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DOE OSTI2026

Data and scripts associated with a manuscript modeling microbial regulation of priming effects

This data package is associated with the publication “Modeling Microbial Regulatory Feedback in Organic Matter Decomposition Identifies Copiotrophic Traits as Key Drivers of Positive Priming” published as a preprint on BioRXiv by Ahamed et al. (2026); https://doi.org/10.1101/2024.08.11.607483. The package contains MATLAB scripts and saved simulation outputs used to implement a cybernetic model of microbial regulation during complex organic matter (OM) decomposition governing priming effects. It includes models of (i) single microbial functional groups (copiotrophic or oligotrophic degraders) and (ii) binary consortia composed of degraders and non-degraders with contrasting or common growth traits. Simulation results were generated using Monte Carlo analyses, with randomized key model parameters across a range of environmental mixing fractions of complex and labile OM. The dataset was created to provide a transparent and reusable computational framework for systematically exploring how microbial growth traits, metabolic regulation, and community composition influence OM decomposition dynamics and priming effects. For details on how to navigate data packages generated by this project, see https://data.ess-dive.lbl.gov/portals/PNNLRiverCorridorSFA/About. In addition to a readme, this data package also includes a file-level metadata (FLMD) file that describes each file and a data dictionary (DD) that describes the variable definitions. This package includes: (1) annotated MATLAB code implementing the system of ordinary differential equations and cybernetic control laws; (2) saved output files containing data (e.g., biomass, substrates, enzyme levels, priming metrics); and (3) scripts for processing saved outputs and regenerating figures. Specifically, the data package contains three main MATLAB scripts: runPrimingModel.m, runPlotData.m, and runPlotSuppFigS1.m, along with this readme and supporting documentation. Users should begin with runPrimingModel.m, which contains the annotated code implementing the system of ordinary differential equations and cybernetic control laws. This script runs the Monte Carlo simulations of microbial OM decomposition and allows users to modify microbial trait definitions, adjust parameter distributions, or define new community configurations. Simulation outputs are automatically saved as .mat files in the folder named SavedData, which stores all pre-generated results included in this package. The second script, runPlotData.m, reads files from the SavedData folder and processes them to regenerate the figures presented in the manuscript. The third script, runPlotSuppFigS1.m, specifically generates Figure S1 in the Supplementary Material of the manuscript. The package also includes the aforementioned files in non-proprietary .txt format. If users intend to use them, they should first save the files in their respective .m or .mat formats prior to execution in MATLAB.

Biomass concentration↗

NASA NTRS1980

Flexible materials technology

A survey of all presently defined or proposed large space systems indicated an ever increasing demand for flexible components and materials, primarily as a result of the widening disparity between the stowage space of launch vehicles and the size of advanced systems. Typical flexible components and material requirements were identified on the basis of recurrence and/or functional commonality. This was followed by the evaluation of candidate materials and the search for material capabilities which promise to satisfy the postulated requirements. Particular attention was placed on thin films, and on the requirements of deployable antennas. The assessment of the performance of specific materials was based primarily on the failure mode, derived from a detailed failure analysis. In view of extensive on going work on thermal and environmental degradation effects, prime emphasis was placed on the assessment of the performance loss by meteoroid damage. Quantitative data were generated for tension members and antenna reflector materials. A methodology was developed for the representation of the overall materials performance as related to systems service life. A number of promising new concepts for flexible materials were identified.

Steurer, W. H.↗

DOE OSTI2022

Effect of bacterial cell addition on Fe(III) reduction and soil organic matter transformation in a farmland soil

The coupled biogeochemical processes of Fe(III) reduction and organic matter transformation profoundly impact terrestrial carbon cycling. However, little is known about the microbial role in soil organic matter (SOM) transformation during Fe(III) bio-reduction. Here we investigated the bio-reduction behavior of a black farmland soil and corresponding SOM transformation under circumneutral and anoxic conditions. A model dissimilatory Fe reducing bacterium, Geobacter sulfurreducens, was added in either live or dead form in order to enhance Fe(III) reduction in soil and to evaluate the accompanying transformation of SOM. The progress of Fe reduction was monitored and SOM transformation was characterized by various spectroscopy methods. Results showed that addition of either dead or live G. sulfurreducens cells increased the Fe(III) reduction rate and extent. Without cell addition, SOM transformation was insignificant within 13 days of incubation, only with some consumption of aliphatic/protein compounds, apparently due to their higher bio-degradability. Addition of dead or live cells resulted in more drastic SOM transformation, but through different mechanisms. With dead cell amendment, cell necromass and debris stimulated the activity of indigenous soil microbes by serving as extra carbon/energy sources. During Fe(III) reduction, the aliphatic/protein compounds were preferentially consumed by indigenous microbial communities, similar to the treatment without cell addition but with a greater extent of consumption. In comparison, addition of live G. sulfurreducens cells stimulated degradation of less bioavailable compounds, including more saturated and higher molecular weight SOM. It is possible that fast depletion of labile SOM by live G. sulfurreducens cells favored utilization of less bioavailable molecules (such as those with more aromatic structures) by the originally dormant species in native microbial community, suggesting an active role of live Fe(III)-reducing bacteria in affecting SOM transformation. In addition, fast assimilation of microbial related carbon, such as aromatic proteins and microbial byproducts, into SOM pools was also observed. Overall, our results suggest that addition of Fe(III)-reducing bacteria to a native soil can enhance Fe(III) reduction and accelerate the turnover of SOM through various mechanisms. The study provides new insights into coupled Fe(III) reduction and SOM transformation, as well as the “priming effect” of SOM using microbial cells as substrate.

54 ENVIRONMENTAL SCIENCES↗

NASA NTRS1983

Weight trends for a fully reusable advanced single-stage shuttle

The rate at which subsystem weights grow with vehicle gross weight is assessed and is shown to be critical to the efficiency of large Earth to orbit transports. The overall trend, however, is a reduction in the inerts as a percentage of gross weight as the vehicle size is increased. For this reason, the larger the vehicle, the greater the payload weight delivered per pound of vehicle manufactured. Other critical issues addressed include the effects of wing loading and wing size on wing weight, the effect of entry planform loading on thermal protection system weight, the impact of power demand on cooling system and prime power weight, and tank fineness ratio on insulation weight. The effects of body shape and various internal packaging arrangements on weight and balance are also discussed. The greatest impact on overall vehicle weight is body shape and internal packaging, and could account for weight savings of up to 30 percent in body structure. Other subsystems are important, but the savings are much smaller in relation to overall vehicle weight--individually less than one percent.

Macconochie, I. O.↗

NASA NTRS1983

Weight trends for a fully reusable advanced single-stage Shuttle

The rate at which subsystem weights grow with vehicle gross weight is assessed and is shown to be critical to the efficiency of large earth-to-orbit transports. Some subsystems grow as the square of vehicle size, others grow as the cube of vehicle size, and still others remain nearly constant irrespective of vehicle size. The overall trend, however, is a reduction in the inerts as a percentage of gross as the vehicle size is increased. For this reason, the larger the vehicle, the greater the payload weight delivered per pound of vehicle manufactured. Other critical issues addressed include the effects of wing loading and wing size on wing weight, the effect of entry planform loading on thermal protection system weight, the impact of power demand on cooling system and prime power weight, and tank fineness ratio on insulation weight. The effects of body shape and various internal packaging arrangements on weight and balance are also discussed. Of greatest impact on overall vehicle weight is body shape and internal packaging and could account for weight savings of up to 30 percent in body structure. Other subsystems are important but the savings are much smaller in relation to overall vehicle weight - individually less than one percent.

Macconochie, I. O.↗

DOE OSTI2024

Evaluating the Impact of Off-Design CHP Performance on the Optimal Sizing and Dispatch on Hybrid Renewable-CHP Distributed Energy Resources

The maturation of distributed energy resources (DER) has prompted the exploration of their deployment in commercial building applications due to their potential to supply energy at lower costs and emissions rates compared to centralized generation. While several software tools exist for evaluating the techno-economic potential of integrated renewable energy and combined heat and power (CHP) systems for distributed generation applications, many suffer from poor accuracy in capturing off-design (part load and changes in ambient air temperature and pressure) performance characteristics of microturbines, combustion turbines, or internal combustion engines. Thus, this paper presents a methodology for integrating these off-design characteristics in the mixed-integer linear program within REopt, a hybrid DER screening tool. The economic impact of the CHP off-design performance is observed through several application studies of various hybrid system configurations in different climates. Each study indicates how CHP off-design performance influences optimal sizing and dispatch decisions and therefore overall system economic value. We observe through case studies that modeling without the off-design effects, depending on the CHP prime mover and site, can result in Net Present Value predictions of hybrid systems that can be overoptimistic in frequently hot climates (up to 52%), too conservative in frequently cold climates (up to 11%), or unaffected (+/-1%) in temperate climates. Cases also highlight several advantages of hybrid systems relative to non-hybrid systems such as total economic value and the systems' ability to mitigate potentially negative consequences attributed to off-design performance.

ambient de-rate↗

DOE OSTI2021

Phenolic acid-degrading Paraburkholderia prime decomposition in forest soil

Abstract Plant-derived phenolic acids are catabolized by soil microorganisms whose activity may enhance the decomposition of soil organic carbon (SOC). We characterized whether phenolic acid-degrading bacteria enhance SOC mineralization in forest soils when primed with 13C-labeled p-hydroxybenzoic acid (pHB). We further tested whether pHB-induced priming could explain differences in SOC content among mono-specific tree plantations in a 70-year-old common garden experiment. pHB addition primed significant losses of SOC (3–13 µmols C g−1 dry wt soil over 7 days) compared to glucose, which reduced mineralization (-3 to -8 µmols C g−1 dry wt soil over 7 days). The principal degraders of pHB were Paraburkholderia and Caballeronia in all plantations regardless of tree species or soil type, with one predominant phylotype (RP11ASV) enriched 23-fold following peak pHB respiration. We isolated and confirmed the phenolic degrading activity of a strain matching this phylotype (RP11T), which encoded numerous oxidative enzymes, including secretion signal-bearing laccase, Dyp-type peroxidase and aryl-alcohol oxidase. Increased relative abundance of RP11ASV corresponded with higher pHB respiration and expression of pHB monooxygenase (pobA), which was inversely proportional to SOC content among plantations. pobA expression proved a responsive measure of priming activity. We found that stimulating phenolic-acid degrading bacteria can prime decomposition and that this activity, corresponding with differences in tree species, is a potential mechanism in SOC cycling in forests. Overall, this study highlights the ecology and function of Paraburkholderia whose associations with plant roots and capacity to degrade phenolics suggest a role for specialized bacteria in the priming effect.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2022

Techno-Economic Implications of Electrical Machine Scaling for Wave Energy Converters: Preprint

The sizing of an electrical machine for a Wave Energy Converter (WEC) can have a substantial impact on the overall sizing, cost, and rating of the device. An electrical generator is typically part of the power take-off (PTO) system, which is the mechanism by which the energy absorbed by the prime mover is transformed into useable electrical. For practically all WECs, the rate of change of actuation is predominantly determined by the wave resource (i.e., the wave height and frequency) and devices will see a sinusoidally varying velocity according to the wave conditions. The same can then be said for both directly and indirectly coupled PTOs with electrical generators. This techno-economic study investigates electrical machine scaling and associated cost implications through core machine design theory, manufacturer data, supporting literature, and the DOE sponsored Reference Model Project (RMP). The RMP was a partnered effort to develop open-source marine energy (ME) point designs as reference models (RMs) to benchmark ME technology performance and costs, methods for design and analysis of ME technologies, estimations for capital costs, operational costs, and levelized costs of energy (LCOE). The results from this study show torque is directly related to (1) the physical size of the machine required to increase the airgap sheer stresses, (2) the amount of active material, (3) the support structure, (4) bearing size and rating, and (5) offshore cable rating, all of which have a significant effect on overall system costs in terms of both CAPEX and OPEX. This paper aims to be a critical benchmark in helping determine an “optimal” nameplate rating for wave energy devices and their associated PTO. With an optimized rating and sizing process, WEC costs can be reduced, and overall performance can be improved.

cables↗

DOE OSTI2022

Techno-Economic Implications of Electrical Machine Scaling for Wave Energy Converters

cables↗

DOE OSTI2022

Techno-Economic Implications of Electrical Machine Scaling for Wave Energy Converters

The sizing of an electrical machine for a Wave Energy Converter (WEC) can have a substantial impact on the overall sizing, cost, and rating of the device. An electrical generator is typically part of the power take-off system, which is the mechanism by which the energy absorbed by the prime mover is transformed into usable electrical energy. For practically all WECs, the rate of change of actuation is predominantly determined by the wave resource (i.e., the wave height and frequency), and devices will see a sinusoidal varying velocity according to the wave conditions. The same can then be said for both directly and indirectly coupled power take-offs with electrical generators. This techno-economic study investigates electrical machine scaling and associated cost implications through core machine design theory, manufacturer data, supporting literature, and the Reference Model Project sponsored by the U.S. Department of Energy. The Reference Model Project was a partnered effort to develop open-source marine energy point designs as reference models to benchmark marine energy technology performance and costs, methods for design and analysis of marine energy technologies, estimations for capital costs, operational costs, and levelized cost of energy. The results from this study show torque is directly related to (1) the physical size of the machine required to increase the air-gap sheer stresses, (2)the amount of active material, (3) the support structure, (4) bearing size and rating, and (5) offshore cable rating, all of which have a significant effect on overall system costs in terms of both capital and operational expenditures. This paper aims to be a critical benchmark in helping determine an "optimal" nameplate rating for wave energy devices and their associated power take-offs. With an optimized rating and sizing process, WEC costs can be reduced and overall performance can be improved.

cables↗

DOE OSTI2025

Tracing priming effects in palsa peat carbon dynamics using a stable isotope-assisted metabolomics approach

Introduction: Peatlands store up to a third of global soil carbon, and in high latitudes their litter inputs are increasing and changing in composition under climate change. Although litter significantly influences peatland carbon and nutrient dynamics by changing the overall lability of peatland organic matter, the physicochemical mechanisms of this impact—and thus its full scope—remain poorly understood. Methods: We applied multimodal metabolomics (UPLC-HRMS, 1 H NMR) paired with 13 C Stable Isotope-Assisted Metabolomics (SIAM) to track litter carbon and its potential priming effects on both existing soil organic matter and carbon gas emissions. Through this approach, we achieved molecule-specific tracking of carbon transformations at unprecedented detail. Results: Our analysis revealed several key findings about carbon dynamics in palsa peat. Microbes responded rapidly to litter addition, producing a short-term increase in CO 2 emissions, fueled nearly exclusively by transformations of litter carbon. Litter inputs significantly contributed to the organic nitrogen pool through amino acids and peptide derivatives, which served as readily accessible nutrient sources for microbial communities. We traced the fate of plant-derived polyphenols including flavonoids like rutin, finding evidence of their degradation through heterocyclic C-ring fission, while accumulation of some polyphenols suggested their role in limiting overall decomposition. The SIAM approach detected subtle molecular changes indicating minimal and transient priming activity that was undetectable through conventional gas measurements alone. This transient response was characterized by brief microbial stimulation followed by rapid return to baseline metabolism. Pre-existing peat organic matter remained relatively stable; significant priming of its consumption was not observed, nor was its structural alteration. Discussion: This suggests that while litter inputs temporarily increase CO 2 emissions, they don’t sustain long-term acceleration of stored carbon decomposition or substantially decrease peat’s carbon store capacity. Our findings demonstrate how technological advancements in analytical tools can provide a more detailed view of carbon cycling processes in complex soil systems.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2024

Homologous mutations in human β, embryonic, and perinatal muscle myosins have divergent effects on molecular power generation

Mutations at a highly conserved homologous residue in three closely related muscle myosins cause three distinct diseases involving muscle defects: R671C in β-cardiac myosin causes hypertrophic cardiomyopathy, R672C and R672H in embryonic skeletal myosin cause Freeman–Sheldon syndrome, and R674Q in perinatal skeletal myosin causes trismus-pseudocamptodactyly syndrome. It is not known whether their effects at the molecular level are similar to one another or correlate with disease phenotype and severity. To this end, we investigated the effects of the homologous mutations on key factors of molecular power production using recombinantly expressed human β, embryonic, and perinatal myosin subfragment-1. We found large effects in the developmental myosins but minimal effects in β myosin, and magnitude of changes correlated partially with clinical severity. The mutations in the developmental myosins dramatically decreased the step size and load-sensitive actin-detachment rate of single molecules measured by optical tweezers, in addition to decreasing overall enzymatic (ATPase) cycle rate. In contrast, the only measured effect of R671C in β myosin was a larger step size. Our measurements of step size and bound times predicted velocities consistent with those measured in an in vitro motility assay. Finally, molecular dynamics simulations predicted that the arginine to cysteine mutation in embryonic, but not β, myosin may reduce pre-powerstroke lever arm priming and ADP pocket opening, providing a possible structural mechanism consistent with the experimental observations. This paper presents direct comparisons of homologous mutations in several different myosin isoforms, whose divergent functional effects are a testament to myosin’s highly allosteric nature.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2019

Audibility of Multiple, Low-Frequency Tonal Signals in Noise

The main purpose of this study is to examine the audibility of multiple, low-frequency tones that are placed in distinct auditory channels. Three experiments are described, the goals of which are to determine if the presence of sound in multiple channels results in enhanced audibility and to assess the applicability of the Statistical Summation Model (SSM) to this frequency range. This model predicts that for the case of multiple signals that are in separate auditory channels, implying statistical independence, each with sensitivity value d prime of i, the resulting total sensitivity is given by the square root of the sum of the squares of the individual d prime of i values. In common with previous studies conducted at higher frequencies, the signals are pure tones and the maskers are broadband noise. The requirement that low frequency tones be placed in separate auditory filters limited the number of tones to a maximum of three. The first of the three experiments measured the change in masked thresholds for two- and three-tone signals relative to the level of the equally-detectable single tones. The multiple tone signals were composed of combinations of 55, 120 and 200 Hz tones. The measured changes in thresholds exceeded those predicted by the SSM, although they did not differ statistically from the model predictions. The second experiment employed the same overall approach but acquired more data and concentrated on the three-tone signal. Once again, the measured changes in masked threshold exceeded the model predictions, this time to a statistically-significant degree. Two issues were postulated with the potential to yield inflated changes in masked threshold: interaction between tones resulting in perceptible intermodulation/difference tones, and the assumption that the tones were in distinct auditory filters and statistically independent of one another. The third experiment used two sets of three-tone signals to address these latter concerns. The first set of three tones was composed of harmonically related tone frequencies of 55, 110 and 165 Hz, which was an attempt to reduce effects of intermodulation difference tones. The second set of three tones was chosen to be 110, 220 and 330 Hz, again reducing effects of difference tones, but also providing greater separation between tones. Results for the first set of three tones compared to those of the earlier experiments indicated that intermodulation was not an important effect. The second set of three tones (110, 220, 330 Hz) yielded changes in masked thresholds that, on average, were in good agreement with the SSM, although intersubject variability was large and prohibited a definitive conclusion regarding the concern that tone spacing was inadequate. The results of the three experiments showed that the masked threshold of sounds with multiple (two or three) equally-detectable low frequency tones was lower than those of the single tones. In other words, it is clear that audibility is enhanced by the presence of signals in multiple auditory filters. This finding is consistent with most previous research conducted at higher frequencies. In contrast with previous research, test subjects were, on average, able to detect multitone sounds at lower levels than those predicted using the SSM. Analyses that included Monte Carlo simulations showed that normally distributed errors in the single tone thresholds result in biased estimates of the thresholds of multitone sounds. This phenomenon is likely responsible for at least a substantial fraction of the unexpected deviation of measurements from SSM predictions.

Rafaelof, Menachem↗

NASA NTRS2022

Ideas on the NASA Senior Review and Mission Extension Process

Extended mission budgets fund a significant part of our community, particularly young scientists who in a relatively short time are exposed to a broad range of experiences and opportunities available in space science and other STEM fields. In particular, mission-funded science offers a relatively stable funding stream that enables meaningful mentoring efforts and interactions, including apprenticeships ranging from project management, to mission operations, to data processing and analysis, to hardware and software development, to STEM-related education and outreach. It moreover provides professional contacts that fuel diversity in mission leadership and in the science and technical workforce at-large. Last but not least, it sustains the scientific productivity of NASA’s missions, which constitute major national investments meriting their full exploitation. Budgets cuts, however, are exercising pressure on the science and operations of the extended mission fleet despite being only a small portion of the overall Division budget. The history of Heliophysics has demonstrated that maintaining the operation of missions beyond their prime phase invariably leads to a much deeper knowledge of the original mission science goals, together with new applications often not foreseen during their original design. Their operation as part of the Heliophysics System Observatory (HSO)---a coordinated, 1eliophysics, multipoint observing system of the heliosphere, is a prime example. Some of the first activities undertaken by the Parker Solar Probe and Solar Orbiter missions required recreating the larger context of their new observations by using the diversity of HSO mission capabilities. The marginal cost of maintaining this valuable, working science resource, in the form of both continuing high quality data access and expertise, is small compared to the cost of the original missions or of new missions to achieve similar objectives, making the operation of extended missions, the best “science per dollar” investment in Heliophysics. We argue that reducing both extended mission operations and science budgets will have a ripple effect across the community, including reducing employment and training opportunities for young scientists, to lowering the quality/impact of the Heliophysics Systems Observatory. Instead, we suggest that the proposal process for mission extension should be re-examined. We offer several ideas for reducing costs of extended missions toward maintaining science output and streamlining the management process.

A. Vourlidas↗

NASA NTRS2003

Scale Model Experiments on Sound Propagation From a Mach 2.5 Cold Nitrogen Jet Flowing Through a Rigid-Walled Duct With a J-Deflector

The Launch Systems Testbed (LST) represents the evolution of vibroacoustics research and development work performed at NASA John F. Kennedy Space Center (KSC) over the last 15 years. The LST is located at the Launch Equipment Test Facility (LETF) in the KSC industrial complex. The LETF is operated by Sierra Lobo, Inc., as a member of University-Affiliated Technology Development Contract (USTDC) to KSC Spaceport and Engineering and Technology Directorate (YA), with ASRC Aerospace Corporation as a the prime contractor. Trajectory Simulation Mechanism (TSM) is a major component of the LST, developed specifically to simulate nonstationary acoustic loads on launch pad structures, vehicles, and payloads. TSM enhances the capabilities within LST for simulating launch environments of future vehicles. The scaled launch environments will be used to predict the full-scale launch environment via an appropriate scaling procedure. Air Force Research Laboratory (AFRL) has tasked NASA KSC to perform a basic technology test program in support of developing a low-cost clean pad (incorporating passive mitigation techniques) for future launch vehicles. The overall goal of the program is to develop innovative launch exhaust management systems, which effectively reduce launch acoustic environment with innovative duct designs, while eliminating traditional sound suppression water systems. Passive techniques, such as nontraditional duct geometries, resonators, and diffusers, etc., will be investigated. The overall goals are to advance innovative concepts for a clean pad while developing ideas to reduce transmitted sound via investigation and modeling of jet exhaust acoustic and flow field characteristics. The series of tests outlined in this report represent baseline tests and are geared towards defining the acoustic load environment on the TSM pad for open and closed duct configurations. This report summarizes the cold jet acoustic testing for Mach 2.5 supersonic nitrogen jet issuing from a nozzle with 1-inch exit diameter. Acoustic data, including spectral sound power and Overall Sound Pressure Level (OASPL), are obtained both for a free jet and with the jet flowing through a rigid-walled duct with a J-deflector. The relative performance of closed duct and open duct is evaluated. The results show that the closed duct is superior to the partially open duct, and results in about 3-decibel (dB) noise reduction (near the duct axis) relative to the free jet. The location of the nozzle exit plane (NEP) relative to the duct inlet plane (DIP) has a significant effect on the acoustic field. The results suggest that the location of NEP at 10 inches above the DIP results in reduced acoustic loads relative to 5 inches above the duct inlet and 1 inch into the duct inlet.

Kandula, Max↗

DOE OSTI2021

Differential localization patterns of pyruvate kinase isoforms in murine naïve, formative, and primed pluripotent states

Highlights: • PKM1/2 protein abundance is greater in formative mEpiLCs compared to naïve mESCs or primed mEpiSCs. The ratio of PKM1/2 is maintained. • PKM1/2 are localized in both nuclear and cytoplasmic regions relative to GAPDH and OCT4 across the pluripotent continuum. • PKM1 localization is strongly correlated to OCT4 localization, and moderately to GAPDH in formative mEpiLCs. • PKM1/2 localization is moderately correlated to OCT4 and GAPDH localization in naïve mESCs. • PKM1/2 localization are moderately correlated to GAPDH localization in primed mEpiSCs. Mouse embryonic stem cells (mESCs) and mouse epiblast stem cells (mEpiSCs) represent opposite ends of the pluripotency continuum, referred to as naïve and primed pluripotent states, respectively. These divergent pluripotent states differ in several ways, including growth factor requirements, transcription factor expression, DNA methylation patterns, and metabolic profiles. Naïve cells employ both glycolysis and oxidative phosphorylation (OXPHOS), whereas primed cells preferentially utilize aerobic glycolysis, a trait shared with cancer cells referred to as the Warburg Effect. Until recently, metabolism has been regarded as a by-product of cell fate, however, evidence now supports metabolism as being a driver of stem cell state and fate decisions. Pyruvate kinase muscle isoforms (PKM1 and PKM2) are important for generating and maintaining pluripotent stem cells (PSCs) and mediating the Warburg Effect. Both isoforms catalyze the final, rate limiting step of glycolysis, generating adenosine triphosphate and pyruvate, however, the precise role(s) of PKM1/2 in naïve and primed pluripotency is not well understood. The primary objective of this study was to characterize the cellular expression and localization patterns of PKM1 and PKM2 in mESCs, chemically transitioned epiblast-like cells (mEpiLCs) representing formative pluripotency, and mEpiSCs using immunoblotting and confocal microscopy. The results indicate that PKM1 and PKM2 are not only localized to the cytoplasm, but also accumulate in differential subnuclear regions of mESC, mEpiLCs, and mEpiSCs as determined by a quantitative confocal microscopy employing orthogonal projections and airyscan processing. Importantly, we discovered that the subnuclear localization of PKM1/2 changes during the transition from mESCs, mEpiLCs, and mEpiSCs. Finally, we have comprehensively validated the appropriateness and power of the Pearson's correlation coefficient and Manders's overlap coefficient for assessing nuclear and cytoplasmic protein colocalization in PSCs by immunofluorescence confocal microscopy. We propose that nuclear PKM1/2 may assist with distinct pluripotency state maintenance and lineage priming by non-canonical mechanisms. These results advance our understanding of the overall mechanisms controlling naïve, formative, and primed pluripotency.

60 APPLIED LIFE SCIENCES↗

NASA NTRS2006

Embedded Sensors for Measuring Surface Regression

The development and evaluation of new hybrid and solid rocket motors requires accurate characterization of the propellant surface regression as a function of key operational parameters. These characteristics establish the propellant flow rate and are prime design drivers affecting the propulsion system geometry, size, and overall performance. There is a similar need for the development of advanced ablative materials, and the use of conventional ablatives exposed to new operational environments. The Miniature Surface Regression Sensor (MSRS) was developed to serve these applications. It is designed to be cast or embedded in the material of interest and regresses along with it. During this process, the resistance of the sensor is related to its instantaneous length, allowing the real-time thickness of the host material to be established. The time derivative of this data reveals the instantaneous surface regression rate. The MSRS could also be adapted to perform similar measurements for a variety of other host materials when it is desired to monitor thicknesses and/or regression rate for purposes of safety, operational control, or research. For example, the sensor could be used to monitor the thicknesses of brake linings or racecar tires and indicate when they need to be replaced. At the time of this reporting, over 200 of these sensors have been installed into a variety of host materials. An MSRS can be made in either of two configurations, denoted ladder and continuous (see Figure 1). A ladder MSRS includes two highly electrically conductive legs, across which narrow strips of electrically resistive material are placed at small increments of length. These strips resemble the rungs of a ladder and are electrically equivalent to many tiny resistors connected in parallel. A substrate material provides structural support for the legs and rungs. The instantaneous sensor resistance is read by an external signal conditioner via wires attached to the conductive legs on the non-eroding end of the sensor. The sensor signal can be transmitted from inside a high-pressure chamber to the ambient environment, using commercially available feedthrough connectors. Miniaturized internal recorders or wireless data transmission could also potentially be employed to eliminate the need for producing penetrations in the chamber case. The rungs are designed so that as each successive rung is eroded away, the resistance changes by an amount that yields a readily measurable signal larger than the background noise. (In addition, signal-conditioning techniques are used in processing the resistance readings to mitigate the effect of noise.) Hence, each discrete change of resistance serves to indicate the arrival of the regressing host material front at the known depth of the affected resistor rung. The average rate of regression between two adjacent resistors can be calculated simply as the distance between the resistors divided by the time interval between their resistance jumps. Advanced data reduction techniques have also been developed to establish the instantaneous surface position and regression rate when the regressing front is between rungs.

Gramer, Daniel J.↗