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All sourcesNASA NTRSDOE OSTI
DOE OSTI2024

Large-Scale Hydrogen Storage Cyber Risk Assessment

Hydrogen storage systems are becoming more widely deployed throughout the country, and as their presence continues to grow, it is possible that individual and interconnected systems will be exposed to cyber-attacks. These events can cause physical and financial harm to employees, people in the vicinity, and to the company that owns the facility. The two main mechanisms malicious actors may access information or control from a hydrogen storage facility are through information technology and operations technology devices, the former of which refers to data and information from networked devices and the latter of which refers to onsite controls for the physical system. Both types of entryways into the system should be considered when facility managers conduct cyber risk assessments and when regulators develop or revise relevant codes and standards. This report analyzes cybersecurity risks applicable to a wide variety of hydrogen storage systems by outlining the system's purpose and the importance of its cybersecurity. The hydrogen storage system architecture and communication protocols are provided to understand potential cyber vulnerabilities. Later, an event tree analysis is performed on hydrogen operation to identify system weaknesses by outlining potential attack scenarios. This report also identifies critical cyber assets related to different hydrogen operations followed by an examination of potential threats, and the impact of cyber assets on those operational assets.

08 HYDROGEN↗
DOE OSTI2025

Proposed Classifications of Remote Operations for Nuclear Reactors Based on Physical and Cybersecurity Considerations

The incorporation of remote operations into reactor operations is a topic of high interest among advanced and small modular reactor (A/SMR) vendors, with some considering it essential to the success of their business models. However, remote operations are a concept novel to the nuclear industry. While various technical aspects of remote operations have been explored, a significant gap remains in understanding the security implications of integrating remote operations into reactor designs, particularly concerning the security requirements for remote-operations facilities and infrastructure. This report aims to address this gap by first defining classes of remote operation based on the extent of remote access to reactor control systems and grounded in the existing regulatory framework with compatible terminology. Secondly, the report outlines the physical and cybersecurity requirements applicable to remote-operations facilities and infrastructure at each defined class. These requirements are based on existing licensing frameworks provided by 10 Code of Federal Regulations (CFR) Part 50 and 10 CFR Part 52, as well as the upcoming A/SMR licensing framework in the proposed Part 53. The assessment focuses specifically on security regulations, such as 10 CFR Part 73, which includes provisions for both cybersecurity (§ 73.54) and physical security (§ 73.55). This report proposes five classes of remote reactor operations. Class 1 involves remote monitoring only, with no control over reactor systems. Class 2 allows for the remote issuance of allowlisted commands to the reactor facility. Class 3 extends control to non-safety-significant, non-safety-related, or not important to safety systems and equipment. Class 4 permits remote control of safety-significant systems. Finally, Class 5 allows remote control of safety-related systems. It is important to note that these classes were defined purely with functionality in mind, without considering the practicality or feasibility of implementation for each class under current or upcoming regulatory guidance. The intention behind this approach is to enable an assessment of which security requirements apply to each class, allowing readers to evaluate the implementation possibilities for their specific use cases. Following the definition of remote-operation classes, the report assesses the specific physical and cybersecurity requirements applicable to the remote-operations facility and infrastructure within each defined class. This includes defining the types and locations of operators that are possible at each class of operation and, based on operator type and location, as well as functionality within each class, outlining the physical and cybersecurity requirements. By detailing the security requirements by class, the report provides readers with the information needed to determine the type of security program they may need to implement for their desired concept of operation. The next contribution of this report was to assess the practicality of implementing each proposed class of remote operations based upon the security requirement assessment. In short, three of the five proposed remote-operation classes were found to possibly have a practical path forward to implementation under the U.S. regulatory framework. Class 1 remote operations are currently in use in the U.S. while Class 2 and 3 remote operations may be logistically possible to implement under the U.S. regulatory framework. The final two Classes, 4 and 5, would likely be logistically difficult, if not infeasible to implement within the current U.S. physical- and cybersecurity regulatory framework. Given the results of the feasibility assessment, an example architecture is proposed for both Class 2, remote allowlisted commands, and Class 3, remote control of non-safety systems as well as security implication assessments of each architecture. These example implementations are not meant to be prescriptive in terms of how Class 2 or Class 3 remote operations should be deployed; instead, they are intended to be informative to stakeholders on how Class 2 or Class 3 could potentially be applied in order to inform their system design. An example architecture for Class 1 remote monitoring was not provided as Class 1 in already in use in U.S. nuclear operations. Example architectures for Class 4 and Class 5 were not provided due to their assessment of being likely infeasible to implement. The final contribution is an assessment of the physical- and cybersecurity implications of introducing autonomous operations into an A/SMR. What was found was that the security implications can be separated into two cases. Autonomous operations supported by SSCs located only at the reactor site, and autonomous operations supported by SSCs outside of the reactor site. For the first case, the introduction of autonomous systems will likely not change the facility’s requirement to comply with existing cyber and physical security regulation

22 - GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2024

"Traffic Control via Connected and Automated Vehicles: An Open-Road Field Experiment with 100 CAVs"

The CIRCLES project aims to reduce instabilities in traffic flow, which are naturally occurring phenomena due to human driving behavior. These "phantom jams" or "stop-and-go waves,"are a significant source of wasted energy. Toward this goal, the CIRCLES project designed a control system referred to as the MegaController by the CIRCLES team, that could be deployed in real traffic. Our field experiment leveraged a heterogeneous fleet of 100 longitudinally-controlled vehicles as Lagrangian traffic actuators, each of which ran a controller with the architecture described in this paper. The MegaController is a hierarchical control architecture, which consists of two main layers. The upper layer is called Speed Planner, and is a centralized optimal control algorithm. It assigns speed targets to the vehicles, conveyed through the LTE cellular network. The lower layer is a control layer, running on each vehicle. It performs local actuation by overriding the stock adaptive cruise controller, using the stock on-board sensors. The Speed Planner ingests live data feeds provided by third parties, as well as data from our own control vehicles, and uses both to perform the speed assignment. The architecture of the speed planner allows for modular use of standard control techniques, such as optimal control, model predictive control, kernel methods and others, including Deep RL, model predictive control and explicit controllers. Depending on the vehicle architecture, all onboard sensing data can be accessed by the local controllers, or only some. Control inputs vary across different automakers, with inputs ranging from torque or acceleration requests for some cars, and electronic selection of ACC set points in others. The proposed architecture allows for the combination of all possible settings proposed above. Most configurations were tested throughout the ramp up to the MegaVandertest.

Lee, Jonathan↗
DOE OSTI2019

TRANSP-based Optimization Towards Tokamak Scenario Development

An optimization approach that incorporates the predictive transport code TRANSP is proposed for tokamak scenario development. Optimization methods are often employed to develop open-loop control strategies to aid access to high performance tokamak scenarios. In general, the optimization approaches use control-oriented models, i.e. models that are reduced in complexity and prediction accuracy as compared to physics-oriented transport codes such as TRANSP. In the presented approach, an optimization procedure using the TRANSP code to simulate the tokamak plasma is considered for improved predictive capabilities. As a test case, the neutral beam injection (NBI) power is optimized to develop a control strategy that maximizes the non-inductive current fraction during the ramp-up phase for NSTX-U. Simulation studies towards the achievement of non-inductive ramp up in NSTX-U have already been carried out with the TRANSP code. The optimization-based approach proposed in this work is used to maximize the non-inductive current fraction during ramp-up in NSTX-U, demonstrating that the scenario development task can be automated. An additional test case considers optimization of the current ramp rate in DIII-D for obtaining a stationary plasma characterized bya flat loop voltage profile in the flattop phase.

DIII-D↗
DOE OSTI2025

Development of an FGPA-Based Cavity Simulator for Testing RF Controls

LLRF is used to precisely control the amplitude and phase of the RF field in cavities. Often times, access to test the control algorithms with RF equipment, especially in the presence of beam, is limited or beyond reach. In such cases, testing must be done through computer modeling or simulations. Computer modeling is often too slow and difficult to interface with the LLRF hardware. Analog or digital cavity simulators are preferred as they allow for interaction with the LLRF controls platform in real-time, and compared to their analog counterparts, FPGA-based digital cavity simulators allow for a more adjustable and sophisticated implementation. The newly developed FPGA-based cavity simulator includes the cavity electrical model, the cavity mechanical model including Lorentz Force Detuning and microphonics, an amplifier model which can simulate real amplifier nonlinearities, and a beam model. The simulator has been validated using measurements from BNL’s CeC 704 MHz 5-cell SRF cryomodule.

43 PARTICLE ACCELERATORS↗
DOE OSTI2015

Baseline Risk Assessment Supporting Closure at Waste Management Area C at the Hanford Site Washington

The Office of River Protection under the U.S. Department of Energy is pursuing closure of the Single-Shell Tank (SST) Waste Management Area (WMA) C under the requirements of the Hanford Federal Facility Agreement and Consent Order (HFFACO). A baseline risk assessment (BRA) of current conditions is based on available characterization data and information collected at WMA C. The baseline risk assessment is being developed as a part of a Resource Conservation and Recovery Act (RCRA) Facility Investigation (RFI)/Corrective Measures Study (CMS) at WMA C that is mandatory under Comprehensive Environmental Response, Compensation, and Liability Act and RCRA corrective action. The RFI/CMS is needed to identify and evaluate the hazardous chemical and radiological contamination in the vadose zone from past releases of waste from WMA C. WMA C will be under Federal ownership and control for the foreseeable future, and managed as an industrial area with restricted access and various institutional controls. The exposure scenarios evaluated under these conditions include Model Toxics Control Act (MTCA) Method C, industrial worker, maintenance and surveillance worker, construction worker, and trespasser scenarios. The BRA evaluates several unrestricted land use scenarios (residential all-pathway, MTCA Method B, and Tribal) to provide additional information for risk management. Analytical results from 13 shallow zone (0 to 15 ft. below ground surface) sampling locations were collected to evaluate human health impacts at WMA C. In addition, soil analytical data were screened against background concentrations and ecological soil screening levels to determine if soil concentrations have the potential to adversely affect ecological receptors. Analytical data from 12 groundwater monitoring wells were evaluated between 2004 and 2013. A screening of groundwater monitoring data against background concentrations and Federal maximum concentration levels was used to determine vadose zone contamination impacts on groundwater. Waste Management Area C is the first of the Hanford tank farms to begin the closure planning process. The current baseline risk assessment will provide valuable information for making corrective actions and closure decisions for WMA C, and will also support the planning for future tank farm soil investigation and baseline risk assessments.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗
DOE OSTI2025

The git based ATLAS data acquisition configuration service in LHC Run 3

The ATLAS experiment at the LHC at CERN uses a large, distributed trigger and data acquisition system composed of many computing nodes, networks, and hardware modules. Its configuration service is used to provide descriptions of control, monitoring, diagnostic, recovery, dataflow and data quality configurations, interconnections, and parameters for modules, chips, and channels of various online systems, detectors, and the whole ATLAS experiment. Those descriptions have historically been stored in more than one thousand interconnected XML files, which are updated by various experts many times per day. Maintaining error-free and consistent sets of such files and providing reliable and fast access to current and historical configurations is a major challenge. This paper gives details of the configuration service upgrade on the modern Git version control system backend for LHC Run 3 and its exploitation experience. It may be interesting for developers using human-readable file formats, where consistency of the files, performance, access control, traceability of modifications, and effective archiving are key requirements.

Soloviev, Igor [Univ. of California, Irvine, CA (U↗
DOE OSTI2011

Network Upgrade for the SLC: PEP II Network

The PEP-II control system required a new network to support the system functions. This network, called CTLnet, is an FDDI/Ethernet based network using only TCP/IP protocols. An upgrade of the SLC Control System micro communications to use TCP/IP and SLCNET would allow all PEP-II control system nodes to use TCP/IP. CTLnet is private and separate from the SLAC public network. Access to nodes and control system functions is provided by multi-homed application servers with connections to both the private CTLnet and the SLAC public network. Monitoring and diagnostics are provided using a dedicated system. Future plans and current status information is included.

43 PARTICLE ACCELERATORS↗
DOE OSTI2013

Apparatus and method supporting wireless access to multiple security layers in an industrial control and automation system or other system

A method includes receiving a message at a first wireless node. The first wireless node is associated with a first wired network, and the first wired network is associated with a first security layer. The method also includes transmitting the message over the first wired network when at least one destination of the message is located in the first security layer. The method further includes wirelessly transmitting the message for delivery to a second wireless node when at least one destination of the message is located in a second security layer. The second wireless node is associated with a second wired network, and the second wired network is associated with the second security layer. The first and second security layers may be associated with different security paradigms and/or different security domains. Also, the message could be associated with destinations in the first and second security layers.

42 ENGINEERING↗
DOE OSTI2020

Method and apparatus for maintaining data coherence in a non-uniform compute device

A data processing apparatus includes one or more host processors with first processing units, one or more caches with second processing unit, a non-cache memory having a third processing unit and a reorder buffer operable to maintain data order during execution of a program of instructions. An instruction scheduler routes instructions to the processing units. Data coherence is maintained by control logic that blocks access to data locations in use by a selected processing unit other than the selected processing unit until data associated with the data locations are released from the reorder buffer. Data stored in the cache is written to the memory if it is already in a modified state, otherwise the state is set to the modified state. A memory controller may be used to restrict access to memory locations to be operated on.

97 MATHEMATICS AND COMPUTING↗
DOE OSTI2018

Accessing the Intrinsic Spin Transport in a Topological Insulator by Controlling the Crossover of Bulk-to-Surface Conductance

Here we report a method to control contributions of bulk and surface states in the topological insulator Bi 2 Te 2 Se that allows accessing the spin-polarized transport endowed by topological surface states. An intrinsic surface dominant transport is established when cooling the sample to low temperature or reducing the conduction channel length, both achieved in situ in the transport measurements with a four-probe scanning tunneling microscope without the need of further tailoring the sample. The topological surface states show characteristic transport behaviors with mobility about an order of magnitude higher than reported before, and a spin polarization approaching the theoretically predicted value. Our result demonstrates accessibility to the intrinsic high mobility spin transport of topological surface states, which paves a way to realizing topological spintronic devices.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗
DOE OSTI2025

Scalable fabrication of an array-type fixed-target device for automated room temperature X-ray protein crystallography

X-ray crystallography is one of the leading tools to analyze the 3-D structure, and therefore, function of proteins and other biological macromolecules. Traditional methods of mounting individual crystals for X-ray diffraction analysis can be tedious and result in damage to fragile protein crystals. Furthermore, the advent of multi-crystal and serial crystallography methods explicitly require the mounting of larger numbers of crystals. To address this need, we have developed a device that facilitates the straightforward mounting of protein crystals for diffraction analysis, and that can be easily manufactured at scale. Inspired by grid-style devices that have been reported in the literature, we have developed an X-ray compatible microfluidic device that can be used to trap protein crystals in an array configuration, while also providing excellent optical transparency, a low X-ray background, and compatibility with the robotic sample handling and environmental controls used at synchrotron macromolecular crystallography beamlines. At the Stanford Synchrotron Radiation Lightsource (SSRL), these capabilities allow for fully remote-access data collection at controlled humidity conditions. Furthermore, we have demonstrated continuous manufacturing of these devices via roll-to-roll fabrication to enable cost-effective and efficient large-scale production.

chemical engineering↗
DOE OSTI2021

Automated Cyber Security Testing Platform for Industrial Control Systems

Nuclear Power Plants (NPPs) are a complex system of coupled physics controlled by a network of Programmable Logic Controllers (PLCs). These PLCs communicate process data across the network to coordinate control actions with each other and inform the operators of process variables and control decisions. Networking the PLCs allows more effective process control and provides the operator more information which results in more efficient plant operation. This interconnectivity creates new security issues, as operators have more access to the plant controls, so will bad actors. As plant networks become more digitized and encompass more sophisticated controllers, the network surface exposed to cyber interference grows. Understanding the dynamics of these coupled systems of physics, control logic, and network communications is critical to their protection. The research into the cybersecurity of the Operational Technologies of NPPs is developing and requires a platform that can allow high fidelity physics simulations to interact with digital networks of controllers. This will require three main components: a network simulation environment, a physics simulator, and virtual PLCs (vPLC) that represent typical industry hardware. A platform that incorporates these three components to provide the most accurate representation of actual NPP networks and controllers is developed in this paper.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗
DOE OSTI2017

CRISPR-Cpf1 mediates efficient homology-directed repair and temperature-controlled genome editing

Cpf1 is a novel class of CRISPR-Cas DNA endonucleases, with a wide range of activity across different eukaryotic systems. Yet, the underlying determinants of this variability are poorly understood. Here, we demonstrate that LbCpf1, but not AsCpf1, ribonucleoprotein complexes allow efficient mutagenesis in zebrafish and Xenopus. We show that temperature modulates Cpf1 activity by controlling its ability to access genomic DNA. This effect is stronger on AsCpf1, explaining its lower efficiency in ectothermic organisms. We capitalize on this property to show that temporal control of the temperature allows post-translational modulation of Cpf1-mediated genome editing. Finally, we determine that LbCpf1 significantly increases homology-directed repair in zebrafish, improving current approaches for targeted DNA integration in the genome. Together, we provide a molecular understanding of Cpf1 activity in vivo and establish Cpf1 as an efficient and inducible genome engineering tool across ectothermic species.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Project 57 Air Monitoring Report: (Jan 1 - Dec 31 2018)

During the late 1950s, the Atomic Energy Commission (AEC) (now the U.S. Department of Energy [DOE]) conducted a series of safety experiments to determine if a nuclear device subjected to a large conventional explosives detonation would result in a nuclear yield. The AEC obtained temporary use of a large portion of western Emigrant Valley from the U.S. Department of Defense (DOD) for one of these experiments: Project 57. Following the Project 57 safety experiment, the AEC fenced the contaminated area and returned control of the surrounding land to the DOD. Emigrant Valley is part of the Nevada Test and Training Range (NTTR). For safety and security reasons, access to the NTTR is controlled using both physical (i.e., fences) and administrative (e.g., signs and postings) controls. Therefore, the public cannot access the Project 57 site and there are no known human receptors that routinely access the site. Project 57 was detonated on April 24, 1957, in Emigrant Valley approximately 13 mi (21 km) northeast of the north end of Yucca Flat. This test was undertaken to develop: (1) a means of estimating immediate distribution and long-term redistribution of plutonium dispersed during a nonnuclear detonation; (2) biomedical evaluation techniques for use in likely plutonium-laden environments; (3) methods of decontaminating ground areas, pavements, and building materials; and (4) alpha survey instruments and field monitoring procedures to promptly estimate contaminant deposition. Although the test did not result in the fission of nuclear materials, it did disseminate plutonium across the ground surface. Various radiological surveys have been performed in the area since Project 57 was conducted. The AEC delineated the area containing radioactive material based on radioactivity surveys performed shortly after the test was conducted. A designated Contamination Area (CA), marked by a fence, was established later. The distribution of americium-241 (Am-241) in the area was determined again in a 1997 flyover, which showed Am-241 ranging from as much as 70,000 counts per second (cps) at ground zero to background (<70 cps). This survey also documented Am-241 radioactivity on the ground surface beyond the east side CA fence at levels of up to 150 cps. In 2007, the DOE expanded the CA by posting “Contamination Area” signs 200 ft to 400 ft (60 m to 120 m) beyond the original fence, which formed a new CA boundary.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2019

Increasing access to microfluidics for studying fungi and other branched biological structures

Background: Microfluidic systems are well-suited for studying mixed biological communities for improving industrial processes of fermentation, biofuel production, and pharmaceutical production. The results of which have the potential to resolve the underlying mechanisms of growth and transport in these complex branched living systems. Microfluidics provide controlled environments and improved optical access for real-time and high-resolution imaging studies that allow high-content and quantitative analyses. Studying growing branched structures and the dynamics of cellular interactions with both biotic and abiotic cues provides context for molecule production and genetic manipulations. To make progress in this arena, technical and logistical barriers must be overcome to more effectively deploy microfluidics in biological disciplines. A principle technical barrier is the process of assembling, sterilizing, and hydrating the microfluidic system; the lack of the necessary equipment for the preparatory process is a contributing factor to this barrier. To improve access to microfluidic systems, we present the development, characterization, and implementation of a microfluidics assembly and packaging process that builds on self-priming point-of-care principles to achieve “ready-to-use microfluidics.”Results: We present results from domestic and international collaborations using novel microfluidic architectures prepared with a unique packaging protocol. We implement this approach by focusing primarily on filamentous fungi; we also demonstrate the utility of this approach for collaborations on plants and neurons. In this work we (1) determine the shelf-life of ready-to-use microfluidics, (2) demonstrate biofilm-like colonization on fungi, (3) describe bacterial motility on fungal hyphae (fungal highway), (4) report material-dependent bacterial-fungal colonization, (5) demonstrate germination of vacuum-sealed Arabidopsis seeds in microfluidics stored for up to 2 weeks, and (6) observe bidirectional cytoplasmic streaming in fungi.Conclusions: This pre-packaging approach provides a simple, one step process to initiate microfluidics in any setting for fungal studies, bacteria-fungal interactions, and other biological inquiries. This process improves access to microfluidics for controlling biological microenvironments, and further enabling visual and quantitative analysis of fungal cultures.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Oil and Natural Gas Development near and beneath Uranium Tailings Cells and Other Remediated Sites - 20358

DOE LM is responsible for maintaining remedies at more than 50 remediated sites that require active long-term surveillance and maintenance (LTS and M). Institutional controls (ICs) were established to limit human and environmental exposures to residual contamination by controlling land use, restricting access to potential hazards, and making the public aware of potential dangers from the residual contamination. ICs for each site were developed based on current and foreseeable conditions; however, these controls should evolve to mitigate potential human health and other environmental risks from unanticipated changes in activities or site conditions. Activities conducted beyond the DOE long-term care boundaries by non-DOE parties may impact the effectiveness of ICs or even the long-term stability of disposal cells and groundwater remedies. Several sites that LM manages or that will transfer to LM for LTS and M are in active oil and gas production basins. The sites are associated with uranium ore milling or nuclear test sites and have been remediated or are in the process of remediation and have no ongoing DOE mission. Particularly since 2005, hydraulic fracturing ('fracking') combined with directional drilling has dramatically increased oil and natural gas (oil/gas) production in the United States, now making it the number one hydrocarbon-producing country. Fracking has allowed production from shale and other rocks of low natural permeability that were historically not considered viable oil/gas resources. Also, because wellbores can now be drilled as many as 5 kilometers or more horizontally, hydrocarbons can be recovered from zones without having access to the surface above them. Drilling activity near Rulison, Colorado, the location of an underground nuclear detonation managed by LM, has raised public concerns that remnant radioactivity in the detonation zone could migrate to producing wells and enter the natural gas distribution system. At sites like the Falls City, Texas, Disposal site, transitioned to LM with a split estate, increased production in the region has resulted in wellbores beneath the long-term care boundary of the site. While the DOE license for Falls City was approved by the NRC in 1997 without the acquisition of the subsurface rights, NRC has raised concerns on whether this activity impacts the integrity of the cell and/or groundwater remedies. In Wyoming, LM is anticipating receiving the license for the Bear Creek site and manages the Spook site, both locations of uranium mill tailings disposal cells. These Wyoming sites are in an active region of major oil and gas development. This change in surrounding land use will require LM to work collaboratively with NRC to address these concerns. At several DOE sites radioactive waste, and often intermixed nonradioactive constituents, is permanently isolated in engineered disposal cells. At other sites, residual contamination can be present in groundwater, blast cavities, and infinite amounts within soil. Areas targeted for oil and gas development at the Texas and Wyoming sites range from depths of 2440 to 3350 meters (8000-11000 feet) below ground surface. When most site remedies were designed, oil and gas development was not considered to be an issue within the site boundary. With the more recent uses of directional drilling, fracking, and associated wastewater injections, the evaluative criteria for incoming sites and ICs for managing long-term protectiveness and regulatory compliance are changing. Although DOE surface ownership was previously thought to be a robust IC to prevent surface and subsurface development, particularly at disposal sites, that may no longer be the case. Whereas the subsurface development at Texas and Wyoming sites is currently occurring at thousands of meters beneath the surface, shallower and direct onsite development would be more of a concern for LM. The energy introduced when a well is hydraulically fractured causes microseismic events (magnitude -2 to 1.0) as the fractures propagate but does not directly cause seismic events (earthquakes) of sufficient magnitude to damage surface structures. Additionally, the extent of the hydraulic fractures is limited to the targeted depths, which are far below the surface. However, earthquakes of sufficient magnitude to damage surface structures have been attributed to the injection of wastewater into disposal wells. States that have experienced these effects have enacted regulations that limit the rate and pressures that wastewater can be injected into disposal wells. Items LM may consider are the need for monitoring the volume of water injected into nearby disposal wells and possibly installing seismic stations at sites most susceptible to damage. While the need to demonstrate that no impact from oil and gas extraction activities on a site groundwater remedy or disposal cells will vary from site to site, LM is assessing risk criteria for this activity. Though LM has not experienced an issue yet at a uranium tailings disposal site, it cannot assume that there will be none, and must address NRC concerns. LM is building on past experiences at other sites and beginning to form new processes with the hope that these concepts can be applied for other future sites. (authors)

04 OIL SHALES AND TAR SANDS↗
DOE OSTI2022

An active site loop toggles between conformations to control antibiotic hydrolysis and inhibition potency for CTX-M β-lactamase drug-resistance enzymes

Abstract β-lactamases inactivate β-lactam antibiotics leading to drug resistance. Consequently, inhibitors of β-lactamases can combat this resistance, and the β-lactamase inhibitory protein (BLIP) is a naturally occurring inhibitor. The widespread CTX-M-14 and CTX-M-15 β-lactamases have an 83% sequence identity. In this study, we show that BLIP weakly inhibits CTX-M-14 but potently inhibits CTX-M-15. The structure of the BLIP/CTX-M-15 complex reveals that binding is associated with a conformational change of an active site loop of β-lactamase. Surprisingly, the loop structure in the complex is similar to that in a drug-resistant variant (N106S) of CTX-M-14. We hypothesized that the pre-established favorable loop conformation of the N106S mutant would facilitate binding. The N106S substitution results in a ~100- and 10-fold increase in BLIP inhibition potency for CTX-M-14 and CTX-M-15, respectively. Thus, this indicates that an active site loop in β-lactamase toggles between conformations that control antibiotic hydrolysis and inhibitor susceptibility. These findings highlight the role of accessible active site conformations in controlling enzyme activity and inhibitor susceptibility as well as the influence of mutations in selectively stabilizing discrete conformations.

59 BASIC BIOLOGICAL SCIENCES↗