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At least 253 records · Page 14

VESGEN Mapping of Bioactive Protection against Intestinal Inflammation: Application to Human Spaceflight and ISS Experiments

Challenges to successful space exploration and colonization include adverse physiological reactions to micro gravity and space radiation factors. Constant remodeling of the microvasculature is critical for tissue preservation, wound healing, and recovery after ischemia. Regulation of the vascular system in the intestine is particularly important to enable nutrient absorption while maintaining barrier function and mucosal defense against micro biota. Although tremendous progress has been made in understanding the molecular circuits regulating neovascularization, our knowledge of the adaptations of the vascular system to environmental challenges in the intestine remains incomplete. This is in part because of the lack of methods to observe and quantify the complex processes associated with vascular responses in vivo. Developed by GRC as a mature beta version, pre-release research software, VESsel GENeration Analysis (VESGEN) maps and quantifies the fractal-based complexity of vascular branching for novel insights into the cytokine, transgenic and therapeutic regulation of angiogenesis, lymphangiogenesis and microvascular remodeling. Here we demonstrate that VESGEN can be used to characterize the dynamic vascular responses to acute intestinal inflammation and mucosal recovery from in vivo confocal microscopic 3D image series. We induced transient intestinal inflammation in mice by DSS treatment and investigated whether the ability of the pro biotic yeast Saccharomyces boulardii (Sb) to protect against intestinal inflammation was due to regulation of vascular remodeling. A primary characteristic of inflammation is excessive neovascularization (angiogenesis) resulting in fragile vessels prone to bleeding. Morphological parameters for triplicate specimens revealed that Sb treatment greatly reduced the inflammatory response of vascular networks by an average of 78%. This resulted from Sb inhibition of vascular endothelial growth factor receptor signaling, a major angiogenesis signaling pathway. It needs to be determined whether pro biotic yeast represents a promising approach to GI protection in space. GRC performed only the VESGEN post-testing analysis.

Parsons-Wingerter, P. A.↗

Oxidative Stress and Autophagy Responses of Osteocytes Exposed to Spaceflight-like Radiation.

Weightlessness and radiation, two of the unique elements of the space environment, causes a profound decrement in bone mass that mimics aging. This bone loss is thought to result from increased activity of bone-resorbing osteoclasts and functional changes in bone-forming osteoblasts, cells that give rise to mature osteocytes. Our current understanding of the signaling factors and mechanisms underlying bone loss is incomplete. However, it is known that oxidative stress, characterized by the excess production of free radicals, is elevated during radiation exposure. The goals of this study is to examine the response of osteocytes to spaceflight-like radiation and to identify signaling processes that may be targeted to mitigate bone loss in scenarios of space exploration, earth-based radiotherapy and accidental radiation exposure. We hypothesize that (1) oxidative stress, as induced by radiation, decreases osteocyte survival and increases pro-osteoclastogenic signals and that (2) autophagy is one of the key cellular defenses against oxidative stress. Autophagy is the process by which cellular components including organelles and proteins are broken down and recycled. To test our hypothesis, we exposed the osteocyte-like cell line, MLO-Y4, to 0.5, 1, and 2 Gy of simulated space radiation (Iron-56 radiation at 600 MeV/n) and assessed cell numbers, cell growth-associated molecules as well as markers of autophagy and oxidative stress at various time points post-irradiation. We observed a reduction in cell numbers in the groups exposed to 1 and 2 Gy of Iron-56 radiation. Collectively, flow cytometry and gene expression analysis revealed that radiation caused a shift in cell cycle distribution consistent with growth arrest. Compared to sham-treatment, 2 Gy of Iron-56 increased FoxO3, SOD1, and RANKL gene expression yet unexpectedly decreased LC3B-II protein levels at 4 and 24 hours post-IR. Taken together, these findings suggest that simulated space radiation invoke antioxidant, pro-osteoclastogenic, and growth arrest responses in osteocytes. The implications of reduced autophagy flux at the time points examined remain to be elucidated.

Biological Sciences↗

Rotary Balance Wind Tunnel Testing for the FASER Flight Research Aircraft

Flight dynamics research was conducted to collect and analyze rotary balance wind tunnel test data in order to improve the aerodynamic simulation and modeling of a low-cost small unmanned aircraft called FASER (Free-flying Aircraft for Sub-scale Experimental Research). The impetus for using FASER was to provide risk and cost reduction for flight testing of more expensive aircraft and assist in the improvement of wind tunnel and flight test techniques, and control laws. The FASER research aircraft has the benefit of allowing wind tunnel and flight tests to be conducted on the same model, improving correlation between wind tunnel, flight, and simulation data. Prior wind tunnel tests include a static force and moment test, including power effects, and a roll and yaw damping forced oscillation test. Rotary balance testing allows for the calculation of aircraft rotary derivatives and the prediction of steady-state spins. The rotary balance wind tunnel test was conducted in the NASA Langley Research Center (LaRC) 20-Foot Vertical Spin Tunnel (VST). Rotary balance testing includes runs for a set of given angular rotation rates at a range of angles of attack and sideslip angles in order to fully characterize the aircraft rotary dynamics. Tests were performed at angles of attack from 0 to 50 degrees, sideslip angles of -5 to 10 degrees, and non-dimensional spin rates from -0.5 to 0.5. The effects of pro-spin elevator and rudder deflection and pro- and anti-spin elevator, rudder, and aileron deflection were examined. The data are presented to illustrate the functional dependence of the forces and moments on angle of attack, sideslip angle, and angular rate for the rotary contributions to the forces and moments. Further investigation is necessary to fully characterize the control effectors. The data were also used with a steady state spin prediction tool that did not predict an equilibrium spin mode.

Denham, Casey↗

Superoxide Dismutase Protects Osteoprogenitors from Irradiation with Low-LET but Not High-LET Species

Ionizing radiation-induced bone loss appears to be a two-stage process: first an early increase in pro-resorption cytokines and increased bone resorption by osteoclasts, followed by a decrease in bone formation by osteoblasts. This results in a net loss of mass in mineralized bone tissue. The molecular mechanisms underlying the imbalance in bone remodeling caused by exposure to radiation are not fully understood. We hypothesized that the radiation-induced rise in reactive oxygen species (ROS) damages osteoblast progenitors, leading to a decrease in number and activity of differentiated progeny. We have shown that a diet high in antioxidant capacity prevents radiation-induced bone loss in adult mice (Schreurs et al. 2016) by reducing the early increase in pro-resotption cytokines. Here, we investigated the damaging effects of radiation exposure on cells in the osteoblast lineage, testing if addition of the exogenous antioxidant enzyme, superoxide dismutase (SOD) can mitigate radiation damage. Osteoprogenitors were grown in vitro from the marrow of 16wk old, male C57Bl/6 mice. Cells were irradiated 3 days after plating (day 0) with either gamma (Cs-137, 0.1-5Gy) or iron (Fe-56, 600 MeV/n, 0.5-2Gy), and then grown until day 10. SOD or vehicle was added 2 hours before irradiation (SOD at 200U/ml), twice a day and up to day 5, for a total of 2 days treatment. Cell behavior was assessed by: (a) colony number (counted on day 7), (b) DNA content (surrogate for cell number) to assess cell growth (percent change between day 3 and day 10) and (c) alkaline phosphatase activity (osteoblast differentiation marker). Results show that SOD protected cells from the adverse effects of low-LET ionizing radiation, but not high-LET radiation. These novel results provide an interesting platform to explore further diverse effects and damages caused by low-LET and high-LET, pointing toward different mechanisms and possible intervention strategies for radiation-induced bone loss.

superoxide dismutase↗

Dietary Supplement Attenuates Radiation-Induced Osteoclastogenic and Oxidative Stress-Related Responses and Protects Adult Mice from Radiation-Induced Bone Loss

Our central hypothesis is that oxidative stress plays a key role in cell dysfunction and progressive bone loss caused by radiation exposure during spaceflight. In animal studies, excess free radical formation is associated with pathological changes in bone structure, enhanced bone resorption, reduced bone formation and decreased bone mineral density, which can lead to skeletal fragility. We previously reported that exposure to low or high-LET radiation rapidly increases expression levels of pro-osteoclastogenic and oxidative stress-related genes in bone and marrow, followed by pathological changes in skeletal structure. To screen various antioxidants for radioprotective effects on bone, 4 month old, male C57Bl6/J mice were treated with a dietary antioxidant cocktail, injectable alpha-lipoic acid, or a dried plum-enriched diet (DP). Mice were then exposed to 2Gy 137Cs total body radiation and one day later marrow cells were collected and the relevant genes analyzed for expression levels. Of the candidates tested, DP was most effective in reducing bone resorption-related gene expression. Microcomputed tomography revealed that DP also prevented the radiation-induced deterioration of skeletal microarchitecture, as indicated by percent bone volume, trabecular spacing and trabecular number. DP had similar protective effects on skeletal structure after sequential exposure to protons (0.5 Gy, 150MeV/n) and 56Fe 0.5Gy, 600 MeV/n). When cultured ex vivo under osteogenic conditions, bone marrow-derived cells from DP-fed animals exhibited increased colony numbers compared to control diet-fed animals. These findings suggest that DP exerted pro-osteogenic effects apart from previously identified anti-resorptive actions, which may contribute to radioprotection of skeletal tissue. In conclusion, a diet enriched in certain types of antioxidants and polyphenols such as DP may be useful as an intervention to protect tissues from degenerative effects of ionizing radiation.

Globus, Ruth↗

Skeletal Responses to Long-Duration Simulated Weightlessness in Rats

Damaging effects due to spaceflight and long-duration weightlessness are seen in the musculoskeletal system, specifically with regards to bone loss, bone resorption, and changes in overall bone structure. These adverse effects are all seen with indicators of oxidative stress and a variation in the levels of oxidative gene expression. Once gravity is restored, however, the recovery is slow and incomplete. Despite this, few reports have investigated the correlation between oxidative damage and general modifications within the bone. In this project, we will make use of a ground-based model of simulated weightlessness (hindlimb unloading, HU) in order to observe skeletal changes in response to induced microgravity due to changes in oxidative pressures. With this model we will analyze samples at 14-day and 90-day time points following HU for the determination of acute and chronic effects, each with corresponding controls. We hypothesize that simulated microgravity will lead to skeletal adaptations including time-dependent activation of pro-oxidative processes and pro-osteoclastogenic signals related to the progression, plateau, and recovery of the bone. Microcomputed tomography techniques will be utilized to measure skeletal changes in response to HU. With the results of this study, we hope to further the understanding of skeletal affects as a result of long-duration weightlessness and develop countermeasures to combat bone loss in spaceflight and osteoporosis on Earth.

hindlimb unloading↗

Superoxide Dismutase Protects Osteoprogenitors from Irradiation with Low-LET but Not High-LET Species

Ionizing radiation-induced bone loss appears to be a two-stage process: first an early increase in pro-resorption cytokines and increased bone resorption by osteoclasts, followed by a decrease in bone formation by osteoblasts. This results in a net loss of mass in mineralized bone tissue. The molecular mechanisms underlying the imbalance in bone remodeling caused by exposure to radiation are not fully understood. We hypothesized that the radiation-induced rise in reactive oxygen species (ROS) damages osteoblast progenitors, leading to a decrease in number and activity of differentiated progeny. We have shown that a diet high in antioxidant capacity prevents radiation-induced bone loss in adult mice (Schreurs et al. 2016) by reducing the early increase in pro-resotption cytokines. Here, we investigated the damaging effects of radiation exposure on cells in the osteoblast lineage, testing if addition of the exogenous antioxidant enzyme, superoxide dismutase (SOD) can mitigate radiation damage. Osteoprogenitors were grown in vitro from the marrow of 16-week-old, male C57Bl6 mice. Cells were irradiated 3 days after plating (day 0) with either gamma (137Cs, 0.1-5Gy) or iron (56Fe, 600 MeVn, 0.5-2Gy), and then grown until day 10. SOD or vehicle was added 2 hours before irradiation (SOD at 200Uml), twice a day and up to day 5, for a total of 2 days treatment. Cell behavior was assessed by: (a) colony number (counted on day 7), (b) DNA content (surrogate for cell number) to assess cell growth (percent change between day 3 and day 10) and (c) alkaline phosphatase activity (osteoblast differentiation marker). Results show that SOD protected cells from the adverse effects of low-LET(Linear Energy Transfer) ionizing radiation, but not high-LET radiation. These novel results provide an interesting platform to explore further diverse effects and damages caused by low-LET and high-LET, pointing toward different mechanisms and possible intervention strategies for radiation-induced bone loss.

osteoblasts↗

Overview of the Digitization Workflow Post Image Acquisition of Apollo Lunar and Antarctic Meteorite Samples Using Agisoft Photoscan for the NASA 3D Astromaterials Virtual Samples Collection

The 3D Virtual Astromaterials Samples (3DVAS) collection is a multi-year funded project to create a digital database of sixty Apollo Lunar and Antarctic Meteorite samples following non-destructive documentation conservation protocols. After initial image processing, the photos are evaluated and processed using unique structure-from-motion photogrammetric techniques in a high performance modelling software designed to create a 3D model from 2D images: Agisoft Photoscan Pro. Agisoft Photoscan Pro uses image processing algorithms and techniques originating in computer vision to resolve 3D models for accurate and detailed visualization of a subject. The software provides a stepwise process that is tailored per model based on spatial and specular reflectance properties, for example. The process includes: photo alignment, creation of a dense point cloud, mesh, and finally texture. Photo alignment is dependent on model properties. The 3DVAS process requires a special rotation platform with calibrated photogrammetric targets, specific distance rotation protocols, and a contrasting background for alignment and scale accuracy. As a result of the photographic process, alignment will complete with two mirrored hemispheres that, in a sense, represent the 2D images overlapping to create a 3D model. Each dense point cloud is analyzed with provided statistical measures in a gradual selection process to eliminate outliers. The point cloud is reduced to include only data valuable to the final model. When a precise dense point cloud is achieved, a mesh and texture are applied. Each model is scaled with scale bar accuracies within 100 microns. Each sample has its own intimate process for modelling; there is no standard for the parameters required in the final creation of a high resolution model. By processing multiple samples, a skill is gained in practice to allow a close definition of the original sample and will result in the most detailed version of the sample shell. This process completes one-fifth of the 3DVAS protocol for providing accurate digital documentation. Each model shell is merged with X-ray Computed Tomography data to create a full volumetric sample. All 3DVAS data will be served on NASA's Astromaterials Acquisition and Curation website with an early subset of data available in 2019 and the 3D Virtual Astromaterials Samples Collection launch in 2020.

Thomas, Andi B.↗

Reflective Coatings for the Future X-Ray Mirror Substrates

We present the development of the reflective coating by magnetron sputtering deposition onto precisely-fabricated thin X-ray mirrors. Our goal is to remove distortion induced by the coating and then keep their surface pro les. We first addressed the uniform coating to minimize the distortion by introducing a mask to control the spatial distribution of the coating thickness. The uniformity was finally achieved within 1%. We next tried a platinum single-layer coating on a glass substrate with a dimension of 200 mm 125 mm. The distortion caused by the frontside coating with a thickness of 320 A was found to be at most 1 m, smaller than the previous results obtained from the non-uniform coating. We then carried out the platinum coating with the same amount of the thickness on the backside surface of the glass substrate. The surface pro le of the glass substrate was fully recovered, indicating that the residual stress was successfully balanced by the backside coating. Furthermore, we tried to an iridium single-layer coating with a thickness of 150 Aon the silicon mirrors. The frontside coating caused the degradation of the imaging quality by 7:5 arcsec in half-power width. However, the backside coating with the same amount of the thickness reduced this degradation to be 3:4 arcsec. Finally, an additional backside coating with a thickness of 100 A and the annealing to relax the residual stress were found to eliminate the distortion completely; the final degradation of the imaging quality was only 0:4 arcsec.

Mori, Hideyuki↗

Does Simulated Weightlessness Alter the Methylation Status of Gene Promoters During Bone Remodeling?

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.

Stevick, Rebecca J.↗

Hazardous Product Detection and Environmental Clean-up Removal in Spacecraft Vehicles from Fire Induced Li-Ion Fires

A fire on-board the International Space Station (ISS) resulting from a commercial Surface Pro tablet Lithium-Ion (Li-ion) battery can be detrimental to the spacercraft and the astronauts. The Spacecraft Fire Safety Demonstration (Saffire) program is focused on identifying and quantifying the risks that potentially arise inside a space vehicle by conducting large fire experiments inside the Cygnus cargo vehicle upon re-entry to Earth. The potential candidate that will be flown on the ISS is a 4- cell Li-ion battery pack Surface Pro tablet (42 Wh). The tablets were tested at the White Sands Test Facility (WSTF) using a localized heating method to emulate the failing mechanism that leads the unit into thermal runaway. Measurements inside the test chamber were performed on aerosol mass concentrations and for specific toxic products (i.e. from CO, HCN, HCl and CO2). These toxic products depend on the size of the fire and energy content of the tablet. Comparisons will be made with the Dell XPS (97 Wh). The concentrations are then used to extrapolate to laptop fires on a vehicle at the approximate volumetric size of the Orion spacecraft. The presentation provides an analysis on the detection capability and the response time to trigger fire alarms aboard a vehicle by using the mass concentration levels. These results will consider the rate at which the life support system is able to filter the atmosphere in order to provide a hazardous free environment.

Padilla, Rosa E.↗

Laboratory Investigation of the Effect of Venusian Weathering on Mineral Spectra

Introduction: The recent selection of two missions to Venus has renewed the importance of deter-mining weathering reactions between minerals and the Venusian atmosphere, and the spectral signatures of minerals before, during, and after these reactions. The rate at which weathering reactions progress also constrain show long unstable minerals will be present on the surface (e.g., [1]), and enables the use of mineralogy as a constraint on surface age(e.g., [2-3]).In order to gain an understanding of how mineral compositions and spectra change with weathering, we have begun conducting experiments in a 1 atm experimental setup at Wesleyan University. This setup exposes minerals to the temperature and most abundant gases of the Venus atmosphere (CO2, SO2, N2).We conducted initial experiments using biotite, calcite, and montmorillonite in order to test our methodology with minerals that may be relevant to recording the history of water on Venus. Methods: Experiments were conducted in Thermo Fisher Scientific Lindberg/Blue M Mini-Mite horizontal tube furnaces at Wesleyan University. Experiments used natural mineral chips and powders, and were conducted at 1 atmosphere and 460 °C under pre-mixed gases provided by Air Gas(Table 1). The furnaces are set up in a flow through configuration so that solid samples are exposed to a fixed gas composition, and quartz glass process tubes were used in all experiments (1/4” diameter for experiment V4, ½” diameter for all others). These conditions were maintained for the du-rations listed in Table 1, at which point the furnace was turned off with gas flowing until the sample was cool enough to extract under N2and be placed in a desiccator for storage. Run products were carbon coated and examined using a Hitachi SU5000 Field Emission Gun Scanning Electron Microscope (SEM) equipped with an EDAX Octane Pro EDS detector located at Wesley-an University. Visible-Near Infrared Spectroscopy (VNIR) analysis was performed on powdered samples under a nitrogen atmosphere using an ASD Fieldspec Proover the 350-2500 nm range. Powdered mineral samples were milled to a particle size of < 45μm and were spiked with an internal standard (Al2O3, corundum) to obtain quantitative mineralogy. Samples were analyzed using a Panalytical X’Pert pro X-ray Diffractometer (XRD), with an X’Celerator high speed detector and Co Kα radiation, with data collected at a step size of 0.02 ̊/minute step counting rate from 2 to 80 degrees 2θ at 45 mA/40kV in the X-ray Diffraction Laboratory located at NASA Johnson Space Center. Materials Data Inc (MDI) software suite, JadeTMv9 was used for Rietveld refinement to determine phase abundances and mineral identification by comparing XRD patterns to International Center for Diffraction Data (ICDD) database patterns. Table 1: Experimental matrix. Experiment Name Duration (Days)Gas Com-position (trace gas)MineralsV487SO2/N2(1.4%)Montmorillonite, biotiteV519SO2/N2(1.4%)CalciteV619CO2/SO2/N2(1.4% SO2, 2.1% N2)Montmorillonite, biotiteV828SO2/N2(1.4%)Biotite, calcite Results: Calcite. In both experiments, calcite was exposed to the SO2/N2gas mixture, and in both experimental run products, XRD analysis detected anhydrite, which is consistent with EDS measurements conducted in the SEM. The XRD analyses show greater amounts of anhydrite present after 28 days than 19, suggesting the calcite reaction progressed further given longer duration. Grain surface morphology as seen in the SEM also shows secondary mineral growth (Fig. 1). VNIR spectra show no change, as expected since anhydrite lacks spectral features in this wavelength range. Montmorillonite. Montmorillonite was exposed to two different gas mixtures, the SO2/N2mix and CO2/SO2/N2mix over different durations (87 and 19 days, respectively). VNIR analyses of both run products show a reduction in the 1441and1910 nm water features as well as a shift of the 1411 and 2011 nm features to shorter wavelengths that may indicate re-structuring in the crystal lattice and the production of amorphous phases [4]. XRD results are consistent with this, showing a shift of the 001 peak from 15Åto 10Åin both run products. The amount of X-ray amorphous material in the montmorillonite run products was greater than that present in the unreacted clay, being the greatest in the 87 day V4 experiment. No other secondary phases were detected in the run products, however sulfur was present in EDS analyses of powder samples.

A R Santos↗

Using X-Ray Computed Tomography to Catalog Rock Fragments in Apollo Drive Tube 73002

Overview: The Apollo missions collected 382 kg of rock, regolith, and core samples from six locations on the nearside of the Moon. Approximately 84% by mass of the Apollo collection remains in pristine condition within the curation facility at Johnson Space Center (i.e., never allocated, continuously stored in dry-N2 purged cabinets, exposure history restricted to Teflon, stainless steel, and Al-metal). Although most Apollo samples have been well characterized, there are several types of samples that have remained wholly or largely unstudied since their return, and/or that have been cu-rated under special conditions, e.g., frozen samples, samples stored in He-purged environment, and previously unopened drive tubes. NASA solicited proposals for the Apollo Next Generation Sample Analysis Pro-gram (ANGSA), and 9 teams were selected to study a subset of the unopened and frozen samples [1]. The first sample opened as part of the ANGSA pro-gram was drive tube 73002. This was originally a ~30 cm long, 4 cm diameter drive tube collected on a land-slide deposit near Lara Crater at the Apollo 17 landing site. It was part of a ~60 cm long double drive tube collected, and the bottom half of the tube (73001) was sealed under vacuum on the Moon [2]. Prior to opening sample 73002, the sample was imaged with a high resolution X-ray Computed Tomography (XCT) scan of the entire tube [3], which provided invaluable information during the dissection process [4]. In addition to the pre-dissection XCT scans, individual >4 mm particles were separated from the 73002 regolith during processing and scanned at high resolution by XCT. Here we pre-sent the initial lithologic classification of 134 individual >4 mm rock fragments separated from the 73002 core during the dissection process. Methodology: Drive tube 73002 was manually dissected in 0.5 cm depth intervals in three passes (Fig. 1) [4,5]. Each interval from pass 1 and 2 was sieved to <1 mm and >1 mm size fractions, and >1 mm particles were further manually subdivided into 1-2, 2-4, 4-10, and >10 mm size fractions. Pass 3 was not sieved, but >10 mm clasts were separated manually. Each 4-10 mm and >10 mm fragment was individually weighed, triply bagged in Teflon, and scanned by XCT. There are 60 rock fragments in Pass 1, 64 rock fragments in Pass 2 (from the 4-10 mm and >10 mm size fractions), and 8 rock fragments from Pass 3 (>10 mm size fraction). Each individually bagged rock fragment was scanned using the 180 kV nano-focus transmission source on the Nikon XTH 320 XCT system at NASA Johnson Space Center [6]. Scanning conditions varied considerably for individual particles, in large part be-cause of the large variation in size (0.008-19.623 g). All scans fell within the following range of scan conditions: 2.8-20.6 um voxel size; 90-155 kV voltage; 18-39 uA current; 1891-3141 projections; and 902-2000 slices. Results and Discussion: The 132 rock fragments from sample 73002 fall into the following general cat-gories: agglutinates (n = 6); basalts (13); impact melts (5); impact melt breccias (IMB; 42); regolith breccias (62); and soil breccias (4); see Figure 2 for representative examples of each lithology. Within most of these broad lithologic groups are recognizable subgroups. For example, a significant portion of regolith breccia fragments contain some agglutinate-like glass (n = 9) or are dilithologic (7) because they contain a single large clast (~50% or more by volume). Subgroups can be based on similarities to previously identified lunar lithologies, such as high-Ti basalts (9) and VLT basalts (4; Fig. 3), or based on commonly seen features within the fragments, such as poikilitic ilmenite IMB (10), ilmenite-lath IMB (12), or vesicular IMB (11) “groups”. Particles in the same “group” are not necessarily intended to be genetically related, but rather identify particles that are similar and that follow-up studies can classify in more detail [7]. Conclusion: Identification of lithologies based on XCT is a powerful tool, but a more absolute classification will sometimes require additional textural information from thin sections (e.g., glassy-matrix regolith vs. impact-melt vs. granulitic breccia) or quantitative mineral compositions (e.g., basalt vs. monomict breccia).

R A Zeigler↗

Applicability of Micro X-Ray Fluorescence Spectroscopy to Astromaterials Curation and Research

Introduction: The Astromaterials Acquisition and Curation Office at NASA’s Johnson Space Center (JSC) curates NASA’s astromaterial sample collections which includes: Apollo samples, Luna samples, Ant-arctic meteorites, cosmic dust particles, microparticle impacts into space-flown materials, Genesis solar wind atoms, Stardust comet Wild-2 particles, Stardust inter-stellar particles, Hayabusa asteroid Itokawa particles, Hayabusa 2 asteroid Ryugu particles, and future OSIRIS-Rex asteroid Bennu particles (landing in Sep-tember, 2023) [1–3]. To enhance JSC’s advanced cu-ration capabilities, we have recently installed a high-performance micro-X-ray fluorescence (µXRF) spec-trometer to assist in sample characterization through rapid, non-destructive, in-situ elemental analyses that do not require the sample preparation protocols (i.e., polishing and carbon-coating) commonly needed for electron beam analyses. With this new instrument, we are capable of detecting all elements down to carbon in a variable-pressure or He-purged chamber for anal-ysis of a wide range of sample types. Here we describe the instrumental set-up, capabilities, and applicability of µXRF analysis to astromaterials curation and re-search. Instrumentation and Methodology: The X-ray fluorescence and computed tomography lab (X-FaCT) lab at JSC is now equipped with a Bruker M4 Tornado Plus µXRF (Fig. 1). This system is an energy-dispersive x-ray spectrometer equipped with two 60 mm2 silicon drift detectors (SDD) that are able to be used simulta-neously for output count rates ~500,000 cps. New light element windows allow detecting and analyzing the entire elemental range from carbon to americium. Two x-ray tubes (micro-focus Rh with polycapillary lenses and W with collimators of 0.5, 1.0, 2.0, and 4.5 mm) with max excitation parameters of 50 kV, 30 W and 50 kV, 40 W, respectively, allow for more flexibility of the analysis of high energy lines. The motorized X-Y-Z stage has a mapping range of 190 x 160 mm and can support samples up to 7 kg (~15.5 lbs) and a height of 120 mm [4]. Analytical modes include elemental analysis (down to ~20 µm spot size) via point, line, or area of bulk materials (rock surfaces, thin sections, thick sections, etc.) as well as coating analysis (determination of thickness and composition) of samples. This system has a variable vacuum chamber (1 mbar to 1 atm) that is also equipped with a He-purge system which accommodates vacuum sensitive samples while still allowing detection of light elements at atmospheric pressure. Utility and Applicability of µXRF in Astro-materials research and exploration science (ARES): Elemental analysis using µXRF is commonly em-ployed for both terrestrial and planetary geological science disciplines [5]. It is especially useful for analy-sis of astromaterials given the limited sample prepara-tion required, which is not feasible for certain materi-als. Here we show select applications of µXRF anal-yses of astromaterials that can, have, and will be done at JSC’s X-FaCT lab. Point analysis: In-situ spot analyses (~20 µm spot size) on a cut slab of Martian meteorite NWA 10922 allowed for the discovery, qualitative elemental analy-sis and determination of different feldspar minerals [6]. These point analyses served as an effective prelim-inary step for subsequent quantitative analyses. Ana-lytical standards can be employed for more accurate quantification of µXRF spot analyses. Area analysis: This analytical mode measures all detectable elements (from C to Am) at each pixel (>5 µm pixel size) in a user-defined area. The results are shown as elemental maps which can be extracted as 16-bit TIFF’s for further data processing. In Fig. 2. we show elemental distribution maps of the high-Ti basalt 73001,531 that have been processed using ImageJ software. From these maps you can accurately and quickly (this map took ~50 mins.) identify mineral components, such as pyroxene, plagioclase, oxides, and phosphates, compositional zoning, and mineral textures. Detection of high-Z phases: µXRF techniques are es-pecially effective at analyzing trace minerals with high-atomic-number (high-Z) elements because the high-energy characteristic X-rays used (relative to SEM EDS) allow for mapping of K lines in elements up to La (typically SEM maps use L X-ray lines for elements >Zn, and these can often have interferences). Thus, µXRF is especially suited for identifying minerals like zircon, baddeleyite, REE-rich phosphates, Fe-rich met-als, oxides, sulfides, and phosphides [4]. In Fig. 3 we show elemental distribution maps for 73001,530 where we are able to correlate the original video image with, Zr, Si, Y and Hf elemental maps together identi-fying the location of a zircon. In this location you would expect lower Si compared to surrounding mate-rial, as well as higher Zr, Y, and Hf content compared to surrounding material, all of which is confirmed by our XRF ele-mental distribution maps (Figure 2.) Conclusions: The new M4 Tornado Plus µXRF within the Astromaterials Acquisition and Curation office at NASA JSC allows for rapid and non-destructive elemental analysis of astromaterials with limited or no sample preparation. µXRF analyses pro-vide crucial compositional knowledge for the prelimi-nary examination and curation of astromaterials. This instrument enhances the advanced curation capabili-ties in the X-FaCT laboratory at JSC by allowing pro-ductive, cohesive, and non-destructive multi-modal x-ray analyses on astromaterial samples, which is neces-sary for the comprehensive curation and study of our current and future astromaterial collections. Addition-ally, µXRF can provide complimentary information to researchers for studies on astromaterials. References: [1] Allen, C. et al., (2011). Chemie De Erde Geochemistry, 71, 1-20. [2] McCubbin, F. M. et al., (2016) 47th LPSC, abstract #2668 [3] Zeigler, R. A. et al., (2017) 48th LPSC, abstract #2772 [4] Bruker User Manual [5] Young et al., (2016) Appl. Geochemistry, 72, 77-87 [6] Mor-ris, R. V. et al., (2023) 54th LPSC.

E W O'Neal↗

ON THE EFFECTIVENESS OF LLMS IN UNIT TEST GENERATION FOR STRUCTURED TEXT PROGRAMS

The reliability of industrial automation systems heavily depends on the correctness of Programmable Logic Controller (PLC) programs, which are often written in Structured Text (ST). While Large Language Models (LLMs) have shown promise in automating test generation for mainstream programming languages, their effectiveness for the syntactically strict ST language remains underexplored. This thesis presents a systematic empirical evaluation of three state-of-the-art LLMs—GPT-4o, Gemini 2.5 Pro, and Claude Sonnet 4.5—for generating ST unit tests. We examine three prompting strategies: Natural Language (NL), Code Language (CL), and Chain-of-Thought (CoT), across a curated set of 11 ST function blocks. The quality of the generated tests is assessed using Compilation Success Rate (CSR), Statement Coverage (SC), and Branch Coverage (BC). In the zero-shot setting, Claude Sonnet 4.5 achieves the highest CSR, while Gemini 2.5 Pro consistently delivers the best statement and branch coverage, particularly under CL prompts. By incorporating a one-shot CL prompt, all models exhibit substantial improvements—most notably GPT-4o, whose CSR increases from 45.45% to 90.91%, with substantial gains in both SC and BC. To further contextualize these findings, we compare GPT-4o’s one-shot results with PLCAutoTester, a state-ofthe- art ST unit test generation tool, on an additional benchmark dataset. While LLMgenerated tests approach competitive coverage levels, PLCAutoTester maintains significantly higher and more stable coverage across programs. This study provides the first comprehensive benchmark of modern LLMs for ST unit testing, highlighting their strengths, limitations, and improvements through one-shot prompting, and positioning their performance relative to specialized automated testing tools in industrial automation.

42 ENGINEERING↗

Enhanced Surgical Decision-Making Tools in Breast Cancer: Predicting 2-Year Postoperative Physical, Sexual, and Psychosocial Well-Being following Mastectomy and Breast Reconstruction (INSPiRED 004)

Background: We sought to predict clinically meaningful changes in physical, sexual, and psychosocial well-being for women undergoing cancer-related mastectomy and breast reconstruction 2 years after surgery using machine learning (ML) algorithms trained on clinical and patient-reported outcomes data. Patients and Methods: We used data from women undergoing mastectomy and reconstruction at 11 study sites in North America to develop three distinct ML models. We used data of ten sites to predict clinically meaningful improvement or worsening by comparing pre-surgical scores with 2 year follow-up data measured by validated Breast-Q domains. We employed ten-fold cross-validation to train and test the algorithms, and then externally validated them using the 11th site’s data. We considered area-under-the-receiver-operating-characteristics-curve (AUC) as the primary metric to evaluate performance. Results: Overall, between 1454 and 1538 patients completed 2 year follow-up with data for physical, sexual, and psychosocial well-being. In the hold-out validation set, our ML algorithms were able to predict clinically significant changes in physical well-being (chest and upper body) (worsened: AUC range 0.69–0.70; improved: AUC range 0.81–0.82), sexual well-being (worsened: AUC range 0.76–0.77; improved: AUC range 0.74–0.76), and psychosocial well-being (worsened: AUC range 0.64–0.66; improved: AUC range 0.66–0.66). Baseline patient-reported outcome (PRO) variables showed the largest influence on model predictions. Conclusions: Machine learning can predict long-term individual PROs of patients undergoing postmastectomy breast reconstruction with acceptable accuracy. This may better help patients and clinicians make informed decisions regarding expected long-term effect of treatment, facilitate patient-centered care, and ultimately improve postoperative health-related quality of life.

60 APPLIED LIFE SCIENCES↗

Model results and data for Nontechnical Barriers to Geothermal Development

Data included in this submission support the analysis conducted for the report "Nontechnical Barriers to Geothermal Development" which is linked bellow. These data include information about the power purchase agreements (PPAs) analyzed for the report, inputs and model results for the pro forma economic analysis, and outputs from the regression analysis conducted on PPAs comparing geothermal and other power generation technologies.

15 GEOTHERMAL ENERGY↗

Southern Idaho Ecological Conservation: Investigating the Impact of Targeted Grazing to Improve Wetland Habitat in the Sterling Wildlife Management Area

Wetland ecosystems are vital for biodiversity conservation and ecosystem services. The Sterling Wildlife Management Area in Bingham County, Idaho, has management concerns about decadent and accumulated vegetation growth encroaching on wetland habitat, which presents challenges for wildlife, decreases biodiversity, and limits public access. Targeted grazing has been proposed as a sustainable alternative to chemical herbicides or burning. Land managers introduced targeted cattle grazing in January 2021 to reduce biomass. NASA DEVELOP partnered with the Idaho Department of Fish and Game to determine the impact of grazing using NASA Earth observations from Landsat 8 Operational Land Imager (OLI) in Google Earth Engine (GEE). Images were processed with TerrSet’s Land Change Modeler and ArcGIS Pro’s Change Detection Wizard to understand land changes following grazing. A Normalized Difference Vegetation Index (NDVI) analysis was performed to assess impacts on vegetation productivity and compare variance in biomass before and after grazing. A Normalized Difference Water Index (NDWI) was used to compare changes in the wetland and its vegetation content to evaluate the suitability of the area for migratory birds post-grazing. Results showed a decrease in the vegetation index and an increase in the water index postgrazing. The DEVELOP team’s analysis suggests that grazing helps break down thick, senesced vegetation and increase soil moisture. Providing a workflow model will aid partners in continuing to monitor this management area and other management areas across the state.

change detection↗