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At least 163 records · Page 9

Development of genetic tools for heterologous protein expression in a pentose‐utilizing environmental isolate of Pseudomonas putida

Abstract Pseudomonas putida has emerged as a promising host for the conversion of biomass‐derived sugars and aromatic intermediates into commercially relevant biofuels and bioproducts. Most of the strain development studies previously published have focused on P. putida KT2440, which has been engineered to produce a variety of non‐native bioproducts. However, P. putida is not capable of metabolizing pentose sugars, which can constitute up to 25% of biomass hydrolysates. Related P. putida isolates that metabolize a larger fraction of biomass‐derived carbon may be attractive as complementary hosts to P. putida KT2440. Here we describe genetic tool development for P. putida M2, a soil isolate that can metabolize pentose sugars. The functionality of five inducible promoter systems and 12 ribosome binding sites was assessed to regulate gene expression. The utility of these expression systems was confirmed by the production of indigoidine from C6 and C5 sugars. Chromosomal integration and expression of non‐native genes was achieved by using chassis‐independent recombinase‐assisted genome engineering (CRAGE) for single‐step gene integration of biosynthetic pathways directly into the genome of P. putida M2. These genetic tools provide a foundation to develop hosts complementary to P. putida KT2440 and expand the ability of this versatile microbial group to convert biomass to bioproducts.

59 BASIC BIOLOGICAL SCIENCES↗

Scaling Equitable Finance

Driven by dramatic declines in up-front cost, the U.S. solar photovoltaics (PV) industry has taken off over the past decade, growing from 1 gigawatt of installed capacity in 2009 to 89 gigawatts in 2020—or enough capacity to power roughly 19 million homes. The industry is expected to double in size over just the next 5 years.1 Much of the growth has been driven by large, utility-scale projects that can produce 5 mega- watts or more of power—enough to power at least 1,000 homes. The cost of electricity produced by these projects has decreased by more than 70 percent since 2010. As of Q3 2020, development costs of large, util- ity-scale solar PV power plants were under $1 per watt, down by more than 70 percent from 2010.2 A robust array of investors has come forward to efficiently deliver capital to these kinds of utility-scale projects including large banks, insurance companies, pension funds, and others. But low- and moderate-income communities, including communities of color, are at risk of being left behind in the transition to clean energy. Mission- driven solar project developers and financial institu- tions have been working alongside energy justice advocates to open up solar access for these communi- ties, using strategies ranging from community solar, to solar installations on affordable multifamily housing, to distributed solar and storage programs, and more. Their goals go beyond simply generating more green energy to advancing social equity by: • empowering communities to control their energy future • stabilizing energy prices, saving money, and build- ing wealth for low-income families • creating quality jobs • improving health by reducing pollution • providing energy resilience for vulnerable communities Mission-driven actors are successfully deploying a wide variety of strategies to meet these goals, from helping low-income homeowners get solar—and some- times battery storage, to developing solar projects serv- ing affordable rental housing and community facilities, to building larger “shared solar” projects to which households from across the community can subscribe. However, the financing ecosystem does not work nearly as well for these “mission driven” solar proj- ects as it does for utility-scale projects. For home rooftop solar, even if low-income consumers have a home and suitable roof, they may fail to qualify for federal tax incentives, lack adequate credit to qualify for a loan—or the mission-driven lenders seeking to serve them may not be adequately capitalized to make long-term loans. For mission-driven commercial or community-scale projects, assembling nearly every component of the project capital stack—whether bridging early-stage costs, attracting tax credit equity investors, securing long-term debt, or coming up with sponsor equity and filling gaps—can present challenges. A variety of obstacles contribute to the scarcity of financing for low-income solar, including small project sizes, lack of developer balance sheet capacity, both real and perceived issues with credit risk, elevated technical assistance needs, and greater subsidy requirements to pursue goals such as deep energy affordability, climate resilience, or job creation. Still other obstacles are regulatory: for example, not all states allow community solar projects or Power Purchase Agreements, common strategies used for providing low-income solar—and the potential for regulations to shift over time creates risks that mission-driven projects can ill afford. This report synthesizes information garnered from 47 key informant interviews, four focus group discus- sions involving 60 stakeholders, and a review of the substantial existing literature on low-income solar finance to assess the current landscape of mission- driven solar development in the United States, examine the roles that community-based financial institutions could play, and recommend public invest- ments and policy changes that could help to scale the provision of equitable solar finance. Key recommen- dations for policymakers and funders in the renew- able energy and community development fields that emerge from this process include the following: • Help to capitalize and support community-based lenders to provide flexible, low-cost, and long- term financing to mission-driven solar projects— including providing guarantees or other forms of credit enhancement. • Provide federal support for equitable solar, including a grant-in-lieu-of-credits option for the Investment Tax Credit to improve access to this critical government subsidy. • Develop pools of government and philanthropic support that can complement financing from community-based lenders to complete the capi- tal stack for mission-driven projects, as well as to support education and technical assistance to both consumers and potential project sponsors. • Create a national Renewable Energy Credits pro- gram that includes social equity targets to provide a baseline of support for clean energy generation. • Change utility regulations to remove barriers to low-income solar projects; lower permitting costs; provide greater certainty for developers, consumers and owners; and measure progress toward equity in renewable energy policy implementation.

14 SOLAR ENERGY↗

Expanding the Scope of Bacterial CRISPR Activation with PAM-Flexible dCas9 Variants

CRISPR-Cas transcriptional tools have been widely applied for programmable regulation of complex biological networks. In comparison to eukaryotic systems, bacterial CRISPR activation (CRISPRa) has stringent target site requirements for effective gene activation. While genes may not always have an NGG protospacer adjacent motif (PAM) at the appropriate position, PAM-flexible dCas9 variants can expand the range of targetable sites. Here we systematically evaluate a panel of PAM-flexible dCas9 variants for their ability to activate bacterial genes. We observe that dxCas9-NG provides a high dynamic range of gene activation for sites with NGN PAMs while dSpRY permits modest activity across almost any PAM. Similar trends were observed for heterologous and endogenous promoters. For all variants tested, improved PAM-flexibility comes with the trade-off that CRISPRi-mediated gene repression becomes less effective. Weaker CRISPR interference (CRISPRi) gene repression can be partially rescued by expressing multiple sgRNAs to target many sites in the gene of interest. Furthermore, our work provides a framework to choose the most effective dCas9 variant for a given set of gene targets, which will further expand the utility of CRISPRa/i gene regulation in bacterial systems.

59 BASIC BIOLOGICAL SCIENCES↗

Assessing the energy equity benefits of energy storage solutions

Safety, reliability, efficiency, and affordability are no longer the sole tenets of electric grid planning. The evolving social and policy climate have placed new explicit requirements on the electric grid, including resilience, decarbonization, and energy equity and justice. Integrating energy equity strategies into modern grid design is intended to achieve a fair and just distribution of benefits within the energy system. This study aims to characterize the energy equity and community benefits of energy storage systems (ESS) under the following three use-case models: utility ESS that are operated within the distribution system; community-owned ESS; and behind-the-meter ESS that are customer-owned to serve the household. The goal of this energy equity analysis is to characterize the environmental, economic, and social benefits of ESS through applied metrics such as energy burden, energy poverty, energy vulnerability, job creation, and more. A 13.8 kV, 265 node representative feeder corresponding to the hot-humid climate of Louisiana, chosen for its high energy burden, frequent hurricanes and outage events, was used to perform a storage adequacy analysis for six different outage scenarios to determine energy storage access for each use-case. The results of this analysis are then used to characterize both the grid and equity benefits of storage solutions to inform a prioritization framework matching community needs with system preferences for utility planning processes, market regulations, and the wider network of energy system stakeholders.

Energy Equity, energy justice, equity, reliability↗

Light Water Reactor Sustainability Program Materials Research Pathway Technical Program Plan

The Materials Research (MR) Pathway within the Light Water Reactor Sustainability (LWRS) Program is charged with performing the research and development (R&D) to develop the scientific basis for understanding and predicting long-term environmental degradation behavior of materials in nuclear reactors. Furthermore, it is essential to use the mechanistic understanding of degradation phenomena in materials to develop mitigation, repair, and new material alternatives for existing components. The work will provide data and methods to assess performance of systems, structures, and components (SSCs) essential to safe and sustained reactor operations. The R&D products developed from the LWRS Program will be used by utilities, industry groups, and regulators to inform operational and regulatory requirements for materials in reactor SSCs subjected to long-term operation conditions, providing key input to both regulators and industry. The intent of this research is to help in reducing the operating costs, which may be in the form of offset maintenance costs due to better predictive models for component lifetimes, improved analysis of materials through nondestructive evaluation, reduced costs for repairs, or extended performance of plants through the selection of improved replacement materials. To best achieve this, industry experience and guidance are important as is their role in coordinated or collaborative research projects. The objectives of this report are to describe the motivation and organization of the MR Pathway within the LWRS Program, provide details on the individual research tasks within the MR Pathway, describe the outcomes and deliverables of the MR Pathway, including recent technical highlights and progress, and list the requirements for performing the research.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Light Water Reactor Sustainability: Program Materials Research Pathway (FY22 Technical Program Plan)

The Materials Research (MR) Pathway within the Light Water Reactor Sustainability (LWRS) program is charged with performing R&D to develop the scientific basis for understanding and predicting the long-term environmental degradation behavior of materials in nuclear reactors. Furthermore, the mechanistic understanding of degradation phenomena in materials must be leveraged to develop mitigation and repair strategies and new material alternatives for existing components. This research will provide data and methods to assess the performance of systems, structures, and components essential to safe and sustained reactor operations. The R&D products developed from the LWRS program will be used by stakeholders—including utilities, industry groups, and regulators—to inform operational and regulatory requirements for materials in reactor systems, structures, and components subjected to long-term operation conditions, providing key inputs to regulators and industry. Therefore, the intent of this research is to provide options to reduce the operating costs, which may be in the form of offset maintenance costs due to better predictive models for component lifetimes, improved analyses of materials through nondestructive evaluation, reduced costs for repairs, or extended performance of plants through the selection of improved replacement materials. To provide the best options, industry experience and guidance are important because of their role in coordinated or collaborative research projects. The objectives of this report are to describe the motivation and organization of the MR Pathway within the LWRS program; provide details on the individual research tasks within the MR Pathway; describe the outcomes and deliverables of the MR Pathway, including recent technical highlights and progress; and describe the requirements for performing this critically important research.

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

Comparing four heat-inducible promoters in stably transformed sugarcane regarding spatial and temporal control of transgene expression reveals candidates to drive stem-preferred transgene expression

Small heat shock protein (sHSP) promoters contain cis-regulatory elements that facilitate transcription in response to heat stress, making them valuable tools for functional studies through controlled gene expression and the precise regulation of gene-editing tools or morphogenic regulators. To evaluate their utility, GUS reporter gene expression driven by four plant-sourced HSP promoters (p GmHSP17.5 , p HvHSP17 , p ZmHSP17.7 , and p ZmHSP26 ) was compared across various tissues of stably transformed sugarcane before and after heat treatment. At 22°C, all promoters showed minimal activity in leaves and roots, although p ZmHSP17.7 and p HvHSP17 displayed moderate expression in stems. Following heat treatment, all promoters exhibited their highest activity in stems, followed by leaves and roots. In stem tissues, p GmHSP17.5 displayed heat-induced uidA expression comparable to the constitutive p ZmUbi promoter. Notably, heat-induced reporter gene activity in stem middle sections of single-copy transgenic lines containing p ZmHSP17.7 , p HvHSP17 , or p ZmHSP26 exceeded p ZmUbi -derived uidA activity by 9.7-fold, 3.8-fold, and 3.0-fold, respectively, with 346- to 3,672-fold induction compared to control conditions. Most promoters showed peak expression in the middle sections of the stem, while p HvHSP17 was the most active in the stem apices. Histochemical analysis revealed that p ZmHSP17.7 and p HvHSP17 were active in both parenchyma cells and vascular bundles within sugarcane stems. Among leaf tissues, mature leaves exhibited greater expression than senescing or immature leaves, while root activity remained consistently minimal across all promoters. Temperature-course experiments identified distinct activation thresholds: 34°C–36°C for p ZmHSP17.7 , 36°C for p ZmHSP26 , 36°C–38°C for p HvHSP17 , and 40°C–42°C for p GmHSP17.5 . Drought stress also induced reporter gene transcription in stems under HSP promoters, although with lower fold induction than heat treatment. These findings provide valuable tools for gene function studies and biotechnology applications, including heat stress tolerance research, controlled transgene expression in metabolic engineering, precision gene editing, and developmental biology studies.

60 APPLIED LIFE SCIENCES↗

Grid Value Investigation of Medium-Voltage Back-to-Back Converters: Preprint

This paper presents the work which investigates the value that the medium voltage back to back (MVB2B) converter can bring to the grid, through two potential applications: voltage regulation powered by the reactive power adjustment function, and the distributed energy resource (DER) adoption enhancement powered by the quantitative power transfer function. Two groups of distribution systems have been prepared for the value investigation, including IEEE 123-bus system for voltage regulation study and two utility provided realistic distribution systems for DER adoption enhancement study. The analysis results have demonstrated that the MVB2B converter can bring significant value to the grid through the two aforementioned applications. Particularly, with MVB2B converter transferring power between two feeders, a substantial amount of backfeeding solar power can be avoided, and the size of the energy storage which is used to store the extra solar power can be significantly reduced.

28 EE - Advanced Manufacturing Office (EE-5A)↗

Tuning Catalyst Activation and Utilization Via Controlled Electrode Patterning for Low-Loading and High-Efficiency Water Electrolyzers

An anode electrode concept of thin catalyst-coated liquid/gas diffusion layers (CCLGDLs), by integrating Ir catalysts with Ti thin tunable LGDLs with facile electroplating in proton exchange membrane electrolyzer cells (PEMECs), is proposed. The CCLGDL design with only 0.08 mg Ir cm -2 can achieve comparative cell performances to the conventional commercial electrode design, saving ~97% Ir catalyst and augmenting a catalyst utilization to ~24 times. CCLGDLs with regulated patterns enable insight into how pattern morphology impacts reaction kinetics and catalyst utilization in PEMECs. A specially designed two-sided transparent reaction-visible cell assists the in situ visualization of the PEM/electrode reaction interface for the first time. Oxygen gas is observed accumulating at the reaction interface, limiting the active area and increasing the cell impedances. In this work, it is demonstrated that mass transport in PEMECs can be modified by tuning CCLGDL patterns, thus improving the catalyst activation and utilization. The CCLGDL concept promises a future electrode design strategy with a simplified fabrication process and enhanced catalyst utilization. Furthermore, the CCLGDL concept also shows great potential in being a powerful tool for in situ reaction interface research in PEMECs and other energy conversion devices with solid polymer electrolytes.

08 HYDROGEN↗

Polysaccharide utilization loci in Bacteroides determine population fitness and community-level interactions

Polysaccharide utilization loci (PULs) are co-regulated bacterial genes that sense nutrients and enable glycan digestion. Human gut microbiome members, notably Bacteroides, contain numerous PULs that enable glycan utilization and shape ecological dynamics. To investigate the role of PULs on fitness and inter-species interactions, we develop a CRISPR-based genome editing tool to study 23 PULs in Bacteroides uniformis (BU). BU PULs show distinct glycan-degrading functions and transcriptional coordination that enables the population to adapt upon loss of other PULs. Exploiting a BU mutant barcoding strategy, we demonstrate that in vitro fitness and BU colonization in the murine gut are enhanced by deletion of specific PULs and modulated by glycan availability. PULs mediate glycan-dependent interactions with butyrate producers that depend on the degradation mechanism and glycan utilization ability of the butyrate producer. Thus, PULs determine community dynamics and butyrate production and provide a selective advantage or disadvantage depending on the nutritional landscape.

59 BASIC BIOLOGICAL SCIENCES↗

A Comprehensive Survey of Coal Ash Law and Commercialization: Its Environmental Risks, Disposal Regulation, and Beneficial Use Markets

This white paper was commissioned by the National Association of Regulatory Utility Commissioners (NARUC) under a grant from the U.S. Department of Energy (DOE), in response to a major rule, Hazardous and Solid Waste Management System; Disposal of Coal Combustion Residuals From Electric Utilities; Final Rule (Final Rule) issued by the U.S. Environmental Protection Agency (EPA) on April 17, 2015, on coal ash disposal and remediation of disposal sites, which has significant implications to states. It subjects state environmental agencies and public utility commissions to regulatory responsibilities to implement federal regulations under the Final Rule and oversee the cost implications on electric customers. To better evaluate the regulatory issues underlying the choices facing utilities and independent power producers (IPPs) regarding coal ash management options under the Final Rule, an overview of coal ash production and disposal practices is helpful. This comprehensive survey provides background information on the following topics: (a) coal ash production by a coal combustion generation plant, (b) the historical practices of utility waste management practices, (c) coal ash toxicity and related constituents, (d) further understanding of potential impacts to groundwater, and (e) the aging structures and outdated designs of coal ash surface impoundments and landfills.

01 COAL, LIGNITE, AND PEAT↗

Dynamic Economic Dispatch Considering Transmission–Distribution Coordination and Automatic Regulation Effect

With the rapid increase of renewable energy sources, the variability and uncertainty in power systems are significantly increased, and the power system operation is facing more and more challenges. In the traditional dynamic economic dispatch (DED) of transmission systems, the frequency and voltage regulation effects are not actively utilized. In addition, the transmission systems and distribution systems are optimized separately; the transmission–distribution coordination effect is not considered. In this article, a new DED is proposed. The optimal scheduling results are obtained by making a balance between operating cost and reliability cost. Both the automatic regulation effect and transmission–distribution coordination effect are explored and exploited in terms of the equivalent reserve, which can reduce the reserve provided by generating units, and can further enhance the security and economics of power system operation. The validity and effectiveness of the proposed model are illustrated by case studies.

24 POWER TRANSMISSION AND DISTRIBUTION↗

From Modular ADMS to Plug-and-Play Ops: Distribution Grid Operations with Platform-Level Orchestration to Enable Ambitious App Hosting

The core function of the distribution grid is to provide electricity to consumers affordably, reliably, and securely. In pursuing these core objectives, distribution utilities are accountable to customers, regulators, and in some cases, shareholders. Other third parties such as aggregators and microgrids can also have a stake in the smooth operation of the grid. Each of these stakeholders has economic, business, and/or governance objectives that inform their expectations of the distribution grid. This multi-objective, multi-stakeholder environment creates tension that must be reconciled to successfully design and operate the distribution grid. Innovative companies are competing to bring high-tech solutions to electric utilities and their customers that address each of these objectives. Many developers of advanced distribution management systems (ADMS) and distributed energy resource management systems (DERMS) have adopted a modular architecture that allows grid operators to select functions and features according to their individual system needs. A modular platform also allows the solution provider to develop and integrate specific new product modules; however, the need to pursue multiple objectives with a fixed set of controllable devices makes integration expensive whether it is done at the product development stage or the deployment stage. This cost creates a significant barrier to adoption and can lengthen the product to market time of new solutions. To fundamentally address the complexity of system integration for distribution grid operations, the U.S. Department of Energy Office of Electricity has funded the GridAPPS-D project at PNNL, which streamlines integration by contributing to standards development, defining system architecture, applying advanced mathematics, and developing open-source software to demonstrate the concept of an open data-integration platform for distribution operations. The open data-integration platform concept enables system operators and solution providers to deploy ambitious, best-of-breed applications (or apps) without continually reengineering for integration. Ambitious apps developed by different solution providers will inevitably attempt to achieve different control objectives with the same set of controllable devices. If the open platform itself can resolve these conflicts in a way that achieves the best available outcomes for all apps, doesn’t restrict the ambitious design of apps, and ensures safe and secure operations, apps will be able to plug-and-play with the platform at the same time as other ambitious apps. In this paper, we describe a framework called App Deconfliction that empowers a platform to assign setpoints to controllable devices based on the values preferred by different apps (and even external stakeholder entities like customers or aggregators). The App Deconfliction framework is compatible with several methods for determining setpoint values. We present two methods based on game theory that provide a subtle built-in incentive structure for developers to adapt their apps to the fact that they will be operating in a moderated multi-app environment and to favor device setpoints that have the most effect on their objectives over those that have the least effect. Our simulation-based demonstrations have shown that game-theory-based deconfliction can lead to a 7% improvement in control space utilization compared to design-based methods.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Interface morphogenesis with a deformable secondary phase in solid-state lithium batteries

Here, the complex morphological evolution of lithium metal at the solid-state electrolyte interface limits performance of solid-state batteries, leading to inhomogeneous reactions and contact loss. Inspired by biological morphogenesis, we developed an interfacial self-regulation concept in which a deformable secondary phase dynamically aggregates at the interface in response to local electro-chemo-mechanical stimuli, enhancing contact. The stripping of a lithium electrode that contains 5 to 20 mole % electrochemically inactive sodium domains causes spontaneous sodium accumulation across the interface, with the sodium deforming to attain intimate electrical contact without blocking lithium transport. This process, characterized with operando x-ray tomography and electron microscopy, mitigates voiding and improves cycling at low stack pressures. The counterintuitive strategy of adding electrochemically inactive alkali metal to improve performance demonstrates the utility of interfacial self-regulation for solid-state batteries.

Yoon, Sun Geun [Georgia Institute of Technology, A↗

Optimal assignment for the single-household shared autonomous vehicle problem

Autonomous vehicles have the potential to transform the way people are transported. While driverless technology may mean fewer vehicles are required to transport people to and from their daily activities, such changes may result in increased congestion or total miles traveled. In this study, we solve the single-household shared autonomous vehicle problem to identify cost-optimal routings of vehicles throughout the day. Such a tool will be useful for consumers seeking to minimize cost and for regulators seeking to understand and predict how people may behave in different scenarios. Here, we provide a thorough literature review and construct a mixed-integer linear program to minimize the daily travel cost of a household attending a given set of activities. Since solution time is a determinant for applicability of such a model, we present the model in a component-wise fashion. This approach allows us to understand which features most affect the problem complexity and solution time. We note that modeling carpooling is the feature that most increases time to find an optimal solution, and we therefore propose a novel modeling technique for carpooling two people. We illustrate the performance of our model by comparing it with other models from the literature and note that our model can solve significantly larger problem instances and in a time that is short enough to facilitate real-time scheduling. We also highlight the utility of our model for regulators, who can use it to analyze quickly produced optimal routes under different cost/tax scenarios.

33 ADVANCED PROPULSION SYSTEMS↗

Manipulating symmetry-breaking charge separation employing molecular recognition

The exploration of symmetry-breaking charge separation (SB-CS) is imperative when designing functional light-harvesting materials. Past explorations, however, have been confined to covalent systems, more often than not requiring complicated/demanding syntheses and facing inconvenient regulation of charge transfer processes. Here, in this work, we present a concept that regulates the efficiency of SB-CS through molecular recognition utilizing a pyridinium-based cyclophane as a host. This host undergoes photo-driven excited-state SB-CS. By employing different guests with distinct frontier molecular orbital energy levels, we have achieved comprehensive control of electron transfer pathways in the cyclophane, modulating between accelerated (>10-fold) intramolecular SB-CS involving superexchange and direct intermolecular electron transfer between the host and guest. The improvement in SB-CS efficiency results in catalytic activity for the photo-oxidation of a sulfur-mustard simulant. This research offers an opportunity for tuning SB-CS by utilizing molecular recognition, which holds the potential for achieving precise regulation without complicated organic syntheses.

charge transfer↗

Round Robin Measurements of Molten Salt Properties for LiF-NaF-KF (FLiNaK) and NaCl-KCl Mixtures

The development, operation, and regulation of nuclear reactors that utilize molten salts as fuel or as heat transfer media require knowledge of the thermal properties of the salt systems and quantification of the corresponding uncertainties. Knowledge of molten salt properties is also necessary for applications in material synthesis, processing, separations, solar thermal power generation, and energy storage. A round robin was conducted with national laboratory and university participants from twenty-one laboratories in five countries to compare property measurements, to better understand uncertainties, and to identify possible best practices. Two salt mixtures, each from a common batch, were distributed to participants for evaluation: equimolar NaCl-KCl and 45.0LiF-13.7NaF-41.3KF mol % (FLiNaK). Measurements were performed to determine the major constituent composition, oxygen content, density, thermal expansivity, melting point, and thermal conductivity. Error analysis was performed on each measurement for uncertainty quantification for each type of property that was explored. Finally, the resulting discussion of the methodologies used in this work is meant to lay the groundwork for the development of standard methods and reference materials for future high-temperature property measurements on halide melts.

11 - NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗