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Wilson Corners SWMU 001 2014 Annual Long Term Monitoring Report Kennedy Space Center, Florida

This document presents the findings of the 2014 Long Term Monitoring (LTM) that was completed at the Wilson Corners site, located at the National Aeronautics and Space Administration (NASA) John F. Kennedy Space Center (KSC), Florida. The goals of the 2014 annual LTM event were to evaluate the groundwater flow direction and gradient and to monitor the vertical and downgradient horizontal extent of the volatile organic compounds (VOCs) in groundwater at the site. The LTM activities consisted of an annual groundwater sampling event in December 2014, which included the collection of water levels from the LTM wells. During the annual groundwater sampling event, depth to groundwater was measured and VOC samples were collected using passive diffusion bags (PDBs) from 30 monitoring wells. In addition to the LTM sampling, additional assessment sampling was performed at the site using low-flow techniques based on previous LTM results and assessment activities. Assessment of monitoring well MW0052DD was performed by collecting VOC samples using low-flow techniques before and after purging 100 gallons from the well. Monitoring well MW0064 was sampled to supplement shallow VOC data north of Hot Spot 2 and east of Hot Spot 4. Monitoring well MW0089 was sampled due to its proximity to MW0090. MW0090 is screened in a deeper interval and had an unexpected detection of trichloroethene (TCE) during the 2013 LTM, which was corroborated during the March 2014 verification sampling. Monitoring well MW0130 was sampled to provide additional VOC data beneath the semi-confining clay layer in the Hot Spot 2 area.

Monitoring↗

Wilson Corners, Solid Waste Management Unit 001(SWMU 01) 2023 Annual Long-Term Monitoring Report

This report presents a summary of the long-term monitoring (LTM) activities that occurred in 2023 at Wilson Corners, Solid Waste Management Unit 001, at Kennedy Space Center (KSC), Florida. Annual LTM of groundwater is being conducted at the site. Based on results from groundwater sampling activities performed during the 2019 through 2020 LTM reporting period and the 2020 and 2021 DPT groundwater sampling, it was determined that the LTM sampling plan was no longer meeting the goal of LTM because delineation was not verified and the installation of an air sparge (AS) system to treat the area of the High Concentration Plume was recommended. The AS System was installed in late 2022 and early 2023. System start-up activities were initiated in April 2023. Following system startup, several site wells required retrofitting to equip wellheads for withstanding the air pressure released from air sparge wells during system operation. Some site wells also required repair or abandonment, and replacement. Survey of location and top-of-casing of newly installed monitoring wells was combined with scheduled AS system survey activities and was completed in January 2024. The activities presented in this report include the February and April 2023 LTM monitoring well installations; March and April 2023 LTM and performance monitoring well water level gauging and sampling; November 2023 LTM well retrofits and repairs; a summary of December 2023 LTM well abandonments and installations (complete site well abandonment activities will be presented under a separate cover); and January 2024 LTM well survey. During the March and April 2023 sampling events, the low-flow sampling method was used, and samples were analyzed for a select list of volatile organic compounds. In March 2023, groundwater flow for the site was generally to the west was generally consistent with historical observations at the site. The Low Concentration Plume (LCP) continues to extend both horizontally and vertically beyond the terminal depth of the current monitoring well network. Data, inclusive of the 2023 LTM and baseline performance monitoring sampling events, indicate that the LCP encompasses an estimated 19.5 acres, compared to the 2021 LCP footprint, inclusive of the 2020 and 2021 DPT sampling events of 20.7 acres. The vertical extent of VOCs was historically delineated by monitoring wells screened greater than 48 feet below land surface (bls). The results from the three vertical extent monitoring wells screened below 48 feet bls that were sampled during the 2023 LTM indicate that groundwater vinyl chloride concentrations in these three wells are greater than the GCTL. As presented in the 2021 Long-Term Monitoring Report (NASA 2022), the KSCRT agreed to delay deeper investigations in this area to prevent the creation of additional pathways for vertical migration. Based on groundwater sampling activities performed in 2023, recommendations are to perform the next annual LTM sampling event, scheduled for April 2024 and to conduct quarterly performance monitoring of the AS System. The current selection of monitoring wells in the recommended 2024 LTM plan will provide an adequate data set for monitoring groundwater plume behavior; however, the LTM monitoring well network will be evaluated and refined based on 2024 LTM and year one performance monitoring data.

King Linnea↗

Machine Learning Vacancy Formation Energy in Nickel-Based Superalloys

Thermal vacancies play a critical role in high-temperature Ni-based superalloys and influence various properties such as creep resistance, oxidation, etc. This study systematically investigates the impact of commonly used transition metals (Cr, Co, Fe), refractory metals (Nb, Ta, Mo, W) and other elements (Al, Cu, Ti, Mn) on the thermodynamic stability of 36 binary, 20 ternary, 11 quaternary, 9 quinary, and 3 senary FCC Ni-based alloys covering various elemental combinations. Density functional theory-based studies on Ni-X binary alloys show that higher concentrations of Cr, Nb, Ta, Al, and Ti introduce significant lattice distortions and broaden the distribution of vacancy formation energies (standard deviation up to 0.15 eV). These elements partially donate electrons, reducing their self-consistent chemical potentials relative to single-element reference values and lowering vacancy formation energies, while Co, Fe, Mo, and W show lower charge localization. These trends extend from 3-6 element alloys, where Cr, Nb, and Ta-rich compositions have low-energy states (~0.5 eV) that increase vacancy concentrations. Finally, graph neural network models are developed to screen over 5000 virtual alloys. Eleven leading compositions are identified with mean vacancy formation energy higher than 1.75 eV and vacancy concentration ~2 orders of magnitude lower than pure Ni at 1000 K. These results provide valuable guidelines to achieve controlled defect engineering in structural alloys.

DFT↗

The effect of composition on helium bubble nucleation in complex concentrated alloys

Molecular dynamics simulations of helium implantation have been performed in the MoNbTaTi complex concentrated alloy (CCA) to determine the impact of composition on helium clustering and bubble nucleation. It was found that compositions with lower interstitial atomic volume resulted in smaller cluster sizes, higher helium-to-vacancy ratios, and larger migration barriers. As the atomic volume decreases, it is harder to accommodate additional helium in the lattice and for helium to diffuse between interstitial sites, resulting in smaller cluster sizes. In particular, increasing molybdenum directly correlated with a decreased interstitial atomic volume and increased migration barrier. Niobium exhibited the opposite trend, where more niobium in the material resulted in larger interstitial atomic volumes and cluster sizes but lower migration barriers. The atomic volume was determined to be an indicator of the susceptibility of the material to helium bubble growth and could potentially be used as a metric to screen compositions for resistance to helium damage. In this way, properties such as available interstitial volumes could be used to screen materials more rapidly given the vast compositional space of CCAs.

McCarthy, Megan Jeanne [Sandia National Laboratori↗

Groundwater level elevation and temperature data, Oct 2018-Dec 2021, Slate River Floodplain, Crested Butte, CO

This data package includes a time series of water level and temperature measurements from October 2018 to December 2021 in groundwater and surface water from the Slate River floodplain outside Crested Butte, CO, a focus field site for the SLAC Floodplain Hydro-Biogeochemistry SFA. The data was generated as part of the work targeting the overarching research question for the SLAC SFA: How do ubiquitous subsurface interfaces mediate molecular-scale biogeochemical processes and groundwater quality in floodplains and watersheds? The data were recorded by pressure transducers were installed in four types of piezometers: 1) a network of gravel bed ("GB") drive-point piezometers with a 6-inch screen interval installed all at ~330 cm below ground surface. 2) a network of piezometers screened across the water table, used to measure evapotranspiration ("ET") using the White method. Each of these piezometers is screened along almost its entire length.3) a suite nested piezometers used the measure the vertical hydraulic gradient ("VHG") across the fines-cobble interface. Each of these piezometers uses a 6-inch screen length.4) a group of piezometers scattered across the boundaries of the floodplain, used to monitor boundary conditions ("BC") flowing into and out of the floodplain. With the exception of "SR-BD-WT", each of these piezometers is screened along its entire length. Within the data package, "FLMD.csv" describes file-level metadata and "dd.csv" defines column headers and universal terms across the dataset. The data package includes 11 "*data.csv" files, one for each piezometer type for each calendar year. Because piezometers have been added over time, not every sensor has data dating back to Oct 2018. Each "*data.csv" file has a corresponding "*_InstallationMethods.csv" file that describes the location, elevation, screen depth, sediment type and sensor metadata for each piezometer and pressure transducer.

54 ENVIRONMENTAL SCIENCES↗

Enhancement of Optical Efficiency of CSP Mirrors for Reducing O&M Cost via Near-Continuous Operation of Self-Cleaning Electrodynamic Screens (EDS). Final Report

Over the past decade, techno-economical advancements in solar energy systems, particularly the improvement in the conversion efficiency of PV modules made with mono-crystalline silicon solar cells from 12% to 20%, as well as the cost reduction in manufacturing by a factor of about 10%, have made it possible to achieve a levelized cost of electricity (LCOE) in PV plants that is comparable to, or less than, the cost of deriving electricity from fossil fuels. Operating PV plants in mid-latitude sun-belt regions, where the solar irradiance level is highest, provides a high annual energy-yield (kWh/kWp) due to two factors: (1) the availability of predictable high solar irradiance throughout the year with the fewest interruptions in solar flux from clouds and rain, and (2) the increased conversion efficiency of crystalline solar cells, as recombination loss has decreased with increased intensity of the sunlight that illuminates the silicon solar cells. Semi-arid and desert regions, however, are plagued by high atmospheric dust concentrations and frequent sand storms. The deposition of a layer of dust on the optical surfaces of solar collectors such as PV modules and concentrating mirrors reduces the transmission efficiency of sunlight that actually reaches the solar cells or receivers, resulting in high energy-yield soiling loss. There are two major cost components to operating a solar plant: (1) installation costs, and (2) operation-and-maintenance (O&M) costs. There is no fuel cost; hence operating a solar plant in a semi-arid or desert region provides high returns on investment if soiling losses are mitigated via efficient cleaning methods and optimized cleaning frequency. If solar collectors are not cleaned, the accumulation of dust layers on solar collectors may cause the operation of such plants in arid regions to become economically unviable. Washing solar collectors with water and detergent, as is most commonly done now, is an efficient method for cleaning. The conventional approach in utility-scale solar plants is to use a large truck with a water tank and pump system for spraying deionized water on the surface of the solar collectors. Robotic cleaning with brushes, used for many solar plants, requires lesser water for cleaning. The water consumed using semi-automated cleaning of PV modules in utility-scale solar plants is approximately 2 liters/m 2 per cleaning cycle. The total optical surface area of the solar collectors in 1 TW-scale solar installation will be more than 3 × 109 m 2 ; hence an enormous amount of water be needed for cleaning. There simply is not enough fresh water in the sun-belt areas of the world for predicted cleaning needs. In solar power plants, the estimated cost of cleaning solar collectors includes expenses related to the equipment used, labor, cost of transportation of water, the energy required for cleaning, plus ancillary costs whereas the cost of water is not considered. The water used is obtained from sources located close to plant sites, unmindful of the environmental and societal impact as the power plants are oftentimes located in regions that face severe drought. This practice is very similar to the cost calculations in deriving the levelized cost of electricity (LCOE) in conventional power plants based on burning fossil fuel such as coal and gas, while disregarding the cost of climate change and health effects. Unless a water-free or low-water cleaning method is established, the expansion of solar plants may lose public support in areas suffering long intervals of drought. The goal of this research project has been the development and application of the Electrodynamic Screen (EDS) as a means for a water-free, scalable cleaning process applicable to solar-power installations, including rooftop applications. We describe here the development of an EDS film-based cleaning process as an emerging method for use on PV modules, parabolic troughs, and heliostats. This report aims to show the feasibility of integrating or retrofitting EDS films onto the optical surfaces of solar collectors (both PV and CSP) while maintaining high transmission or reflection efficiency. The cleaning action provided by the EDS film is an active method to remove dust deposits by electrodynamic force. Current lab-scale prototype EDS films, retrofitted onto solar panels and mirrors, have shown to be capable of maintaining optical transmission or specular-reflection efficiencies higher than 90% of initial values under clean conditions. The optical surfaces of solar collectors laminated with EDS films can remove more than 90% of deposited dust when the EDS is activated for less than two minutes. As an electrodynamic dust removal process, the EDS film-based method is designed primarily for the removal of dust in solar installations located in semi-arid and desert areas, where the atmosphere is often dry and dusty and rainfall is infrequent. While EDS film application minimizes water consumption and facilitates cleaning as frequently as needed, it has limitations in removing contaminants such as soot, organic pollutants deposited as fine films on the surface, and bird droppings. The dust removal efficiency of the EDS is maximum at relative humidity RH is 40 to 50% and decreases at when RH > 65%. Many solar plant sites undergo diurnal and seasonal cycles of high ambient, early afternoon temperatures and an RH that reaches the dew point early in the morning. These variations in atmospheric conditions do not limit the operation of the EDS film. This report presents a brief review of the progress and the potential of EDS film technology for mitigating the impact of dust on solar collectors via water-free cleaning, as well as current technical challenges regarding efficiency and durability. Our experimental data on the performance of EDS films show that: (1) the dust-removal efficiency (DRE) can reach levels higher than 90%, (2) the specular reflectivity (SR) of EDS film-laminated second-surface mirrors reach levels in excess of 90%, (3) the specular reflectivity restoration (SRR) can exceed 90%, (4) the output-power restoration (OPR) of PV modules can exceed 95%, and (5) the optical transmission efficiency (TE) of the EDS films can be greater than 90%. Working with Sandia National Laboratories (NM), Corning Research and Development Corporation (NY), Eastman Kodak (NY), Tomark-Worthen Industries (NH), and EDS Chile SPA (Chile), we have produced EDS film-laminated PV modules and demonstrated their self-cleaning functions without requiring water. We have demonstrated that the operational range of EDS films will cover the expected ambient temperature of solar fields at RH cycling varying from 20 to 95% as long as the EDS films are activated in the RH range 20 to 50%. (Typical solar-field climates in deserts and semi-arid lands often reach near dew point in coastal areas.) Our experiments on the application of hydrophobic-fluorinated nanoparticle coatings on EDS film surfaces show that the EDS operational range can be extended to higher RH levels that approach the dew point. At Eastman Kodak, as one of our industrial partners, we were able to establish a process for manufacturing EDS films using flexographic printing of the electrodes onto transparent polymer films. This process utilizes an existing manufacturing line at Eastman Kodak that allows fabrication of medium-scale EDS films (26 cm × 30 cm). The manufacturing process has the capacity to produce EDS films at high production speeds. The medium-scale EDS films that have been printed at Kodak were evaluated in the lab at Boston University. The EDS films produced at Kodak were laminated at Tomark-Worthen using an industrial scale vacuum laminator to produce EDS film stacks, which can be affixed onto the optical surfaces of PV modules or concentrating mirrors. The EDS film stack consists of the EDS films that have Willow® Glass (WG) which has a thickness of 100 μm as the front surface. The WG sheets obtained from Corning Research and Development Corporation are customized in size and shape to cover the active area of the EDS films. The back surface of the EDS film stack is integrated onto the optical surface of the solar panel or mirror using optically clear adhesive (OCA) films or silicone adhesives. The OCA films (thickness 25 μm) are produced by 3M. The EDS film stacks have the architecture: WG/OCA/EDS Film/OCA/ over PV module or solar mirror. The power-supply units needed for activating the electrodes of the EDS film were designed and produced at Boston University. These power supply units provide three-phase, 1.2 kV voltage pulses at a very low current (micro-ampere) level and at a low frequency (≈ 5 Hz). The voltage pulses are applied to the electrodes in a sequence such that the train of pulses resembles a unidirectional traveling wave of electrical field on the surface of the EDS film. The dust particles on the surface become charged electrostatically and are levitated by the Coulomb force. The lateral sweeping action of the traveling electric field created by the three-phase voltages pulses then sweeps the dust off the surface. The energy consumed by the EDS electrodes is less than 0.2 Wh/m 2 /cleaning cycle, enabling energy-efficient restoration of output power (OPR) of PV module or specular reflectivity restoration (SRR) for solar mirrors. The EDS system consists of (1) an EDS film stack laminated onto the solar collectors, (2) connection of the EDS film to its power supply unit and (3) Interconnection of the power supply to PV modules or solar mirrors. Design, construction and assessment of field-testing units that have EDS stack laminated PV modules for evaluating the performance of the EDS films in solar fields is being carried out at BU. Our progress under this project, aimed at the advancement of EDS film technology, has reached DOE Technology Readiness Level (TRL) 6. Based on the extensive laboratory evaluations and limited field trials, as well as contacts with potential users, we believe that the technology has reached its commercial stage. We are conducting a cost analysis using the National Renewable Energy Laboratory (NREL) System Advisory Model (SAM) and are preparing for field trials of EDS films in different solar fields in the US, Chile, India and in the Middle East. A brief description of EDS film performance, construction and testing of the field-test unit, autonomous operation of the field-test unit for evaluating EDS performance in increasing energy yield, associated revenue savings, and water conservation is presented.

14 SOLAR ENERGY↗

Cancer Incidence Trends in Successive Social Generations in the US

Importance: The incidence of some cancers in the US is increasing in younger age groups, but underlying trends in cancer patterns by birth year remain unclear. Objective: To estimate cancer incidence trends in successive social generations. Design, Setting, and Participants: In this cohort study, incident invasive cancers were ascertained from the Surveillance, Epidemiology, and End Results (SEER) program’s 13-registry database (November 2020 submission, accessed August 14, 2023). Invasive cancers diagnosed at ages 35 to 84 years during 1992 to 2018 within 152 strata were defined by cancer site, sex, and race and ethnicity. Exposure: Invasive cancer. Main Outcome and Measures: Stratum-specific semiparametric age-period-cohort (SAGE) models were fitted and incidence per 100 000 person-years at the reference age of 60 years was calculated for single-year birth cohorts from 1908 through 1983 (fitted cohort patterns [FCPs]). The FCPs and FCP incidence rate ratios (IRRs) were compared by site for Generation X (born between 1965 and 1980) and Baby Boomers (born between 1946 and 1964). Results: A total of 3.8 million individuals with invasive cancer (51.0% male; 8.6% Asian or Pacific Islander, 9.5% Hispanic, 10.4% non-Hispanic Black, and 71.5% non-Hispanic White) were included in the analysis. In Generation X vs Baby Boomers, FCP IRRs among women increased significantly for thyroid (2.76; 95% CI, 2.41-3.15), kidney (1.99; 95% CI, 1.70-2.32), rectal (1.84; 95% CI, 1.52-2.22), corpus uterine (1.75; 95% CI, 1.40-2.18), colon (1.56; 95% CI, 1.27-1.92), and pancreatic (1.39; 95% CI, 1.07-1.80) cancers; non-Hodgkins lymphoma (1.40; 95% CI, 1.08-1.82); and leukemia (1.27; 95% CI, 1.03-1.58). Among men, IRRs increased for thyroid (2.16; 95% CI, 1.87-2.50), kidney (2.14; 95% CI, 1.86-2.46), rectal (1.80; 95% CI, 1.52-2.12), colon (1.60; 95% CI, 1.32-1.94), and prostate (1.25; 95% CI, 1.03-1.52) cancers and leukemia (1.34; 95% CI, 1.08-1.66). Lung (IRR, 0.60; 95% CI, 0.50-0.72) and cervical (IRR, 0.71; 95% CI, 0.57-0.89) cancer incidence decreased among women, and lung (IRR, 0.51; 95% CI, 0.43-0.60), liver (IRR, 0.76; 95% CI, 0.63-0.91), and gallbladder (IRR, 0.85; 95% CI, 0.72-1.00) cancer and non-Hodgkins lymphoma (IRR, 0.75; 95% CI, 0.61-0.93) incidence decreased among men. For all cancers combined, FCPs were higher in Generation X than for Baby Boomers because gaining cancers numerically overtook falling cancers in all groups except Asian or Pacific Islander men. Conclusions and Relevance: In this model-based cohort analysis of incident invasive cancer in the general population, decreases in lung and cervical cancers in Generation X may be offset by gains at other sites. Generation X may be experiencing larger per-capita increases in the incidence of leading cancers than any prior generation born in 1908 through 1964. On current trajectories, cancer incidence could remain high for decades.

60 APPLIED LIFE SCIENCES↗

An in vitro BRAF activation assay elucidates molecular mechanisms driving disassembly of the autoinhibited BRAF state

The RAF kinases (ARAF, BRAF, and CRAF) are essential components of the RAS-ERK signaling pathway, which controls vital cellular processes and is frequently dysregulated in human disease. Notably, mutations that alter BRAF function are prominent drivers of human cancer and certain RASopathy disorders, making BRAF an important target for therapeutic intervention. Despite extensive research, several aspects of BRAF regulation remain unclear. In this study, we developed an in vitro BRAF activation assay using purified autoinhibited BRAF:14-3-3 2 :MEK complexes. Our results show that fully processed, active-state KRAS alone can promote dimer-dependent BRAF activation. Moreover, we found that phosphatidylserine (PS)-containing liposomes synergized with KRAS to promote BRAF activation, achieving activity levels comparable to those observed with BRAF proteins that constitutively dimerize. In contrast, the SMP phosphatase complex had only a minimal effect on BRAF catalytic activity in this system but mediated the dephosphorylation of the negative regulatory pS365 14-3-3 binding site in a manner that was accelerated by the presence of KRAS alone or KRAS and 30% PS liposomes. Finally, we show that inhibitors blocking the BRAF RBD:KRAS interaction were able to suppress the in vitro activation of BRAF, underscoring the critical role of RAS binding in initiating the disassembly of the BRAF autoinhibited state. Thus, this assay provides valuable insights into the steps required for BRAF activation and can serve as an effective screening tool for identifying compounds that may inhibit this process and have therapeutic potential.

BRAF↗

Characterization testing of a 40 Ahr bipolar nickel hydrogen battery

In a continuing effort to develop NiH2 bipolar technology to a point where it can be used efficiently in space flight, testing of a second 40 Ahr, 10-cell bipolar battery has begun. This battery has undergone extensive characterization testing to determine the effects of such operating parameters as charge and discharge rates, temperature, and pressure. The fundamental design of this actively cooled bipolar battery is the same as the first battery. Most of the individual components, however, are from different manufacturers. Different testing procedures as well as certain unique battery characteristics make it difficult to directly compare the two sets of results. In general, the performance of this battery throughout characterization produced expected results. The main differences seen between the first and second batteries occurred during the high-rate discharge portion of the test matrix. The first battery also had poor high-rate discharge results, although better than those of the second battery. Minor changes were made to the battery frame design used for the first battery in an attempt to allow better gas access to the reaction sites for the second build and hopefully improve performance. The changes, however, did not improve the performance of the second battery and could have possibly contributed to the poorer performance that was observed. There are other component differences that could have contributed to the poorer performance of the second battery. The H2 electrode in the second battery was constructed with a Goretex backing which could have limited the high-rate current flow. The gas screen in the second battery had a larger mesh which again could have limited the high-rate current flow. Small scale 2 x 2 batteries are being tested to evaluate the effects of the component variations.

Brewer, Jeffrey C.↗

Bifunctional Electrocatalysts with High-Entropy Alloys: Bridging Hydrogen Evolution and Oxygen Reduction

High-entropy alloys (HEAs) have emerged as a promising class of bifunctional electrocatalysts capable of simultaneously driving the hydrogen evolution reaction (HER) and the oxygen reduction reaction (ORR) with high activity and durability. Their near-equiatomic multicomponent compositions give rise to unique physicochemical characteristics, including lattice distortion, sluggish diffusion, high-entropy stabilization, and pronounced electronic heterogeneity, that collectively generate diverse and synergistic active sites inaccessible in conventional alloys. This review summarizes recent progress in HEA-based bifunctional electrocatalysis, with a focus on the fundamental mechanisms governing HER and ORR activity, stability, and selectivity. We discuss advances in synthesis strategies, ranging from confined growth and step-alloying to scalable continuous-flow methods, that enable precise control over composition, size, and surface structure. Complementary computational and data-driven approaches, including density functional theory, machine-learning-assisted screening, and descriptor development, are highlighted as essential tools for navigating the vast HEA design space and establishing structure−property relationships. Particular attention is paid to adsorption-energy distributions, multisite cooperativity, and environmental effects under realistic electrochemical conditions. Finally, we outline current challenges and future opportunities for integrating mechanistic understanding with AI-guided, closed-loop design frameworks to accelerate the discovery of next-generation HEA bifunctional electrocatalysts for sustainable energy conversion.

Alloys↗

Genome dependent Cas9/gRNA search time underlies sequence dependent gRNA activity

Abstract CRISPR-Cas9 is a powerful DNA editing tool. A gRNA directs Cas9 to cleave any DNA sequence with a PAM. However, some gRNA sequences mediate cleavage at higher efficiencies than others. To understand this, numerous studies have screened large gRNA libraries and developed algorithms to predict gRNA sequence dependent activity. These algorithms do not predict other datasets as well as their training dataset and do not predict well between species. Here, to better understand these discrepancies, we retrospectively examine sequence features that impact gRNA activity in 44 published data sets. We find strong evidence that gRNA sequence dependent activity is largely influenced by the ability of the Cas9/gRNA complex to find the target site rather than activity at the target site and that this drives sequence dependent differences in gRNA activity between different species. This understanding will help guide future work to understand Cas9 activity as well as efforts to identify optimal gRNAs and improve Cas9 variants.

59 BASIC BIOLOGICAL SCIENCES↗

Evaluation of Triage Methods for Criticality Accidents

Studies indicate that early identification of persons involved in and receiving high doses of radiation in accidents is key to providing life-saving medical treatment. Although the risk of criticality accidents is low the potential impact to workers is significant. For facilities that employ large numbers of workers a key element in the response to a radiological emergency is identifying personnel that received significant, and potentially harmful, doses. Also important is having the ability to screen large numbers of workers to identify persons that did not receive significant exposure so as to reduce the impact on emergency response efforts. At the Y-12 National Security Complex the focus on criticality accident response is the rapid triage of personnel in order to identify persons exposed to large radiation doses and to prioritize those persons receiving the highest exposures. Once identified personnel are transported to local medical facilities including the Radiation Emergency Assistance Center/Training Site (REAC/TS) for medical evaluation and treatment. The Y-12 external dosimetry program utilizes a number of techniques to identify and prioritize workers and these methods were evaluated at a criticality dosimetry intercomparison exercise. Finally, the methods used were shown to perform as intended and other sites may consider incorporating these methods into their accident dosimetry response procedures.

61 RADIATION PROTECTION AND DOSIMETRY↗

The Micro-Pulse Lidar Network (MPLNET): A Federated Network of Micro-pulse Lidars and AERONET Sunphotometers

We present the formation of a new global-ground based eye-safe lidar network, the NASA Micro-Pulse Lidar Network (MPLNET). The aim of MPLNET is to acquire long- term observations of aerosol and cloud vertical profiles at unique geographic sites within the NASA Aerosol Robotic Network (AERONET). MPLNET utilizes standard instrumentation and data processing algorithms for efficient network operations and direct comparison of data between each site. The micro-pulse lidar is eye-safe, compact, and commercially available, and most easily allows growth of the network without sacrificing standardized instrumentation goals. Network growth follows a federated approach, pioneered by AERONET, wherein independent research groups may join MPLNET with their own instrument and site. MPLNET sites produce not only vertical profile data, but also column-averaged products already available from AERONET (aerosol optical depth, sky radiance, size distributions). Algorithms are presented for each MPLNET data product. Real-time Level 1 data products (next-day) include daily lidar signal images from the surface to -2Okm, and Level 1.5 aerosol extinction profiles at times co-incident with AERONET observations. Quality assured Level 2 aerosol extinction profiles are generated after screening the Level 1.5 results and removing bad data. Level 3 products include continuous day/night aerosol extinction profiles, and are produced using Level 2 calibration data. Rigorous uncertainty calculations are presented for all data products. Analysis of MPLNET data show the MPL and our analysis routines are capable of successfully retrieving aerosol profiles, with the strenuous accounting of uncertainty necessary for accurate interpretation of the results.

Welton, Ellsworth J.↗

Wellbore integrity assessment with casing-based advanced sensing

Wellbore integrity is of paramount importance to subsurface resource extraction, energy storage 1 and waste disposal. After installation, well casing and cement are subject to mechanical stress due to near-well pressure changes and fluid induced corrosion. This is exacerbated for geothermal wells where produced fluid is at high temperature and corrosive. The current state-of-the-art technologies for wellbore integrity assessments are an array of cased hole logging tools. Wireline deployed acoustic, electromagnetic and mechanical tools are all available to inspect steel casing corrosion and casing-cement bond and these tools can provide high-resolution assessment of borehole conditions. They are intrusive, however, in terms of borehole preparation and interruption to the normal operation of the wells, and not suitable for high temperature or highly deviated well deployments. In addition, these measurements are performed infrequently due to high cost, and are therefore incapable of providing frequent data to better predict borehole degradation trajectory, which can help provide early warning of potential borehole failures. For this project we are developing a suite of novel, non-invasive, casing based tools for wellbore integrity assessment, combining fast/low cost screening with higher-precision investigation. Our approach is based on monitoring the response of the casing when energized at the wellhead, thereby interrogating the casing without well intervention. Lab, field and numerical approaches are used in our study. During the early stage of the research, we focus on numerical simulations, which have shown the sensitivity of the low frequency electromagnetic (EM) signals to changes in borehole depths and have successfully tested the concept at a field site with different length well casings. Initial seismic modeling efforts have also demonstrated our capability to simulate seismic tube wave and seismic field alterations due to borehole breakage and associated fluid leakage. Further numerical, laboratory and field experiments are underway for additional technology sensitivity analysis, particularly the transient EM/Seismic reflectometry methods, data acquisition optimization, and numerical simulation improvements.

Wilt, Michael↗

Potent transmission-blocking monoclonal antibodies from naturally exposed individuals target a conserved epitope on Plasmodium falciparum Pfs230

Pfs230 is essential for Plasmodium falciparum transmission to mosquitoes and is the protein targeted by the most advanced malaria-transmission-blocking vaccine candidate. Prior understanding of functional epitopes on Pfs230 is based on two monoclonal antibodies (mAbs) with moderate transmission-reducing activity (TRA), elicited from subunit immunization. Here, we screened the B cell repertoire of two naturally exposed individuals possessing serum TRA and identified five potent mAbs from sixteen Pfs230 domain-1-specific mAbs. Structures of three potent and three low-activity antibodies bound to Pfs230 domain 1 revealed four distinct epitopes. Highly potent mAbs from natural infection recognized a common conformational epitope that is highly conserved across P. falciparum field isolates, while antibodies with negligible TRA derived from natural infection or immunization recognized three distinct sites. Our study provides molecular blueprints describing P. falciparum TRA, informed by contrasting potent and non-functional epitopes elicited by natural exposure and vaccination.

60 APPLIED LIFE SCIENCES↗

Monoclonal antibody to a cancer-specific and drug-responsive hydroquinone (NADH) oxidase from the sera of cancer patients

Monoclonal antibodies were generated in mice to a 34-kDa circulating form of a drug-responsive hydroquinone (NADH) oxidase with a protein disulfide-thiol interchange activity specific to the surface of cancer cells and the sera of cancer patients. Screening used Western blots with purified 34-kDa tNOX from HeLa cells and the sera of cancer patients. Epitopes were sought that inhibited the drug-responsive oxidation of NADH with the sera of cancer patients, but which had no effect on NADH oxidation with the sera of healthy volunteers. Two such antisera were generated. One, designated monoclonal antibody (mAb) 12.1, was characterized extensively. The NADH oxidase activity inhibited by mAb 12.1 also was inhibited by the quinone site inhibitor capsaicin (8-methyl- N-vanillyl-6-noneamide). The inhibition was competitive for the drug-responsive protein disulfide-thiol interchange activity assayed either by restoration of activity to scrambled RNase or by cleavage of a dithiodipyridine substrate, and was uncompetitive for NADH oxidation. Both the mAb 12.1 and the postimmune antisera immunoprecipitated drug-responsive NOX activity and identified the same 34-kDa tNOX protein in the sera of cancer patients that was absent from sera of healthy volunteers, and was utilized as immunogen. Preimmune sera from the same mouse as the postimmune antisera was without effect. Both mouse ascites containing mAb 12.1 and postimmune sera (but not preimmune sera) slowed the growth of human cancer cell lines in culture, but did not affect the growth of non-cancerous cell lines. Immunocytochemical and histochemical findings showed that mAb 12.1 reacted with the surface membranes of human carcinoma cells and tissues.

NASA Discipline Cell Biology↗

Thorium Bis‐Salophen Trimers as Anion Detectors and Binding Agents

Bis‐salophen ligands are the condensation product of a tetramine and a salicylaldehyde derivative. They feature two binding sites, both of which are tetradentate with mixed O/N donor atoms. Reaction with the ligand precursor and thorium nitrate tetrahydrate forms a 3:3 metal‐to‐ligand trimer with a ΔΔΔ‐chirality confirmed by X‐ray crystallography of a racemic single crystal. The structure has a pore in the center of the triangular structure measuring 6.22 Å at its narrowest point. This compound is air‐ and water‐stable as well as soluble in organic solvents. Here, this compound was screened with a series of tetrabutyl ammonium halide salts, and tetrabutylammonium (TBA) chloride showed the strongest binding to the complex. After the addition of 2 equivalents of TBACl, the complex and salt precipitate out of solution.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Molecular characterization of ebselen binding activity to SARS-CoV-2 main protease

There is an urgent need to repurpose drugs against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Recent computational-experimental screenings have identified several existing drugs that could serve as effective inhibitors of the virus’ main protease, M pro , which is involved in gene expression and replication. Among these, ebselen (2-phenyl-1,2-benzoselenazol-3-one) appears to be particularly promising. Here, we examine, at a molecular level, the potential of ebselen to decrease Mpro activity. We find that it exhibits a distinct affinity for the catalytic region. Our results reveal a higher-affinity, previously unknown binding site localized between the II and III domains of the protein. A detailed strain analysis indicates that, on such a site, ebselen exerts a pronounced allosteric effect that regulates catalytic site access through surface-loop interactions, thereby inducing a reconfiguration of water hotspots. Together, these findings highlight the promise of ebselen as a repurposed drug against SARS-CoV-2.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗