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At least 1,351 records · Page 75

DNA Strand Displacement Driven Molecular Additive Manufacturing (DSD-MAM)

The goal of this project was to validate two-dimensional molecular printers, initially selfassembled from DNA and then actuated by externally driven cycles of DNA strand displacement, as prototype integrated nanosystems for molecular additive manufacturing. Novel functionalities of these nanomachines were explored during this project, including the following: nanometer-precision positioning mechanisms based on DNA strand displacement with multivalent interactions for discrete stepping or else diffusive capture; integration of independently moving layers of DNA origami to achieve 2D controllable motion; integration of spatial positioning with deposition functionality. The principal importance of this project was to provide an essential step in the development of a new technology for atomically precise manufacturing. Our first generation molecular 2D printer offers several advantages over conventional DNA-origami patterning, such as faster prototyping, faster dynamic rearrangement of patterns, and the ability to respond with feedback. We anticipate that our first-generation molecular printers may inspire future generations of molecular printers with iterative improvements in robustness and throughput. Potential applications of atomically precise manufacturing include the following: photovoltaics; photosynthetic and fuel cells; thermoelectrics and anisotropic heat spreaders; solid-state lighting; molecular electronic and plasmonic circuits; selectively preamble membranes; self-repairing materials with high strength-to-weight and fracture resistance.

36 MATERIALS SCIENCE↗

FY24 LLNL Laboratory Directed Research and Development (LDRD) Project Accomplishments

Large duration energy storage is the key to couple renewable energy generation with power supply. This project aimed to advance the durability of a low-cost and eco-friendly flow battery technology based on iron chemistry to speed up the technology readiness level rapidly and radically. A novel device which is called “an artificial kidney” was innovated through disruptive research to integrate with the flow battery for rebalancing capacity. Key results at 50 cm2 scale demonstrates that artificial kidney enabled retaining the storage capacity of the flow battery over 100 cycles. Comparison of outcome of this work showing 0% capacity degradation to the state-of-the-art technology corroborates that this novel system is a game changing innovation.

99 GENERAL AND MISCELLANEOUS↗

SCCOUT: Surveying for Categorization and Control of Organism in Unwelcome Territory

Advancements in the fields of science, technology, and medicine have contributed largely to the growth of society over the last century. Thanks to these discoveries, all corners of the planet are connected both digitally and physically. Progress in the field of medicine has led to people living longer, fuller lives and a general increase in global population. Awareness of our world’s changing climate and the effects that has on local and global ecosystems has spurred great efforts to protect the environment. These discoveries along with many others have helped our world grow in a myriad of ways; however, certain efforts in agriculture, natural resource management, and ecosystem management have not scaled to accommodate this growth. Increasing prevalence and ease of dissemination of invasive plants and animals, diseases, and changing ecological conditions pose significant challenges to biodiversity, ecosystem health, and agricultural productivity. Moreover, as the world population grows, there are more people to feed and less viable farmland to produce food for them. Despite the increasing prevalence of these issues, methods for addressing and tracking them have stayed relatively the same for at least the last several decades.

54 ENVIRONMENTAL SCIENCES↗

Investigation of encapsulin nanocompartment systems as a scaffold for biomaterials synthesis in Rhodococcus species (Annual Report 2025)

Engineered protein compartmentalization systems hold significant promise to enhance reaction efficiencies through co-localization, concentration, and sequestration of biosynthetic pathways. As such, they have the potential to enable the bioproduction of next generation bioproducts and biomaterials in genetically engineered microbes in support of DOE’s mission to build a strong bioeconomy. Among systems of particular interest are protein nanocompartment systems called encapsulins that are natively produced by a variety of bacteria including those with a high potential for bioproduction. This ECRP project is focused on understanding how encapsulins can be used to enhance the biosynthesis of next-generation biomaterials in Rhodococcus species. Specifically, we seek: (1) to probe the mechanistic basis for how these compartments are regulated, biosynthesized, and maintained, and (2) to engineer these systems to achieve new biosynthetic functions (e.g., alkene, inorganic nanoparticle biosynthesis). We anticipate that this work will establish encapsulin compartmentalization systems as a means of improving yields and enabling biosynthetic routes toward new biomaterials, thus advancing the U.S. bioeconomy.

60 APPLIED LIFE SCIENCES↗

CFD Simulation of Aerobic Gas Fermentation to Enable Commercial Conversion of CO 2 into Aquaculture and Animal Feed: Cooperative Research and Development (Final Report)

NovoNutrients’ fermentation technology uses energy from hydrogen to transform industrial CO2 emissions into premium animal feed ingredients and other valuable products. A single NovoNutrients’ commercial manufacturing plant will capture and convert over 200,000 tons/yr of CO2 into over 100,000 tons/yr of high-protein feed. Key to the rapid and widespread deployment of the technology is maximization of its productivity and energy efficiency. Robust, physically based computational models of the technology will significantly increase productivity and efficiency, accelerating NovoNutrients’ technology to manufacturing scale. NREL has unique capabilities for creating and running such computational models. NREL's existing aerobic bioreaction computational fluid dynamics (CFD) models will be adapted to NovoNutrients’ gas fermentation (CO2, H2, O2) technology. The multiphysics CFD simulations require thousands of high-performance computing (HPC) node hours to simulate the complex geometries and contents of NovoNutrients’ industrial bioreactors. The experimentally validated CFD models were used to identify optimally efficient and productive bioreactor designs and operating conditions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Assessing kelp nutrient bioextraction capacity in aquaculture farms in the US with implications for conservation and management

This proposal aimed to assess the carbon and nitrogen bioextraction capabilities of sugar kelp farms across a broad geographic range encompassing both the East and West Coasts of the United States. To meet this objective, the project team first developed a cost-effective sampling toolkit using readily available materials and an illustrated protocol to guide kelp farmers in standardized sample collection. Farmers were trained to use the toolkit and prepare samples for shipment to the University of Alaska Fairbanks, where laboratory analyses were conducted to quantify carbon and nitrogen removal. Results were then shared with collaborating farmers, farm managers, regulatory agencies, and the general public to disseminate findings and gauge interest in the bioextraction services offered by kelp aquaculture. The project also investigated temporal and species-specific differences in carbon and nitrogen removal as a complementary objective.

54 ENVIRONMENTAL SCIENCES↗

Identifying Hot Spots and Hot Moments of Metabolic Activity in Salt Marsh Sediments through BONCAT-FISH Microscale Mapping (Final Technical Report)

Understanding the biogeography and timing of microbial metabolic activity is a key priority for microbial ecologists. With such knowledge, the cumulative biogeochemical contributions of microbial communities become more predictable, and our ability to both understand the effects of environmental change on microbial activity and build synthetic communities with desired functions will increase substantially. In this project, we advanced this broad, ambitious goal in two substantial ways: we developed a multiplexed Fluorescence In Situ Hybridization (FISH) approach that links microbial identity with metabolic function, and we established a novel Bio-Orthogonal Non-Canonical Amino acid Tagging (BONCAT) technique that can resolve the timing of anabolic activity, providing new resolution of when microbial constituents are growing. We deployed both of these techniques in multiple environmental settings to demonstrate their versatility. At the Little Sippewissett Salt Marsh, we combined a novel dual-BONCAT technique with fluorescence activated cell sorting to determine which population of cells was metabolically active during the day and which was active during the night. We found that Methylobacterium was active during daylight hours, potentially feeding on carbon-rich molecules released from plant roots. Sulfur-cycling microbes dominate the population active during the dark night-time hours. Overall, the work conducted under the auspices of this project developed two promising new techniques for identifying the “hot spots” and “hot moments” of microbial activity in complex communities with taxonomic and functional resolution. We focused largely on a salt marsh sediment context, but are confident that microbial ecologists seeking to understand the microbial role in biogeochemical cycles in a wide range of settings will find our newly developed techniques useful in future work.

54 ENVIRONMENTAL SCIENCES↗

Partial Support of the Fast-Track Consensus Study on Foundational Research Gaps and Future Directions for Digital Twins (Final Report)

This study from the National Academies of Sciences, Engineering, and Medicine was launched to explore the foundational research gaps and opportunities for digital twins. As part of the information gathering process, the committee organized three targeted workshops—in engineering, climate sciences, and biomedical sciences—to better understand domain-specific nuances and barriers to developing digital twins. These sessions enabled cross-sector experts to surface field-specific needs, challenges, and open questions related to digital twins. Thousands of participants across multiple domains engaged in the discussions, which workshops were summarized in three separate Proceedings-in-Brief.

42 ENGINEERING↗

Molecular mechanisms of mycorrhizal‐decomposer interactions and impacts on terrestrial biogeochemistry (Final Technical Report)

Soil microorganisms make nutrients available for plants and control a large portion of greenhouse gas emissions from terrestrial ecosystems by respiring CO 2 to the atmosphere, yet our understanding of how they do this is not clear enough to predict the rate of carbon (C) and nutrient flow through soils and the ecosystems they support. A common and abundant type of soil microorganism that associates with the roots of live plants, known as ectomycorrhizal fungi (EMF), is often able to control the amount of nutrients and C that are released from soil (e.g., as CO 2 respired to the atmosphere) during decomposition by microorganisms.

54 ENVIRONMENTAL SCIENCES↗

EvoNet: A phylogenomic and systems biology approach to identify genes underlying plant survival in marginal, low‐N soils

The DOE‐BER “EvoNet” project investigates the genetic and molecular basis of plant resilience in extreme environments. We do this by identifying key genes that enable “extreme survivor” species to thrive in the nitrogen-poor soils of Chile’s hyper-arid Atacama Desert. Our collections focus on 32 Atacama extremophile species, including seven grass species with potential biofuel applications. To identify genes-of-importance to survival we compared genomic and transcriptomic profiles of extremophile species that thrive in the Atacama to those of closely related “sister” species from nitrogen-rich arid and mesic regions of California. Deep RNA sequencing and de novo transcriptome assembly across these triplet species sets supported a phylogenomic framework for identifying positively selected genes associated with adaptive divergence. Our integrative analysis combined ecological and environmental data, metagenomics, evolutionary and systems biology, and metabolomics. This enabled us to create an unprecedented framework for systematically understanding how non-model plants have adapted to survive in extreme conditions. Our resulting database of positively selected ortholog groups in the extremophile plants offers promising targets for engineering crop and biofuel species with enhanced resilience to drought and extreme weather. Additionally, our newest dataset explores and exploits a complementary metabolomic approach. This new aspect provides innovative strategies to manipulate plant cell metabolism, further supporting efforts to improve agricultural productivity in the face of extreme climates. Importantly, our combined evolutionary- and metabolomic-based strategies focused on convergent patterns of adaptation, providing a genetic and metabolomic toolkit for improving crop and biofuel resilience across diverse plant species. Finally, our novel exploration of ecological and evolutionary dynamics delivered to the community a phylogenomic computational pipeline called “PhyloGeneious.” Our continued adaptations of this pipeline are publicly available to expedite evolutionary genomic research for future scientific discoveries. In total, our DOE-BER has provided genomic, metabolomic, and computational strategies to understand how extremophile plants provide evolutionary and physiological targets for improving agricultural and biofuel production.

59 BASIC BIOLOGICAL SCIENCES↗

Mapping Nitrogen Pathways in Organic Manure through GCAM

As the world has become more and more globalized, a need for a comprehensive and extensive modelling system that maps out the vast majority of processes and items around the world and can display how interdependent they are. Furthermore, such a model should be able to predict future processes and values such that it can guide scientific thinking and policy. This is where the importance of GCAM becomes so apparent. GCAM, the Global Change Analysis Model, that incorporates many different fields and can predict such a wide variety of processes. As the model expands and encompasses more fields and goes into greater detail, there is a need for previously incorporated processes to be further fleshed out. My project is dedicated to creating the organic nitrogen fertilizer pathway, such that there are two nitrogen fertilizer pathways, synthetic and organic (manure). Previously, only synthetic fertilizer was included, with all organic fertilizer (manure) being omitted from the model. Developing the synthetic path is relatively easy, as the process for creating synthetic fertilizer is an industrial process that has been perfected over decades, and thus has a very precise input/output coefficient of nitrogen, but determining that same coefficient for organic manure is much harder, because, well, digestion is not an industrial process, especially considering the many different types of animals have different processes, and a good amount of the manure they produce won’t be applied to the soil. I obtained data from the UN’s FAO on manure nitrogen applied to soil, and filled in the gaps and created many different visualizations to help comprehend the data. These findings will foster a greater understanding of nitrogen flows, which is crucial to fields like industrial agriculture, polluting runoff, and nitrifying bacteria and archaea in the soil.

54 ENVIRONMENTAL SCIENCES↗

Remote Sensing Approach for Monitoring Tree Health Adjacent to Transmission Corridors

This study presents an initial proof-of-concept for a satellite-based remote sensing approach to identify and monitor potential areas of poor tree health across the entire BPA service territory on an annual basis. We tested three variants of “delta peak NDVI” ( ΔPN ) change detection metrics that express interannual variation in primary productivity relative to a baseline by comparing ΔPN values for known insect/disease disturbances and nearby reference locations. All three metrics showed promise for detecting poor tree health in the year during disturbance, but the metric based on the difference from the long-term (2016-2024) median ( Δ Med PN ) was preferred due to its responsiveness to change in the years during and after disturbance, resilience to interannual variation, and ease of interpretation as being above or below normal. Comparison of Δ Med PN grouped by relative severity of disturbance indicated it was not sensitive enough to detect “low” severity disturbances, as mapped by USGS’s LANDFIRE program, but could distinguish “moderate” and “high” severity disturbances from reference locations. These findings informed selection of a threshold for Δ Med PN , which was combined with areas exhibiting negative NDVI to map potential areas of concern. Visual inspection of before/after high-resolution imagery and NDVI time series showed that many areas of concern aligned with visible signs of defoliation and die-off as well as other types of disturbance (e.g., landslides, logging, road grading, flooding). Some areas of concern are thought to be false detections caused by persistent shadow, and some could not be explained with visual inspection due to spatiotemporal limitations of before/after imagery. In summary, our approach shows promise for large-scale monitoring of tree health adjacent to BPA transmission lines, but additional work is recommended to improve model sophistication and remove noise.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Microbial Influence on Radionuclide Transport through the Culebra

The potential release of radionuclides into the Culebra dolomite formation is modeled extensively in the Waste Isolation Pilot Plant’s Performance Assessment (WIPP PA) calculations. Uranium is of chief concern in these modeled scenarios, as it shows the greatest release. Because of this, a series of experiments was undertaken to determine if microbial activity in the Culebra formation could transform uranium, either directly or indirectly, to a state that could be immobilized.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Effects of Salt Concentration, Ionic Strength, and Water Activity on the Growth of a WIPP Archaeal Isolate, Halobacterium sp.

An extremely halophilic archaeon ( Halobacterium sp., WIPP strain), isolated from WIPP halite, was tested for its ability to grow in media containing various combinations of sodium and magnesium chloride that influenced overall ionic strength and water activity. Halobacterium sp. grew well in most tested salt combinations, and its growth appeared to depend more upon the relative contribution of sodium to overall ionic strength than on other parameters. Growth rates were highest when media consisted of at least 2 M NaCl and lowest when NaCl dropped below 20% of the ionic strength (usually less than 0.5 M). High magnesium concentrations ( 1 and 1.5 M MgCh 2 ) did not prevent Halobacterium sp. growth, as long as the sodium contribution was sufficient. Water activity was not a definitive factor in determining growth, because this value did not reach an extreme lower limit in the experimental media.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Evaluating Tools and Technologies for Monitoring Baleen Whales During Offshore Wind Foundation Installation

Impacts to baleen whales are of particular concern in the context of offshore wind development because of their conservation status, the overlap of their habitat with current and planned offshore wind energy facilities, and their expected sensitivity to low-frequency sounds, such as those produced during pile driving. Therefore, this report focuses on monitoring technologies capable of detecting this species group. The scope of this report is to assess the current state of technologies and methods for monitoring baleen whales in low- and no-light conditions in conjunction with offshore wind foundation installation. This work considers the performance of various sensing technologies that can be used to observe the presence of baleen whales near offshore construction activities. The scope of work includes: 1. Technology Performance Metrics – Identify metrics that can be used to assess technology performance. 2. Technology Evaluation – Synthesize the past performance of technologies with respect to the performance metrics based on available literature. 3. Technology Characterization Framework – Describe a potential framework to characterize technology performance with a standardized approach. 4. Technology Research and Development Needs – Identify future needs and opportunities for research, development, and deployment to improve technologies.

17 WIND ENERGY↗

Technical Background and Validation Report on the Residential Water Inhalation Risk Calculator Presented in the Risk Assessment Information System

Indoor air quality (IAQ) is critical for human health. Poor IAQ is linked to respiratory issues, cardiovascular diseases, and cancer. Indoor pollutants are emitted by typical household items such as cleaning products, personal care items, building materials, and tap water - an understudied volatile organic compound (VOC) source. This document presents the Residential Water Inhalation Risk Calculator (RWIRC), which estimates daily VOC exposure concentrations from various household water uses, such as showering and dishwashing, to assess exposure risks for the most vulnerable occupant. Integrated into the Risk Assessment Information System (RAIS) and sponsored by the US Department of Energy (DOE), the calculator divides a house into three compartments: shower, bathroom, and other spaces, accounting for daily water usage patterns and calculating VOC concentrations. Exposure data generated using the calculator can assist health assessors in estimating excess lifetime cancer risk (ELCR) and hazard index (HI) from VOC inhalation. Unlike traditional exposure models that utilize Andelman’s constant, the RWIRC continuously assesses variability in VOC concentrations and environmental conditions using differential equations to track VOC concentrations and air exchange between compartments. The calculator also provides unique volatilization fractions for each chemical and appliance, enhancing accuracy of the exposure concentration estimation. The RWIRC is accessible online and allows users to customize parameters (i.e., number of bathrooms, water temperature, and exhaust fan conditions) and input VOC characteristics (i.e., tap water and ambient air concentrations). This document provides a step-by-step guide on implementing the calculator. It also provides comparisons with the ATSDR-SHOWER calculator, using eight VOCs with varying physicochemical properties to reveal differences in algorithms and output concentrations. Simulations also assess how bathroom door positions and exhaust fan usage affect VOC exposure. The calculator results can enhance EPA risk screening levels for inhalation exposure to VOCs from tap water, offering a sophisticated tool for assessing inhalation risks and improving public health protection.

54 ENVIRONMENTAL SCIENCES↗

Burrowing Owl Conservation Report: 2020

The Western Burrowing Owl (Athene cunicularia) is a small migratory species of owl found throughout the Western United States. Burrowing Owls are found throughout a variety of habitats including shrubsteppe; and during the breeding season the Hanford Site is situated at the center of the predicted distribution of Burrowing Owls in Washington State (Smith et al. 1997). The Western Burrowing Owl is declining over much of its range, including on the Hanford Site and throughout the Columbia Basin (HNF-59375, Conway 2006). These declines have been attributed to habitat loss and a subsequent loss in burrowing mammals that the owls depend on for nesting habitat (Conway et al. 2006, Johnson et al. 2010). The Washington State Department of Fish and Wildlife lists Western Burrowing Owls as a Candidate Species, meaning the species is a candidate for listing as Sensitive, Threatened, or Endangered in Washington (WDFW 2020). Without intervention, continued declines may result in the extirpation of Burrowing Owls from the Hanford Site and the Columbia Basin.

54 ENVIRONMENTAL SCIENCES↗

Hanford Site Raptor Nest Monitoring Report for Calendar Year 2022

The U.S. Department of Energy, Richland Operations Office (DOE-RL), conducts ecological monitoring on the Hanford Site to collect and track data needed to ensure compliance with an array of environmental laws, regulations, and policies governing DOE-RL activities. Ecological monitoring data provide baseline information about the plants, animals, and habitats under DOERL stewardship at the Hanford Site required for decision making under the National Environmental Policy Act of 1969 (NEPA) and Comprehensive Environmental Response, Compensation, and Liability Act of 1980. DOE/EIS-0222, Final Hanford Comprehensive Land Use Plan Environmental Impact Statement, (CLUP) evaluates the potential environmental impacts associated with implementing a comprehensive land-use plan for the Hanford Site for at least the next 50 years and ensures that DOE-RL, its contractors, and other entities conduct activities on the Hanford Site in compliance with NEPA.

54 ENVIRONMENTAL SCIENCES↗