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At least 181 records · Page 10

Non-Resource Conservation and Recovery Act Corrective Action Unit (CAU) Post-Closure Inspection Report, Nevada National Security Site, Nevada For Calendar Year 2022

This report includes visual inspection results, descriptions of maintenance and repair activities, and recommendations for calendar year (CY) 2022 for use restrictions (URs) at corrective action sites (CASs) located on the Nevada National Security Site (NNSS) and on the Nevada Test and Training Range (NTTR) that are accessed through the NNSS Main Gate. This document is formatted by project activity (i.e., Industrial Sites, Soils, and Defense Threat Reduction Agency [DTRA]); then by corrective action unit (CAU); and then by CAS.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Non-Resource Conservation and Recovery Act Corrective Action Unit (CAU) Post-Closure Inspection Report, Nevada National Security Site, Nevada: For Calendar Year 2023

This report includes visual inspection results, descriptions of maintenance and repair activities, and recommendations for CY 2023 for URs at CASs located on the NNSS and on the NTTR that are accessed through the NNSS Main Gate. This document is formatted by project activity (i.e., Industrial Sites, Soils, and DTRA]; then by CAU; and then by CAS.

54 ENVIRONMENTAL SCIENCES↗

Microbial Community Analysis & Functional Evaluation in Soils

The overall objective of this proposal was to develop technologies to alter the composition and function of important members of microbial communities. In particular, the overall objective of the microbial community editing portion of the proposal focuses on developing foundational tools and understanding required to predict, alter and design grass rhizosphere communities impacting DOE missions. Specifically, the project is centered on the Microbial Community Analysis & Functional Evaluation in Soils (m-CAFES) to manipulate microbial consortia associated with plants of interest for the bioenergy sector, under the presumption that bacterial communities can be manipulated to enhance plant health. For tasks of specific interest to us, we are focusing on developing novel Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) based technologies (primarily focusing on Aim 1) and their delivery modalities (notably subaim 1.2) to edit specific bacterial genomes of interest to enhance their functionalities, and programmably ablate specific undesirable members of bacterial communities for plant health. We are focusing on engineering bacteriophages (bacterial viruses, for subaim 1.2) to carry programmable CRISPR-Cas systems (subaim 1.1) to target (ablate) or alter (edit) genomes of interest. This will enable us to carry out microbial perturbations that will impact community composition and function and ultimately plant growth and health, to enable the next phase of the project by deploying them in situ (subaims 1.3 and 1.4).

59 BASIC BIOLOGICAL SCIENCES↗

Evaluating Probability of Assessment for Central Alarm Station Operators

This project focused on developing and analyzing a method that could be employed by operational nuclear facilities to evaluate their CAS operators and determine a probability of assessment for each operator. This method is agnostic of alarm communication and display system (AC&D), sensors and cameras in place in the field, and any other technical security measures. This security-measure-agnostic method enables operational facilities to adopt it to determine the effectiveness of their alarm station operators.

42 ENGINEERING↗

Non-Resource Conservation and Recovery Act Corrective Action Unit (CAU) Post-Closure Inspection Report, Nevada National Security Site, Nevada For Calendar Year 2025

This report includes visual inspection results, descriptions of maintenance and repair activities, and recommendations for CY 2025 for use restrictions (URs) at corrective action sites (CASs) located on the Nevada National Security Site and on the Nevada Test and Training Range (NTTR) that are accessed through the NNSS Main Gate. This document is arranged by project activity (i.e., Industrial Sites, Soils, and Defense Threat Reduction Agency [DTRA]); then by corrective action unit (CAU); and then by CAS. Post-closure UR inspections are performed on an annual basis, unless as noted in text. Post-closure UR inspections verify the condition of physical controls required by each UR, which may include the following activities: • Visual verification of the legibility of signs • Visual inspection of fencing, signs, monuments, and/or markers for indications of wear • Visual inspection of soil covers for indications of subsidence, erosion, or unauthorized use Deficiencies in UR requirements that were identified during CY 2024 inspections and not addressed in the CY 2024 report are addressed in this CY 2025 report.

54 ENVIRONMENTAL SCIENCES↗

CRISPR-Cas9/Cas12a systems for efficient genome editing and large genomic fragment deletions in Aspergillus niger

CRISPR technology has revolutionized fungal genetic engineering by accelerating the pace and expanding the feasible scope of experiments in this field. Among various CRISPR-Cas systems, Cas9 and Cas12a are widely used in genetic and metabolic engineering. In filamentous fungi, both Cas9 and Cas12a have been utilized as CRISPR nucleases. In this work we first compared efficacies and types of genetic edits for CRISPR-Cas9 and -Cas12a systems at the polyketide synthase (albA) gene locus in Aspergillus niger. By employing a tRNA-based gRNA polycistronic cassette, both Cas9 and Cas12a have demonstrated equally remarkable editing efficacy. Cas12a showed potential superiority over Cas9 protein when one gRNA was used for targeting, achieving an editing efficiency of 86.5% compared to 31.7% for Cas9. Moreover, when employing two gRNAs for targeting, both systems achieved up to 100% editing efficiency for single gene editing. In addition, the CRISPR-Cas9 system has been reported to induce large genomic deletions in various species. However, its use for engineering large chromosomal segments deletions in filamentous fungi still requires optimization. Here, we engineered Cas9 and -Cas12a-induced large genomic fragment deletions by targeting various genomic regions of A. niger ranging from 3.5 kb to 40 kb. Our findings demonstrate that targeted engineering of large chromosomal segments can be achieved, with deletions of up to 69.1% efficiency. Furthermore, by targeting a secondary metabolite gene cluster, we show that fragments over 100 kb can be efficiently and specifically deleted using the CRISPR-Cas9 or -Cas12a system. Overall, in this paper, we present an efficient multi-gRNA genome editing system utilizing Cas9 or Cas12a that enables highly efficient targeted editing of genes and large chromosomal regions in A. niger.

59 BASIC BIOLOGICAL SCIENCES↗

The High Energy X-ray Probe (HEX-P): supernova remnants, pulsar wind nebulae, and nuclear astrophysics

HEX-P is a probe-class mission concept that will combine high spatial resolution X-ray imaging (< 10" full width at half maximum) and broad spectral coverage (0.2–80 keV) with an effective area far superior to current facilities (including XMM-Newton and NuSTAR) to enable revolutionary new insights into a variety of important astrophysical problems. HEX-P is ideally suited to address important problems in the physics and astrophysics of supernova remnants (SNRs) and pulsar wind nebulae (PWNe). For shell SNRs, HEX-P can greatly improve our understanding via more accurate spectral characterization and localization of non-thermal X-ray emission from both non-thermal-dominated SNRs and those containing both thermal and non-thermal components, and can discover previously unknown non-thermal components in SNRs. Multi-epoch HEX-P observations of several young SNRs (e.g., Cas A and Tycho) are expected to detect year-scale variabilities of X-ray filaments and knots, thus enabling us to determine fundamental parameters related to diffusive shock acceleration, such as local magnetic field strengths and maximum electron energies. For PWNe, HEX-P will provide spatially-resolved, broadband X-ray spectral data separately from their pulsar emission, allowing us to study how particle acceleration, cooling, and propagation operate in different evolution stages of PWNe. HEX-P is also poised to make unique and significant contributions to nuclear astrophysics of Galactic radioactive sources by improving detections of, or limits on, 44Ti in the youngest SNRs and by potentially discovering rare nuclear lines as evidence of double neutron star mergers. Throughout the paper, we present simulations of each class of objects, demonstrating the power of both the imaging and spectral capabilities of HEX-P to advance our knowledge of SNRs, PWNe, and nuclear astrophysics.

79 ASTRONOMY AND ASTROPHYSICS↗

eYGFPuv-Assisted Transgenic Selection in Populus deltoides WV94 and Multiplex Genome Editing in Protoplasts of P. trichocarpa × P. deltoides Clone ‘52-225’

Although CRISPR/Cas-based genome editing has been widely used for plant genetic engineering, its application in the genetic improvement of trees has been limited, partly because of challenges in Agrobacterium-mediated transformation. As an important model for poplar genomics and biotechnology research, eastern cottonwood (Populus deltoides) clone WV94 can be transformed by A. tumefaciens, but several challenges remain unresolved, including the relatively low transformation efficiency and the relatively high rate of false positives from antibiotic-based selection of transgenic events. Moreover, the efficacy of CRISPR-Cas system has not been explored in P. deltoides yet. Here, we first optimized the protocol for Agrobacterium-mediated stable transformation in P. deltoides WV94 and applied a UV-visible reporter called eYGFPuv in transformation. Our results showed that the transgenic events in the early stage of transformation could be easily recognized and counted in a non-invasive manner to narrow down the number of regenerated shoots for further molecular characterization (at the DNA or mRNA level) using PCR. We found that approximately 8.7% of explants regenerated transgenic shoots with green fluorescence within two months. Next, we examined the efficacy of multiplex CRISPR-based genome editing in the protoplasts derived from P. deltoides WV94 and hybrid poplar clone ‘52-225’ (P. trichocarpa × P. deltoides clone ‘52-225’). The two constructs expressing the Trex2-Cas9 system resulted in mutation efficiency ranging from 31% to 57% in hybrid poplar clone 52-225, but no editing events were observed in P. deltoides WV94 transient assay. The eYGFPuv-assisted plant transformation and genome editing approach demonstrated in this study has great potential for accelerating the genome editing-based breeding process in poplar and other non-model plants species and point to the need for additional CRISPR work in P. deltoides.

59 BASIC BIOLOGICAL SCIENCES↗

Prime Editing Technology and Its Prospects for Future Applications in Plant Biology Research

Many applications in plant biology requires editing genomes accurately including correcting point mutations, incorporation of single-nucleotide polymorphisms (SNPs), and introduction of multinucleotide insertion/deletions (indels) into a predetermined position in the genome. These types of modifications are possible using existing genome-editing technologies such as the CRISPR-Cas systems, which require induction of double-stranded breaks in the target DNA site and the supply of a donor DNA molecule that contains the desired edit sequence. However, low frequency of homologous recombination in plants and difficulty of delivering the donor DNA molecules make this process extremely inefficient. Another kind of technology known as base editing can perform precise editing; however, only certain types of modifications can be obtained, e.g., C/G-to-T/A and A/T-to-G/C. Recently, a new type of genome-editing technology, referred to as “prime editing,” has been developed, which can achieve various types of editing such as any base-to-base conversion, including both transitions (C→T, G→A, A→G, and T→C) and transversion mutations (C→A, C→G, G→C, G→T, A→C, A→T, T→A, and T→G), as well as small indels without the requirement for inducing double-stranded break in the DNA. Because prime editing has wide flexibility to achieve different types of edits in the genome, it holds a great potential for developing superior crops for various purposes, such as increasing yield, providing resistance to various abiotic and biotic stresses, and improving quality of plant product. In this review, we describe the prime editing technology and discuss its limitations and potential applications in plant biology research.

59 BASIC BIOLOGICAL SCIENCES↗

Advancing How We Learn from Biodesign to Mitigate Risks with Next-Generation Genome Engineering

In the last decade, the unprecedented simplicity and flexibility of the CRISPR-Cas system has made it the dominant transformative tool in gene and genome editing. However, this democratized technology is both a boon and a bane, for which we have yet to understand the full potential to investigate and rewrite genomes (also named “genome biodesign”). Rapid CRISPR advances in a range of applications in basic research, agriculture, and clinical applications pose new risks and raise several biosecurity concerns. In such a fast-moving field of research, we emphasize the importance of properly communicating the quality and accuracy of results and recommend new reporting requirements for results derived from next-generation genome engineering.

59 BASIC BIOLOGICAL SCIENCES↗

Reflection on the Challenges, Accomplishments, and New Frontiers of Gene Drives

Ongoing pest and disease outbreaks pose a serious threat to human, crop, and animal lives, emphasizing the need for constant genetic discoveries that could serve as mitigation strategies. Gene drives are genetic engineering approaches discovered decades ago that may allow quick, super-Mendelian dissemination of genetic modifications in wild populations, offering hopes for medicine, agriculture, and ecology in combating diseases. Following its first discovery, several naturally occurring selfish genetic elements were identified and several gene drive mechanisms that could attain relatively high threshold population replacement have been proposed. This review provides a comprehensive overview of the recent advances in gene drive research with a particular emphasis on CRISPR-Cas gene drives, the technology that has revolutionized the process of genome engineering. Herein, we discuss the benefits and caveats of this technology and place it within the context of natural gene drives discovered to date and various synthetic drives engineered. Later, we elaborate on the strategies for designing synthetic drive systems to address resistance issues and prevent them from altering the entire wild populations. Lastly, we highlight the major applications of synthetic CRISPR-based gene drives in different living organisms, including plants, animals, and microorganisms.

59 BASIC BIOLOGICAL SCIENCES↗

NS 1987A in SN 1987A

The possible detection of a compact object in the remnant of SN 1987A presents an unprecedented opportunity to follow its early evolution. The suspected detection stems from an excess of infrared emission from a dust blob near the compact object’s predicted position. The infrared excess could be due to the decay of isotopes like 44 Ti, accretion luminosity from a neutron star or black hole, magnetospheric emission or a wind originating from the spin down of a pulsar, or to thermal emission from an embedded, cooling neutron star. In this work it is shown that the last possibility is the most plausible as the other explanations are disfavored by other observations and/or require fine-tuning of parameters. Not only are there indications that the dust blob overlaps the predicted location of a kicked compact remnant, but its excess luminosity also matches the expected thermal power of a 30 yr old neutron star. Furthermore, models of cooling neutron stars within the minimal cooling paradigm readily fit both NS 1987A and Cas A, the next-youngest known neutron star. If correct, a long heat transport timescale in the crust and a large effective stellar temperature are favored, implying relatively limited crustal n- 1 S 0 superfluidity and an envelope with a thick layer of light elements, respectively. If the locations do not overlap, then pulsar spin down or accretion might be more likely, but the pulsar’s period and magnetic field or the accretion rate must be rather finely tuned. In this case, NS 1987A may have enhanced cooling and/or a heavy-element envelope.

79 ASTRONOMY AND ASTROPHYSICS↗

Nucleosynthetic Analysis of Three-dimensional Core-collapse Supernova Simulations

Abstract We study in detail the ejecta conditions and theoretical nucleosynthetic results for 18 three-dimensional core-collapse supernova (CCSN) simulations done by F ornax . Most of the simulations are carried out to at least 3 s after bounce, which allows us to follow their longer-term behaviors. We find that multidimensional effects introduce many complexities into the ejecta conditions. We see a stochastic electron fraction evolution, complex peak temperature distributions and histories, and long-tail distributions of the time spent within nucleosynthetic temperature ranges. These all lead to substantial variation in CCSN nucleosynthetic yields and differences from 1D results. We discuss the production of lighter α -nuclei, radioactive isotopes, heavier elements, and a few isotopes of special interest. Comparing pre-CCSN and CCSN contributions, we find that a significant fraction of elements between roughly Si and Ge are generally produced in CCSNe. We find that 44 Ti exhibits an extended production timescale as compared to 56 Ni, which may explain its different distribution and higher than previously predicted abundances in supernova remnants such as Cas A and SN1987A. We also discuss the morphology of the ejected elements. This study highlights the high-level diversity of ejecta conditions and nucleosynthetic results in 3D CCSN simulations and emphasizes the need for additional long-term (∼10 s) 3D simulations to properly address such complexities.

Astronomy & Astrophysics↗

Simulation to a Newborn Supernova Remnant from a Low-mass Iron Core Star

Supernova remnant observations show a high degree of asymmetry, mixing, and inhomogeneity. These asymmetries are seeded during the early seconds of the explosion and are further enhanced and modified as the shock and ejecta move through the stellar progenitor and into the circumstellar medium. We present simulations of a 9.6 M⊙ zero-metallicity progenitor initialized after shock revival and evolved for several years when the ejecta is in the circumstellar medium. A suite of 1D and 2D simulations examines the effects of neutron-star wind and radioactive decay heating. In 1D, decay heating forms a low-density bubble that suppresses the reverse shock. While in 2D, the heating is localized to metal-rich pockets, inflating them and compressing the surrounding material into dense shells. In 3D, the neutron-star wind and decay heating modify the plume morphology, producing more large-scale structures. The extended plume morphology leads to an asymmetrical shock breakout. After breakout, the leading plumes cannot keep up with the shock front, resulting in deceleration and fragmentation by the reverse shock while retaining the large-scale asymmetry. The projected ejecta morphology and velocities are strongly viewing angle dependent. The relatively uniform metal-rich distribution does not resemble the strongly inhomogeneous ejecta structure of Cas A. The 160-isotope decay network shows that 24.4% of the radioactive heating comes from decay chains other than the canonical 56Ni chain. The low explosion energy, low 56Ni yield, and Ni/Fe ratio greater than unity suggest an observational signature similar to an electron capture supernova.

Neopane, Sudarshan [University of Tennessee (UT)]↗

Evaluation of four ground-based retrievals of cloud droplet number concentration in marine stratocumulus with aircraft in situ measurements

Abstract. Cloud droplet number concentration (Nd) is crucial for understanding aerosol–cloud interactions (ACI) and associated radiative effects. We present evaluations of four ground-based Nd retrievals based on comprehensive datasets from the Atmospheric Radiation Measurement (ARM) Aerosol and Cloud Experiments in the Eastern North Atlantic (ACE-ENA) field campaign. The Nd retrieval methods use ARM ENA observatory ground-based remote sensing observations from a micropulse lidar, Raman lidar, cloud radar, and the ARM NDROP (Droplet Number Concentration) value-added product (VAP), all of which also retrieve cloud effective radius (re). The retrievals are compared against aircraft measurements from the fast cloud droplet probe (FCDP) and the cloud and aerosol spectrometer (CAS) obtained from low-level marine boundary layer clouds on 12 flight days during summer and winter seasons. Additionally, the in situ measurements are used to validate the assumptions and characterizations used in the retrieval algorithms. Statistical comparisons of the probability distribution function (PDF) of the Nd and cloud re retrievals with aircraft measurements demonstrate that these retrievals align well with in situ measurements for overcast clouds, but they may substantially differ for broken clouds or clouds with low liquid water path (LWP). The retrievals are applied to 4 years of ground-based remote sensing measurements of overcast marine boundary layer clouds at the ARM ENA observatory to find that Nd (re) values exhibit seasonal variations, with higher (lower) values during the summer season and lower (higher) values during the winter season. The ensemble of various retrievals using different measurements and retrieval algorithms such as those in this paper can help to quantify Nd retrieval uncertainties and identify reliable Nd retrieval scenarios. Of the retrieval methods, we recommend using the micropulse lidar-based method. This method has good agreement with in situ measurements, less sensitivity to issues arising from precipitation and low cloud LWP and/or optical depth, and broad applicability by functioning for both daytime and nighttime conditions.

54 ENVIRONMENTAL SCIENCES↗

Evaluation of Four Ground-based Retrievals of Cloud Droplet Number Concentration in Marine Stratocumulus with Aircraft In Situ Measurements

Cloud droplet number concentration (N d ) is crucial for understanding aerosol-cloud interactions (ACI) and associated radiative effects. We present evaluations of four ground-based N d retrievals based on comprehensive datasets from the Atmospheric Radiation Measurements (ARM) Aerosol and Cloud Experiments in the Eastern North Atlantic (ACE-ENA) field campaign. The N d retrieval methods use ARM ENA observatory ground-based remote sensing observations from a Micropulse lidar, Raman lidar, cloud radar, and the ARM NDROP Value-added Product (VAP), all of which also retrieve cloud effective radius (r e ). The retrievals are compared against aircraft measurements from the Fast-Cloud Droplet Probe (FCDP) and the Cloud and Aerosol Spectrometer (CAS) obtained from low-level marine boundary layer clouds on 12 flight days during summer and winter seasons. Additionally, the in situ measurements are used to validate the assumptions and characterizations used in the retrieval algorithms. Statistical comparisons of the probability distribution function (PDF) of the N d and cloud r e retrievals with aircraft measurements demonstrate that these retrievals align well with in situ measurements for overcast clouds, but they may substantially differ for broken clouds or clouds with low liquid water path (LWP). The retrievals are applied to four years of ground-based remote sensing measurements of overcast marine boundary layer clouds at the ARM ENA observatory to find that N d (r e ) values exhibit seasonal variations, with higher (lower) values during the summer season and lower (higher) values during the winter season. The ensemble of various retrievals using different measurements and retrieval algorithms such as those in this paper can help to quantify N d retrieval uncertainties and identify reliable N d retrieval scenarios. Of the retrieval methods, we recommend using the using the Micropulse lidar-based method given its good agreement with in situ measurements, it has less sensitivity to issues arising from precipitation and low cloud LWP/optical depth, and it has broad applicability by functioning for both day and nighttime conditions.

54 ENVIRONMENTAL SCIENCES↗

Fiscal Year 2020 Annual Summary Report for the Area 3 and Area 5 Radioactive Waste Management Sites at the Nevada National Security Site, Nye County, Nevada

U.S. Department of Energy Standard DOE-STD-5002-2017, “Disposal Authorization Statement and Tank Closure Documentation,” requires preparation of an Annual Summary Report comparing low-level radioactive waste disposal facilities operations with the Disposal Authorization Statement (DAS) and its supporting technical documents including the performance assessment (PA) and the composite analysis (CA). The fiscal year (FY) 2020 Annual Summary Report for the Area 3 and Area 5 Radioactive Waste Management Sites (RWMSs) was prepared by reviewing planned and discovered changes, waste receipts, monitoring results, research and development (R&D) results, and the status of DAS conditions and issues to assess the validity of the DASs, PAs, and CAs.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗