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DOE OSTI2023

Characterizing inhibitors of human AP endonuclease 1

AP endonuclease 1 (APE1) processes DNA lesions including apurinic/apyrimidinic sites and 3´-blocking groups, mediating base excision repair and single strand break repair. Much effort has focused on developing specific inhibitors of APE1, which could have important applications in basic research and potentially lead to clinical anticancer agents. We used structural, biophysical, and biochemical methods to characterize several reported inhibitors, including 7-nitroindole-2-carboxylic acid (CRT0044876), given its small size, reported potency, and widespread use for studying APE1. Intriguingly, NMR chemical shift perturbation (CSP) experiments show that CRT0044876 and three similar indole-2-carboxylic acids bind a pocket distal from the APE1 active site. A crystal structure confirms these findings and defines the pose for 5-nitroindole-2-carboxylic acid. However, dynamic light scattering experiments show the indole compounds form colloidal aggregates that could bind (sequester) APE1, causing nonspecific inhibition. Endonuclease assays show the compounds lack significant APE1 inhibition under conditions (detergent) that disrupt aggregation. Thus, binding of the indole-2-carboxylic acids at the remote pocket does not inhibit APE1 repair activity. Myricetin also forms aggregates and lacks APE1 inhibition under aggregate-disrupting conditions. Two other reported compounds (MLS000552981, MLS000419194) inhibit APE1 in vitro with low micromolar IC50 and do not appear to aggregate in this concentration range. However, NMR CSP experiments indicate the compounds do not bind specifically to apo- or Mg 2+ -bound APE1, pointing to a non-specific mode of inhibition, possibly DNA binding. Our results highlight methods for rigorous interrogation of putative APE1 inhibitors and should facilitate future efforts to discover compounds that specifically inhibit this important repair enzyme.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

DNA asymmetry promotes SUMO modification of the single-stranded DNA-binding protein RPA

Repair of DNA double-stranded breaks by homologous recombination (HR) is dependent on DNA end resection and on post-translational modification of repair factors. In budding yeast, single-stranded DNA is coated by replication protein A (RPA) following DNA end resection, and DNA–RPA complexes are then SUMO-modified by the E3 ligase Siz2 to promote repair. Here, we show using enzymatic assays that DNA duplexes containing 3' single-stranded DNA overhangs increase the rate of RPA SUMO modification by Siz2. The SAP domain of Siz2 binds DNA duplexes and makes a key contribution to this process as highlighted by models and a crystal structure of Siz2 and by assays performed using protein mutants. Enzymatic assays performed using DNA that can accommodate multiple RPA proteins suggest a model in which the SUMO-RPA signal is amplified by successive rounds of Siz2-dependent SUMO modification of RPA and dissociation of SUMO-RPA at the junction between single- and double-stranded DNA. Our results provide insights on how DNA architecture scaffolds a substrate and E3 ligase to promote SUMO modification in the context of DNA repair.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2017

Coordination of the Ser2056 and Thr2609 Clusters of DNA-PKcs in Regulating Gamma Rays and Extremely Low Fluencies of Alpha-Particle Irradiation to G 0 /G 1 Phase Cells

The catalytic subunit of DNA dependent protein kinase (DNA-PKcs) and its kinase activity are critical for mediation of non-homologous end-joining (NHEJ) of DNA double-strand breaks (DSB) in mammalian cells after gamma-ray irradiation. Additionally, DNA-PKcs phosphorylations at the T2609 cluster and the S2056 cluster also affect DSB repair and cellular sensitivity to gamma radiation. Previously we reported that phosphorylations within these two regions affect not only NHEJ but also homologous recombination repair (HRR) dependent DSB repair. In this study, we further examine phenotypic effects on cells bearing various combinations of mutations within either or both regions. Effects studied included cell killing as well as chromosomal aberration induction after 0.5–8 Gy gamma-ray irradiation delivered to synchronized cells during the G0/G1 phase of the cell cycle. Blocking phosphorylation within the T2609 cluster was most critical regarding sensitization and depended on the number of available phosphorylation sites. It was also especially interesting that only one substitution of alanine in each of the two clusters separately abolished the restoration of wild-type sensitivity by DNA-PKcs. Similar patterns were seen for induction of chromosomal aberrations, reflecting their connection to cell killing. To study possible change in coordination between HRR and NHEJ directed repair in these DNA-PKcs mutant cell lines, we compared the induction of sister chromatid exchanges (SCEs) by very low fluencies of alpha particles with mutant cells defective in the HRR pathway that is required for induction of SCEs. Finally, levels of true SCEs induced by very low fluence of alpha-particle irradiation normally seen in wild-type cells were only slightly decreased in the S2056 cluster mutants, but were completely abolished in the T2609 cluster mutants and were indistinguishable from levels seen in HRR deficient cells. Again, a single substitution in the S2056 together with a single substitution in the T2609 cluster abolished SCE formation and thus also effectively interferes with HRR.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2018

A New Standard DNA Damage (SDD) Data Format

Our understanding of radiation-induced cellular damage has greatly improved over the past few decades. Despite this progress, there are still many obstacles to fully understand how radiation interacts with biologically relevant cellular components, such as DNA, to cause observable end points such as cell killing. Damage in DNA is identified as a major route of cell killing. One hurdle when modeling biological effects is the difficulty in directly comparing results generated by members of different research groups. Multiple Monte Carlo codes have been developed to simulate damage induction at the DNA scale, while at the same time various groups have developed models that describe DNA repair processes with varying levels of detail. These repair models are intrinsically linked to the damage model employed in their development, making it difficult to disentangle systematic effects in either part of the modeling chain. These modeling chains typically consist of track-structure Monte Carlo simulations of the physical interactions creating direct damages to DNA, followed by simulations of the production and initial reactions of chemical species causing so-called “indirect” damages. After the induction of DNA damage, DNA repair models combine the simulated damage patterns with biological models to determine the biological consequences of the damage. To date, the effect of the environment, such as molecular oxygen (normoxic vs. hypoxic), has been poorly considered. We propose a new standard DNA damage (SDD) data format to unify the interface between the simulation of damage induction in DNA and the biological modeling of DNA repair processes, and introduce the effect of the environment (molecular oxygen or other compounds) as a flexible parameter. Such a standard greatly facilitates intermodel comparisons, providing an ideal environment to tease out model assumptions and identify persistent, underlying mechanisms. Through inter-model comparisons, this unified standard has the potential to significantly advance our understanding of the underlying mechanisms of radiation-induced DNA damage and the resulting observable biological effects when radiation parameters and/or environmental conditions change.

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTAN↗

DOE OSTI2020

Cellular Response to Proton Irradiation: A Simulation Study with TOPAS-nBio

The cellular response to ionizing radiation continues to be of significant research interest in cancer radiotherapy, and DNA is recognized as the critical target for most of the biologic effects of radiation. Incident particles can cause initial DNA damages through physical and chemical interactions within a short time scale. Initial DNA damages can undergo repair via different pathways available at different stages of the cell cycle. The misrepair of DNA damage results in genomic rearrangement and causes mutations and chromosome aberrations, which are drivers of cell death. This work presents an integrated study of simulating cell response after proton irradiation with energies of 0.5–500 MeV (LET of 60–0.2 keV/µm). A model of a whole nucleus with fractal DNA geometry was implemented in TOPAS-nBio for initial DNA damage simulations. The default physics and chemistry models in TOPAS-nBio were used to describe interactions of primary particles, secondary particles, and radiolysis products within the nucleus. The initial DNA double-strand break (DSB) yield was found to increase from 6.5 DSB/Gy/Gbp at low-linear energy transfer (LET) of 0.2 keV/µm to 21.2 DSB/Gy/Gbp at high LET of 60 keV/µm. A mechanistic repair model was applied to predict the characteristics of DNA damage repair and dose response of chromosome aberrations. It was found that more than 95% of the DSBs are repaired within the first 24 h and the misrepaired DSB fraction increases rapidly with LET and reaches 15.8% at 60 keV/µm with an estimated chromosome aberration detection threshold of 3 Mbp. The dicentric and acentric fragment yields and the dose response of micronuclei formation after proton irradiation were calculated and compared with experimental results.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2022

PyPVRPM: Photovoltaic Reliability and Performance Model in Python

The ability to perform accurate techno-economic analysis of solar photovoltaic (PV) systems is essential for bankability and investment purposes. Most energy yield models assume an almost flawless operation (i.e., no failures); however, realistically, components fail and get repaired stochastically. This package, PyPVRPM, is a Python translation and improvement of the Language Kit (LK) based PhotoVoltaic Reliability Performance Model (PVRPM), which was first developed at Sandia National Laboratories in Goldsim software (Granata et al., 2011) (Miller et al., 2012). PyPVRPM allows the user to define a PV system at a specific location and incorporate failure, repair, and detection rates and distributions to calculate energy yield and other financial metrics such as the levelized cost of energy and net present value (Klise, Lavrova, et al., 2017). Our package is a simulation tool that uses NREL’s Python interface for System Advisor Model (SAM) (National Renewable Energy Laboratory, 2020b) (National Renewable Energy Laboratory, 2020a) to evaluate the performance of a PV plant throughout its lifetime by considering component reliability metrics. Besides the numerous benefits from migrating to Python (e.g., speed, libraries, batch analyses), it also expands on the failure and repair processes from the LK version by including the ability to vary monitoring strategies. These failures, repairs, and monitoring processes are based on user-defined distributions and values, enabling a more accurate and realistic representation of cost and availability throughout a PV system’s lifetime.

97 MATHEMATICS AND COMPUTING↗

DOE OSTI2014

Post-Closure Inspection Report for the Tonopah Test Range, Nevada, for Calendar Year 2013

This report provides the results of the annual post-closure inspections conducted at the closed Corrective Action Units (CAUs) located on the Tonopah Test Range (TTR), Nevada. This report covers calendar year 2013 and includes inspection and repair activities completed at the following CAUs: • CAU 400: Bomblet Pit and Five Points Landfill (TTR) • CAU 407: Roller Coaster RadSafe Area (TTR) • CAU 424: Area 3 Landfill Complexes (TTR) • CAU 453: Area 9 UXO Landfill (TTR) • CAU 487: Thunderwell Site (TTR) Inspections were conducted according to the post-closure plans in the approved Closure Reports and subsequent correspondence with the Nevada Division of Environmental Protection. The post-closure inspection plans and subsequent correspondence modifying the requirements for each CAU are included in Appendix B. The inspection checklists are included in Appendix C. Field notes are included in Appendix D. Photographs taken during inspections are included in Appendix E. The annual post-closure inspections were conducted on May 14, 2013. Maintenance was performed at CAU 400, CAU 424, and CAU 453. At CAU 400, animal burrows were backfilled. At CAU 424, erosion repairs were completed at Landfill Cell A3-3, subsidence was repaired at Landfill Cell A3-4, and additional lava rock was placed in high-traffic areas to mark the locations of the surface grade monuments at Landfill Cell A3-3 and Landfill Cell A3-8. At CAU 453, two areas of subsidence were repaired and animal burrows were backfilled. Vegetation monitoring was performed at the CAU 400 Five Points Landfill and CAU 407 in June 2013. The vegetation monitoring report is included in Appendix F.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2015

Evaluation of Hydrogen Sensors: Cooperative Research and Development Final Report, CRADA Number CRD-14-547

In preparation for the projected 2015 release of commercial hydrogen fuel cell vehicles, KPA has been contracted by Toyota Motors to develop a hydrogen safety system for vehicle repair facilities. Repair facility safety designs will include hydrogen sensors. KPA will identify critical sensor specifications for vehicle repair facilities. In collaboration with NREL, KPA will select and purchase commercial hydrogen sensors that meet or nearly meet requirements for deployment in vehicle repair facility. A two-phase field deployment plan to verify sensor performance has been developed.

30 DIRECT ENERGY CONVERSION↗

DOE OSTI2017

Validation of PV-RPM Code in the System Advisor Model.

This paper describes efforts made by Sandia National Laboratories (SNL) and the National Renewable Energy Laboratory (NREL) to validate the SNL developed PV Reliability Performance Model (PV - RPM) algorithm as implemented in the NREL System Advisor Model (SAM). The PV - RPM model is a library of functions that estimates component failure and repair in a photovoltaic system over a desired simulation period. The failure and repair distributions in this paper are probabilistic representations of component failure and repair based on data collected by SNL for a PV power plant operating in Arizona. The validation effort focuses on whether the failure and repair dist ributions used in the SAM implementation result in estimated failures that match the expected failures developed in the proof - of - concept implementation. Results indicate that the SAM implementation of PV - RPM provides the same results as the proof - of - concep t implementation, indicating the algorithms were reproduced successfully.

97 MATHEMATICS AND COMPUTING↗

DOE OSTI2017

Complete Report on the Development of Welding Parameters for Irradiated Materials

The advanced welding facility at the Radiochemical Engineering Development Center of Oak Ridge National Laboratory, which was conceived to enable research and development of weld repair techniques for nuclear power plant life extension, is now operational. The development of the facility and its advanced welding capabilities, along with the model materials for initial welding trials, were funded jointly by the U.S. Department of Energy, Office of Nuclear Energy, Light Water Reactor Sustainability Program, the Electric Power Research Institute, Long Term Operations Program and the Welding and Repair Technology Center, with additional support from Oak Ridge National Laboratory. Welding of irradiated materials was initiated on November 17, 2017, which marked a significant step in the development of the facility and the beginning of extensive welding research and development campaigns on irradiated materials that will eventually produce validated techniques and guidelines for weld repair activities carried out to extend the operational lifetimes of nuclear power plants beyond 60 years. This report summarizes the final steps that were required to complete weld process development, initial irradiated materials welding activities, near-term plans for irradiated materials welding, and plans for post-weld analyses that will be carried out to assess the ability of the advanced welding processes to make repairs on irradiated materials.

36 MATERIALS SCIENCE↗

DOE OSTI2021

Comprehensive Total Cost of Ownership Quantification for Vehicles with Different Size Classes and Powertrains

In order to accurately compare the costs of two vehicles, the total cost of ownership (TCO) should consist of all costs related to both purchasing and operating the vehicle. This TCO analysis builds on previous work to provide a comprehensive perspective of all relevant vehicle costs of ownership. In this report, we present what we believe to be the most comprehensive explicit financial analysis of the costs that will be incurred by a vehicle owner. This study considers vehicle cost and depreciation, financing, fuel costs, insurance costs, maintenance and repair costs, taxes and fees, and other operational costs to formulate a holistic total cost of ownership and operation of multiple different vehicles. For each of these cost parameters that together constitute a comprehensive TCO, extensive literature review and data analysis were performed to find representative values in order to build a holistic TCO for vehicles of all size classes. The light- and heavy-duty vehicles selected for analysis in this report are representative of those that are on the road today and expected to be available in the future. Important additive analyses in this study include systematic analysis of vehicle depreciation, in-depth examination of insurance premium costs, comprehensive maintenance and repair estimates, analysis of all relevant taxes and fees, and considerations of specific costs applicable to commercial vehicles. We find that cars depreciate faster than light trucks and that older plug-in electric vehicles have a greater depreciation rate than newer electric vehicles. Light-duty vehicle (LDV) insurance costs show comparable costs for different powertrains, and lower costs for larger size classes. Medium- and heavy-duty vehicle (MHDV) insurance costs vary significantly by vocation. Electric and electrified powertrains have lower maintenance and repair costs than internal combustion engine (ICE) powertrains for all vehicle sizes, relative to vehicle price. MHDV maintenance and repair costs depend heavily on vocation and duty cycle. LDV taxes and fees are comparable across powertrain types and size classes, though marginally higher registration fees exist for alternative fuel vehicles. MHDV fees depend on the vocation, weight rating, and state. Many electric tractor trailers would be affected by additional battery weight, reducing the available payload capacity, and this cost can be substantial. Electric vehicle charging for commercial vehicles can be time-consuming; labor rates can cause this cost to dominate TCO. With improved knowledge of each of the cost components, we calculate a lifetime TCO for comparison across vehicles of different types and attributes. For a simulated small sport utility vehicle in 2025, modeled using Autonomie, the hybrid electric vehicle (HEV) has the lowest cost, followed by the conventional ICE vehicle. For MHDV, TCO can be drastically different depending on the vocation. Long-haul vehicles typically have the lowest per-mile costs. Excluding labor costs, the class 4 delivery has a comparable TCO to the day cab. Vocational trucks, refuse trucks, and transit buses have a high per-mile cost of ownership due to maintenance and insurance. For all of these vehicles, the cost of operating the vehicle is heavily weighted by the labor of the driver, followed by the fuel costs. While the HEV begins as the lowest cost powertrain for passenger vehicles, fuel cells are forecast to reach cost parity by 2030 when hydrogen prices reach $\$ 5$/kg while battery electric vehicles (BEV) reach cost parity by 2035 at a battery cost of $\$ 98$ per usable kWh of capacity, with these two technologies being the lowest cost in 2050. For the class 8 day cab tractor, the HEV and ICE vehicle begin as the lowest cost powertrains, and the 250-mile BEV reduces in cost from the most expensive to the least expensive by 2030.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

DOE OSTI2021

Smart Methane Emission Detection System Development Final Report

03 NATURAL GAS↗

DOE OSTI2021

Smart Methane Emission Detection System Development (Final Report)

Working with the Department of Energy's National Energy Technology Laboratory, Southwest Research Institute® (SwRI®) developed a system to identify methane leaks reliably, accurately, and autonomously at critical midstream sections of the natural gas distribution network in real-time for the purpose of mitigating methane emissions using Optical Gas Imaging (OGI) cameras. SwRI's Smart Leak Detection – Methane (SLED/M) adds a high degree of automation to the process of methane leak detection to minimize sources of human error, minimize response time to a leak event, and maximize midstream visibility. Furthermore, SwRI has been working towards integrating Quantitative OGI (QOGI) capabilities into this existing technology. By leveraging Deep Learning, SwRI now has the capability to estimate fugitive emission leak rates quickly and reliably, which allows operators to detect emissions, quantify leak rate, prioritize repairs, and validate the repairs in a single instrument. The next generation QOGI technology leverages the same cameras used in Leak Detection and Repair (LDAR) programs, with improvements in safety and speed for traditional quantification-based repairs, ultimately leading to less overhead cost for the operators. The goals for this research were to develop two types of models with the following goals: Run in real-time on the edge (≥ 12 Hz), Classification: Achieve less than 5% false positive detection, Classification: Achieve ≥ 95% methane plume detection rate, Regression: achieve ≤ 10 standard cubic feet per hour (scfh) prediction > 70% of the time. In order to achieve these results, multiple infrared (IR) and other sensors were investigated in tandem with the midwave IR (MWIR) OGI to provide additional information to train the underlying models. Information on atmospheric conditions including humidity, temperature, pressure, and solar radiation was provided by a weather station. Several machine learning and deep learning architectures and methods, including looking at quantized classification networks and regressions networks, were explored. As further data was collected, curated, and labeled, it allowed for more refined regressive networks to be adequately trained, leading to better insight into the true flow rates being observed. An important valuable deliverable of this research effort was the development of an advanced network which underwent multiple iterations capable of giving a continuous output. The current network has a predicted mean average percentage error (MAPE) of 12.3% just outside our target goal of 10.00%, but an accuracy of 97.78% at ±50 scfh, well within the overall goal for the Department of Energy (DOE) program. Upon closer inspection, it was observed that more than 10% of datapoints contributing to the MAPE predictions were the result of low flow rate predictions and are beyond the sensitivity of instrument measurement as a result of normal operational variation and noise.

03 NATURAL GAS↗

DOE OSTI2021

Post-Closure Inspection Report for the Tonopah Test Range and Nevada Test and Training Range, Nevada for Calendar Year 2020 (Rev. 0)

This report provides the results of the annual post-closure use restriction (UR) inspections at corrective action sites (CASs) located on the Tonopah Test Range (TTR) and Nevada Test and Training Range (NTTR). This report covers post-closure UR inspections for calendar year 2020, and includes visual inspections and repair activities completed at the following corrective action units (CAUs): • CAU 407, Roller Coaster RadSafe Area (TTR) • CAU 424, Area 3 Landfill Complexes (TTR) • CAU 453, Area 9 UXO Landfill (TTR) • CAU 487, Thunderwell Site (TTR). Note: CAU 400, Bomblet Pit and Five Points Landfill (TTR) (CASs TA-19-001-05PT, Ordnance Disposal Pit; and TA-55-00-TAB2, Ordnance Disposal Pit) no longer requires inspection and reporting, as described in the Post-Closure Inspection Report for the Tonopah Test Range and Nevada Test and Training Range, Nevada for Calendar Year 2019. Site figures are included in Appendix A. The Post-Closure Inspection Plans and Record of Technical Change modifying the requirements for each UR are included in Appendix B. The Post-Closure Inspection Checklists are included in Appendix C. Field Notes are included in Appendix D. Appendix E, Inspection Photographs, does not include photographs because none were taken during the 2020 post-closure inspections. Appendix F, Post-Closure Vegetation Monitoring Report, does not include any data because vegetation monitoring was not conducted or required at any CAU in 2020. Visual inspections were conducted according to the post-closure requirements outlined in specific CAU and CAS URs, and approved by NDEP. The annual post-closure inspections for CASs associated with CAUs 407, 424, 453, and 487 were conducted on May 20, 2020. No maintenance or repair issues were noted at CAUs 487, or at CASs associated with CAU 424 (except CASs 03-08-002-A304 and 03-08-002-A308). Maintenance items and subsequent repairs include the following: • CAU 407, CAS TA-23-001-TARC (Roller Coaster RadSafe Area): A UR sign was loose and was rehung during the inspection. • CAU 424, CAS 03-08-002-A304 (Landfill Cell A3-4): The northeast monument needed a UR sign. The UR sign was attached to the monument on August 11, 2020. • CAU 424, CAS 03-08-002-A308 (Landfill Cell A3-8): The southeast monument needed additional lava rock. The lava rock was added during the inspection. • CAU 453, CAS 09-55-001-0952 (Area 9 Landfill): Two animal burrows were noted in the western portion of the northern and southern trenches (one burrow per trench). The animal burrows were repaired on August 11, 2020.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2023

Resilient Hydrogels from the Nanoscale to the Macroscale

Biological systems illustrate how a material composed of fragile molecular components can collectively be highly resilient. While the average protein, cell, or even tissue may not last more than a few weeks, many animals and plants live for more than a century. Continual component regeneration and multiple systems to resist mechanical and chemical damage together make this longetivity possible. This project sought to develop biomimetic methods to enable a specific type of material, a hydrogel, to resist damage across multiple scales using distinct, modular damage protection mechanisms. Because hydrogels share many features with biological tissues, they are an ideal substrate for exploring biomimetic strategies for designing resilience and self-repair. We specifically focused in this study on DNA-crosslinked hydrogels, which contain DNA strands that can serve as material to link it together or to control its current state or properties. We investigated how new tools from dynamic DNA nanotechnology could make it possible to actively recover from damage by continually growing and forming a materials shape, or by identifying damage and directing an adaptive response to that damage involving chemical synthesis to reconstruct a structure. We developed embedded molecular sensors able to detect and strain of the gel before damage occurred and react to counteract damage. We also developed methods to continually create shapes and patterns using chemical processes that will allow those patterns to reform when they are damaged. The ability to design resilient materials capable of self-repair has important implications for materials engineering. Instead of designing a material to withstand the worst stresses it may encounter, we could instead design a material to survive under average conditions, but self-repair or reconfigure to resist impending damage. Resilient, self-repairing hydrogels will also have diverse applications such as sensors or actuators.

36 MATERIALS SCIENCE↗

DOE OSTI2015

Arabidopsis DNA polymerase lambda mutant is mildly sensitive to DNA double strand breaks but defective in integration of a transgene

The DNA double-strand break (DSB) is a critical type of damage, and can be induced by both endogenous sources (e.g., errors of oxidative metabolism, transposable elements, programmed meiotic breaks, or perturbation of the DNA replication fork) and exogenous sources (e.g., ionizing radiation or radiomimetic chemicals). Although higher plants, like mammals, are thought to preferentially repair DSBs via nonhomologous end joining (NHEJ), much remains unclear about plant DSB repair pathways. Our reverse genetic approach suggests that DNA polymerase λ is involved in DSB repair in Arabidopsis . The Arabidopsis T-DNA insertion mutant (atpolλ-1) displayed sensitivity to both gamma-irradiation and treatment with radiomimetic reagents, but not to other DNA damaging treatments. The (atpolλ-1) mutant showed a moderate sensitivity to DSBs, while Arabidopsis Ku70 and DNA ligase 4 mutants (atku70-3 and atlig4-2) , both of which play critical roles in NHEJ, exhibited a hypersensitivity to these treatments. The atpolλ-1/atlig4-2 double mutant exhibited a higher sensitivity to DSBs than each single mutant, but the atku70/atpolλ-1 showed similar sensitivity to the atku70-3 . We showed that transcription of the DNA ligase 1, DNA ligase 6 , and Wee1 genes was quickly induced by BLM in several NHEJ deficient mutants in contrast to wild-type. Finally, the T-DNA transformation efficiency dropped in NHEJ deficient mutants and the lowest transformation efficiency was scored in the atpolλ-1/atlig4-2 double mutant. These results imply that AtPolλ is involved in both DSB repair and DNA damage response pathway.

Plant Sciences↗

DOE OSTI2024

Investigation of Debonding Effect in Internal Replacement Pipe System Under Lateral Loading

The aging pipeline infrastructure around the world necessitates immediate rehabilitation. Internal replacement pipe (IRP) is a trenchless system offering a versatile and cost-effective solution across a variety of industries, including oil, natural gas, water, and wastewater. As a structural pipeline repair system, IRPs are subject to lateral deformation because of surface traffic loading. The present study evaluates the impact of adhesion between the host pipe and the IRP, with a focus on assessing the debonding effect on the behavior of the repair system under lateral deformation and bending. This was achieved using a comprehensive approach, including experimental, numerical, and analytical techniques. Varying levels of adhesive strength resulting from different methods of surface preparation were considered. The effectiveness of the IRP system on both discontinuous host pipes with various crack widths and continuous host pipes was also investigated. The results demonstrate that adhesive strength exerts a significant influence on the repair system, especially in the case of narrow circumferential cracks, while its impact on the continuous system is minimal. For optimal performance, it is essential to choose adhesives that possess sufficient shear strength while also accounting for the required debonding length. This approach ensures that minor discontinuities are effectively controlled, thereby enhancing the system′s fatigue life. The reliable determination of the maximum allowable shear strength for the adhesive or the debonding length can ensure that it does not negatively affect fatigue life. The findings presented in this study offer new insights into the development of trenchless repair techniques that can enhance system performance and extend service life.

Tien, Tri C. M.↗

NASA NTRS1976

Aerospike thrust chamber program

An existing, but damaged, 25,000-pound thrust, flightweight, oxygen/hydrogen aerospike rocket thrust chamber was disassembled and partially repaired. A description is presented of the aerospike chamber configuration and of the damage it had suffered. Techniques for aerospike thrust chamber repair were developed, and are described, covering repair procedures for lightweight tubular nozzles, titanium thrust structures, and copper channel combustors. Effort was terminated prior to completion of the repairs and conduct of a planned hot fire test program when it was found that the copper alloy walls of many of the thrust chamber's 24 combustors had been degraded in strength and ductility during the initial fabrication of the thrust chamber. The degradation is discussed and traced to a reaction between oxygen and/or oxides diffused into the copper alloy during fabrication processes and the hydrogen utilized as a brazing furnace atmosphere during the initial assembly operation on many of the combustors. The effects of the H2/O2 reaction within the copper alloy are described.

Campbell, J., Jr.↗