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Regulatory interventions improve the biosynthesis of limiting amino acids from methanol carbon to improve synthetic methylotrophy in Escherichia coli

Synthetic methylotrophy aims to engineer methane and methanol utilization pathways in platform hosts like Escherichia coli for industrial bioprocessing of natural gas and biogas. While recent attempts to engineer synthetic methylotrophs have proved successful, autonomous methylotrophy, that is, the ability to utilize methane or methanol as sole carbon and energy substrates, has not yet been realized. Here, we address an important limitation of autonomous methylotrophy in E. coli: the inability of the organism to synthesize several amino acids when grown on methanol. We targeted global and local amino acid regulatory networks. Those include removal of amino acid allosteric feedback inhibition (argA H15Y , ilvA L447F , hisG E271K , leuA G462D , proB D107N , thrA S345F, trpE S40F ), knockouts of transcriptional repressors (ihfA, metJ); and overexpression of amino acid biosynthetic operons (hisGDCBHAFI, leuABCD, thrABC, trpEDCBA) and transcriptional regulators (crp, purR). Compared to the parent methylotrophic E. coli strain that was unable to synthesize these amino acids from methanol carbon, these strategies resulted in improved biosynthesis of limiting proteinogenic amino acids (histidine, leucine, lysine, methionine, phenylalanine, threonine, tyrosine) from methanol carbon. In several cases, improved amino acid biosynthesis from methanol carbon led to improvements in methylotrophic growth in methanol minimal medium supplemented with a small amount of yeast extract. Here, this study addresses a key limitation currently preventing autonomous methylotrophy in E. coli and possibly other synthetic methylotrophs and provides insight as to how this limitation can be alleviated via global and local regulatory modifications.

60 APPLIED LIFE SCIENCES↗

A pangenomic analysis of the Nannochloropsis organellar genomes reveals novel genetic variations in key metabolic genes

Background: Microalgae in the genus Nannochloropsis are photosynthetic marine Eustigmatophytes of significant interest to the bioenergy and aquaculture sectors due to their ability to efficiently accumulate biomass and lipids for utilization in renewable transportation fuels, aquaculture feed, and other useful bioproducts. To better understand the genetic complement that drives the metabolic processes of these organisms, we present the assembly and comparative pangenomic analysis of the chloroplast and mitochondrial genomes from Nannochloropsis salina CCMP1776. Results: The chloroplast and mitochondrial genomes of N. salina are 98.4% and 97% identical to their counterparts in Nannochloropsis gaditana. Comparison of the Nannochloropsis pangenome to other algae within and outside of the same phyla revealed regions of significant genetic divergence in key genes that encode proteins needed for regulation of branched chain amino synthesis (acetohydroxyacid synthase), carbon fixation (RuBisCO activase), energy conservation (ATP synthase), protein synthesis and homeostasis (Clp protease, ribosome). Conclusions: Many organellar gene modifications in Nannochloropsis are unique and deviate from conserved orthologs found across the tree of life. Implementation of secondary and tertiary structure prediction was crucial to functionally characterize many proteins and therefore should be implemented in automated annotation pipelines. The exceptional similarity of the N. salina and N. gaditana organellar genomes suggests that N. gaditana be reclassified as a strain of N. salina.

59 BASIC BIOLOGICAL SCIENCES↗

Radiotracers For Lipid Signaling Pathways In Biological Systems

The primary focus of this project continues to be the development of radiotracers and radiotracer methodology for studying physiology and biochemistry. The compounds that have been labeled areacylethanolamines and acylglycerols that are, as classes, represented in both in plants and in animals. In the latter, some of these act as ligands for cannabinoid receptors and they are therefore known as endocannabinoids. Cannabinoid receptors are not found in plant genomes so that plants must contain other receptors and signaling systems that use acylethanolamines. Relatively little work has been done on that issue, though acylethanolamines do modulate plant growth and stress resistance, thus possessing obvious relevance to agriculture and energy production. Progress has been described in five peer-reviewed papers and seven meeting abstracts. Preparation of 2-acylglycerol lipid messengers in high purity. A novel enzymatic synthesis was developedthat gave pure 2-acylglycerols free of any rearrrangement to the thermodynamically more stable 1(3)-acylglycerol byproducts. The method utilized 1,3-dibutyryl-2-acylglycerol substrate ethanolysis by a resinimobilized lipase. Thus, pure radiolabeled 2-acylglycerols can now be conveniently prepared just prior to their utilization. These synthetic studies were published in the Journal of Medicinal Chemistry, 2011. Diacylglycerol lipase assay methodology. Diacylglycerol lipases (DAGLs) generate 2- acylglycerols, and are thus potential targets for disease- or growth-modifying agents, by means of reducing formation of 2-acylglycerols. A radioTLC assay of the hydrolysis of radiolabeled diglyceride substrate [1''-carbon-14]2-arachidonoyl-1-stearoyl-sn-glycerol has been implemented, and used to validate a novel, potentially highthroughput fluorescence resonance energy transfer (FRET) based assay. A number of new DAGL inhibitors that have selectivity for DAGLs were synthesized and screened. This work was very recently published in Bioorganic and Medicinal Chemistry Letters.Acylethanolamines. The work initially focused on myristoylethanolamine (MEA) labeled in the acyl moiety with tritium and with carbon-14. Standard coupling conditions were used and gave the desired radiotracers in decent yields. Tritiated product was needed for the search for binding sites of this signaling messenger, and C-14 labeled product for imaging using phosphorimaging screens. Additionally, C-14 MEA was prepared labeled in the ethanolamine moiety. Fluorine-18 labeled acylethanolamines. A multi-step synthesis was performed to obtain unlabeled N-(16- fluorohexadecanoyl)ethanolamide for an HPLC standard via the precursor 16-bromo-N- [2[(tetrahydro-2Hpyran-2-yl)oxy]ethyl]hexadecanoylamide. Near-anhydrous F-18 and the bromo precursor (approximately 2-3 mg) in acetonitrile (0.5 ml) were heated with a PETWave microwave reactor for 10 min at 80 °C. F-18 incorporation was checked using silica-gel TLC (8:92 methanol/chloroform). Deprotection was carried out using TFA. The crude product was purified by semi-preparative ODS HPLC (80:20 AcN/ H2O). The product peak was diluted in water, concentrated on an SPE column, and eluted in 1 mL EtOH. F-18 incorporation to form the THP protected product was >90% with negligible side products observed. Deprotection and HPLC purification proceeded successfully with >99% radiochemical l purity. Brain distribution studies were done in mice. This work was presented at the Society of Nuclear Medicine meeting in June 2012. Subsequent more detailed studies including whole body tissue distribution studies, microPET experiments and radiochromatographic studies were published in ACS Chemical Neurosciences in 2014. Iodine-125 labeled acylethanolamines. The precursor 12-bromo-N-[2[(tetrahydro-2H-pyran-2-yl)oxy]ethyl]dodecanoylamide was prepared and a 1 mg sample was subject to exchange labeling withradioiodide in refluxing acetonitrile for 1 hour. HPLC analysis as above indicated incorporation of about10% of the radioactivity into a compound with the expected retention time of the corresponding iodocompound. Treatment with TFA converted the radioactivity into a compound with the expected retentiontime of 12-iodododenanoyl ethanolamine. Radiotracer studies in plants. One of the aims of this funding was to follow up the studies of Tripathy et al. (Plant Physiol., 2003) who first reported high affinity binding sites for tritiated myristoylethanolamine in plants, and also reported blockage of these sites by the cannabinoid receptor antagonist AM281. Because plant genomes do not contain genes for cannabinoid receptors, this was an intriguing report. I-125 labeled AM281 was therefore prepared, to facilitate identification of binding sites for this compound in Arabodopsis thaliana plants. However, such sites could not be found, the binding studies were repeated in tobacco plants. Again, binding sites for AM281 were not found. Additionally, it has become evident that clear demonstration of binding sites for tritiated MEA is obscured by metabolic incorporation of radioactivity into plant tissues. Studies on this issue are being aggressively pursued. Binding methodologies used in experiments with animal tissues require modification for their optimal use with plant tissues. Detailed studies of [14C]myristoylethanolamine isotopomers and of [14C]arachidonoylethanolamine isotopomers Since the labeled compounds being prepared for experiments with plants may have utility in Nuclear Medicine, studies were conducted with some of these compounds in mice. The brain uptake of C-14 MEA labeled in either acyl or ethanolamine moiety was 3-4 fold higher than uptake of C-14 myristic acid or C-14 ethanolamine. Carrier MEA increased brain uptake. Autoradiographs of MEA showed regionally specific uptake, with somewhat different patterns for acyl and ethanolamine isotopomers. It is hypothesized that these labeled compounds might form the basis of autoradiographic imaging of regional activity of enzymes such as fatty acid amide hydrolase, which may regulate endocannabinoid tone in animals. Early results were presented at the 2011 ICRS meeting, and at the 2012 Society for Neurosciences. Narachidonoylethanolamine is an endocannabinoid signaling messenger in animals and is known as “anandamide”. It is one of several families of signaling molecules derived from arachidonic acid, the principal omega-6 polyunsaturated fatty acids (PUFA’s) in animal species. Other derivatives of arachidonic acid include thromboxanes and prostaglandins. Full details of the studies with the ethanolamide isotopomers were a part of the PhD dissertation of Kun Hu (nee Qian), and were submitted for publication to Nuclear Medicine and Biology in August 2016. Syntheses of [14C]docosahexanoylethanolamine isotopomers and preliminary biological investigations Docosahexaenoic acid (DHA) is the omega-3 PUFA that can be regarded in some respects as the counterpart of arachidonic acid in the omega-6 series. While arachidonic acid is proinflammatory, DHA is anti-inflammatory, and foods high in DHA (or artificially enriched in DHA) are commonly regarded as promoting health. In contrast to the large literature on the Nethanolamide of arachidonic acid (i.e. the endocannabinoid anandamide) as of now (9/25/2016) there are only six papers on the corresponding ethanolamide of DHA, and when our studies under this grant began there were none. Beneficial actions of endogenously produced DHAethanolamine (“synaptamide”) have been indicated, and to help elucidate the possible roles of synaptamide, we have synthesized this molecule for the first time labeled with C-14 in either the ethanolamine moiety or the fatty acid moiety. Studies of the disposition of endogenously administered isotopomers of DHA-ethanolamine are in progress, to complement tissue culture experiments evaluation hypothesized protective effects of this DHA derivative.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

Thermostatic Valves Containing Silicone-Oil Actuators

Flow-splitting and flow-mixing thermally actuated spool valves have been developed for controlling flows of a heat-transfer fluid in a temperature-regulation system aboard the Mars Science Laboratory (MSL) rover. Valves like these could also be useful in terrestrial temperature-regulation systems, including automobile air-conditioning systems and general refrigeration systems. These valves are required to provide smoother actuation over a wider temperature range than the flow-splitting, thermally actuated spool valves used in the Mars Explorer Rover (MER). Also, whereas the MER valves are unstable (tending to oscillate) in certain transition temperature ranges, these valves are required not to oscillate. The MER valves are actuated by thermal expansion of a wax against spring-loaded piston rods (as in common automotive thermostats). The MSL valves contain similar actuators that utilize thermal expansion of a silicone oil, because silicone-oil actuators were found to afford greater and more nearly linear displacements, needed for smoother actuation, over the required wider temperature range. The MSL valves also feature improved spool designs that reflect greater understanding of fluid dynamics, consideration of pressure drops in valves, and a requirement for balancing of pressures in different flow branches.

Bhandari, Pradeep↗

A New Gold Mine? Achieving HVAC Energy Efficiency Through a System Metric

Washington State's Commercial Energy Code adopted a new energy metric called HVAC Total System Performance Ratio (TSPR) in 2019, a first in the codes world to regulate HVAC system efficiency. TSPR is a ratio of annual heating and cooling loads to the annual carbon emissions associated with the energy consumed by the HVAC system. TSPR provides a performance-based solution to evaluate and improve the overall HVAC design. The TSPR metric and its companion calculation tool were developed by Pacific Northwest National Laboratory (PNNL) with support from U.S. Department of Energy (DOE), Northwest Energy Efficiency Alliance (NEEA) and the City of Seattle. The new metric represents a significant shift in how HVAC design will meet code requirements. Utility programs can also leverage TSPR as a measure to determine energy savings and incentive amounts for HVAC retrofits. This paper describes the efforts by NEEA and its collaborators to prepare the market for TSPR adoption in code. This paper provides the pilot projects led by University of Washington Integrated Design Lab (IDL) to understand potential issues that could be faced by early adopters. This paper also covers how training and outreach provide engagement opportunities that can streamline code compliance, help address issues faced by early adopters and promote participation in utility programs. As Washington State works on the goals of 70% energy reduction and zero fossil-fuel greenhouse gas emission homes and buildings by the year 2031 , system level performance metrics will likely become increasingly more necessary and prevalent. This paper concludes that the HVAC TSPR requirement helps familiarize the HVAC industry with this approach and helps Washington achieve its long-term goals.

Liu, Bing↗

Progress in Monitoring Strategies for Risk Reduction in Geologic CO2 Storage

Abstract: This geophysical monitoring review is intended to lay the ground work for an approach to integrating monitoring strategies with risk assessment and risk reduction and to summarize work conducted by U.S. Department of Energy's National Risk Assessment Partnership (NRAP) participants that addresses key questions related to monitoring strategies. Commercial, demonstration and pilot geologic CO2 storage (GCS) projects have provided a wealth of practical experience in monitoring technologies. In this manuscript we review a few key projects, specific geophysical methods, well-based methods, geophysical workflows and some suggested best practices. These practices are often driven and governed by regulatory requirements (e.g. U.S. Environmental Protection Agency [US-EPA] Class VI underground injection regulation) and the need for effective reservoir management of pressures and volumes as well as the site-specific risks to containment, plume migration, and underground sources of drinking water. Reducing uncertainty in monitoring is likely to lead to reducing uncertainty in risk. One of our conclusions is the need for field experiment testing that is risk-driven. It is notable that GCS pilot projects have been designed for safe storage, and to date they have been successful. Technical challenges at all geologic CO2 storage sites include assuring injectivity and capacity predictions are reliable, the fate and subsurface topology of the CO2 plume is acceptable, regulatory permitting and compliance are accomplished in a timely manner, needed business assurances are supported by quantitative technical data, and the storage operation is acceptable to stakeholders. There is risk associated with all of these challenges necessitating a monitoring program utilizing both direct and indirect monitoring methods. Because of the direct association of risk and monitoring in GCS, geophysical monitoring is viewed as a key component in approaching risk assessment. To address monitoring in the context of risk assessment the U.S. Department of Energy's National Risk Assessment Partnership (NRAP) has undertaken a review of work related to monitoring storage sites. In this review we provide a framework for monitoring and then present some specific monitoring tools that may be useful for regulatory compliance, as well as technologies that are applicable to leakage detection.

Harbert, William↗

Oxysterols selectively promote short-term apoptosis in tumor cell lines

Highlights: • Several cholesterol oxides were evaluated for their capacity to induce cell death. • Short-term cytotoxicity of oxysterols varied with their type and concentration. • Short-term cytotoxicity of oxysterols varied with the cell lineage. • Cytotoxic oxysterols induced apoptosis in human breast cancer and mouse melanoma. • They also inhibited the S phase and stimulated either the G0/G1 or the G2/M phase. Oxysterols are 27-carbon oxidation products of cholesterol metabolism. Oxysterols possess several biological actions, including the promotion of cell death. Here, we examined the ability of several oxysterols to induce short-term death in cancerous (human breast cancer and mouse skin melanoma cells) and non-cancerous (human endothelial cells and lung fibroblasts) cell lines. We determined cell viability, Ki67 expression, cell cycle regulation, and apoptosis after 24-h incubations with oxysterols. We found that different oxysterols had different effects on the studied parameters. Moreover, the effects depended on cell type and oxysterol concentration. Three cytotoxic oxysterols (7-ketocholesterol, cholestane-3β-5α-6β-triol, and 5α-cholestane-3β,6β-diol) inhibited the S phase and stimulated the G0/G1 or G2/M phases. These oxysterols promoted apoptosis, determined with Annexin V and propidium iodide assays. These results showed that different oxysterols have cytotoxic effects depending on the cell line. The findings suggest a potential pharmacological utility of cytotoxic oxysterols.

60 APPLIED LIFE SCIENCES↗

A sucrose-binding site provides a lead towards an isoform-specific inhibitor of the cancer-associated enzyme carbonic anhydrase IX

Human carbonic anhydrase (CA; EC 4.2.1.1) isoform IX (CA IX) is an extracellular zinc metalloenzyme that catalyzes the reversible hydration of CO 2 to HCO 3 $-$ , thereby playing a role in pH regulation. The majority of normal functioning cells exhibit low-level expression of CA IX. However, in cancer cells CA IX is upregulated as a consequence of a metabolic transition known as the Warburg effect. The upregulation of CA IX for cancer progression has drawn interest in it being a potential therapeutic target. CA IX is a transmembrane protein, and its purification, yield and crystallization have proven challenging to structure-based drug design, whereas the closely related cytosolic soluble isoform CA II can be expressed and crystallized with ease. Therefore, we have utilized structural alignments and site-directed mutagenesis to engineer a CA II that mimics the active site of CA IX. In this paper, the X-ray crystal structure of this CA IX mimic in complex with sucrose is presented and has been refined to a resolution of 1.5 Å, an R cryst of 18.0% and an R free of 21.2%. Finally, the binding of sucrose at the entrance to the active site of the CA IX mimic, and not CA II, in a non-inhibitory mechanism provides a novel carbohydrate moiety binding site that could be further exploited to design isoform-specific inhibitors of CA IX.

59 BASIC BIOLOGICAL SCIENCES↗

Stacked Low-Inertia Converter or Solid-State Transformer: Modeling and Model Predictive Priority-Shifting Control for Voltage Balance

This paper presents control challenges of stacked low-inertia converter (SLIC) or cascaded reduced dc-link solid-state transformer (SST) and proposes a novel model predictive priority-shifting (MPPS) control with implicit modulator and a discrete-time large-signal model for voltage balancing and dc-link regulation. Low-inertia converters, featuring small electrolytic capacitor-less dc links, dramatically reduce cost, size, and weight compared to conventional solutions. However, without a large dc-link buffer, the input and output are tightly coupled, leading to significant control challenges. The control becomes even more challenging with these converters stacked input-series output-parallel (ISOP) for medium-voltage (MV) grid, which causes coupling between the modules besides the coupling within each module. This paper analyzes the multi-objective, multi-degree of freedom control problem, using the modular soft-switching solid-state transformer (M-S4T) as an example of the SLIC. First, distribution of control efforts under controller saturation is critical because multiple control objectives can be conflicting, especially when the module voltages are unbalanced and are being restored. The MPPS can shift the priorities to address this issue. Second, due to the low inertia and high dc-link ripple, classic space vector pulse-width modulation (SVPWM), average model with small-ripple assumption, and control design based on small-signal model cannot accurately modulate, model, and control the nonlinear reduced dc link. Therefore, a discrete-time large-signal model of the M-S4T is established to derive the predictive control in the MPPS. The MPPS and the PI control are compared in MV simulations to show the issue of applying the PI to the SLIC and the effectiveness of the MPPS for voltage balancing and dc-link regulation in a deadbeat manner. Finally, the proposed control is tested on a 5 kV ISOP SiC SST prototype to verify priority shifting to address controller saturation issue and fast and robust voltage balancing.

14 SOLAR ENERGY↗

Genomic and Secretomic Analyses Reveal Unique Features of the Lignocellulolytic Enzyme System of Penicillium decumbens

Many Penicillium species could produce extracellular enzyme systems with good lignocellulose hydrolysis performance. However, these species and their enzyme systems are still poorly understood and explored due to the lacking of genetic information. Here, we present the genomic and secretomic analyses of Penicillium decumbens that has been used in industrial production of lignocellulolytic enzymes in China for more than fifteen years. Comparative genomics analysis with the phylogenetically most similar species Penicillium chrysogenum revealed that P. decumbens has evolved with more genes involved in plant cell wall degradation, but fewer genes in cellular metabolism and regulation. Compared with the widely used cellulase producer Trichoderma reesei, P. decumbens has a lignocellulolytic enzyme system with more diverse components, particularly for cellulose binding domain-containing proteins and hemicellulases. Further, proteomic analysis of secretomes revealed that P. decumbens produced significantly more lignocellulolytic enzymes in the medium with cellulose-wheat bran as the carbon source than with glucose. The results expand our knowledge on the genetic information of lignocellulolytic enzyme systems in Penicillium species, and will facilitate rational strain improvement for the production of highly efficient enzyme systems used in lignocellulose utilization from Penicillium species.

59 BASIC BIOLOGICAL SCIENCES↗

Stabilizing Metallic Na Anodes via Sodiophilicity Regulation: A Review

This review focuses on the Na wetting challenges and relevant strategies regarding stabilizing sodium-metal anodes in sodium-metal batteries (SMBs). The Na anode is the essential component of three key energy storage systems, including molten SMBs (i.e., intermediate-temperature Na-S and ZEBRA batteries), all-solid-state SMBs, and conventional SMBs using liquid electrolytes. We begin with a general description of issues encountered by different SMB systems and point out the common challenge in Na wetting. We detail the emerging strategies of improving Na wettability and stabilizing Na metal anodes for the three types of batteries, with the emphasis on discussing various types of tactics developed for SMBs using liquid electrolytes. We conclude with a discussion of the overlooked yet critical aspects (Na metal utilization, N/P ratio, critical current density, etc.) in the existing strategies for an individual battery system and propose promising areas (anolyte incorporation and catholyte modifications for lower-temperature molten SMBs, cell evaluation under practically relevant current density and areal capacity, etc.) that we believe to be the most urgent for further pursuit. Comprehensive investigations combining complementary post-mortem, in situ, and operando analyses to elucidate cell-level structure-performance relations are advocated.

25 ENERGY STORAGE↗

Automation of Data Collection Methods for Online Monitoring of Nuclear Power Plants

Maintenance activities in the United States nuclear power industry are mainly centered on preventive and corrective maintenance. Predictive maintenance, which is a variable time basis maintenance strategy that is dependent on the condition of the equipment, is less utilized but potentially more effective. In the current maintenance practices, the nuclear power plants rely on manual processes to supply information to a human decision making process. The plants have several processes that generate information that is not typically used beyond the intended target for collecting that information. This information is a source of data that is manually collected by labor intensive tasks. The data is therefore expensive to collect, yet limited in use. The industry has realized the opportunity present in reducing the labor intensive tasks by automating them and increasing the uses of the data collected by either developing advanced predictive maintenance methods using data-driven decision making that could span from trending to machine learning and artificial intelligence. This shift in maintenance strategy may result in high cost saving impact. In addition to reducing labor requirements and frequency of maintenance activities, this shift in strategy will reduce the materials portion of the operations and maintenance cost as well. The aim of this specific effort is to focus on the automation of data collection process. It is part of a series of planned efforts to target multiple elements of migrating the current preventive maintenance approach in nuclear power plants to a data-driven approach. These elements include data collection, management, analysis, visualization, in addition to value analysis and change enabling. The other five elements of migration were not considered because they are planned to be explicitly studied in multiple parallel efforts. Challenges including regulations requirements or limitations are planned in a future effort that focuses on change enabling. The risk and benefit/cost of the approach shift is planned in another effort targeted towards value. The storage, communication, computational power, data architecture, and protection are part of the information infrastructure and are planned in dedicated data management effort. The best means to present the data for assisted human decision making is planned in a data visualization effort. The data interpretation and patterns recognition for machine decision making is planned through multiple efforts in data analytics over the following years. This work identified seventeen data sources collection methods in nuclear power plants that could be automated for an integrated data platform that enables comprehensive and informed decision making. For each of the identified seventeen data sources, the base, modern, and state of the art states were described. The current state of data collection used by most utilities was identified as the "base state". The current achievable state was identified as the "modern state", a state that has either been achieved by at least some US utilities or in other industries. The future state that requires research and development to reach was identified as the "state of the art" state. The described states used an out-of-the-box thinking approach. This effort found that there is a hug potential for task automation and data fidelity improvements by evolving the current industry data collection methods. The majority of the processes analyzed were found to be manual and labor intensive and fell in the base state. As a result, the modern state often targeted reducing the manual logging efforts by using electronic and semi-intelligent means or complementing the work with sensors and mobile or fixed technologies that can reduce the demand of labor activities. The state of the art state involved applications advanced methods of data correlation, data mining, and machine learning along with increasing the spatial distribution and fidelity of data, coupling multiple data sources, and enabling the use of sophisticated mobile devices and machine comprehension methods.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Nanoscale control of LaAlO 3 /SrTiO 3 metal–insulator transition using ultra-low-voltage electron-beam lithography

In this work, we describe a method to control the insulator–metal transition at the LaAlO 3 /SrTiO 3 interface using ultra-low-voltage electron beam lithography. Compared to previous reports that utilize conductive atomic force microscope (c-AFM) lithography, this approach can provide comparable resolution (~10 nm) at write speeds (10 mm/s) that are up to 10 000× faster than c-AFM. The writing technique is nondestructive, and the conductive state is reversible via prolonged exposure to air. Transport properties of representative devices are measured at milli-Kelvin temperatures, where superconducting behavior is observed. We also demonstrate the ability to create conducting devices on graphene/LaAlO 3 /SrTiO 3 heterostructures. The underlying mechanism is believed to be closely related to the same mechanism regulating c-AFM-based methods.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Reverse β-oxidation pathways for efficient chemical production

Abstract Microbial production of fuels, chemicals, and materials has the potential to reduce greenhouse gas emissions and contribute to a sustainable bioeconomy. While synthetic biology allows readjusting of native metabolic pathways for the synthesis of desired products, often these native pathways do not support maximum efficiency and are affected by complex regulatory mechanisms. A synthetic or engineered pathway that allows modular synthesis of versatile bioproducts with minimal enzyme requirement and regulation while achieving high carbon and energy efficiency could be an alternative solution to address these issues. The reverse β-oxidation (rBOX) pathways enable iterative non-decarboxylative elongation of carbon molecules of varying chain lengths and functional groups with only four core enzymes and no ATP requirement. Here, we describe recent developments in rBOX pathway engineering to produce alcohols and carboxylic acids with diverse functional groups, along with other commercially important molecules such as polyketides. We discuss the application of rBOX beyond the pathway itself by its interfacing with various carbon-utilization pathways and deployment in different organisms, which allows feedstock diversification from sugars to glycerol, carbon dioxide, methane, and other substrates.

59 BASIC BIOLOGICAL SCIENCES↗

Least-cost Optimal Distribution Grid Expansion (LODGE) v1

LODGE finds the least-cost portfolio of traditional distribution system upgrades to integrate new loads and behind-the-meter DERs in combination with advanced non-wire alternative (NWA) planning solutions, such as utility-owned storage and distributed PV. Working with a set of least-cost solutions per feeder/substation allows us to benchmark and compare techno-economic performance of different traditional and NWA planning solutions. An important aspect of the LODGE model is that it includes power flow operation constraints. This means that the least-cost planning solutions delivered by LODGE are technically feasible. Available projects in LODGE portfolio include: 1) feeder reconductoring; 2) transformers upgrades; 3) non-wire alternatives, such as strategic siting and sizing of storage and distributed PV; 4) voltage regulators.

Heleno, Miguel↗

Transitioning from Cobalt-60 to X-Ray or E-Beam for Medical Sterilization: Filling Data and Education Gaps

The Office of Radiological Security (ORS) within the U.S. DOE’s National Nuclear Security Administration (NNSA) is charged to “work with government, law enforcement, and businesses across the globe to protect radioactive sources used for medical, research, and commercial purposes; remove and dispose of disused radioactive sources; and reduce the global reliance on high activity radioactive sources through the promotion of viable non-radioisotopic alternative technologies.” In this effort, the NNSA/ORS, along with the Office of Defense Nuclear Nonproliferation R&D, has been working with government and private entities that utilize gamma-ray irradiators in areas that include blood irradiation, radiotherapy, biology research, well logging, and radiation processing to consider alternative technologies that do not depend upon radioactive materials. In the medical device industry, nearly 50% of products are sterilized with ethylene oxide, 40.5% with cobalt-60 gamma-rays, 4.5% with electron-beam (e-beam), and <1% with X-ray.(1,2) The cobalt-60 irradiation facilities use an estimated 150 MCi of isotope to sterilize approximately 200 million cubic feet of product annually.(3) Due to growing safety regulations governing cobalt-60 use, continued advancements in e-beam and X-ray accelerator technology, and the greater irradiation durations and costs for cobalt-60 irradiation compared to e-beam and X-ray, switching to these non-radionuclide modalities is increasingly economically competitive. However, this breakdown of 40.5% cobalt-60 and 5.5% accelerator for product sterilization illustrates the difficulty that market forces and industry players have had in overcoming barriers in transitioning to non-nuclide alternatives. Besides those associated with construction of new e-beam and X-ray facilities, significant financial and regulatory obstacles are also associated with the irradiation and comprehensive testing of product constituent polymers and elastomers. The standards associated with product material performance testing are the Association for the Advancement of Medical Instrumentation (AAMI) Technical Information Report 17 (TIR 17), and its referenced American Society for Testing and Materials (ASTM) polymer testing standards. In the study now underway, and the subject of this article, the NNSA/ORS provided R&D funding to the Pacific Northwest National Laboratory (PNNL) in partnership with university researchers, leading medical product manufacturers, accelerator manufacturers, and contract sterilizers. The resulting Team Nablo (in memory of Sam Nablo) includes individuals representing PNNL, Becton Dickinson (BD), Johnson & Johnson, Stryker, Texas A&M University (TAMU), Steri-Tek, Mevex, Ion Beam Applications (IBA), and Sterigenics. The goals of this study are to 1) Identify specific polymer and/or elastomer-based medical products that are currently sterilized with gamma-rays, and for which there are data gaps and would be of greatest industry impact, 2) Work with relevant ASTM subcommittees for guidance and for potential standard revision, 3) Work with an appropriate Food & Drug Administration (FDA) office for potential collaboration on a guidance document for device manufacturers, 4) Measure the physical effects that product materials exhibit when they are given sterilization-level radiation doses from cobalt-60, e-beam or X-ray, and determine whether these effects would preclude the use of e-beam or X-ray for sterilization, 5) Communicate the resulting scientific datasets and findings via industry conferences, peer-reviewed journals, and relevant trade magazines, and 6) Post the resulting datasets on a publicly accessible website, which can be built on in future years. The end objective is to identify technical barriers preventing the adoption of X-ray and e-beam technologies, and share the results to encourage X-ray and e-beam adoption where feasible.

sterilization, alternate technologies, gamma, X-ra↗

Power Hardware-in-the-Loop Testing of Distribution Solid State Transformers

Savannah River National Laboratory and Resilient Power Systems have developed a utility grade solid state transformer and are planning on deploying it for testing in January 2018. The solid state transformer is the next generation solution to implementation of distributed energy resources such as solar and batteries. They can directly connect the solar and battery systems to DC ports and rout power between these DC ports and load ports. The solid state transformer can revolutionize current grid modernization initiatives by supplying grid support and building support services at the edge of the grid. These services consist of volt-VAR optimization, power factor correction, voltage regulation, frequency regulation, fault current support, demand side management, DC bus for AC/DC capable houses, and power routing between multiple ports for transactive control. For this fiscal year a test plan for testing the SST capabilities was develop. Along with the FPGA controls for the SST output ports. These controls will allow for any waveform to be produced by each port of the SST. Finally, a hybrid distribution SST was tested at the eGRID resulting in medium voltage testing experience for SRNL engineering team. DOE is currently engaged, through the Building Technology Office and Infrastructure Security and Energy Restoration in future work building from this project.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Development and Assessment of a Novel Training Package for Basic Maneuvering Tasks on a Flight Simulator Using Self Instruction Methods and Above Real Time Training (ARTT)

One result of the relatively recent advances in computing technology has been the decreasing cost of computers and increasing computational power. This has allowed high fidelity airplane simulations to be run on personal computers (PC). Thus, simulators are now used routinely by pilots to substitute real flight hours for simulated flight hours for training for an aircraft type rating thereby reducing the cost of flight training. However, FAA regulations require that such substitution training must be supervised by Certified Flight Instructors (CFI). If the CFI presence could be reduced or eliminated for certain tasks this would mean a further cost savings to the pilot. This would require that the flight simulator have a certain level of 'intelligence' in order to provide feedback on pilot performance similar to that of a CFI. The 'intelligent' flight simulator would have at least the capability to use data gathered from the flight to create a measure for the performance of the student pilot. Also, to fully utilize the advances in computational power, the simulator would be capable of interacting with the student pilot using the best possible training interventions. This thesis reports on the two studies conducted at Tuskegee University investigating the effects of interventions on the learning of two flight maneuvers on a flight simulator and the robustness and accuracy of calculated performance indices as compared to CFI evaluations of performance. The intent of these studies is to take a step in the direction of creating an 'intelligent' flight simulator. The first study deals with the comparisons of novice pilot performance trained at different levels of above real-time to execute a level S-turn. The second study examined the effect of out-of-the-window (OTW) visual cues in the form of hoops on the performance of novice pilots learning to fly a landing approach on the flight simulator. The reliability/robustness of the computed performance metrics was assessed by comparing them with the evaluations of the landing approach maneuver by a number of CFIs.

Ali, Syed Firasat↗