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At least 19 records

Role of karyopherin nuclear transport receptors in nuclear transport by nuclear trafficking peptide

Nuclear trafficking peptide (NTP), a cell-penetrating peptide (CPP) composed of 10 amino acids (aa) (RIFIHFRIGC), has potent nuclear trafficking activity. Recently, we established a protein-based cell engineering system by using NTP, but it remained elusive how NTP functions as a CPP with nuclear orientation. In the present study, we identified importin subunit β1 (IMB1) and transportin 1 (TNPO1) as cellular proteins underlying the activity of NTP. These karyopherin nuclear transport receptors were identified as candidate molecules by liquid chromatography/mass spectrometry analysis, and downregulation of each protein by small interfering RNA significantly reduced NTP activity (P < 0.01). Biochemical analyses revealed that NTP bound directly to both molecules, and the forced expression of an IMB1 fragment (296–516 aa) or TNPO1 fragment (1–297 aa), which both contain binding sites to NTP, reduced nuclear NTP-green fluorescent protein (GFP) levels when it was added to cell culture medium. NTP is derived from viral protein R (Vpr) of human immunodeficiency virus-1, and Vpr enters the nucleus and exerts pleiotropic functions. Notably, Vpr bound directly to IMB1 and TNPO1, and its function was significantly impaired by the forced expression of the 296–516-aa fragment of IMB1 and 1–297-aa fragment of TNPO1. Interestingly, NTP completely blocked the physical association of Vpr with IMB1 and TNPO1. Although the nuclear localization mechanism of Vpr remains unknown, our data suggest that NTP functions as a novel nuclear localization signal of Vpr.

60 APPLIED LIFE SCIENCES↗

Applicability of the Milestones Approach to Deployments of Transportable Nuclear Power Plants (TNPPs)

Transportable nuclear power plants (TNPPs) can provide potential benefits to countries embarking on nuclear programs, offering reduced infrastructure requirements, shorter timeframes for implementation, cost savings and greater deployment flexibility than larger conventional reactors. However, the deployment of a TNPP in a Host State comes with the obligation to establish sufficient regulatory, institutional, and technical infrastructure, which, among others, includes a legal and regulatory framework and a competent regulatory body to implement a State’s safeguards obligations. This paper considers how the unique technical and deployment features of TNPPs may affect the process of preparing for and implementing safeguards in nuclear newcomer countries. Evaluating this issue through the lens of the IAEA’s Milestones Approach, this paper discusses some potential implications arising from the shortening of some milestones phases due to reduced construction or licensing time for TNPPs, and the need for increased cooperation between Host States and Supplier States in preparing for and meeting certain safeguards obligations. These considerations are potentially relevant to various stakeholders: newcomer States considering TNPP deployment; the States and companies that supply such reactors; as well as organizations that support international safeguards capacity building.

Siserman-Gray, Ioana-Cristina↗

SURVEILLANCE DETECTION FOR TRANSPORTATION OPERATIONS PERSONNEL TO PREVENT HIJACKING, THEFT, SABOTAGE, AND MALICIOUS SECURITY EVENTS DURING TRANSPORTING NUCLEAR MATERIAL

The secure transportation of high-consequence materials, including nuclear and radiological assets, is a critical global priority in the face of escalating terrorism, security threats, and violent protests targeting these operations. Effective surveillance detection—the ability to identify, assess, and respond to potential threats across a continuum of scenarios—is paramount in addressing these challenges. This paper outlines a phased, multi-tiered training program designed to strengthen the surveillance detection capabilities of organizations responsible for nuclear material transport. The proposed training program adopts a progressive approach, gradually increasing in technical complexity to provide participants with comprehensive knowledge and tools for implementing robust security strategies. It targets a wide spectrum of stakeholders, including competent authorities, regulators, inspectors, shippers, carriers, law enforcement, and emergency response personnel, equipping them to plan, evaluate, and safeguard nuclear material transportation effectively. Each phase of the program emphasizes distinct elements of the surveillance detection continuum and transport security, focusing on critical topics such as threat identification, adversary task timelines, protective methodologies, and attack mitigation strategies. The training framework is anchored in technical exchanges and scenario-driven courses that reflect real-world complexities and challenges. By addressing the surveillance detection continuum comprehensively—from early threat assessment to active countermeasures—the program reinforces global efforts to secure nuclear assets. It aligns with international security objectives and fosters a strong security culture within participating organizations, ensuring personnel are prepared to counter potential threats and maintain the safe, secure movement of these materials. Ultimately, this initiative aims to enhance preparedness, security, and response capabilities, supporting the global mission to safeguard high-consequence materials against evolving threats.

Zineddin, Dr. Z. [ORNL] (ORCID:0009000848740725)↗

Plan for the Development and Application of a Risk Assessment Approach for Transportation Package Approval of a Transportable Nuclear Power Plant for Maritime Shipment (Rev. 1)

This plan proposes a strategy for licensing a transportable nuclear power plant, containing its unirradiated and irradiated fuel, as a transportation package for maritime transport using a probabilistic risk assessment framework in order to meet the regulatory requirements of 10 CFR Part 71. It is anticipated that initially, the deterministic transportation package licensing approach will be challenging due to the functional requirements of the transportable nuclear power plant. As such, a partial exemption request from the U.S. Nuclear Regulatory Commission coupled with applied compensatory measures are anticipated in support of providing equivalent safety to the public, worker, and environment. A plan is outlined as to how this would be demonstrated and a pathway is primarily outlined using the accompanying low accident rates which in turn support very low probabilities of release for a maritime application. Wherever possible, the proposed plan is supported by actual accident (casualty) data. Additional supporting information regarding a highway transportation probabilistic risk assessment in support of the Project Pele demonstration reactor that is currently under development is also referenced and discussed.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Development and Demonstration of a Risk Assessment Approach for Approval of a Transportation Package of a Transportable Nuclear Power Plant for Domestic Highway Shipment

The U.S. Department of Defense (DoD) Strategic Capabilities Office (SCO) has tasked PNNL to address the regulatory challenges associated with confirming the safe transport of Transportable Nuclear Power Plants (TNPPs) containing irradiated nuclear fuel. A previous report—Proposed Risk-Informed Regulatory Framework for Approval of Microreactor Transportation Packages (PNNL-31867)—determined that the expected radioactive inventory in the irradiated fuel of a TNPP would likely require shipment in an NRC-approved Type B package (or spent nuclear fuel cask) but that a TNPP “package” is unlikely to meet the entire suite of NRC requirements set forth in Part 71 of Title 10 of the Code of Federal Regulations (CFR) for a Type B package. It was therefore concluded that shipment of this initial TNPP transportation package, as well as possibly others, under existing regulations would likely require NRC approval using the 10 CFR 71.12 (“Specific exemptions”) process that relies on risk-informed decision making supported by quantitative risk assessment (i.e., Probabilistic Risk Assessment).

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

CRISPR–Cas9-mediated nuclear transport and genomic integration of nanostructured genes in human primary cells

DNA nanostructures are a promising tool to deliver molecular payloads to cells. DNA origami structures, where long single-stranded DNA is folded into a compact nanostructure, present an attractive approach to package genes; however, effective delivery of genetic material into cell nuclei has remained a critical challenge. Here, we describe the use of DNA nanostructures encoding an intact human gene and a fluorescent protein encoding gene as compact templates for gene integration by CRISPR-mediated homology-directed repair (HDR). Our design includes CRISPR–Cas9 ribonucleoprotein binding sites on DNA nanostructures to increase shuttling into the nucleus. We demonstrate efficient shuttling and genomic integration of DNA nanostructures using transfection and electroporation. These nanostructured templates display lower toxicity and higher insertion efficiency compared to unstructured double-stranded DNA templates in human primary cells. Furthermore, our study validates virus-like particles as an efficient method of DNA nanostructure delivery, opening the possibility of delivering nanostructures in vivo to specific cell types. Together, these results provide new approaches to gene delivery with DNA nanostructures and establish their use as HDR templates, exploiting both their design features and their ability to encode genetic information. This work also opens a door to translate other DNA nanodevice functions, such as biosensing, into cell nuclei.

59 BASIC BIOLOGICAL SCIENCES↗

Integrative mapping reveals molecular features underlying the mechanism of nucleocytoplasmic transport

Nuclear pore complexes (NPCs) enable rapid, selective, and robust nucleocytoplasmic transport. To explain how transport emerges from the system components and their interactions, we used experimental data and theoretical information to construct an integrative Brownian dynamics model of transport through an NPC, coupled to a kinetic model of transport in the cell. The model recapitulates key aspects of transport for a wide range of molecular cargoes, including preribosomes and viral capsids. Our model quantifies how flexible phenylalanine-glycine (FG) repeat proteins create an entropic barrier to passive diffusion and how this barrier is selectively lowered in facilitated diffusion by the many transient interactions of nuclear transport receptors with the FG repeats. Selective transport is enhanced by “fuzzy” multivalent interactions, redundant FG repeat mass, coupling to the energy-dependent RanGTP concentration gradient, and exponential dependence of transport kinetics on the transport barrier. Our model will facilitate rational modulation of the NPC and its artificial mimics.

Raveh, Barak [School of Computer Science and Engin↗

Cask RT-100 Type B(U) Self-Certification for DOT 7A, Type A Nuclear Waste Transportation - 20098

The purpose of this Type A certification is to diversify the use of the RT-100 cask by allowing the end user to transport either Type A or Type B radioactive waste in accordance with the handling procedures for each waste category. The project's scope included the development of a matrix that contains all applicable requirements for a Type A package from the Code of Federal Regulations (CFR), Title 49 for the Department of Transportation (DOT) with a justification for each requirement. Moreover, the scope included the writing of conditions and restrictions list for the transportation of Type A nuclear waste, the writing of a compliance statement, and the design and fabrication of a Type A placard to be permanently placed on the RT-100 cask. Type A packages are self-certified by the end user using one of three methods as permitted by 49 CFR 173.461, either by performance of tests with prototypes or samples of the specimens representing the packaging, or reference to a previous satisfactory demonstration of compliance of a sufficiently similar nature, or calculations or reasoned evaluation using reliable and conservative procedures and parameters. Since the RT-100 cask was licensed by the Nuclear Regulatory Commission (NRC) as a Type B cask, Robatel relied on the method of reference to a previous satisfactory demonstration of compliance of a sufficiently similar nature for Type A certification. This method was determined to be acceptable, since Type B packages are evaluated for more demanding transport conditions than those required for Type A packages. This paper presents Robatel's process for certifying the RT-100 cask for Type A shipments, an overview of the DOT regulations related to Type A packages, and the documents that were generated for the end user. This paper is concluded with a discussion on the advantages of having the option to transport both Type A and Type B nuclear waste, and possible future projects to further improve the operation of the cask when making Type A shipments. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Influence of pre-existing defects on thermal transport in nuclear graphite

Nuclear graphite is a critical material in high-temperature nuclear reactors due to its superior thermal and mechanical properties. The manufacturing process leaves multi-scale ‘pre-existing’ defects that can affect thermal transport characteristics. Because these defects are remnant of graphitization temperature, they cannot be thermally annealed. This study employs a non-thermal electron wind force (EWF) annealing technique to avoid this obstacle. 2 min of EWF treatment of the as-received graphite IG-110 at temperatures below 100 °C led up to 67% increase in thermal diffusivity and ~ 35% decrease in electrical resistivity in average. Differential scanning calorimetry also showed similar outcome for specific heat. X-ray diffraction characterization was performed by fitting a bi-modal crystallite size distribution model to reveal the enhancement in crystallinity after the EWF treatment. The findings emphasize the potential of EWF annealing for optimizing thermal performance in nuclear graphite and its implications for reactor efficiency and safety.

Annealing↗

AWC for Nuclear Material Transportation

Presentation to the Forefront22 Workshop in DC to NNSA DNN. Generalized presentation on Advanced Wireless Communications and areas where Nuclear Material Transportation may find interesting to investigate further for security and cybersecurity interests. Summary of the Forefront22 workshop Traditional wireless communication technologies, such as Wi-Fi and Cellular, have modernized industries worldwide. The Nuclear industry has also started to modernize its operations by utilizing wireless connectivity and is well poised to expand the use of wireless to an increasing number of use cases. The most recent cellular technology, called 5G for Fifth Generation, brings a transformation not only to wireless; but to industries worldwide by fueling the growth of Internet of Things (IoT) and Industrial IoT (IIoT). Availability of additional spectrum e.g., use of millimeter wave (mmWave) spectrum bands, is a critical boost to wireless growth. These advanced wireless technologies bring multiplicative gains in data rates, latency reduction, capacity growth, and localization accuracy, that can be harnessed for Nuclear Nonproliferation activities along with other mission critical applications. However, using these functional advances securely is of utmost important specially for these critical applications. The functional capabilities of these advanced technologies including 5G & Wi-Fi 6, and the associated security issues and mitigations will be discussed in this session. The future technologies including 6G which will follow 5G and newer versions of Wi-Fi will be also introduced.

99 GENERAL AND MISCELLANEOUS↗

International Symposium on Transport Security of Nuclear and other Radioactive Material; Tokyo, Japan, November 2019

As an outcome of the Fourth Nuclear Security Summit of 2016, held in Washington D.C., the Governments of Australia, Canada, Czech Republic, Finland, France, Hungary, Italy, Japan, Kazakhstan, Morocco, Spain, the Republic of Korea, Thailand, the United Kingdom, and the United States reaffirmed, through INFCIRC/909, their will to further improve the overall security in the transport of nuclear and other radioactive materials. With this goal in mind, each expressed their commitment to further exchange national practices with other countries through the IAEA and the Global Initiative to Combat Nuclear Terrorism (GICNT). They also committed to actively support the IAEA as the central organization for coordinating activities and developing guidance documents and to support the GICNT and the Global Partnership Against the Spread of Weapons and Materials of Mass Destruction in developing and implementing its nuclear transport security activities. Both of these organizations address the commitments regarding nuclear transport security as outcomes of the Nuclear Security Summit 2016. In an effort to continue the momentum from the Nuclear Security Summit, the Government of Japan, in cooperation with the Integrated Support Center for Nuclear Nonproliferation and Nuclear Security of Japan Atomic Energy Agency (ISCN/JAEA), hosted the International Transport Security Symposium in November 2019. With over fifty countries invited to attend, the event exchanged and promoted good practices in several topical area sessions related to the security of nuclear and other radioactive material in transport. Topical sessions included Laws and Regulatory Framework, Physical Protection Systems, Response Actions, and Insider/Information Security. The Symposium built support for transport security and encouraged IAEA Member States to actively work regionally and internationally with the IAEA’s transport security programme through the practical implementation and incorporation of IAEA Nuclear Security Series recommendations and guidance into their national frameworks for transport security. From these sessions, to the host detailed a recommended path forward for future engagements in the security of nuclear and radioactive material in transport. The paper will discuss the Symposium’s outcomes and highlight the forward momentum made by both signatories and non-signatories for the strengthening of all the critical elements of a State’s transport security regime, including physical protection systems, insider threats to the nuclear and radioactive material supply chains, planning and capabilities for law enforcement response to malicious transport security events, and employment of technology to address critical vulnerabilities to the transport of nuclear and other radioactive materials.

Holt, Katherine↗

Characterizing Binding Interactions That Are Essential for Selective Transport through the Nuclear Pore Complex

Specific macromolecules are rapidly transported across the nuclear envelope via the nuclear pore complex (NPC). The selective transport process is facilitated when nuclear transport receptors (NTRs) weakly and transiently bind to intrinsically disordered constituents of the NPC, FG Nups. These two types of proteins help maintain the selective NPC barrier. To interrogate their binding interactions in vitro, we deployed an NPC barrier mimic. We created the stationary phase by covalently attaching fragments of a yeast FG Nup called Nsp1 to glass coverslips. We used a tunable mobile phase containing NTR, nuclear transport factor 2 (NTF2). In the stationary phase, three main factors affected binding: the number of FG repeats, the charge of fragments, and the fragment density. We also identified three main factors affecting binding in the mobile phase: the avidity of the NTF2 variant for Nsp1, the presence of nonspecific proteins, and the presence of additional NTRs. We used both experimentally determined binding parameters and molecular dynamics simulations of Nsp1FG fragments to create an agent-based model. The results suggest that NTF2 binding is negatively cooperative and dependent on the density of Nsp1FG molecules. Our results demonstrate the strengths of combining experimental and physical modeling approaches to study NPC-mediated transport.

59 BASIC BIOLOGICAL SCIENCES↗

The Floating Chameleon: Floating Nuclear Power Plants and the Nexus of Maritime and Nuclear Security Law

During the 63rd regular session of the International Atomic Energy Agency (IAEA) General Conference, as part of Norway's statement to the General Conference, Mr. Auden Halvorsen, State Secretary for Norway, made the following statement about transportable nuclear power plants (TNPPs): "Deployment of transportable nuclear power plants—TNPPs—demands our attention. The Agency must intensify its conversations of all aspects of the safety and security of such facilities. . . . The scope and applicability of existing requirements and instruments need to be clarified and developed including dialogue with the International Maritime Organization [IMO]." Ensuring the security of TNPPs is essential for their deployment and implementation. TNPPs may be floating nuclear power plants (FNPPs), which bring together elements of nuclear security related to the nuclear power plant with elements of maritime security. Insofar as an FNPP weds the principles of nuclear and maritime security, it also weds the two domains, the ancient traditions and contemporary practices of maritime law with the contemporary practices of nuclear law. Fundamentally, this raises the question, what are the legal requirements for the security of a FNPP? Although both maritime law and nuclear law have established regimes for security through various legal instruments, does an FNPP expose gaps between the current international legal instruments? The previous question of whether international legal regimes define requirements for the security of FNPPs, but a threshold question remains: what is an FNPP? For purposes of nuclear and maritime security, is an FNPP a facility, is an FNPP a vessel and therefore a transport, or does an FNPP change its status based upon temporal and spatial considerations such as when it is docked at a port generating power or in transit to its destination? This set of questions, and others addressed later in this article, highlights that the issue is not merely a question about security but intersects elements of safety whether an FNPP is in uncharted waters of the international regime for both nuclear and maritime law. Here, this paper addresses many of the questions posed above related to the security regulations for an FNPP.

98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗

Biallelic variants in SNUPN cause a limb girdle muscular dystrophy with myofibrillar-like features

Abstract Alterations in RNA-splicing are a molecular hallmark of several neurological diseases, including muscular dystrophies, where mutations in genes involved in RNA metabolism or characterized by alterations in RNA splicing have been described. Here, we present five patients from two unrelated families with a limb-girdle muscular dystrophy (LGMD) phenotype carrying a biallelic variant in SNUPN gene. Snurportin-1, the protein encoded by SNUPN, plays an important role in the nuclear transport of small nuclear ribonucleoproteins (snRNPs), essential components of the spliceosome. We combine deep phenotyping, including clinical features, histopathology and muscle MRI, with functional studies in patient-derived cells and muscle biopsies to demonstrate that variants in SNUPN are the cause of a new type of LGMD according to current definition. Moreover, an in vivo model in Drosophila melanogaster further supports the relevance of Snurportin-1 in muscle. SNUPN patients show a similar phenotype characterized by proximal weakness starting in childhood, restrictive respiratory dysfunction and prominent contractures, although inter-individual variability in terms of severity even in individuals from the same family was found. Muscle biopsy showed myofibrillar-like features consisting of myotilin deposits and Z-disc disorganization. MRI showed predominant impairment of paravertebral, vasti, sartorius, gracilis, peroneal and medial gastrocnemius muscles. Conservation and structural analyses of Snurportin-1 p.Ile309Ser variant suggest an effect in nuclear-cytosol snRNP trafficking. In patient-derived fibroblasts and muscle, cytoplasmic accumulation of snRNP components is observed, while total expression of Snurportin-1 and snRNPs remains unchanged, which demonstrates a functional impact of SNUPN variant in snRNP metabolism. Furthermore, RNA-splicing analysis in patients’ muscle showed widespread splicing deregulation, in particular in genes relevant for muscle development and splicing factors that participate in the early steps of spliceosome assembly. In conclusion, we report that SNUPN variants are a new cause of limb girdle muscular dystrophy with specific clinical, histopathological and imaging features, supporting SNUPN as a new gene to be included in genetic testing of myopathies. These results further support the relevance of splicing-related proteins in muscle disorders.

Iruzubieta, Pablo (ORCID:0000000303316222)↗

Modeling and Measurement of Axial Gas Transport in Nuclear Fuels

This document is an American Nuclear Society (ANS) transaction for the ANS annual conference. This transaction provides an overview of the Modeling and Measurement of Axial Gas Transport Laboratory Directed Research and Development project including preliminary modeling results. The goal is to provide model for axial gas transport in the Bison Fuel Performance code to support industry's desire to achieve higher burnups in existing light water reactors. A brief overview of the experimental approach and associated modeling activities is presented.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Coupled Multiphysics Simulations of Heat Pipe Microreactors Using DireWolf

DireWolf is a multiphysics software driver application designed to simulate heat pipe–cooled nuclear microreactors. Developed under the U.S. Department of Energy, Office of Nuclear Energy Nuclear Energy Advanced Modeling and Simulation (NEAMS) program, the DireWolf software application’s objective is to provide the nuclear community with a design and safety analysis simulation capability. Based upon the NEAMS program Multiphysics Object-Oriented Simulation Environment (MOOSE) computational framework, DireWolf tightly couples nuclear microreactor physics, reactor physics, radiation transport, nuclear fuel performance, heat pipe thermal hydraulics, power generation, and structural mechanics to resolve the interdependent nonlinearities. DireWolf is capable of simulating both steady and transient normal reactor operation and several postulated failure scenarios. We will present the fundamental physics of heat pipe–cooled nuclear microreactors and the MOOSE-based software employed in DireWolf. Both steady and transient results for coupled reactor physics, radiation transport, and nuclear fuel performance are demonstrated.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗