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At least 127 records · Page 7

Structural snapshots of human DNA polymerase μ engaged on a DNA double-strand break

Genomic integrity is threatened by cytotoxic DNA double-strand breaks (DSBs), which must be resolved efficiently to prevent sequence loss, chromosomal rearrangements/translocations, or cell death. Polymerase μ (Polμ) participates in DSB repair via the nonhomologous end-joining (NHEJ) pathway, by filling small sequence gaps in broken ends to create substrates ultimately ligatable by DNA Ligase IV. Here we present structures of human Polμ engaging a DSB substrate. Synapsis is mediated solely by Polμ, facilitated by single-nucleotide homology at the break site, wherein both ends of the discontinuous template strand are stabilized by a hydrogen bonding network. The active site in the quaternary Pol μ complex is poised for catalysis and nucleotide incoporation proceeds in crystallo. These structures demonstrate that Polμ may address complementary DSB substrates during NHEJ in a manner indistinguishable from single-strand breaks.

59 BASIC BIOLOGICAL SCIENCES↗

Elongation factor-Tu can repetitively engage aminoacyl-tRNA within the ribosome during the proofreading stage of tRNA selection

The substrate for ribosomes actively engaged in protein synthesis is a ternary complex of elongation factor Tu (EF-Tu), aminoacyl-tRNA (aa-tRNA), and GTP. EF-Tu plays a critical role in mRNA decoding by increasing the rate and fidelity of aa-tRNA selection at each mRNA codon. Here, using three-color single-molecule fluorescence resonance energy transfer imaging and molecular dynamics simulations, we examine the timing and role of conformational events that mediate the release of aa-tRNA from EF-Tu and EF-Tu from the ribosome after GTP hydrolysis. Our investigations reveal that conformational changes in EF-Tu coordinate the rate-limiting passage of aa-tRNA through the accommodation corridor en route to the peptidyl transferase center of the large ribosomal subunit. Experiments using distinct inhibitors of the accommodation process further show that aa-tRNA must at least partially transit the accommodation corridor for EF-Tu∙GDP to release. aa-tRNAs failing to undergo peptide bond formation at the end of accommodation corridor passage after EF-Tu release can be reengaged by EF-Tu∙GTP from solution, coupled to GTP hydrolysis. These observations suggest that additional rounds of ternary complex formation can occur on the ribosome during proofreading, particularly when peptide bond formation is slow, which may serve to increase both the rate and fidelity of protein synthesis at the expense of GTP hydrolysis.

59 BASIC BIOLOGICAL SCIENCES↗

Engineering a tumor-selective prodrug T-cell engager bispecific antibody for safer immunotherapy

T-cell engaging (TCE) bispecific antibodies are potent drugs that trigger the immune system to eliminate cancer cells, but administration can be accompanied by toxic side effects that limit dosing. TCEs function by binding to cell surface receptors on T cells, frequently CD3, with one arm of the bispecific antibody while the other arm binds to cell surface antigens on cancer cells. On-target, off-tumor toxicity can arise when the target antigen is also present on healthy cells. The toxicity of TCEs may be ameliorated through the use of pro-drug forms of the TCE, which are not fully functional until recruited to the tumor microenvironment. This can be accomplished by masking the anti-CD3 arm of the TCE with an autoinhibitory motif that is released by tumor-enriched proteases. Here, we solve the crystal structure of the antigen-binding fragment of a novel anti-CD3 antibody, E10, in complex with its epitope from CD3 and use this information to engineer a masked form of the antibody that can activate by the tumor-enriched protease matrix metalloproteinase 2 (MMP-2). We demonstrate with binding experiments and in vitro T-cell activation and killing assays that our designed prodrug TCE is capable of tumor-selective T-cell activity that is dependent upon MMP-2. Furthermore, we demonstrate that a similar masking strategy can be used to create a pro-drug form of the frequently used anti-CD3 antibody SP34. This study showcases an approach to developing immune-modulating therapeutics that prioritizes safety and has the potential to advance cancer immunotherapy treatment strategies.

60 APPLIED LIFE SCIENCES↗

A Two-Stage Approach for PV Inverter Engagement in Power Factor Correction and Voltage Regulation

The rapid integration of distributed energy resources, like solar photovoltaics (PVs), can lead to overvolt-age challenges due to reverse power flow and a noticeable decrease in power factor at the substation interface. While existing literature extensively explores utilizing smart inverter capabilities for reactive power flexibility using a volt-var curve (VVC), obtaining time-varying operating points of such curves in real-time is challenging due to computational demands and communication requirements. Similarly, employing optimization-based approaches for reactive power control and active voltage regulation in large-scale distribution feeders is difficult due to the complexity of the problem and the challenges in effectively engaging customer-owned resources. This paper proposes a two-stage strategy to harness smart inverters for reactive power support. The first stage formulates short-term planning by optimally designing VVCs (on a daily or hourly basis) for large-scale solar PVs based on projected system needs and communicating optimal curves to smart inverters in advance. Subsequently, the second stage employs a transactive-based method to involve customer-owned PVs for reactive power support, effectively enhancing overall system performance and addressing real-time demands. In conclusion, the efficacy of this approach will be demonstrated using real-world distribution circuits provided by Vermont Electric Power Company (VELCO) and Vermont Electric Cooperative (VEC).

Poudel, Shiva [Pacific Northwest National Laborato↗

A protective and broadly binding antibody class engages the influenza virus hemagglutinin head at its stem interface

ABSTRACT Influenza infection and vaccination impart strain-specific immunity that protects against neither seasonal antigenic variants nor the next pandemic. However, antibodies directed to conserved sites can confer broad protection. Here, we identify and characterize a class of human antibodies that engage a previously undescribed, conserved epitope on the influenza hemagglutinin (HA) protein. Prototype antibody S8V1-157 binds at the normally occluded interface between the HA head and stem. Antibodies to this HA head–stem interface epitope are non-neutralizingin vitrobut protect against lethal influenza infection in mice. These antibodies bind to most influenza A subtypes and seasonal human variants, and are present at low frequencies in the memory B cell populations of multiple human donors. Vaccines designed to elicit these antibodies might contribute to “universal” influenza immunity. IMPORTANCE Antibodies to the influenza virus hemagglutinin (HA) protein confer the strongest protection against infection. Human antibodies elicited by infection and/or vaccination fail to protect against antigenically novel animal, pandemic, or human seasonal viruses. Improved vaccines are needed. We identify a novel class of antibodies that bind most divergent HA subtypes and all seasonal human HA antigenic variants tested. These antibodies confer protection from lethal influenza challenge in animal models. The corresponding epitope on the HA head is occluded by its interaction with the stem and is inaccessible in the well-resolved prefusion state. The immunogenicity of this head–stem interface indicates that poorly understood conformations of HA presenting widely conserved surfaces are explored in biochemical, cell-based, andin vivoassays. Head–stem interface antibodies warrant further investigation as an avenue to improve influenza vaccines and therapeutics.

Microbiology↗

EPOC Deep Dive Retrospective: A Brief Overview of 7 years of Science Engagement Discussions

Understanding the appropriate ways cyberinfrastructure can be designed, implemented, and executed for scientific use cases requires a deep understanding of the way that researchers and educators interact with technology, and how it may be best implemented to suit their needs. The Engagement and Performance Operations Center (EPOC) has conducted a series of scientific “Deep Dives” of use cases at partner institutions to better understand the requirements for modern scientific innovation across the United States research complex. The results of these activities have revealed gaps in the way that technology has been used to foster research activities. This gap in cyberinfrastructure support has impacts for the overall productivity and innovation possibilities for scientific users.

Zurawski, Jason↗

Remote Target Engagement System (RTES) Generation 4 ("Gen 4")

RTES is designed to provide teleoperation of remote weapon platform capability at defensive physical protection locations requiring active lethal capabilities for final access denial. Multiple operator consoles allow for coordinated engagement of many weapon platforms by one or more operator consoles. This results in faster response times, increased effectiveness, and increased survivability from removing response force from harm's way increasing survivability. This software is developed against DoD requirements and has been safety and functionally reviewed and approved for operational use. Sandia National Laboratories is a multimission laboratory managed and operated by National Technology & Engineering Solutions of Sandia, LLC, a wholly owned subsidiary of Honeywell International Inc., for the U.S. Department of Energy’s National Nuclear Security Administration under contract DE-NA0003525. SAND2021-1935 O

Klingler, KristopherR.↗

NLR Engage™: A Calliope Web Environment for Collaborative Energy Systems Modeling and Planning [SWR-20-16]

NLR Engage™ is a free, open-access energy system planning tool that allows multiple users/stakeholders to develop and collaborate on capacity expansion models. The tool, developed at the National Laboratory of the Rockies (NLR), provides a collaborative and easy-to-use interface built on Calliope, a multi-scale energy systems modeling framework. Visit the NLR hosted webtool at https://engage.nlr.gov/en/login/?next=/en/

Harris, Tom↗

Informing and Engaging the Future Workforce on Generation IV International Forum Reactor Systems

Generation IV reactors are six nuclear reactor designs, Sodium Fast reactor (SFR), Very High Temperature Reactor (VHTR), Gas cooled Fast Reactor (GFR), Molten Salt Reactor (MSR), Lead Fast Reactor (LFR), and Supercritical Water Reactor (SCWR), that are considered to be the most promising in light of various criteria based on the following objectives: • continuation of the progress made by Generation III water reactors in terms of competitiveness and safety; • more effective use of uranium resources; • less radioactive waste, especially high-level, long-lived waste; • greater protection against malicious acts and the diversion or theft of nuclear materials. The GIF Education and Training Working Group (ETWG) launched two major initiatives in 2016 and in 2021 to not only inform and educate but also to engage the future workforce in support of these reactor systems. This paper will present an overview of the GIF EWTG webinar series as well as the 2021 Pitch your Gen IV Research Competition which is followed by the 2023 Pitch your Gen IV Research. Competition

Paviet, Patricia D.↗

Partnership and Community Engagement [Slides]

Partnerships ensure a strong Laboratory and support our missions. This presentation introduces partnerships then discusses institutes and university engagement, industrial partnerships, and regional pueblos.

96 KNOWLEDGE MANAGEMENT AND PRESERVATION↗

Learning Management System User Requirements for the National Nuclear Security Administration's International Nuclear Safeguards Engagement Program

The National Nuclear Security Administration's (NNSA) International Nuclear Safeguards Engagement Program (INSEP) is considering investing in new tools that would allow the program to support its partner states from a distance. At the same time, the program is considering approaches that would allow several organizations, including NNSA, IAEA, national laboratories and contractor staff, to collaborate in the development and maintenance of instructional content. Software systems known as Learning Management Systems (LMSs) might represent a mechanism through which INSEP could accomplish these goals (collaborative development and remote support). To assess the usefulness of an LMS, INSEP has specified its needs for delivering online training and compared those needs to the capability of a range of LMSs. This comparison will allow INSEP to determine whether an LMS would be a useful tool and may set the stage for a "make-buy" decision in the future. The study team concluded that INSEP's content development and delivery needs align well with the capabilities of the leading LMSs on the market today and that that INSEP performance requirements allow for a customized approach using existing training portals that are already available to NNSA. Additional work would be required to specify the desired processes for developing online training and outreach materials, structuring the databases, specifying the data that should be collected, and detailing the desired system reports and documentation.

96 KNOWLEDGE MANAGEMENT AND PRESERVATION↗

Barriers and Opportunities for U.S. Hydropower Industry Engagement in Brazil and Argentina

This Barriers and Opportunities for U.S. Hydropower Industry Engagement in Brazil and Argentina project seeks to enhance the understanding of hydropower resources, existing installations, and opportunities for U.S. company involvement in hydropower development in Brazil and Argentina. It covers conventional hydropower, pumped storage hydropower, run-of-river-hydropower, in-conduit generation, hydropower upgrades to existing facilities, and similar systems. This work recognizes that each of these countries, and even regions within the countries, have varied topography, weather, existing grid infrastructure, social, economic, and other factors, leading to relatively unique opportunities and challenges for hydropower development.

13 HYDRO ENERGY↗

INECP Country Engagement Plans and Transformational Events [Slides]

The International Nonproliferation Export Control Program (INECP) harnesses the expertise of the U.S. Department of Energy (DOE) national laboratories to assist partners internationally in implementing and adopting strategic trade control measures that can effectively counter WMD procurement efforts. This presentation includes discussion of work with LANL INECP technical leads to update the engagement plan for Turkey, which involved researching Turkey’s WMD related legal framework, relevant export control agencies, and WMD-relevant industries (e.g. advanced manufacturing and aerospace).

98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗

Science and Solutions for a Planet Under Pressure: Synthetic Research, Analytics, and Community Engagement on Five Priority Theme Areas

Support from the Department of Energy proved instrumental in supporting the work of Global Council for Science and the Environment (GCSE) on synthetic research, analysis, and community engagement, oriented around the themes and discussions from the GCSE Drawdown 2021 Conference — Research to Action: Science and Solutions for a Planet Under Pressure. These efforts helped inform and enhance the societal relevance of the knowledge that has been generated through the conference and, at the same time, advance the scientific insights acquired through synthesis and scientific translation efforts. The work centered on three main objectives: 1) documentation of findings from the completed GCSE 2021, culminating in the publication of a synthesis report; 2) development and vetting of integrated use cases that illustrate synthesis methods and enable scientific outreach, communications, and translational discussions among universities and the broader research community; and 3) strengthening the scientific basis for energy education research by sharing findings in university-educator fora.

54 ENVIRONMENTAL SCIENCES↗

A Guide to Engaging Underserved Communities in Commercial Energy Efficiency Field Validations

Underserved communities in the United States often experience the negative impacts of climate change and environmental degradation but enjoy few of the benefits of technological and environmental advances. The White House has addressed this inequity through the Justice40 initiative, which requires 40% of the benefits of select federal investments to be directed to underserved communities (The White House, 2022). Clean energy and energy efficiency are two highlighted investment categories, so the U.S. Department of Energy will guide implementation of the Justice40 initiative by, among other things, decreasing energy burdens, increasing parity in clean energy technology access and adoption, and increasing energy resiliency. A strategy for reaching these goals is to evaluate and validate new energy efficiency technologies in commercial buildings in underserved communities, where buildings may be older, smaller, and have deferred maintenance due to historical underinvestment. This paper develops guidance for researchers pursuing field validations with underserved communities. Historical redlining and past negative experiences with government and large institutions may make residents wary of participating in these field validations. Researchers, therefore, may need to spend more time building relationships and matching technologies to buildings. In this paper, we analyzed technical reports to identify common field validation building characteristics and conducted semi-structured expert conversations to identify key stages and major themes of engaging underserved communities. Results indicate there may be flexibility in site selection and there are steps researchers can take to support collaboration with communities. Results also suggest benefits to both the community and energy efficiency research.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Electrically Engaged UnduLation (EEL) Marine Energy System

Current technologies at the forefront of instrumentation for ocean observation through acoustic mapping are powered by batteries that cannot achieve the lasting durations required for high-resolution measurements. Pyro-E has designed an Electrically Engaged unduLation (EEL) system capable of in situ energy harvest and deployment in sub-sea configurations. By extracting kinetic energy from the ocean, EEL proposes to provide power output ranging from 1 mW to 1,000 mW. Such a magnitude of power can sustain active instrumentation with onboard processing of data and over-the-horizon telemetry. Pyro-E and Oak Ridge National Laboratory have utilized software tools and powerful computing resources to develop a techno-economic design of the EEL capable of harvesting 5 W. The material response of the EEL under simulated hydromechanical loads is described through time-accurate predictions. All physical tests were conducted by Pyro-E to guide simulations at Oak Ridge National Laboratory.

16 TIDAL AND WAVE POWER↗

Louisville Communities LEAP Engagement: Improving Energy Efficiency in Affordable Housing [Slides]

This presentation outlines the initial results from a year-plus engagement between the National Renewable Energy Lab and Louisville, Kentucky through the U.S. Department of Energy's Communities LEAP (Local Energy Action Program) effort. Louisville, as represented by the Louisville Metro Government, Kentuckians for the Commonwealth, and the Metropolitan Housing Coalition, sought technical assistance from the labs to address energy usage in its residential housing stock as part of efforts to reduce community-wide emissions and high energy burdens throughout its community. The Communities LEAP scope of work included an energy efficiency analysis that leveraged ResStock data to identify key efficient technologies to explore for retrofitting Louisville's housing stock; technical support around developing a community benchmarking ordinance to help Louisville track its progress towards reducing energy usage; a workforce development overview to help Louisville ensure a sufficient and equitable distribution of clean-energy-related jobs supported by its energy efficiency efforts; a policy analysis component to track energy efficiency related policies Louisville may be interested in adapting from peer communities across the country; and a financing analysis component to identify opportunities to lower the barrier to clean energy adoption.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗