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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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At least 847 records · Page 47

Does Plant Biomass Manipulation in Static Chambers Affect Nitrous Oxide Emissions from Soils?

One of the most widespread approaches for measurement of greenhouse gas emissions from soils involves the use of static chambers. This method is relatively inexpensive, is easily replicated, and is ideally suited to plot-based experimental systems. Among its limitations is the loss of detection sensitivity with increasing chamber height, which creates challenges for deployment in systems including tall vegetation. It is not always possible to avoid inclusion of plants within chambers or to extend chamber height to fully accommodate plant growth. Thus, in many systems, such as perennial forages and biomass crops, plants growing within static chambers must either be trimmed or folded during lid closure. Currently, data on how different types of biomass manipulation affect measured results is limited. For this study, we compare the effects of cutting vs. folding of biomass on nitrous oxide measurements in switchgrass (Panicum virgatum L.) and alfalfa (Medicago sativa L.) systems. We report only limited evidence of treatment effects during discrete sampling events and little basis for concern that effects may intensify over time as biomass manipulation is repeatedly imposed. However, nonsignificant treatment effects that were consistently present amounted to significant overall trends in three out of the four systems studied. Such minor disparities in flux could amount to considerable quantities over time, suggesting that caution should be exercised when comparing cumulative emission values from studies using different biomass manipulation strategies.

54 ENVIRONMENTAL SCIENCES↗

Structural Mechanisms of Cooperative DNA Binding by Bacterial Single-Stranded DNA-Binding Proteins

Bacteria encode homooligomeric single-stranded (ss) DNA-binding proteins (SSBs) that coat and protect ssDNA intermediates formed during genome maintenance reactions. The prototypical Escherichia coli SSB tetramer can bind ssDNA using multiple modes that differ by the number of bases bound per tetramer and the magnitude of the binding cooperativity. Our understanding of the mechanisms underlying cooperative ssDNA binding by SSBs has been hampered by the limited amount of structural information available for interfaces that link adjacent SSB proteins on ssDNA. Here in this paper we present a crystal structure of Bacillus subtilis SsbA bound to ssDNA. The structure resolves SsbA tetramers joined together by a ssDNA “bridge” and identifies an interface, termed the “bridge interface,” that links adjacent SSB tetramers through an evolutionarily conserved surface near the ssDNA-binding site. E. coli SSB variants with altered bridge interface residues bind ssDNA with reduced cooperativity and with an altered distribution of DNA binding modes. These variants are also more readily displaced from ssDNA by RecA than wild-type SSB. In spite of these biochemical differences, each variant is able to complement deletion of the ssb gene in E. coli. Together our data suggest a model in which the bridge interface contributes to cooperative ssDNA binding and SSB function but that destabilization of the bridge interface is tolerated in cells.

59 BASIC BIOLOGICAL SCIENCES↗

Yeast require redox switching in DNA primase

Eukaryotic DNA primases contain a [4Fe4S] cluster in the C-terminal domain of the p58 subunit (p58C) that affects substrate affinity but is not required for catalysis. We show that, in yeast primase, the cluster serves as a DNA-mediated redox switch governing DNA binding, just as in human primase. Despite a different structural arrangement of tyrosines to facilitate electron transfer between the DNA substrate and [4Fe4S] cluster, in yeast, mutation of tyrosines Y395 and Y397 alters the same electron transfer chemistry and redox switch. Mutation of conserved tyrosine 395 diminishes the extent of p58C participation in normal redox-switching reactions, whereas mutation of conserved tyrosine 397 causes oxidative cluster degradation to the [3Fe4S] + species during p58C redox signaling. Switching between oxidized and reduced states in the presence of the Y397 mutations thus puts primase [4Fe4S] cluster integrity and function at risk. Consistent with these observations, we find that yeast tolerate mutations to Y395 in p58C, but the single-residue mutation Y397L in p58C is lethal. Our data thus show that a constellation of tyrosines for protein-DNA electron transfer mediates the redox switch in eukaryotic primases and is required for primase function in vivo.

59 BASIC BIOLOGICAL SCIENCES↗

Synthetic Graph Generation for Data-Intensive HPC Benchmarking: Scalability, Analysis and Real-World Application

The benchmarking effort within the Extreme Scale Systems Center at Oak Ridge National Laboratory seeks to provide High Performance Computing benchmarks and test suites of interest to the DoD sponsor. The work described in this report is a part of the effort focusing on graph generation. A previously developed benchmark, SystemBurn, allows the emulation of a broad spectrum of application behavior profiles within a single framework. To complement this effort, similar capabilities are desired for graph-centric problems. This report described the in-depth analysis of the generated synthetic graphs' properties at a variety of scales using different generator implementations and examines their applicability to replicating real world datasets.

97 MATHEMATICS AND COMPUTING↗

PINN surrogate of Li-ion battery models for parameter inference, Part II: Regularization and application of the pseudo-2D model

Bayesian parameter inference is useful to improve Li-ion battery diagnostics and can help formulate battery aging models. However, it is computationally intensive and cannot be easily repeated for multiple cycles, multiple operating conditions, or multiple replicate cells. To reduce the computational cost of Bayesian calibration, numerical solvers for physics-based models can be replaced with faster surrogates. A physics-informed neural network (PINN) is developed as a surrogate for the pseudo-2D (P2D) battery model calibration. For the P2D surrogate, additional training regularization was needed as compared to the PINN single-particle model (SPM) developed in Part I. Both the PINN SPM and P2D surrogate models are exercised for parameter inference and compared to data obtained from a direct numerical solution of the governing equations. A parameter inference study highlights the ability to use these PINNs to calibrate scaling parameters for the cathode Li diffusion and the anode exchange current density. By realizing computational speed-ups of ~2250x for the P2D model, as compared to using standard integrating methods, the PINN surrogates enable rapid state-of-health diagnostics. Finally, in the low-data availability scenario, the testing error was estimated to ~2 mV for the SPM surrogate and ~10 mV for the P2D surrogate which could be mitigated with additional data.

25 ENERGY STORAGE↗

Structural basis for antagonizing a host restriction factor by C7 family of poxvirus host-range proteins

Human sterile alpha motif domain-containing 9 (SAMD9) protein is a host restriction factor for poxviruses, but it can be overcome by some poxvirus host-range proteins that share homology with vaccinia virus C7 protein. To understand the mechanism of action for this important family of host-range factors, we determined the crystal structures of C7 and myxoma virus M64, a C7 family member that is unable to antagonize SAMD9. Despite their different functions and only 23% sequence identity, the two proteins have very similar overall structures, displaying a previously unidentified fold comprised of a compact 12-stranded antiparallel β-sandwich wrapped in two short α helices. Extensive structure-guided mutagenesis of C7 identified three loops clustered on one edge of the β sandwich as critical for viral replication and binding with SAMD9. Here, the loops are characterized with functionally important negatively charged, positively charged, and hydrophobic residues, respectively, together forming a unique “three-fingered molecular claw.” The key residues of the claw are not conserved in two C7 family members that do not antagonize SAMD9 but are conserved in distantly related C7 family members from four poxvirus genera that infect diverse mammalian species. Indeed, we found that all in the latter group of proteins bind SAMD9. Taken together, our data indicate that diverse mammalian poxviruses use a conserved molecular claw in a C7-like protein to target SAMD9 and overcome host restriction.

59 BASIC BIOLOGICAL SCIENCES↗

An updated gene atlas for maize reveals organ-specific and stress-induced genes

Maize (Zea mays L.), a model species for genetic studies, is one of the two most important crop species worldwide. The genome sequence of the reference genotype, B73, representative of the stiff stalk heterotic group was recently updated (AGPv4) using long-read sequencing and optical mapping technology. To facilitate the use of AGPv4 and to enable functional genomic studies and association of genotype with phenotype, we determined expression abundances for replicated mRNA-sequencing datasets from 79 tissues and five abiotic/biotic stress treatments revealing 36 207 expressed genes. Characterization of the B73 transcriptome across six organs revealed 4154 organ-specific and 7704 differentially expressed (DE) genes following stress treatment. Gene co-expression network analyses revealed 12 modules associated with distinct biological processes containing 13 590 genes providing a resource for further association of gene function based on co-expression patterns. Presence–absence variants (PAVs) previously identified using whole genome resequencing data from 61 additional inbred lines were enriched in organ-specific and stress-induced DE genes suggesting that PAVs may function in phenological variation and adaptation to environment. Relative to core genes conserved across the 62 profiled inbreds, PAVs have lower expression abundances which are correlated with their frequency of dispersion across inbreds and on average have significantly fewer co-expression network connections suggesting that a subset of PAVs may be on an evolutionary path to pseudogenization. Here, to facilitate use by the community, we developed the Maize Genomics Resource website (maize.plantbiology.msu.edu) for viewing and data-mining these resources and deployed two new views on the maize electronic Fluorescent Pictograph Browser (bar.utoronto.ca/efp_maize).

59 BASIC BIOLOGICAL SCIENCES↗

CD8 + lymphocyte control of SIV infection during antiretroviral therapy

CD8 + lymphocytes play an important role in suppressing in vivo viral replication in HIV infection. However, both the extent to which and the mechanisms by which CD8 + lymphocytes contribute to viral control are not completely understood. A recent experiment depleted CD8 + lymphocytes in simian immunodeficiency virus (SIV)-infected rhesus macaques (RMs) on antiretroviral treatment (ART) to study the role of CD8 + lymphocytes. CD8 + lymphocytes depletion resulted in temporary plasma viremia in all studied RMs. Viral control was restored when CD8 + lymphocytes repopulated. We developed a viral dynamic model to fit the viral load (VL) data from the CD8 depletion experiment. We explicitly modeled the dynamics of the latent reservoir and the SIV-specific effector cell population including their exhaustion and their potential cytolytic and noncytolytic functions. We found that the latent reservoir significantly contributes to the size of the peak VL after CD8 depletion, while drug efficacy plays a lesser role. Our model suggests that the overall CD8 + lymphocyte cytolytic killing rate is dynamically changing depending on the levels of antigen-induced effector cell activation and exhaustion. Based on estimated parameters, our model suggests that before ART or without ART the overall CD8 cytolytic killing rate is small due to exhaustion. However, after the start of ART, the overall CD8 cytolytic killing rate increases due to an expansion of SIV-specific CD8 effector cells. Further, we estimate that the cytolytic killing rate can be significantly larger than the cytopathic death rate in some animals during the second phase of ART-induced viral decay. Lastly, our model provides a new explanation for the puzzling findings by Klatt et al. and Wong et al. that CD8 depletion done immediately before ART has no noticeable effect on the first phase viral decay slope seen after ART initiation Overall, by incorporating effector cells and their exhaustion, our model can explain the effects of CD8 depletion on VL during ART, reveals a detailed dynamic role of CD8 + lymphocytes in controlling viral infection, and provides a unified explanation for CD8 depletion experimental data.

59 BASIC BIOLOGICAL SCIENCES↗

Modeling DOE Standard Canister Configurations with Updated Surface Chemistry

Road-ready and final disposition packaging configurations for the advanced test reactor (ATR) fuel currently specifies storage within helium backfilled DOE sealed standard canisters. The aluminum cladding of the ATR fuel contains an oxyhydroxide layer of boehmite/bayerite that generates hydrogen when subjected to irradiation. Understanding the effect of this hydrogen buildup over time to important for long term storage considerations. Previous modeling efforts have built a coupled CFD-chemical model to simulate the temperature gas phase concentrations within the DOE sealed standard canisters. These models have coupled the temperature conditions to both the gas phase radiolysis chemistry and the surface chemistry associated with the oxyhydroxide layer. Continued experimental work has identified trends for aluminum surrogate samples with oxyhydroxide layers for tests done at higher dose rates. At low initial doses a fast generation rate of hydrogen occurs which starts to roll over to a lower generation rate as the total dose applied in increased. A previous study created a small-scale chemical model was built to replicate a mini-canister surrogate system at SRNL as well as for the smaller capsule tests performed at INL. The previous iteration of the model utilized step function for its G-values for the hydrogen generation over a 200-year period. This model makes an update to the surface chemistry to account for theorized surface chemistry reactions allowing for oxygen to remain bounded to the oxyhydroxide layer. As additional experiments continue, the kinetic fits may be adjusted to adapt to new data. Three canister configurations are modeled – the base 18-inch, 15-foot DOR standard canister with 30 ATR fuel elements, an 18-inch, 10-foot DOE standard canister with 32 ATR fuel elements and a 24-inch, 10-foot DOE standard canister with 40 ATR fuel elements. In previous reports, the primary sensitivity of the canister conditions was identified as the decay heat of the fuel and the dried condition of the fuel. Only these parameters are studied in the report. For the nominal case with dried fuel, the hydrogen generation is only about 2%, so it is even less than the flammability limit if it were exposed to oxygen. For the densely packed 18-inch case the total hydrogen concentration is only around 4% with the nominal decay heat. For the densely packed 24-inch case the total hydrogen concentration is only 2.5%, roughly 20% higher than the original packing design, and still under flammability conditions if exposed to oxygen.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Aggregate attack surface management for network discovery of operational technology

Interconnectivity has become a substratum of technology as the benefits of data-driven functionality are being realized in nearly all industries. Increased connectivity of Operational Technology (OT) exacerbates cyber risks because Industrial Control Systems (ICS) are becoming exposed to the Internet. These exposures are often done inadvertently through misconfigurations as additional network devices come online. Attack surface management (ASM) platforms can be used to identify vulnerabilities by performing external network discovery over the Internet using web spiders. These web spiders enable big data analytics of Internet of Things (IoT) devices as identifiable information of Internet-exposed equipment are archived in searchable databases that are made publicly available. There are a multitude of ASM service providers on the market. Here, this study was conducted to evaluate several commonly known tools to determine the aggregate attack surface of control systems. Queries were crafted by targeting commonly known manufacturers and communication protocols found in OT networks. Identified devices were that categorized based on technology types. Each query was replicated between several tools to target identical ICS equipment. Findings in this paper suggested a significant variance in the exposures discovered by each tool, but unique contributions were identified for each tool when a merged attack surface was derived. Therefore, all tools should be used in aggregate.

97 MATHEMATICS AND COMPUTING↗

A comparison of cost and quality of three methods for estimating density for wild pig (Sus scrofa)

A critical element in efective wildlife management is monitoring the status of wildlife populations; however, resources to monitor wildlife populations are typically limited. We compared cost efectiveness of three common population estimation methods (i.e. non-invasive DNA sampling, camera sampling, and sampling from trapping) by applying them to wild pigs (Sus scrofa) across three habitats in South Carolina, U.S.A where they are invasive. We used mark-recapture analyses for fecal DNA sampling data, spatially-explicit capture-recapture analyses for camera sampling data, and a removal analysis for removal sampling from trap data. Density estimates were similar across methods. Camera sampling was the least expensive, but had large variances. Fecal DNA sampling was the most expensive, although this technique generally performed well. We examined how reductions in efort by method related to increases in relative bias or imprecision. For removal sampling, the largest cost savings while maintaining unbiased density estimates was from reducing the number of traps. For fecal DNA sampling, a reduction in efort only minimally reduced costs due to the need for increased lab replicates while maintaining high quality estimates. For camera sampling, efort could only be marginally reduced before inducing bias. We provide a decision tree for researchers to help make monitoring decisions.

54 ENVIRONMENTAL SCIENCES↗

SU-E-J-258: Inter- and Intra-Fraction Setup Stability and Couch Change Tolerance for Image Guided Radiation Therapy

Purpose: Immobilization devices combine rigid patient fixation as well as comfort and play a key role providing the stability required for accurate radiation delivery. In the setup step, couch re-positioning needed to align the patient is derived via registration of acquired versus reference image. For subsequent fractions, replicating the initial setup should yield identical alignment errors when compared to the reference. This is not always the case and further couch re-positioning can be needed. An important quality assurance measure is to set couch tolerances beyond which additional investigations are needed. The purpose of this work was to study the inter-fraction couch changes needed to re-align the patient and the intra-fraction stability of the alignment as a guide to establish the couch tolerances. Methods: Data from twelve patients treated on the Accuray CyberKnife (CK) system for fractionated intracranial radiotherapy and immobilized with Aquaplast RT, U-frame, F-Head-Support (Qfix, PA, USA) was used. Each fraction involved image acquisitions and registration with the reference to re-align the patient. The absolute couch position corresponding to the approved setup alignment was recorded per fraction. Intra-fraction set-up corrections were recorded throughout the treatment. Results: The average approved setup alignment was 0.03±0.28mm, 0.15±0.22mm, 0.06±0.31mm in the L/R, A/P, S/I directions respectively and 0.00±0.35degrees, 0.03±0.32degrees, 0.08±0.45degrees for roll, pitch and yaw respectively. The inter-fraction reproducibility of the couch position was 6.65mm, 10.55mm, and 4.77mm in the L/R, A/P and S/I directions respectively and 0.82degrees, 0.71degrees for roll and pitch respectively. Intra-fraction monitoring showed small average errors of 0.21±0.21mm, 0.00±0.08mm, 0.23±0.22mm in the L/R, A/P, S/I directions respectively and 0.03±0.12degrees, 0.04±0.25degrees, and 0.13±0.15degrees in the roll, pitch and yaw respectively. Conclusion: The inter-fraction reproducibility should serve as a guide to couch tolerances, specific to a site and immobilization. More patients need to be included to make general conclusions.

60 APPLIED LIFE SCIENCES↗

New Report Highlights LAMP's Role as a Robust and Reliable System for Wave Energy Converter Testing

Harnessing the power of waves requires more than just putting devices in the ocean - it takes tools that can refine and accelerate innovation before the first deployment. That's where NLR's largeamplitude motion platform, or LAMP, comes in. LAMP is a six-degree-of-freedom motion system designed to replicate the response of a wave energy converter (WEC) device in realistic ocean conditions. By letting researchers evaluate WEC performance in a safe, controlled lab environment before moving offshore, LAMP helps accelerate innovation while reducing risk. An NLR report, titled Initial Characterization of the NLR LargeAmplitude Motion Platform (https://www.nrel.gov/docs/fy26osti/93733.pdf), provides a deeper understanding of LAMP's capabilities and also lays the groundwork for future advancements in WEC testing methodologies. Researchers studied LAMP's amplitude and frequency limits across different payloads and collected data from 16 test profiles. Testing demonstrated that platform accuracy remained consistent across all payloads evaluated; researchers expect similar accuracy up to LAMP's 10,000-kilogram capacity limit. The measured position limits for single degrees of freedom experienced by WECs, including surge, sway, heave, roll, pitch, and yaw, also exceeded the original manufacturer specifications, providing new insight into the true boundaries of the system.

16 TIDAL AND WAVE POWER↗

Assessment of a tomography-informed polyhedral discrete element modelling approach for complex-shaped granular woody biomass in stress consolidation

The design of handling equipment for granular biomass primarily requires experiments. Discrete element models (DEM) can provide designers with detailed insight into the behaviour of granular materials. However, granular biomass comprising complex-shaped particles is difficult to model with DEM. A tomography-informed DEM approach and an exhaustive assessment of the ability of the approach to predict the bulk behaviour of milled pines using experimental data is presented. Nano-CT scan was conducted to obtain 3D particle surface geometries as the basis for particle shape approximation by a polyhedral model and a sphero-polyhedral model. These models were applied in the simulation of a compressibility test. Our parametric study showed that particle Young's modulus and restitution coefficient are the two main properties influencing the simulated bulk behaviour of the DEM particles and that the level of approximating DEM particle shape for real particles is critical for accurately replicating the bulk behaviour of milled pines. The polyhedral model demonstrated better suitability than its sphero-polyhedral counterpart for modelling pine particles. The polyhedral model calibrated in a compressibility test was then applied in the simulations of a friction test without additional parameter tuning. Interlocking was dominant in the shear of bulk pine particles. Remarkably, the polyhedral model predicted the frictional behaviour of the pine particles when compared to the experimental results. The limitations of the model, as well as possible ways for enhancement, are discussed. This work provided novel insights into the suitability of complex-shaped DEM models for granular woody biomass.

Biomass flow↗

Scalability analysis of heavy-duty gas turbines using data-driven machine learning

With the increasing integration of variable renewable energy sources into power systems, the role of flexible power generation technologies like gas turbines (GT) in rapid grid balancing remains crucial. This sustained importance underscores the need for scaled and precise modeling of GT to ensure effective integration within evolving energy frameworks. While physics-driven GT models integrate thermodynamics, fluid dynamics, and combustion principles, they often rely on approximate mathematical representations to accommodate scaling that may not capture the actual complex dynamics for GTs and inertial effects associated to GTs with different ratings. In this study, a data-driven model is proposed using machine learning (ML) techniques to conduct GT scalability analysis and performance evaluation with high accuracy. The ML model, trained on data from various operating conditions and performance parameters, aims to uncover intricate relationships and patterns, resembling GT characteristics at different scales (ratings). The model is developed to capture complex system interaction and to adapt to changing operational scenarios at different capacities, providing valuable insights of power system dynamics. In this study, the real-time digital simulator platform was employed to generate training data for the ML model and assess its dynamic characteristics. The ultimate objective was to develop a detailed modeling framework based on governing equations and data-driven ML capable of predicting key performance indicators, in thermal systems such as GTs, including power output, speed, fuel consumption, and exhaust temperature under diverse operating conditions at different scales. The developed ML framework demonstrated high accuracy, with mean relative errors for GT power prediction, reference speed, exhaust temperature, and compressor pressure ratio (CPR) parameters consistently below 0.1% across typical load fluctuation scenarios. Maximum deviations were limited to approximately 0.5 K for exhaust temperature and 0.009 for CPR, underscoring the model’s ability to replicating dynamic GT behavior with high precision. The adaptability of the ML model enables its application across diverse operational conditions and its extension to other thermal systems. By leveraging advanced ML techniques, this study presents a robust and scalable modeling framework that enhances GT simulation precision, facilitating improved integration into evolving power systems.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Enhanced Hanford Low-Activity Waste Glass Property Data Development: Phase 1

This report summarizes and analyzes data collected on a first test matrix of 36 low-activity waste (LAW) glass compositions intended to expand the LAW glass composition region over which glass property models are valid. The 36 LAW glass compositions were selected using statistical layered experimental design to explore a new glass composition region that overlaps and expands beyond the previously explored LAW glass composition region. The layered design of 36 LAW glasses consisted of 18 outer-layer glasses, 13 inner-layer glasses, a center glass (two replicates), and a glass previously tested at another laboratory (three replicates). The outer-layer glass compositions include extrema of the glass compositions that extend across and slightly beyond the compositions that are expected to be targeted for production. The analyses performed on these glasses include chemical composition (for target compositional verification), density, viscosity, electrical conductivity, crystal fraction, Canister Centerline Cooling with crystal identification, Product Consistency Test response, sulfur solubility, iron redox, and Vapor Hydration Test response. This report documents and discusses the results obtained from this testing.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Measurement of Plutonium Ratios for CRM 137A Recertification

This report documents the analytical methodologies used by the Chemical and Isotopics Mass Spectrometry (CIMS) Group at the US Department of Energy’s Oak Ridge National Laboratory (ORNL) for plutonium isotopic ratio characterizations on purified ~1 mg units prepared by Lawrence Livermore National Laboratory (LLNL) from a mother solution created from an existing 250 mg unit of certified reference material (CRM) 137. The analytical processes described are designed to maintain traceability of the final certified attributes and to eliminate sources of systematic bias for the measurements described. The measurands for certification are the plutonium isotope ratios: 238 Pu/ 239 Pu, 240 Pu/ 239 Pu, 241 Pu/ 239 Pu, and 242 Pu/ 239 Pu. The preparation and analytical process included the following activities: Preparation of reference materials to be used as calibrants to establish traceability to the SI unit of mass for the isotopic analyses and quality controls. For the plutonium isotopic analyses, CRM 128 (a CRM comprising equal atoms of 239 Pu and 242 Pu) was used as the primary calibrant; Each CRM 137A unit was reconstituted using dilute high-purity HNO3. From each, seven replicates were distributed for the isotopic characterizations by thermal ionization mass spectrometry (TIMS) using the total evaporation (TE) method for isotope ratio measurements and MC-ICP-MS using a dynamic peak hooping method for secondary isotope ratio measurements; Due to time delay in the analyses from the last purification of the original CRM 137 mother solution by LLNL, an aliquot of the three provided plutonium units was purified using Eichrom TEVA separation methods to remove americium in-growth and uranium impurities; Minimally corrected data were reported from each analysis using reporting templates provided by the New Brunswick Laboratory Program Office (NBL PO) for the specific methods used for isotopic measurements. In addition, a copy of the instrument exports, Excel templates for isotopic data corrections, and the uncertainty budgets completed with the GUM Workbench software package were submitted to NBL PO.

07 ISOTOPE AND RADIATION SOURCES↗

Pol I DNA polymerases stimulate DNA end-joining by Escherichia coli DNA ligase

Highlights: • Pol I DNA polymerases stimulate E. coli DNA ligase activity. • End-joining activity is enhanced by up to 600%, nick closure is slightly enhanced. • T4 ligase is not similarly effected. Klenow and Klentaq are the large fragment domains of the Pol I DNA polymerases from Escherichia coli and Thermus aquaticus, respectively. Herein, we show that both polymerases can significantly stimulate complementary intermolecular end-joining ligations by E.coli DNA ligase when the polymerases are present at concentrations lower than that of the DNA substrates. In contrast, high polymerase concentrations relative to the DNA substrates inhibit the intermolecular ligation activity of DNA ligase. Neither polymerase was able to stimulate the DNA ligase from T4 bacteriophage. Additionally, nick-closure by E. coli DNA ligase (but not T4 ligase) is slightly stimulated by both polymerases, but only at about 10% of the magnitude seen for end-joining enhancement. The data represent one of the first observations of direct polymerase-ligase interactions in prokaryotes, and suggest that the polymerases stabilize the associated DNA ends during intermolecular ligation, and that such a complex can be taken advantage of by some, but not all, DNA ligases.

60 APPLIED LIFE SCIENCES↗