and prove that this miRNA signature actually exists in the circulation of a host. The other systemic factor we uncovered was the impact the mitochondria on the whole body due to spaceflight. We hypothesize that spaceflight may promote a physiologic response driven by systemic mitochondria pathways leading to metabolic disorder stemming from the liver and directly impacting other organs and tissues. A systems biology method was implemented utilizing GeneLab datasets that involved in vitro experiments performed at the low Earth orbit, in vivo experiments involving mice flown to space, and finally human physiological data from astronauts. A comprehensive multi-omics approach was implemented which involved correlating transcriptomic analysis with proteomics, metabolomics, and methylation analysis. This approach led us to confirm our hypothesis that a systemic mitochondrial driven response is responsible for increasing potential health risk and is conserved from the in vitro studies, to the in vivo studies, and finally confirmed in astronauts.

Description: No abstract available

Description: No abstract available

Description: A method of measuring motion blur is disclosed comprising obtaining a moving edge temporal profile r(sub 1)(k) of an image of a high-contrast moving edge, calculating the masked local contrast m(sub1)(k) for r(sub 1)(k) and the masked local contrast m(sub 2)(k) for an ideal step edge waveform r(sub 2)(k) with the same amplitude as r(sub 1)(k), and calculating the measure or motion blur Psi as a difference function, The masked local contrasts are calculated using a set of convolution kernels scaled to simulate the performance of the human visual system, and Psi is measured in units of just-noticeable differences.

Description: A method for fabricating a micro-organ device comprises providing a microscale support having one or more microfluidic channels and one or more micro-chambers for housing a micro-organ and printing a micro-organ on the microscale support using a cell suspension in a syringe controlled by a computer-aided tissue engineering system, wherein the cell suspension comprises cells suspended in a solution containing a material that functions as a three-dimensional scaffold. The printing is performed with the computer-aided tissue engineering system according to a particular pattern. The micro-organ device comprises at least one micro-chamber each housing a micro-organ; and at least one microfluidic channel connected to the micro-chamber, wherein the micro-organ comprises cells arranged in a configuration that includes microscale spacing between portions of the cells to facilitate diffusion exchange between the cells and a medium supplied from the at least one microfluidic channel.

Description: A system detects a plant's exposure to trichloroethylene (TCE) through plant leaf imaging. White light impinging upon a plant's leaf interacts therewith to produce interacted light. A detector is positioned to detect at least one spectral band of the interacted light. A processor coupled to the detector performs comparisons between photonic energy of the interacted light at the one or more spectral bands thereof and reference data defining spectral responses indicative of leaf exposure to TCE. An output device coupled to the processor provides indications of the comparisons.

Description: A method for fabricating a micro-organ device comprises providing a microscale support having one or more microfluidic channels and one or more micro-chambers for housing a micro-organ and printing a micro-organ on the microscale support using a cell suspension in a syringe controlled by a computer-aided tissue engineering system, wherein the cell suspension comprises cells suspended in a solution containing a material that functions as a three-dimensional scaffold. The printing is performed with the computer-aided tissue engineering system according to a particular pattern. The micro-organ device comprises at least one micro-chamber each housing a micro-organ; and at least one microfluidic channel connected to the micro-chamber, wherein the micro-organ comprises cells arranged in a configuration that includes microscale spacing between portions of the cells to facilitate diffusion exchange between the cells and a medium supplied from the at least one microfluidic channel.

Description: Three-dimensional human broncho-epithelial tissue-like assemblies (TLAs) are produced in a rotating wall vessel (RWV) with microcarriers by coculturing mesenchymal bronchial-tracheal cells (BTC) and bronchial epithelium cells (BEC). These TLAs display structural characteristics and express markers of in vivo respiratory epithelia. TLAs are useful for screening compounds active in lung tissues such as antiviral compounds, cystic fibrosis treatments, allergens, and cytotoxic compounds.

Description: Systems and methods for the use of compounds from the Hofmeister series coupled with specific pH and temperature to provide rapid physico-chemical-managed killing of penicillin-resistant static and growing Gram-positive and Gram-negative vegetative bacteria. The systems and methods represent the more general physico-chemical enhancement of susceptibility for a wide range of pathological macromolecular targets to clinical management by establishing the reactivity of those targets to topically applied drugs or anti-toxins.

Description: The present disclosure provides ex vivo-derived mineralized three-dimensional bone constructs. The bone constructs are obtained by culturing osteoblasts and osteoclast precursors under randomized gravity vector conditions. Preferably, the randomized gravity vector conditions are obtained using a low shear stress rotating bioreactor, such as a High Aspect Ratio Vessel (HARV) culture system. The bone constructs of the disclosure have utility in physiological studies of bone formation and bone function, in drug discovery, and in orthopedics.

Description: A method for the production of functional proteins including hormones by renal cells in a three dimensional culturing process responsive to shear stress uses a rotating wall vessel. Natural mixture of renal cells expresses the enzyme 1-.alpha.-hydroxylase which can be used to generate the active form of vitamin D: 1,25-diOH vitamin D.sub.3. The fibroblast cultures and co-culture of renal cortical cells express the gene for erythropoietin and secrete erythropoietin into the culture supernatant. Other shear stress response genes are also modulated by shear stress, such as toxin receptors megalin and cubulin (gp280). Also provided is a method of treating an in-need individual with the functional proteins produced in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel.

Description: The present invention provides chaperonin polypeptides which are modified to include N-terminal and C-terminal ends that are relocated from the central pore region to various different positions in the polypeptide which are located on the exterior of the folded modified chaperonin polypeptide. In the modified chaperonin polypeptide, the naturally-occurring N-terminal and C-terminal ends are joined together directly or with an intervening linker peptide sequence. The relocated N-terminal or C-terminal ends can be covalently joined to, or bound with another molecule such as a nucleic acid molecule, a lipid, a carbohydrate, a second polypeptide, or a nanoparticle. The modified chaperonin polypeptides can assemble into double-ringed chaperonin structures. Further, the chaperonin structures can organize into higher order structures such as nanofilaments or nanoarrays which can be used to produce nanodevices and nanocoatings.

Description: Topics include: Electronic Components and Circuits. Electronic Systems, Physical Sciences, Materials, Computer Programs, Mechanics, Machinery, Fabrication Technology, Mathematics and Information Sciences, and Life Sciences

Description: Cardiac electrical data are received from a patient, manipulated to determine various useful aspects of the ECG signal, and displayed and stored in a useful form using a computer. The computer monitor displays various useful information, and in particular graphically displays various permutations of reduced amplitude zones and kurtosis that increase the rapidity and accuracy of cardiac diagnoses. New criteria for reduced amplitude zones are defined that enhance the sensitivity and specificity for detecting cardiac abnormalities.

Description: Microcapsules prepared by encapsulating an aqueous solution of a protein, drug or other bioactive substance inside a semi-permeable membrane by are disclosed. The microcapsules are formed by interfacial coacervation under conditions where the shear forces are limited to 0-100 dynes/cm.sup.2 at the interface. By placing the microcapsules in a high osmotic dewatering solution, the protein solution is gradually made saturated and then supersaturated, and the controlled nucleation and crystallization of the protein is achieved. The crystal-filled microcapsules prepared by this method can be conveniently harvested and stored while keeping the encapsulated crystals in essentially pristine condition due to the rugged, protective membrane. Because the membrane components themselves are x-ray transparent, large crystal-containing microcapsules can be individually selected, mounted in x-ray capillary tubes and subjected to high energy x-ray diffraction studies to determine the 3-D structure of the protein molecules. Certain embodiments of the microcapsules of the invention have composite polymeric outer membranes which are somewhat elastic, water insoluble, permeable only to water, salts, and low molecular weight molecules and are structurally stable in fluid shear forces typically encountered in the human vascular system.

Description: Session TA2 includes short reports covering: (1) The Interaction of Microgravity and Ethylene on Soybean Growth and Metabolism; (2) Structure and G-Sensitivity of Root Statocytes under Different Mass Acceleration; (3) Extracellular Production of Taxanes on Cell Surfaces in Simulated Microgravity and Hypergravity; (4) Current Problems of Space Cell Phytobiology; (5) Biological Consequences of Microgravity-Induced Alterations in Water Metabolism of Plant Cells; (6) Localization of Calcium Ions in Chlorella Cells Under Clinorotation; (7) Changes of Fatty Acids Content of Plant Cell Plasma Membranes under Altered Gravity; (8) Simulation of Gravity by Non-Symmetrical Vibrations and Ultrasound; and (9) Response to Simulated weightlessness of In Vitro Cultures of Differentiated Epithelial Follicular Cells from Thyroid.

Description: Systems, methods and apparatus are provided through which in some embodiments an autonomic entity manages a system by generating one or more stay alive signals based on the functioning status and operating state of the system. In some embodiments, an evolvable synthetic neural system is operably coupled to one or more evolvable synthetic neural systems in a hierarchy. The evolvable neural interface receives and generates heartbeat monitor signals and pulse monitor signals that are used to generate a stay alive signal that is used to manage the operations of the synthetic neural system. In another embodiment an asynchronous Alice signal (Autonomic license) requiring valid credentials of an anonymous autonomous agent is initiated. An unsatisfactory Alice exchange may lead to self-destruction of the anonymous autonomous agent for self-protection.

Description: A miniature microwave antenna is disclosed which may be utilized for biomedical applications such as, for example, radiation induced hyperthermia through catheter systems. One feature of the antenna is that it possesses azimuthal directionality despite its small size. This directionality permits targeting of certain tissues while limiting thermal exposure of adjacent tissue. One embodiment has an outer diameter of about 0.095'' (2.4 mm) but the design permits for smaller diameters.

Description: The present invention provides a method for production of functional proteins including hormones by renal cells in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel. Natural mixture of renal cells expresses the enzyme 1-a-hydroxylase which can be used to generate the active form of vitamin D: 1,25-diOH vitamin D3. The fibroblast cultures and co-culture of renal cortical cells express the gene for erythropoietin and secrete erythropoietin into the culture supernatant. Other shear stress response genes are also modulated by shear stress, such as toxin receptors megalin and cubulin (gp280). Also provided is a method of treating in-need individual with the functional proteins produced in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel.

Description: A method, simulation, and apparatus are provided that are highly suitable for treatment of benign prostatic hyperplasia (BPH). A catheter is disclosed that includes a small diameter disk loaded monopole antenna surrounded by fusion material having a high heat of fusion and a melting point preferably at or near body temperature. Microwaves from the antenna heat prostatic tissue to promote necrosing of the prostatic tissue that relieves the pressure of the prostatic tissue against the urethra as the body reabsorbs the necrosed or dead tissue. The fusion material keeps the urethra cool by means of the heat of fusion of the fusion material. This prevents damage to the urethra while the prostatic tissue is necrosed. A computer simulation is provided that can be used to predict the resulting temperature profile produced in the prostatic tissue. By changing the various control features of the catheter and method of applying microwave energy a temperature profile can be predicted and produced that is similar to the temperature profile desired for the particular patient.

Description: The present invention relates to telemetry-based sensing systems that continuously measures physical, chemical and biological parameters. More specifically, these sensing systems comprise a small, modular, low-power implantable biotelemetry system capable of continuously sensing physiological characteristics using implantable transmitters, a receiver, and a data acquisition system to analyze and record the transmitted signal over several months. The preferred embodiment is a preterm labor and fetal monitoring system. Key features of the invention include Pulse Interval Modulation (PIM) that is used to send temperature and pressure information out of the biological environment. The RF carrier frequency is 174-216 MHz and a pair of RF bursts (pulses) is transmitted at a frequency of about 1-2 Hz. The transmission range is 3 to 10 feet, depending on the position of the transmitter in the body and its biological environment. The entire transmitter is encapsulated in biocompatible silicone rubber. Power is supplied by on-board silver-oxide batteries. The average power consumption of the current design is less than 30 .mu.W., which yields a lifetime of approximately 6-9 months. Chip-on-Board technology (COB) drastically reduces the size of the printed circuit board from 38.times.28 mm to 22.times.8 mm. Unpackaged dies are flip-chip bonded directly onto the printed circuit board, along with surface mount resistors and capacitors. The invention can monitor additional physiological parameters including, but not limited to, ECG, blood gases, glucose, and ions such as calcium, potassium, and sodium.

Description: A method is presented for determining diastolic intracranial pressure (ICP) in a patient. A first change in the length of a path across the skull of the patient caused by a known change in ICP is measured and used to determine an elasticity constant for the patient. Next, a second change in the length of the path across the patient's skull occurring between systolic and diastolic portions of the patient's heartbeat is measured. The patient's diastolic ICP is a function of the elasticity constant and the second change.

Description: A fetal heart monitoring system preferably comprising a backing plate having a generally concave front surface and a generally convex back surface, and at least one sensor element attached to the concave front surface for acquiring acoustic fetal heart signals produced by a fetus within a body. The sensor element has a shape that conforms to the generally concave back surface of the backing plate. In one embodiment, the at least one sensor element comprises an inner sensor, and a plurality of outer sensors surrounding the inner sensor. The fetal heart monitoring system can further comprise a web belt, and a web belt guide movably attached to the web belt. The web belt guide being is to the convex back surface of the backing plate.

Description: A system for and method of detecting and measuring concentrations of an ultrasonically-reflective microsphere contrast agent involving detecting non-linear sum and difference beat frequencies produced by the microspheres when two impinging signals with non-identical frequencies are combined by mixing. These beat frequencies can be used for a variety of applications such as detecting the presence of and measuring the flow rates of biological fluids and industrial liquids, including determining the concentration level of microspheres in the myocardium.

Description: In accordance with the present invention, biologically active protein fragments can be constructed which contain only those specific portions of the serum albumin family of proteins such as regions known as subdomains IIA and IIIA which are primarily responsible for the binding properties of the serum albumins. The artificial serums that can be prepared from these biologically active protein fragments are advantageous in that they can be produced much more easily than serums containing the whole albumin, yet still retain all or most of the original binding potential of the full albumin proteins. In addition, since the protein fragment serums of the present invention can be made from non-natural sources using conventional recombinant DNA techniques, they are far safer than serums containing natural albumin because they do not carry the potentially harmful viruses and other contaminants that will be found in the natural substances.

Description: A rotary pump for pumping fluids through a patient having a housing with an internal region, a stator member and an impeller positioned within the housing and having impeller blades, wherein the impeller is magnetically suspended and rotated, and wherein the geometric configuration of the rotary pump is sized and proportioned to minimize stagnant and traumatic fluid flow within the rotary pump. The plurality of magnetic impeller blades are preferably rare earth, high-energy-density magnets selected from the group consisting of samarium cobalt and neodymium-iron-boron alloy.

Description: A tissue simulating gel and a method for preparing the tissue simulating gel are disclosed. The tissue simulating gel is prepared by a process using water, gelatin, ethylene glycol, and a cross-linking agent. In order to closely approximate the characteristics of the type of tissue being simulated, other material has been added to change the electrical, sound conducting, and wave scattering properties of the tissue simulating gel. The result of the entire process is a formulation that will not melt at the elevated temperatures involved in hyperthermia medical research. Furthermore, the tissue simulating gel will not support mold or bacterial growth, is of a sufficient mechanical strength to maintain a desired shape without a supporting shell, and is non-hardening and non-drying. Substances have been injected into the tissue simulating gel prior to the setting-up thereof just as they could be injected into actual tissue, and the tissue simulating gel is translucent so as to permit visual inspection of its interior. A polyurethane spray often used for coating circuit boards can be applied to the surface of the tissue simulating gel to give a texture similar to human skin, making the tissue simulating gel easier to handle and contributing to its longevity.

Description: An apparatus for acquiring signals emitted by a fetus, identifying fetal heart beats and determining a fetal heart rate. Multiple sensor signals are outputted by a passive fetal heart rate monitoring sensor. Multiple parallel nonlinear filters filter these multiple sensor signals to identify fetal heart beats in the signal data. A processor determines a fetal heart rate based on these identified fetal heart beats. The processor includes the use of a figure of merit weighting of heart rate estimates based on the identified heart beats from each filter for each signal. The fetal heart rate thus determined is outputted to a display, storage, or communications channel. A method for enhanced fetal heart beat discrimination includes acquiring signals from a fetus, identifying fetal heart beats from the signals by multiple parallel nonlinear filtering, and determining a fetal heart rate based on the identified fetal heart beats. A figure of merit operation in this method provides for weighting a plurality of fetal heart rate estimates based on the identified fetal heart beats and selecting the highest ranking fetal heart rate estimate.

Description: This chapter intends to lay out some essential ethics questions that might frame the next step of space exploration. In this, I undertake two sorts of tasks. The first is to respond to the core ethic question: Is it ethical to travel in space? The second, assuming for the moment that I can convince you that the first premise can be justified, is to comment on what ethical challenges will face us there. It is appropriate to have a philosopher comment on this at the fortieth anniversary celebration, since it was also in 1962 that the National Academy of Science first convened a panel of philosophers to comment on space travel. They asked at that time whether it was indeed a worthwhile pursuit to travel in space and what might be expected of such an effort. What is at stake in any such boundary crossing is how the changing of essential human perimeters changes our own moral status. Will such boundary crossing worsen our human condition, or will it enhance it? In this way, the geopolitical quest is then linked to the quest for ontology, Pisarro hunting for the fountain of youth, for gold, and for territory. What follows are a series of ethical claims that link the problem of discovery in the larger world and the attendant ethical dilemmas of our explorations, as well as how this exploration alters our concepts of life on Earth. In this, the role of the ethicist is to function as both a skeptic and a stranger, aware of the optimism of science and the pessimism of philosophy.

Description: Dr. France Cordova, NASA's Chief Scientist, opened this, the sixth seminar in the Administrator's Seminar Series, by introducing NASA Administrator Daniel S. Goldin. Mr Goldin welcomed the attendees and set the stage for Dr. Cordova's introduction of the first speaker, Dr. Robert Kates of Brown University. Dr. Kates primary concerns are global environmental changes, world hunger, and the size of the population. Human changes, he said, rival the changes of nature. Changes in the size of world population affect the need for more agricultural products, therefore more land for growing food, which leads to deforestation, which affects rainfall, and therefore the water supply which is in increased demand. Human ingenuity can reduce some shortages but generally doesn't keep up with increased demand for life-sustaining essentials. These problems require the concern of intergovernmental organizations, treaties and activities, as well as transnational corporations, and non-governmental and private, volunteer organizations. Next Dr. Diana Liverman of Pennsylvania State University spoke on human interactions regarding climate and society. She considered the effect of changes in land use on climate, using Mexico as an example. Mexicans changed from raising much wheat to raising more fruits and vegetables. This was in response to the demands of the market. The results were more industry, population growth, greater income, drought (because the new crops required more water), and conflicts over water supplies. Dr. Charles Kennel of the Office of Mission to Planet Earth joined Dr.s Cordova, Kates, and Liverman for the question and answer session that followed.

Description: This final rule adopts with changes the interim rule published in the Federal Register on April 5, 2001 (65 FR 18051-18053), which amended the NASA FAR Supplement to implement a Safety and Health (Short Form) clause to address safety and occupational health in all NASA contracts above the micro-purchase threshold where the existing Safety and Health clause did not apply, and amended other safety and health clauses to be consistent with the new NASA Safety and Health (Short Form) clause.

Description: It is hypothesized that the hematopoietic stem cell therapy (HSCT) might countermeasure various space-caused disorders so as to maintain astronauts' homeostasis. If this were achievable, the HSCT could promote human exploration of deep space. Using animal models of disorders (hindlimb suspension unloading system and beta-thalassemia), the HSCT was tested for muscle loss, immunodeficiency and space anemia. The results indicate feasibility of HSCT for these disorders. To facilitate the HSCT in space, growth of HSCs were optimized in the NASA Rotating Wall Vessel (RWV) culture systems, including Hydrodynamic Focusing Bioreactor (HFB).

Description: This document includes information on all peer reviewed projects funded by the Office of Life and Microgravity Sciences and Applications, Life Sciences Division during fiscal year 1995. Additionally, this inaugural edition of the Task Book includes information for FY 1994 programs. This document will be published annually and made available to scientists in the space life sciences field both as a hard copy and as an interactive Internet web page

Description: The topics of agency and enterprise goals, OLMSA organization, life sciences relationship to NASA/HEDS strategic plans, budget allocated by the HEDS strategic plan goals, 1998 successes, exploration and the International Space Station, congressional budgets, OLMSA grants, biomedical research and countermeasures, medical care, biologically inspired technologies, and publication, education and outreach are all presented in viewgraph form.

Description: An eye movement-based methodology and assessment tool may be used to quantify many aspects of human dynamic visual processing using a relatively simple and short oculomotor task, noninvasive video-based eye tracking, and validated oculometric analysis techniques. By examining the eye movement responses to a task including a radially-organized appropriately randomized sequence of Rashbass-like step-ramp pursuit-tracking trials, distinct performance measurements may be generated that may be associated with, for example, pursuit initiation (e.g., latency and open-loop pursuit acceleration), steady-state tracking (e.g., gain, catch-up saccade amplitude, and the proportion of the steady-state response consisting of smooth movement), direction tuning (e.g., oblique effect amplitude, horizontal-vertical asymmetry, and direction noise), and speed tuning (e.g., speed responsiveness and noise). This quantitative approach may provide fast and results (e.g., a multi-dimensional set of oculometrics and a single scalar impairment index) that can be interpreted by one without a high degree of scientific sophistication or extensive training.