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Uranium Mill Tailings Radiation Control Act Title II DOE Due Diligence and Lessons Learned from a Previous Site Transfer - 20352

Title II of the Uranium Mill Tailings Radiation Control Act (UMTRCA) established that a government agency will provide perpetual care for closed uranium and thorium ore-processing sites that were operating under an NRC source material license in 1978 or were licensed thereafter. Commercial owners (licensees) operating under an NRC or agreement state specific license when UMTRCA was passed are responsible for conducting reclamation of any byproduct material remaining from uranium-ore processing operations in accordance with an NRC or agreement state approved reclamation plan. Reclamation includes both surface and groundwater remedies. Upon completion of reclamation and approval by NRC, the site is required to be transferred to either the host state or the DOE for long-term surveillance and maintenance. Since UMTRCA's enactment, six Title II sites have been transferred to DOE; an additional 24 Title II sites are anticipated to be transferred before 2050. DoE's role mandated under UMTRCA Title II as the long-term care custodian is to perform 'monitoring, maintenance, and emergency measures necessary to protect the public health and safety.' UMTRCA requires that the licensee pay a long-term surveillance charge 'sufficient to cover the annual costs of site surveillance.' However, at some sites such as the Bluewater, New Mexico, Disposal Site, this mandate has required additional effort and expense by DOE, beyond the originally anticipated and intended scope within UMTRCA, but within the authority of DOE under UMTRCA. In 1997, the Bluewater site became the second UMTRCA Title II site to be transferred to DOE. The site was the location of a uranium mill operated from 1953 until 1982. The specific licensee began site reclamation in 1991, and by 1995 all tailings and contaminated materials were encapsulated in two tailings disposal cells and other disposal areas. In addition to surface contamination, milling activities impacted groundwater in the two upper aquifers. In 1989, the specific licensee attempted active groundwater remediation; however, no significant reduction in contaminant concentrations was observed. As a result, the specific licensee applied to NRC for alternate concentration limits (ACLs) in 1990, which were approved in 1996 as being protective, after additional corrective actions were performed. Since transfer of the Bluewater site to DOE, unforeseen challenges have occurred, requiring additional actions. The first challenge is the occurrence of surface depressions located on the northern section of the main tailings disposal cell. The depressions were first observed during DoE's initial inspection in 1998; however, evidence of these can be observed on satellite images taken prior to transfer. Since being first observed, the depressions have continued to grow both in depth and areal extent. Due to the design of the main tailings disposal cell, the depressions impede storm water from being effectively shed off the 101- hectare (250-acre) top slope of the main tailings disposal cell. Instead, storm water accumulates in the depressions, forming a large ephemeral pond that has stored up to 16.3 x 10{sup 6} liters (4.3 million gallons) of stormwater. The ponding poses a potential risk to the integrity of the main tailings disposal cell in the case of a large storm event, with the potential to cause the pond to overtop and erode the cover material and underlying waste. DOE has taken a number of short-term actions to monitor, measure, and reduce the ponding and is currently working with the US Army Corps of Engineers to design and construct a repair. Additional challenges are associated with groundwater at the Bluewater site. Nine wells were present on the 1335-hectare (3300-acre) site upon transfer. Groundwater compliance was called into question after the State of New Mexico reduced its uranium groundwater standard from 5.0 to 0.03 milligrams per liter in 2004, and when an ACL for uranium was exceeded in a site monitoring well in 2010. Acquiring historical groundwater data and subsequent evaluations as well as additional DOE groundwater monitoring led to installing 10 new monitoring wells and performing additional site hydrogeology recharacterization. DOE continues to evaluate groundwater conditions at the site and works with the NRC to determine regulatory requirements and a path forward. As a result of lessons learned at the Bluewater site and other Title II sites, improved processes have been implemented at a programmatic level to increase due diligence before site transfer and prevent similar issues from occurring at other UMTRCA Title II sites under long-term management. DoE's due diligence process is documented in the Process for Transition of UMTRCA Title II Disposal Sites to DOE for Long-Term Surveillance and Maintenance and is designed to ensure that DOE has no technical or compliance concerns with regulatory decisions that might compromise protectiveness following site transfer to DOE. Implementation of the due diligence process has increased DoE's role prior to site transfer and has been effective in identifying potential issues. Actions by NRC and specific licensees, in response to enhanced due diligence efforts by DOE, are expected to minimize, if not totally prevent, the need for unanticipated actions by DOE pertaining to the surface and groundwater remedies after site transfer. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Xylanolytic metabolism is regulated by coordination of transcription factors XynR and XylR in extremely thermophilic Caldicellulosiruptorales

ABSTRACT Global transcription factors (TFs) control metabolic processes in bacteria to efficiently utilize available carbon. The orderCaldicellulosiruptoraleshas drawn interest due to the ability of its members to degrade components of lignocellulosic biomass. Regulatory reconstruction ofAnaerocellum (f. Caldicellulosiruptor) besciiidentified two major global transcription factors for xylan utilization, XynR and XylR, and the corresponding putative transcription factor binding sites. Recombinant versions of XynR (LacI family) and XylR (ROK family) were subjected to fluorescence polarization (FP) and biolayer interferometry (BLI) analysis to confirm the predicted binding sites. Four XynR sites and two XylR sites were validated, accounting for 20 of 26 genes regulated by XynR and six of seven genes regulated by XylR. Bioinformatic analysis of the individual genes controlled by the two regulators showed an inter-dependent scheme for xylan conversion; the transport of xylooligosaccharides (XOS) is dependent on XylR, while enzymes responsible for hydrolysis are controlled by both regulators. For xylose catabolism by the xylose isomerase-xylulose kinase pathway, regulation is also split, with XylR controlling xylose isomerase and XynR controlling xylokinase. The XynR/XylR regulator pair withinA. besciiis conserved in all sequenced species ofCaldicellulosiruptorales, suggesting similarities in regulating linear xylan conversion. In other xylanolytic thermophiles, XylR homologs control xylan degradation, compared to just 6 out of 26 genes forA. bescii. These results show that two separate regulatory schemes (dual repression) are coordinated byA. besciito effectively regulate the hemicellulose inventory and xylan catabolism. IMPORTANCE To take full advantage of extreme thermophiles as platform metabolic engineering microorganisms, the tools for genetic manipulation must be further developed, and strategies that exploit a better understanding of metabolic regulation need to be discerned.Anaerocellum bescii, the most studied of the extremely thermophilic fermentative anaerobic bacteria that can utilize microcrystalline cellulose, can degrade microcrystalline cellulose and hemicellulose and has been metabolically engineered to convert the resulting sugars to products such as ethanol and acetone. For xylan, in particular, two major global transcription factors (TFs), XynR and XylR, play a role in sugar metabolism, although their predicted regulatory interdependence from bioinformatics analysis has not been elucidated experimentally. Here, fluorescence polarization (FP) and biolayer interferometry (BLI) were used to explore this issue to support metabolic engineering efforts aimed at improving carbohydrate processing to industrial chemicals.

Biotechnology & Applied Microbiology↗

Safety-Related Instrumentation and Control Pilot Upgrade: Conceptual - Detailed Design Phase Report and Lessons Learned

Historically, regulatory barriers have precluded nuclear plant safety-related (SR) Instrumentation and Control (I&C) system modernizations. These barriers have been largely addressed by industry and United States Nuclear Regulatory Commission (NRC) efforts, including development of the License Amendment Request (LAR) Alternate Review (AR) process for digital I&C upgrades. The U.S. Department of Energy (DOE) and the Idaho National Laboratory (INL) are supporting a SR I&C pilot upgrade at the Limerick Generating Station (LGS) to demonstrate this process and capturing associated lessons learned for the benefit of industry. In 2021, a paper summarizing lessons learned as captured in INL/EXT-20-59809, Safety Related Instrumentation & Control Pilot Upgrade Initial Scoping Phase Implementation Report and Lessons Learned was presented at the 12th NPIC & HMIT conference. It is recommended that readers of this paper review reference 1 or the associated Nuclear Journal paper for context. This paper summarizes activities performed and lessons learned for the conceptual design phase and portions of the detailed design phase as captured in INL/RPT-23-XXXX, “Safety-Related Instrumentation and Control Pilot Upgrade: Conceptual - Detailed Design Phase Report and Lessons Learned.” The portions of the effort covered by reference 2 include conceptual design phase activities from December 2020, through completion in November 2021, and detailed design activities through September 2022. These efforts and associated detailed documentation in reference 3 are intended to support similar industry upgrades as a foundation stone for a Digital Transformation that will improve plant safety, reliability, and operational performance while lowering plant total cost of ownership.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Application and Certification of Comparative Vacuum Monitoring Sensors for Structural Health Monitoring of 737 Wing Box Fittings

Multi-site fatigue damage, hidden cracks in hard-to-reach locations, disbonded joints, erosion, impact, and corrosion are among the major flaws encountered in today's extensive fleet of aging aircraft and space vehicles. The use of in-situ sensors for real-time health monitoring of aircraft structures are a viable option to overcome inspection impediments stemming from accessibility limitations, complex geometries, and the location and depth of hidden damage. Reliable, structural health monitoring systems can automatically process data, assess structural condition, and signal the need for human intervention. Prevention of unexpected flaw growth and structural failure can be improved if on-board health monitoring systems are used to continuously assess structural integrity. Such systems are able to detect incipient damage before catastrophic failures occurs. Condition-based maintenance practices could be substituted for the current time-based maintenance approach. Other advantages of on-board distributed sensor systems are that they can eliminate costly, and potentially damaging, disassembly, improve sensitivity by producing optimum placement of sensors and decrease maintenance costs by eliminating more time- consuming manual inspections. This report presents a Sandia Labs-aviation industry effort to move SHM into routine use for aircraft maintenance. This program addressed formal SHM technology validation and certification issues so that the full spectrum of concerns, including design, deployment, performance and certification were appropriately considered. The Airworthiness Assurance NDI Validation Center (AANC) at Sandia Labs, in conjunction with Boeing, Delta Air Lines, Structural Monitoring Systems Ltd., Anodyne Electronics Manufacturing Corp. and the Federal Aviation Administration (FAA) carried out a certification program to formally introduce Comparative Vacuum Monitoring (CVM) as a structural health monitoring solution to a specific aircraft wing box application. Validation tasks were designed to address the SHM equipment, the health monitoring task, the resolution required, the sensor interrogation procedures, the conditions under which the monitoring will occur, the potential inspector population, adoption of CVM into an airline maintenance program and the document revisions necessary to allow for routine use of CVM as an alternate means of performing periodic structural inspects. To carry out the validation process, knowledge of aircraft maintenance practices was coupled with an unbiased, independent evaluation. Sandia Labs designed, implemented, and analyzed the results from a focused and statistically-relevant experimental effort to quantify the reliability of the CVM system applied to the Boeing 737 Wing Box fitting application. All factors that affect SHM sensitivity were included in this program: flaw size, shape, orientation and location relative to the sensors, as well as operational and environmental variables. Statistical methods were applied to performance data to derive Probability of Detection (POD) values for CVM sensors in a manner that agrees with current nondestructive inspection (NDI) validation requirements and also is acceptable to both the aviation industry and regulatory bodies. This report presents the use of several different statistical methods, some of them adapted from NDI performance assessments and some proposed to address the unique nature of damage detection via SHM systems, and discusses how they can converge to produce a confident quantification of SHM performance An important element in developing SHM validation processes is a clear understanding of the regulatory measures needed to adopt SHM solutions along with the knowledge of the structural and maintenance characteristics that may impact the operational performance of an SHM system. This report describes the major elements of an SHM validation approach and differentiates the SHM elements from those found in NDI validation. The activities conducted in this program demonstrated the feasibility of routine SHM usage in general and CVM in particular for the application selected. They also helped establish an optimum OEM-airline-regulator process and determined how to safely adopt SHM solutions. This formal SHM validation will allow aircraft manufacturers and airlines to confidently make informed decisions about the proper utilization of CVM technology. It will also streamline the regulatory actions and formal certification measures needed to assure the safe application of SHM solutions.

42 ENGINEERING↗

A Functional Survey of the Regulatory Landscape of Estrogen Receptor–Positive Breast Cancer Evolution

Abstract Only a handful of somatic alterations have been linked to endocrine therapy resistance in hormone-dependent breast cancer, potentially explaining ∼40% of relapses. If other mechanisms underlie the evolution of hormone-dependent breast cancer under adjuvant therapy is currently unknown. In this work, we employ functional genomics to dissect the contribution of cis-regulatory elements (CRE) to cancer evolution by focusing on 12 megabases of noncoding DNA, including clonal enhancers, gene promoters, and boundaries of topologically associating domains. Parallel epigenetic perturbation (CRISPRi) in vitro reveals context-dependent roles for many of these CREs, with a specific impact on dormancy entrance and endocrine therapy resistance. Profiling of CRE somatic alterations in a unique, longitudinal cohort of patients treated with endocrine therapies identifies a limited set of noncoding changes potentially involved in therapy resistance. Overall, our data uncover how endocrine therapies trigger the emergence of transient features which could ultimately be exploited to hinder the adaptive process. Significance: This study shows that cells adapting to endocrine therapies undergo changes in the usage or regulatory regions. Dormant cells are less vulnerable to regulatory perturbation but gain transient dependencies which can be exploited to decrease the formation of dormant persisters.

Oncology↗

First STAX detector installation at the National Institute for Radioelements (IRE)

The Source Term Analysis of Xenon (STAX) project has been installing stack detectors at medical isotope production facilities to measure radioxenon emissions to investigate the effect of radioxenon releases on nuclear explosion monitoring. This paper outlines the installation of the first STAX detection system at the National Institute for Radioelements (IRE) in Fleurus, Belgium which has been operating for over three years and transferring collected data to the STAX repository. Information about the equipment installed, the data flow established, and calculations for determination of radioxenon releases from the facility are presented. Further. data quality was investigated to confirm values reported by STAX automated data processing and in a comparison of collected STAX data with data collected by IRE for regulatory reporting.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Review of Software Packages for Local Intense Precipitation Flood Modeling

This report documents the review of the hydrologic and hydraulic modeling approaches used to estimate local intense precipitation (LIP) flooding and their implementation in readily available simulation software packages. The review focused on a few representative simulation software packages because of the similarity of their underlying mathematical bases and the rainfall-runoff and hydraulic processes included. Their availability for use in estimating LIP flood hazards and for U.S. Nuclear Regulatory Commission staff review of these estimates at the nuclear power plant (NPP) site scale were also considered.

54 ENVIRONMENTAL SCIENCES↗

Developing an Equity Framework for State Regulatory Decision-Making

The report presents a framework for states that seek to incorporate equity into regulatory decision-making. Berkeley Lab contextualizes approaches and metrics from various states into example processes to exemplify how this may be done, including topics such as the development of equity goals and definitions, intervenor funding, community engagement, performance-based ratemaking, and utility resource planning. The report offers five takeaways and considerations, supported by these examples: 1. Equity comprises multiple tenets and stages, all of which must be considered in parallel. 2. At the start of designing new equity-related processes, it is critical to establish clear and actionable goals, definitions, roles, and responsibilities to ensure progress. 3. Once goals are established, it is critical to align tools and metrics that bridge the gap between what an intervention may do and how it may impact communities and households. 4. Processes should be stakeholder driven. It is important to not only increase education and outreach, but to actively seek out and incorporate feedback from inclusive public processes and build in accountability mechanisms. Processes should be iterative. Feedback loops between evaluations and program design provide the flexibility to better align existing interventions with community priorities and to incorporate equity into future decision-making.

99 GENERAL AND MISCELLANEOUS↗

ORNL Package Testing Program Overview

When transporting radioactive or hazardous materials, safety and security are top priorities. Packaging and transportation regulations require reliable evidence that containers have passed rigorous performance tests to ensure that the public and the environment are protected from the hazardous nature of the cargo. Because of its long history in energy and security research, Oak Ridge National Laboratory (ORNL) has needed to ship hazardous packages for the past 65 years. Since the 1940s, ORNL has made significant contributions to transportation regulations and has been at the forefront of regulatory testing development. Today, ORNL is a leader in standards development and testing of designs of radioactive material packages. ORNL plays a critical role in ensuring the safe transportation of radioactive materials across the United States by executing rigorous testing campaigns of packages that contain radioactive materials. Early testing activities focused on supporting the development of transportation regulations established by both the International Atomic Energy Agency (IAEA) and the United States. Currently, all package testing activities are performed under the Package Testing Program (PTP) at the Package Evaluation Facility (PEF) located at the National Transportation Research Center (NTRC), which is about 10 miles from the main ORNL campus. The PTP develops and evaluates testing solutions, ensuring that they are safe efficient, and in compliance with regulatory requirements. The vision for the PTP is to be a world-class leader in the evaluation and testing of radioactive and hazardous material packages. The key elements necessary to fulfill this mission are an experienced and professional staff; state-of-the-art facilities, equipment, and instrumentation; and completion of challenging programs and projects important to package transportation. Collaborations with other internal and external organizations play a significant role in building stronger teams and achieving this vision. By adhering to and advancing regulatory standards, ORNL’s PTP supports development of safe and compliant packaging, safeguarding the transportation process from potential risks associated with radioactive material logistics.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Lessons Learned in the Development of Industry Guidance - 20279

The availability of industry guidance on different aspects of waste management forms a valuable reference for waste practitioners. The guidance typically details the principles, processes and practices that should be consulted during each stage of the relevant activities. It also includes relevant regulatory and international requirements, appropriate case studies and examples of best practices to illustrate technical concepts and make the information available to the reader in an accessible manner. Over recent years Wood have developed (or are developing) a number of industry guidance documents on a range of topics including Solid Waste Characterization, Routine Water Quality Monitoring on Nuclear Sites: Pipeline Characterization: Determining Background Radioactivity for Land Quality Assessment: Interim Storage and Aqueous Waste Management. This talk provides an overview of lessons learned along the way including: - the importance of stakeholder engagement through the process: - how to identify 'good practice'; - the importance of iterative development; and - the selection of illustrative and informative case studies. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

SoK: What does it Mean to Benchmark Database Forensics?

Relational Database Management Systems are the backbone of modern enterprises and public-sector services, and are thus frequent targets of security incidents, insider threats, and thorough regulatory audits. Consequently, databases have become key sources of digital evidence, requiring investigators to reconstruct past activity from audit logs, transaction logs, and backups. Although benchmarking frameworks such as those developed by the Transaction Processing Performance Council (TPC) are widely used to evaluate database performance, they do not capture forensic requirements such as evidentiary completeness, tamper-evidence, chain of custody, or regulatory compliance under GDPR and CCPA. This survey examines the emerging domain of forensic database benchmarking. We gathered prior research on database forensics, secure logging, and tamper-evident data structures; we analyze modern forensic-ready features in commercial and open-source systems (SQL Server Ledger, Oracle Blockchain Tables, PostgreSQL pgAudit, Db2 Audit, Aurora Database Activity Streams, Oracle Real Application Security and IBM Guardium) and assess why existing benchmarks are insufficient. We propose forensic workloads, metrics, and methodologies that incorporate adversarial stressors, deleted-record recovery, and backup analysis. We also identify open research problems and call for a community-driven forensic benchmark suite. The result is an idea for evaluating not only database performance but also forensic soundness, bridging the gap between system engineering, compliance, and digital investigations.

Lenard, Ben↗

Lipo-chitooligosaccharides as regulatory signals of fungal growth and development

Lipo-chitooligosaccharides (LCOs) are signaling molecules produced by rhizobial bacteria that trigger the nodulation process in legumes, and by some fungi that also establish symbiotic relationships with plants, notably the arbuscular and ecto mycorrhizal fungi. Here, we show that many other fungi also produce LCOs. We tested 59 species representing most fungal phyla, and found that 53 species produce LCOs that can be detected by functional assays and/or by mass spectroscopy. LCO treatment affects spore germination, branching of hyphae, pseudohyphal growth, and transcription in non-symbiotic fungi from the Ascomycete and Basidiomycete phyla. Our findings suggest that LCO production is common among fungi, and LCOs may function as signals regulating fungal growth and development.

59 BASIC BIOLOGICAL SCIENCES↗

Development of Guidance for the Disposition of Used Oil and Oil Samples at the Columbia Generating Station - 20232

It is a fact that oil can become contaminated during plant operations. When used oil is collected, the water is decanted from the used oil. The separated used oil is typically analyzed for gamma emitting radionuclides and then sent off-site for processing (incineration) or recycling. If there is no detectable radioactivity, then some plants may release the oil for unrestricted use. Many nuclear power plants have no standard process for evaluating Tritium and Carbon-14 in oil; therefore, there may be variability in how plants are determining whether oil contains Tritium and Carbon-14, and at what concentration. Questions may arise whether there is a technically sound basis and evaluation method for used oil and if process knowledge and/or sampling may be appropriate in making that determination. Recently, there has been an increase in regulatory scrutiny regarding these practices to ensure that the utilities are evaluating oil for all plant-produced radionuclides prior to free release. Regulators have indicated that the requirement to conduct adequate surveys should include evaluating oils for Tritium and Carbon-14. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Evolution of the regulatory subunits for the heteromeric acetyl-CoA carboxylase

The committed step for de novo fatty acid (FA) synthesis is the ATP-dependent carboxylation of acetyl-coenzyme A catalysed by acetyl-CoA carboxylase (ACCase). In most plants, ACCase is a multi-subunit complex orthologous to prokaryotes. However, unlike prokaryotes, the plant and algal orthologues are comprised both catalytic and additional dedicated regulatory subunits. Novel regulatory subunits, biotin lipoyl attachment domain-containing proteins (BADC) and carboxyltransferase interactors (CTI) (both three-gene families inArabidopsis) represent new effectors specific to plants and certain algal species. The evolutionary history of these genes in autotrophic eukaryotes remains elusive, making it an ongoing area of research. Analyses of potential protein–protein and co-occurrence interactions, informed by gene network patterns using the STRING database, inArabidopsis thalianaandChlamydomonas reinhardtiiunveil intricate gene associations with ACCase, suggesting a complex interplay between FA synthesis and other cellular processes. Among both species, a higher number of co-expressed genes was identified inArabidopsis, indicating a wider potential regulatory network of ACCase in plants. This review investigates the extent to which these genes arose in autotrophic eukaryotes and provides insights into their evolutionary trajectory. This article is part of the theme issue ‘The evolution of plant metabolism’.

Life Sciences & Biomedicine - Other Topics↗

EVALUATION OF HRA METHODOLOGIES FOR APPLICATION IN SDP WORK

This study critically evaluates human reliability analysis (HRA) methodologies applicable to regulatory probabilistic safety assessment (PSA) model, with a particular focus on their role in supporting the significance determination process (SDP) in nuclear safety assessment. Firstly, three widely utilized HRA methods – IDHEAS-ECA, SPAR-H, and ASEP/THERP – were qualitatively and quantitatively assessed. Qualitative assessments were conducted using attributes from the NEA/CSNI/R(2015)1 report, while quantitative evaluations employed regression and correlation analyses to compare predicted human error probabilities (HEPs) against empirical data. Results reveal distinct strengths, for example, IDHEAS-ECA’s robust predictive accuracy and K-HRA’s alignment with operational practices. In addition, dependency analysis and recovery analysis were critically evaluated. For dependency analysis, the methods’ handling of inter-task dependencies and their impact on HEPs were examined, while recovery analysis highlighted strategies for mitigating failure events. Furthermore, strategies were proposed to evaluate performance-shaping factors under conditions of reduced human performance, such as stress, fatigue, or cognitive overload, addressing specific challenges faced in SDP evaluations. Human errors from KINS’s operational performance information system event reports were evaluated as a case study. This study identifies gaps and provides actionable insights to ensure their validity and applicability in SDP HRA applications. This paper is a part of research conducted by KINS, and it should be noted that this result does not represent the regulatory position of KINS.

99 - GENERAL AND MISCELLANEOUS↗

Inference of Gene Regulatory Network Uncovers the Linkage between Circadian Clock and Crassulacean Acid Metabolism in Kalanchoë fedtschenkoi

The circadian clock drives time-specific gene expression, enabling biological processes to be temporally controlled. Plants that conduct crassulacean acid metabolism (CAM) photosynthesis represent an interesting case of circadian regulation of gene expression as stomatal movement is temporally inverted relative to stomatal movement in C3 plants. The mechanisms behind how the circadian clock enabled physiological differences at the molecular level is not well understood. Recently, the rescheduling of gene expression was reported as a mechanism to explain how CAM evolved from C3. Therefore, we investigated whether core circadian clock genes in CAM plants were re-phased during evolution, or whether networks of phase-specific genes were simply re-wired to different core clock genes. We identified candidate core clock genes based on gene expression features and then applied the Local Edge Machine (LEM) algorithm to infer regulatory relationships between this new set of core candidates and known core clock genes in Kalanchoë fedtschenkoi. We further inferred stomata-related gene targets for known and candidate core clock genes and constructed a gene regulatory network for core clock and stomata-related genes. Our results provide new insight into the mechanism of circadian control of CAM-related genes in K. fedtschenkoi, facilitating the engineering of CAM machinery into non-CAM plants for sustainable crop production in water-limited environments.

59 BASIC BIOLOGICAL SCIENCES↗

Hsa_circ_0000515 is a novel circular RNA implicated in the development of breast cancer through its regulation of the microRNA‐296‐5p/CXCL10 axis

Cancer metastasis is a major cause of death among women afflicted with breast cancer (BC) and understanding the molecular processes involved is a major focus in BC research. Circular RNAs (circRNAs) have emerged as genomic regulatory molecules in carcinogenesis and metastasis; however, their role in BC is unclear. We characterized a novel circRNA, hsa_circ_0000515, in context of BC. We collected 340 cancerous tissues surgically resected from BC patients and found hsa_circ_0000515 was upregulated in BC tissues and associated with poor prognosis of BC. Silencing of hsa_circ_0000515 impaired cell cycle progression, cell proliferation, and invasion, attenuated inflammatory response, and reduced the proangiogenetic potential of BC cells. RNA pull‐down and dual‐luciferase reporter gene assays showed that hsa_circ_0000515 binds miR‐296‐5p, preventing it from repressing CXCL10 expression. We also observed that miR‐296‐5p inhibition or CXCL10 overexpression promoted cell cycle progression, restored proliferative, invasive and proangiogenetic abilities, and increased inflammatory response in MCF‐7 cells in the absence of hsa_circ_0000515. In vivo analyses showed that partial loss of hsa_circ_0000515 reduced the tumor growth of MCF‐7 cells in nude mice. The key findings from this study revealed that targeting hsa_circ_0000515 might be an effective strategy to combat BC.

Cai, Fenglin↗

Capability Building Progression of an Insider Threat Mitigation Program at an International Research Reactor

The nuclear industry recognizes the difficulties involved in developing effective managerial and leadership skills in a highly technical and proficient workforce such as that found in nuclear facilities. Implementing an insider threat mitigation program (ITMP) within the nuclear industry is a complex and ongoing process that demands a comprehensive understanding of human behavior, an organization’s security culture, and rigorous regulatory requirements yet also accounts for facility characteristics, physical security, material flow, and activities involving nuclear material. Given the high-consequence nature of research reactor operations, even minor lapses can lead to safety, security, and reputational risks. An effective ITMP requires a defense-in-depth approach that incorporates behavioral analysis, robust vetting procedures, continuous monitoring, and cross-disciplinary coordination. It must also promote a culture of vigilance and accountability at all levels up to and including executive leadership but be flexible enough to adapt to evolving global threats and technological advances. Insider threat mitigation is not a one-time effort but rather a sustained commitment to excellence in safety and security. Establishing a culture in which personnel proactively report incidents and issues that could affect nuclear safety and security is vital to maintaining a safe and secure operational environment. This document was developed to guide senior management and research reactor organizations in creating comprehensive programs to effectively manage and mitigate insider threat behaviors and actions. It focuses on the key pillars of an effective ITMP, including the national legal framework, security culture, preventive and protective measures, cyber security, and performance evaluation. By using a systematic approach during implementation, facilities can foster environments conducive to insider threat detection and support long-term program sustainability. The document also provides strategies for improving communication across all levels of an organization, helping to eliminate barriers that hinder the development of robust ITMPs and enhance overall security culture. In today’s organizations, the concept of leveraging safety and security culture lessons to facilitate knowledge transfer is rapidly evolving to expedite insider threat management and security culture improvements. This document outlines the rationale for evaluating an ITMP based on national customs, culture, and stakeholders. The elements are all germane to reliability and trustworthiness and relate to security concerns that states may encounter. The document focuses not only on individual perceptions regarding security issues and capability building but also on team building and how to resolve concerns. The implementers of a facility’s ITMP may zero in on indicators of insider threats within their enterprise. This material will benefit organizations when it is applied using a systematic and structured approach as demonstrated throughout the document.

98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗