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DOE OSTI2022

Triton Field Trials - Changes in Habitats, a Literature Review of Monitoring Technologies

Marine energy devices are installed in highly dynamic environments and have the potential to affect benthic and pelagic habitats around them. Regulatory bodies often require baseline characterization and/or post-installation monitoring to determine whether changes in these habitats are being observed. However, a great diversity of technologies is available for surveying and sampling marine habitats. Selecting the most suitable instrument to identify and measure changes in habitats at marine energy sites can become a daunting task. We conducted a thorough review of journal articles, survey reports, and grey literature to extract information about the technologies used, the data collection and processing methods, and the performance and effectiveness of these instruments. We examined documents related to marine energy development, offshore wind farms, oil and gas offshore sites, and other marine industries around the world over the last 20 years, as well as national and international guidelines for surveying habitats around offshore activities. A total of 120 different technologies were identified across six main habitat categories: seafloor, sediment, infauna, epifauna, pelagic, and biofouling. The technologies were organized into 12 broad technology classes: acoustic, corer, dredge, grab, hook and line, net and trawl, plate, remote sensing, scrape samples, trap, visual, and others. Visual was the most common and the most diverse technology class, with applications across all six habitat categories. Sampling designs varied considerably among the reviewed studies but transect was the predominant design for surveying seafloor, epifauna, and pelagic habitats. The most common data analyses were univariate and multivariate statistical analyses aimed at calculating and comparing biodiversity indices, characterizing faunal assemblages or sediment classes, or modeling the distribution of animals related to abiotic parameters. Technologies and sampling methods adaptable and designed to work efficiently in energetic environments have greater success at marine energy sites. In addition, sampling designs and statistical analyses should be carefully thought through to identify differences in faunal assemblages and spatiotemporal changes in habitats.

16 TIDAL AND WAVE POWER↗

DOE OSTI2021

Guidance for Monitoring Passive Groundwater Remedies Over Extended Time Scales

Passive remediation can be appropriate where natural processes and actions such as institutional controls mitigate exposure to contaminated groundwater, achieving remedial action objectives and protectiveness of human health and the environment. Monitored natural attenuation (MNA) is a prevalent passive remediation strategy supported by a regulatory framework and monitoring design guidance. However, long-term passive remedies are usually selected in combination with at least one active remedy, such as source removal, in situ treatment, or pump-and-treat, functioning as a complementary method for achieving remediation objectives and meeting the applicable statutory and regulatory requirements. However, MNA and existing monitoring guidance primarily target situations where the remedial action objectives are met within a few decades. When time scales for passive remediation extend to many decades (50 years or more), a corresponding change in monitoring strategy is needed to adapt to the extended time scale. This document provides guidance for implementing an extended-scale monitoring (ESM) approach appropriate for long-duration passive remediation. Extended-scale is defined in this document with respect to time (i.e., a long duration of remediation) and a large enough physical scale such that downstream receptors will not be impacted within the remediation timeframe. ESM applies to slow-moving groundwater contaminant plumes and emphasizes monitoring primarily for potential exposure pathways. For this approach, the primary monitoring objective is to demonstrate that the plume diminishes before reaching the receptor zone or point of compliance and/or a receptor does not receive concentrations above the compliance limit. While the overall objectives of protecting human health and the environment are the same as for plumes where remediation can occur over a shorter time period, the time scales between decisions are longer and the dynamics of plume evolution are slower. To this end, a scenario-based strategy is described for different plume and source conditions, defining a containment and receptor zones. The containment zone is the area where the risk of exposure to groundwater contamination can be mitigated (e.g., through institutional controls) during the remediation time period. The receptor zone is defined as the area where exposure to groundwater contamination cannot be mitigated and compliance concentration standards must be met. Within the containment zone, slow plume migration may occur, leading to concentrations that exceed compliance standards. However, where distance to the receptor zone is large relative to plume migration and attenuation rate, this approach can be protective of the receptor zone. Selection of a long-duration passive remedy needs to be based on sufficient understanding of contaminant sources, hydrogeology, and contaminant plumes. A strong technical basis, supported by predictive analysis, is recommended to substantiate that contamination is expected to stay within the containment zone during the active remediation and attainment phase of the remedy, and diminish to meet compliance standards within the extended timeframe prior to reaching the receptor zone (e.g., many decades or even centuries). The ESM approach is based on verification of plume behavior and not on detailed plume dynamics. Monitoring is conducted to confirm expected behavior with an emphasis on exposure pathways to verify that plumes remain contained in areas where the protectiveness objectives can be met. ESM should not be adopted if there is significant risk of the plume extending beyond the containment zone. Given the slow movement within the containment zone, less frequent sampling is required relative to approaches used for conventional-scale remediation.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2026

Microbial spies and bloggers: programming cells to convert environmental information into discernible signals

Microbes regulate their dynamic behaviors using the chemical and physical characteristics of their environment. The ability of microbes to continuously convert this physicochemical information into biochemical information and to use organic matter in the environment as a power source makes these organisms attractive as chassis for building sensors. However, most biosensors have severe limitations when considering applications in hard-to-image settings like soils, sediments, and wastewater. Emerging technologies at the interface of biomolecular design, microbiome engineering, and synthetic biology offer new tools to program cells and communities as biosensors for these settings. Here, in this review, we describe innovations in biosensor outputs that are enabling new applications in complex environments, including reporters that are read out using electrochemical, gas chromatography, hyperspectral imaging, and next-generation sequencing methods. We also discuss computational advances that are accelerating the diversification of sensing components by mining metagenomics data for new transcriptional regulators and by designing allosteric protein switches that directly regulate reporter outputs using analytes. We highlight emerging opportunities for programming undomesticated microbes in communities to function as distributed sensors in the environment. Finally, we discuss the need for responsible biosensor development and to modernize regulatory frameworks to support evidence-based assessment of environmental biosensors.

analyte↗

DOE OSTI2023

Genomic characterization of rare earth binding by Shewanella oneidensis

Abstract Rare earth elements (REE) are essential ingredients of sustainable energy technologies, but separation of individual REE is one of the hardest problems in chemistry today. Biosorption, where molecules adsorb to the surface of biological materials, offers a sustainable alternative to environmentally harmful solvent extractions currently used for separation of rare earth elements (REE). The REE-biosorption capability of some microorganisms allows for REE separations that, under specialized conditions, are already competitive with solvent extractions, suggesting that genetic engineering could allow it to leapfrog existing technologies. To identify targets for genomic improvement we screened 3,373 mutants from the whole genome knockout collection of the known REE-biosorbing microorganism Shewanella oneidensis MR-1. We found 130 genes that increased biosorption of the middle REE europium, and 112 that reduced it. We verified biosorption changes from the screen for a mixed solution of three REE (La, Eu, Yb) using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) in solution conditions with a range of ionic strengths and REE concentrations. We identified 18 gene ontologies and 13 gene operons that make up key systems that affect biosorption. We found, among other things, that disruptions of a key regulatory component of the arc system ( hptA ), which regulates cellular response to anoxic environments and polysaccharide biosynthesis related genes ( wbpQ , wbnJ , SO_3183 ) consistently increase biosorption across all our solution conditions. Our largest total biosorption change comes from our SO_4685 , a capsular polysaccharide (CPS) synthesis gene, disruption of which results in an up to 79% increase in biosorption; and nusA, a transcriptional termination/anti-termination protein, disruption of which results in an up to 35% decrease in biosorption. Knockouts of glnA , pyrD , and SO_3183 produce small but significant increases (≈ 1%) in relative biosorption affinity for ytterbium over lanthanum in multiple solution conditions tested, while many other genes we explored have more complex binding affinity changes. Modeling suggests that while these changes to lanthanide biosorption selectivity are small, they could already reduce the length of repeated enrichment process by up to 27%. This broad exploratory study begins to elucidate how genetics affect REE-biosorption by S. oneidensis , suggests new areas of investigation for better mechanistic understanding of the membrane chemistry involved in REE binding, and offer potential targets for improving biosorption and separation of REE by genetic engineering.

Medin, Sean↗

DOE OSTI2021

MACCS Theory Manual

This report describes the models of the MACCS computer code as presented in MACCS Version 3.10.0. The purpose of MACCS is to simulate the impact of severe accidents at nuclear power plants on the surrounding environment. MACCS has been developed by Sandia National Laboratories for the U.S. Nuclear Regulatory Commission. From a given release of radioactive material into the atmosphere, MACCS estimates the extent and magnitude of radiological contamination, offsite doses, protective actions, socioeconomic impacts and costs, and health effects. Since the weather at the time of an accident is not predictable, MACCS supports various sampling options to run a representative set of simulations to evaluate weather variability. MACCS simulates atmospheric transport with a straight-line Gaussian plume segment model. From the estimated air and ground concentrations, MACCS models dose projections through several dose exposure pathways. These exposures can be offset by protective actions during the emergency response and long-term recovery of the accident. MACCS users directly specify the evacuation and sheltering area, while other protective actions (e.g., relocation, farmland restrictions, decontamination) are based on user-specified dose or concentration limits. While protective actions help reduce dose accumulation, they also cause social and economic impacts. MACCS models the extent of displaced individuals and land contamination, and the cost of offsite property damage, economic disruptions, and various accident expenditures caused by protective actions. Finally, from the dose accumulation, MACCS estimates early and stochastic health effects according to dose-response models. The purpose of consequence analyses is to be able to understand and estimate the impact of nuclear accidents. Consequence analysis is an essential tool to inform determinations of adequate protection of the public, to understand nuclear power hazards, to measure the value of regulations, and to help us appreciate the importance of nuclear safety. As such, MACCS has a variety of regulatory uses including environmental analyses (10 CFR 51.53, 52.47), regulatory cost-benefit analyses, backfit analyses (10 CFR 50.109), consequence analysis studies such as SOARCA (NUREG-1935), Level 3 PRA studies, and risk-informing of emergency planning (10 CFR 50 App. E and 50.47). This report updates the previous MACCS theory manual (NUREG/CR-4691 Vol. 2; Chanin, Sprung, Ritchie, & Jow, 1990) and accompanies the MACCS User's Guide (SAND-2021-1588) that describes the use and input requirements of the graphical user interface of MACCS known as WinMACCS. The MACCS User's Guide is also a reference guide that describes data input file formats, describes various software components in the MACCS code suite, and provides a set of example tutorials for running WinMACCS. Also, soon to be published is a MACCS input parameter guidance report (NUREG/CR-7270) that provides technical bases for commonly used MACCS input values. This page left blank

97 MATHEMATICS AND COMPUTING↗

DOE OSTI2023

Differences In High Burnup Fuel Management Strategies to Minimize FFRD and Increase Economic Viability

The nuclear industry is pursuing approval of an increase in the length of the pressurized water reactor (PWR) cycle from 18 months to 24 months to reduce reactor downtime and enhance the economic competitiveness of nuclear energy. Such an increase in reactor cycle length will require that the maximum rod average burnup exceeds the current regulatory limit of 62 GWd/MTU, and it could peak at approximately 75 GWd/MTU, posing potential reactor safety and performance concerns. One such concern is that fuel fragmentation, relocation, and dispersal (FFRD) could occur during a severe loss-of coolant accident (LOCA) in which a fuel rod balloons and bursts, and pulverized fuel fragments are dispersed throughout the reactor’s primary coolant system. Previous analyses have identified which reactor operating conditions leave the core more susceptible to FFRD and have shown that FFRD susceptibility is strongly linked to fuel rod burnup and linear heat rate (LHR) history. The work described in this report uses an optimization strategy known as parallel simulated annealing (PSA) and a coarse mesh Purdue Advanced Reactor Core Simulator (PARCS) reactor physics model to develop two core fuel loading patterns, each with a different optimization objective. One core optimization maximized the core’s cycle length while still respecting regulatory limits on the radial peaking factor and soluble boron concentration with a peak rod average burnup of 75 GWd/MTU. The second optimization was aimed at minimizing FFRD susceptibility while still targeting a 24-month cycle length and respecting regulatory limits. PARCS model predictions were verified using the high-fidelity Virtual Environment for Reactor Applications (VERA). The two core designs were compared to highlight core design strategies to minimize FFRD susceptibility and to maximize economic viability.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

DOE OSTI2026

High-Burnup LOCA Burst Susceptibility BISON Analysis in PWRs and BWRs

Accurately assessing high-burnup fuel behavior during loss-of-coolant accidents (LOCAs) is essential for understanding fuel fragmentation, relocation, and dispersal (FFRD) risks across the US light-water reactor fleet. This work updates previous Nuclear Energy Advanced Modeling and Simulation (NEAMS) Program multiphysics LOCA analyses for a pressurized water reactor (PWR) and a boiling water reactor (BWR) by incorporating recent model and material property advancements in the BISON fuel performance code, including a high-burnup structure (HBS) model, revised cladding burst criteria, and updated thermal–mechanical correlations. This update was needed to support ongoing industry initiatives and upcoming regulatory changes. Full-core, rod-resolved operating histories generated using Virtual Environment for Reactor Analysis (VERA) and system-level LOCA conditions obtained from TRACE were applied to statistically representative rod samples in BISON to evaluate burst behavior and FFRD susceptibility. These calculations used two cladding burst correlations and three fuel pulverization models so that the predictions of these models could be compared. The updated PWR simulations show markedly improved numerical stability as the number of crashed simulations decreased by 95% compared to the previous study, and hence higher confidence in results. The updated PWR simulations predicted cladding bursts exclusively among once-burned, high-power rods, with two different cladding burst models identifying the same burst-susceptible population. Resulting FFRD susceptibility estimates are significantly reduced compared with earlier studies, driven by cooler predicted fuel and plenum temperatures, lower hoop strains, and reduced fission gas release in the updated models. In contrast, none of the BWR rods were predicted to burst under either burst criterion, reaffirming minimal BWR FFRD susceptibility even with updated HBS and material models. Comparisons between the PWR and BWR end-of-cycle predictions are made. Comparison with prior work highlights significant shifts in PWR fuel performance metrics and confirmation of earlier BWR conclusions. Overall, the updated results underscore the importance of having high-resolution detailed modeling capability and continuously integrating evolving material models and physics into high-resolution multiphysics simulations. The unified assessment presented here strengthens confidence in predicting high-burnup LOCA behavior by improving agreement between different cladding burst correlations. These results also provide an improved foundation for future BISON model development, FFRD susceptibility calculations.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

DOE OSTI2022

What’s in My Toolkit? A Review of Technologies for Assessing Changes in Habitats Caused by Marine Energy Development

Marine energy devices are installed in highly dynamic environments and have the potential to affect the benthic and pelagic habitats around them. Regulatory bodies often require baseline characterization and/or post-installation monitoring to determine whether changes in these habitats are being observed. However, a great diversity of technologies is available for surveying and sampling marine habitats, and selecting the most suitable instrument to identify and measure changes in habitats at marine energy sites can become a daunting task. We conducted a thorough review of journal articles, survey reports, and grey literature to extract information about the technologies used, the data collection and processing methods, and the performance and effectiveness of these instruments. We examined documents related to marine energy development, offshore wind farms, oil and gas offshore sites, and other marine industries around the world over the last 20 years. A total of 120 different technologies were identified across six main habitat categories: seafloor, sediment, infauna, epifauna, pelagic, and biofouling. The technologies were organized into 12 broad technology classes: acoustic, corer, dredge, grab, hook and line, net and trawl, plate, remote sensing, scrape samples, trap, visual, and others. Visual was the most common and the most diverse technology class, with applications across all six habitat categories. Technologies and sampling methods that are designed for working efficiently in energetic environments have greater success at marine energy sites. In addition, sampling designs and statistical analyses should be carefully thought through to identify differences in faunal assemblages and spatiotemporal changes in habitats.

16 TIDAL AND WAVE POWER↗

DOE OSTI2022

REGULATORY TESTING AND 3D SCANNING METHODOLOGY OF THE DPP-1 TYPE SHIPPING CONTAINER FOR NCT AND HAC TESTS

The safe transportation of radioactive material using appropriately designed and certified transportation packages is important to protect the public and environment. In the United States of America all transportation of radioactive material is regulated by the Department of Transportation (DOT) along with input by the Nuclear Regulatory Commission (NRC). All packages that contain radioactive material requires that it be transported and regulated by the requirements set by the DOT Class 7 hazardous material transportation. Federal regulations place strict administrative controls on the transport of radioactive materials. The two main goals for transporting radioactive materials are 1. Safety should be the primarily focused on the package because it is the first line of defense. 2. Package integrity should be directly related to the degree of the hazard of the radioactive material contents. The two-part goal ensures that the right package is used. Type B packages are designed to transport radioactive materials with the highest level of radioactivity. Examples of material transported in Type B packagings include spent nuclear fuel, high-level radioactive waste, and high concentrations of other radioactive materials. The purpose of this test protocol is to determine the effects of drop and thermal testing on package performance in a Type B shipping package (DPP-1). Required physical tests for the DPP-1 package include free drop, crush, puncture, penetration, compression, vibration, water spray, water immersion, and thermal. The preparation, regulatory testing, and post-test evaluation of a Type B shipping package tested to the 10 CFR 71.71 and 10 CFR 71.73 requirements will be presented. Six packages were tested and the responses when subjected to the HAC sequence of free drop, puncture, and thermal tests according to 10 CFR 71.73(c)(1), (c)(2), (c)(3) and (c)(4) were measured. The Free Drop, Crush and Puncture tests were performed at the NTRC in Knoxville, TN, and the Thermal tests were performed Southwest Research Institute in San Antonio, TX. The test units were subjected to a pre-operational leak testing before the NCT and HAC tests and a post-operational leak test and a helium leak test once regulatory testing was complete. Finally, all test units were scanned with a handheld blue light scanner to capture a 3D CAD geometry of the test units in the deformed shape. The handheld scanner provided the most effective and reliable way to acquire accurate 3D measurements of the test unit. The scanned geometry was sectioned in areas with deformation and the cross-section profile was measured to determine accurate and repeatable results of the deformed shape of the test units.

Martinez, Oscar↗

DOE OSTI2021

Wyoming CarbonSAFE: Advancing Commercialization of Low-Carbon Energy Technologies in Fossil-Rich Wyoming

Wyoming’s Powder River Basin (PRB) is the most prolific coal producer and exporter in the United States and the State of Wyoming’s largest oil-producing basin. In addition to being a leading energy producer, the PRB is the site of research programs whose aim is to develop and integrate low-carbon technologies into existing fossil fuel energy industries. Much of the PRB’s low-carbon research is focused around Dry Fork Station (DFS), which is the newest coal-fired power station in the western US. Currently, DFS hosts five carbon capture projects, including pilot-scale capture projects that utilize different capture technologies and a full front-end engineering and design (FEED) study, a commercial CO 2 pipeline for nearby CO 2 -enhanced oil recovery industry, and is also co-located with the Wyoming Integrated Test Center, which is a host facility for unconventional carbon utilization research (i.e. flue gas carbon-to-products innovations). DFS is also the host site for the Wyoming CarbonSAFE project. Wyoming CarbonSAFE, funded by the Department of Energy/National Energy Technology Laboratory, is a multi-phased program (currently in Phase II) whose core objective is developing and validating storage sites within a complex capable of storing 50 million metric tonnes of CO 2 over a 30 year project period using carbon capture, utilization and storage (CCUS). This paper will provide an overview of the project to-date, showing that CarbonSAFE goals are achievable with respect to geologic, environmental, regulatory, CO 2 source and economic conditions, and why the State of Wyoming offers one of the more favorable environments to advance the commercialization studies. In 2019, the project team designed and completed a ~10,000 foot stratigraphic test well just south of DFS. From this well, the team collected over 600 feet of core, fluid samples from target injection zones, and a full petrophysical log suite. In addition, legacy 2D seismic lines were acquired and a 3D seismic survey was acquired in the fall of 2020. The objective of these field activities has been to identify and characterize target storage reservoirs and associated caprock. This complex has several reservoirs that could meet commercial injection goals, and over 4000 feet of associated caprock. Simulations of site performance suggest that stacked injection provides the most effective storage strategy, and would necessitate several sites within the greater complex to meet final project injection goals. Other work within this project includes the development of a business-case around proximal fields with CO 2 -enhanced oil recovery potential and existing tax credits, assessment of regulatory conditions, including pore space ownership and Class VI injection well permitting requirements, implementation of a robust public outreach Electronic copy available at: https://ssrn.com/abstract=3821220 GHGT-15 McLaughlin 2 program, and surface site characterization activities that have focused on environmental factors. In the PRB and at DFS, the State of Wyoming and other entities have made carbon management a priority of its future energy industry by providing a regulatory and business framework that is favorable to advancing these technologies. These endeavors will become more realistic with the successful implementation of Wyoming CarbonSAFE, and its ability to secure and validate commercial-scale CCUS at the center of Wyoming’s premier low-carbon research efforts.

McLaughlin, Fred↗

DOE OSTI2020

What does the future hold for utility electricity efficiency programs?

Here, this study develops projections of future spending and savings from electricity efficiency programs funded by electric utility customers in the United States through 2030 based on three scenarios. Our analysis relies on detailed bottom-up modeling of current state energy efficiency policies, demand-side management and integrated resource plans, and regulatory decisions. The three scenarios represent a range of potential outcomes given the policy environment at the time of the study and uncertainties in the broader economic and state policy environment in each state. We project spending to increase to $8.6 billion in 2030 in the medium scenario, about a 45 percent increase relative to 2016 spending. In the high case, annual spending increases to $11.1 billion in 2030 and remains relatively flat in the low case ($6.8 billion in 2030). Our analysis suggests that electricity efficiency programs funded by utility customers will continue to impact load growth significantly at least through 2030, as savings as a percent of retail sales are forecast at 0.7 percent in the medium scenario and 0.98 percent in the high scenario.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

DOE OSTI2025

Populus PtrbHLH011 Is a Transcriptional Co‐Regulator Involved in the Activation of Cell Wall Biosynthesis by Iron Deprivation

The lack of a mechanistic understanding of the environmental plasticity of secondary cell wall (SCW) biosynthesis restricts large-scale biomass and bioenergy production on marginal lands. Using Populus (poplar), a key bioenergy crop, we discovered that iron deprivation, a prevalent abiotic stress on marginal lands, stimulates SCW biosynthesis in stems. We identified the transcription factor PtrbHLH011 as a critical regulator underlying this response. Through integrated analyses involving phenotypic characterisation of PtrbHLH011 knockout and overexpression plants, functional genomics and molecular investigations, we established that PtrbHLH011 functions as a central regulator of SCW biosynthesis, iron homeostasis and flavonoid biosynthesis by directly repressing essential genes in these pathways. Iron deprivation downregulates PtrbHLH011 expression, subsequently activating these biosynthetic pathways. Notably, cytosine base editing-based knockout of PtrbHLH011 significantly enhanced plant growth, yielding up to a 110% increase in stem diameter and a 300% increase in leaf iron content. These findings present a novel regulatory mechanism linking environmental iron availability to SCW biosynthesis and illustrate a practical strategy to improve biomass yield on iron-deficient marginal lands. Furthermore, our mechanistic insights into PtrbHLH011 target recognition and regulation provide a valuable foundation for precise manipulation of gene regulatory networks, facilitating the development of high-performance bioenergy crops adapted to marginal environments.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2023

Simultaneous carbon catabolite repression governs sugar and aromatic co-utilization in Pseudomonas putida M2

ABSTRACT Pseudomonas putida have emerged as promising biocatalysts for the conversion of sugars and aromatic compounds obtained from lignocellulosic biomass. Understanding the role of carbon catabolite repression (CCR) in these strains is critical to optimize biomass conversion to fuels and chemicals. The CCR functioning in P. putida M2, a strain capable of consuming both hexose and pentose sugars as well as aromatic compounds, was investigated by cultivation experiments, proteomics, and CRISPRi-based gene repression. Strain M2 co-utilized sugars and aromatic compounds simultaneously; however, during cultivation with glucose and aromatic compounds ( p- coumarate and ferulate) mixture, intermediates (4-hydroxybenzoate and vanillate) accumulated, and substrate consumption was incomplete. In contrast, xylose-aromatic consumption resulted in transient intermediate accumulation and complete aromatic consumption, while xylose was incompletely consumed. Proteomics analysis revealed that glucose exerted stronger repression than xylose on the aromatic catabolic proteins. Key glucose (Eda) and xylose (XylX) catabolic proteins were also identified at lower abundance during cultivation with aromatic compounds implying simultaneous catabolite repression by sugars and aromatic compounds. Reduction of crc expression via CRISPRi led to faster growth and glucose and p -coumarate uptake in the CRISPRi strains compared to the control, while no difference was observed on xylose+ p -coumarate. The increased abundances of Eda and amino acid biosynthesis proteins in the CRISPRi strain further supported these observations. Lastly, small RNAs (sRNAs) sequencing results showed that CrcY and CrcZ homologues levels in M2, previously identified in P. putida strains, were lower under strong CCR (glucose+ p -coumarate) condition compared to when repression was absent ( p -coumarate or glucose only). IMPORTANCE A newly isolated Pseudomonas putida strain, P. putida M2, can utilize both hexose and pentose sugars as well as aromatic compounds making it a promising host for the valorization of lignocellulosic biomass. Pseudomonads have developed a regulatory strategy, carbon catabolite repression, to control the assimilation of carbon sources in the environment. Carbon catabolite repression may impede the simultaneous and complete metabolism of sugars and aromatic compounds present in lignocellulosic biomass and hinder the development of an efficient industrial biocatalyst. This study provides insight into the cellular physiology and proteome during mixed-substrate utilization in P. putida M2. The phenotypic and proteomics results demonstrated simultaneous catabolite repression in the sugar-aromatic mixtures, while the CRISPRi and sRNA sequencing demonstrated the potential role of the crc gene and small RNAs in carbon catabolite repression.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Guidance for Monitoring Passive Groundwater Remedies Over Extended Time Scales

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2023

Causal CCF Parameter Estimations 2020

This report documents the quantitative results of the causal common-cause failure (CCF) parameter estimations for the failure cause groups “component,” “design,” “environment,” “human,” and “other,” based on CCF data through 2020 in the U.S. Nuclear Regulatory Commission (NRC) CCF database: https://rads.inl.gov/Pages/CCF.aspx. This report utilizes the same data period (2006–2020) and CCF templates as INL/EXT-21-62940, Revision 1, CCF Parameter Estimations, 2020 Update. The 2015 causal CCF prior distributions for the specific failure cause groups (instead of the 2015 generic CCF prior distributions) were used in this report to estimate the associated causal CCF parameters. All the 2015 causal CCF prior distributions and generic CCF prior distributions were developed in INL/EXT-21-43723, Developing Generic Prior Distributions for Common Cause Failure Alpha Factors and Causal Alpha Factors, using CCF data from 1997 to 2015. These quantitative results were developed to support the causal alpha factor model and should be used as appropriate in probabilistic risk assessment (PRA) studies such as the NRC Significance Determination Process for commercial nuclear power plants in the United States.

99 GENERAL AND MISCELLANEOUS↗

DOE OSTI2025

Hands-On, Heads-Up: Blending Cyber T&E with Data Science-Driven Training in Jupyter Notebooks

In an era of increasingly sophisticated threats to critical infrastructure, cybersecurity professionals must be more than just aware; they must be immersed, agile, and equipped to operate in environments where failure is not an option. Nowhere is this truer than in the nuclear sector, where cyber-physical systems, regulatory scrutiny, and insider threat potential demand a new generation of hands-on, technically fluent defenders. This paper presents a unified training approach that integrates Cybersecurity Test and Evaluation (T&E) with data science techniques using Jupyter Notebooks as the interactive lab environment. The program centers on a modular, scenario-driven curriculum designed to build not just knowledge but practical capability in the assessment and defense of radiation detection systems, firmware interfaces, and operational security postures.

98 - NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL↗

DOE OSTI2022

The Promise of Emergent Nanobiotechnologies for In Vivo Applications and Implications for Safety and Security

Nanotechnology, the multi-disciplinary field based on the exploitation of the unique physicochemical properties of nanoparticles (NPs) and nanoscale materials, has opened a new realm of possibilities for biological research and biomedical applications. The development and deployment of mRNA-NP vaccines for COVID-19, for example, may revolutionize vaccines and therapeutics. However, regulatory and ethical frameworks that protect the health and safety of the global community and environment are lagging, particularly for nanotechnology geared towards biological applications (i.e., bio-nanotechnology). Considering this, the following review, while not comprehensive, attempts to illustrate the breadth and promise of bionanotechnology developments, and how they may present future safety and security challenges. Specifically, we address current advancements to streamline the development of engineered NPs for in vivo applications and provide discussion on nano-bio interactions, NP in vivo delivery, nano-enhancement of human performance, nanomedicine, and the impacts of NPs on human health and the environment.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Panel Session 26: Risk-Based Approaches to Environmental Remediation and Adaptive Site Management

This panel discussed the needs and importance of developing risk-based approaches to environmental remediation that will enable complex sites to be adaptively managed by applying cost-effective remedies that are protective of human health and the environment. The discussion emphasized risk-based approaches within decision processes and subsequent incorporation into regulatory decision documents. Remedy approaches, remediation exit strategies, and long-term monitoring plans that support risk-based end-states for difficult-to-remediate sites were discussed Panelists with presentations: Risk-Informed Approaches to Environmental Remediation and Adaptive Site Management (Vicky Freedman); ITRC Adaptive Site Management Perspective (Michael Truex); Optimization During Operations and Maintenance Phase at the Hanford Site 200-ZP-1 Groundwater Operable Unit (Emerald Laija, Kate Amrhein, Michael Truex); Adaptive Management: EPA Overview (Benjamin Simes); Decommissioning and Environmental Remediation Section Division of Nuclear Fuel Cycle and Waste Technology (Horst Monken Fernandes)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗