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DOE OSTI2024

Genome‐wide association identifies a BAHD acyltransferase activity that assembles an ester of glucuronosylglycerol and phenylacetic acid

SUMMARY Genome‐wide association studies (GWAS) are an effective approach to identify new specialized metabolites and the genes involved in their biosynthesis and regulation. In this study, GWAS of Arabidopsis thaliana soluble leaf and stem metabolites identified alleles of an uncharacterized BAHD‐family acyltransferase (AT5G57840) associated with natural variation in three structurally related metabolites. These metabolites were esters of glucuronosylglycerol, with one metabolite containing phenylacetic acid as the acyl component of the ester. Knockout and overexpression of AT5G57840 in Arabidopsis and heterologous overexpression in Nicotiana benthamiana and Escherichia coli demonstrated that it is capable of utilizing phenylacetyl‐CoA as an acyl donor and glucuronosylglycerol as an acyl acceptor. We, thus, named the protein Glucuronosylglycerol Ester Synthase (GGES). Additionally, phenylacetyl glucuronosylglycerol increased in Arabidopsis CYP79A2 mutants that overproduce phenylacetic acid and was lost in knockout mutants of UDP‐sulfoquinovosyl: diacylglycerol sulfoquinovosyl transferase, an enzyme required for glucuronosylglycerol biosynthesis and associated with glycerolipid metabolism under phosphate‐starvation stress. GGES is a member of a well‐supported clade of BAHD family acyltransferases that arose by duplication and neofunctionalized during the evolution of the Brassicales within a larger clade that includes HCT as well as enzymes that synthesize other plant‐specialized metabolites. Together, this work extends our understanding of the catalytic diversity of BAHD acyltransferases and uncovers a pathway that involves contributions from both phenylalanine and lipid metabolism.

09 BIOMASS FUELS↗

DOE OSTI2019

Avista Turner Energy Storage System: Assessment of Battery Technical Performance

This report investigates the technical performance of the 1 MW, 3.2 MWh advanced vanadium flow battery energy storage system (FBESS), consisting of two 0.5 MW, 1.6 MWh strings, based on a number of reference performance and use case tests. The FBESS is located on the utility 12 kV distribution system, between the Turner Substation and SEL’s manufacturing facility in Pullman, Washington. Reference performance tests (RPTs) were intended to assess the general technical capability of the FBESS (e.g., stored energy capacity, ramp rate performance, ability to track varying charge/discharge commands, direct current (DC) battery internal resistance). Use case tests were used to examine the performance of the FBESS while engaged in specific grid services (e.g., arbitrage, frequency regulation, balancing, volt ampere reactive (var) support, dynamic peak shaving). The project determined performance metrics that are important for understanding FBESS performance when subjected to field operation for achieving economic benefits from measurements and/or calculations. These metrics were: • RTE with and without rest, with and without auxiliary loads • Auxiliary power consumption • Reference signal tracking • Temperature excursions beyond normal operating range • Parasitic power loss unaccounted by auxiliary load during rest, and • State of charge (SOC) excursions beyond normal operating range. These RPT and use case performance metrics were analyzed using recorded test results. In addition, the results and lessons presented are also beneficial for any task or effort that needs technical assessment on similar and different types of FBESS, and BESS in general, based on technical specifications and/or field deployment results, since the assessment methodology remains the same.

25 ENERGY STORAGE↗

DOE OSTI2021

Metagenomic Insights Into the Microbial Iron Cycle of Subseafloor Habitats

Microbial iron cycling influences the flux of major nutrients in the environment (e.g., through the adsorptive capacity of iron oxides) and includes biotically induced iron oxidation and reduction processes. The ecological extent of microbial iron cycling is not well understood, even with increased sequencing efforts, in part due to limitations in gene annotation pipelines and limitations in experimental studies linking phenotype to genotype. This is particularly true for the marine subseafloor, which remains undersampled, but represents the largest contiguous habitat on Earth. To address this limitation, we used FeGenie, a database and bioinformatics tool that identifies microbial iron cycling genes and enables the development of testable hypotheses on the biogeochemical cycling of iron. Herein, we survey the microbial iron cycle in diverse subseafloor habitats, including sediment-buried crustal aquifers, as well as surficial and deep sediments. We inferred the genetic potential for iron redox cycling in 32 of the 46 metagenomes included in our analysis, demonstrating the prevalence of these activities across underexplored subseafloor ecosystems. We show that while some processes (e.g., iron uptake and storage, siderophore transport potential, and iron gene regulation) are near-universal, others (e.g., iron reduction/oxidation, siderophore synthesis, and magnetosome formation) are dependent on local redox and nutrient status. Additionally, we detected niche-specific differences in strategies used for dissimilatory iron reduction, suggesting that geochemical constraints likely play an important role in dictating the dominant mechanisms for iron cycling. Overall, our survey advances the known distribution, magnitude, and potential ecological impact of microbe-mediated iron cycling and utilization in sub-benthic ecosystems.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRSDate not supplied

Inlet Radiated Noise Predictions for the NASA Source Diagnostic Test Fan Using Physics-Based Simulations

Commercial supersonic aircraft are expected to produce considerably more noise during landing and take-off operations than existing subsonic aircraft. Establishing noise regulation is challenging because no examples of this aircraft type are currently flying. Existing aircraft noise prediction tools can be used, but the noise source models for these empirical tools are typically based on data from subsonic aircraft components. The present work documents a procedure for using physics-based computational tools to generate the propagating sound field radiating out the nacelle inlet. Although no analytical source modeling is demonstrated, these tools utilize measured data to compare the complex pressure for various propagating modes. The discrepancies found in applying the propagation tools to the subsonic inlet will be ascribed to calculations of supersonic inlets being used to update current empirical noise prediction tools.

Noise Predictions↗

DOE OSTI2019

Development and Calibration of an Online Energy Model for AHU Fan

The model development is necessary for the study of the energy consumption of Heating, Ventilation, and Air Conditioning (HVAC) systems. To predict the HVAC energy consumption accurately, one needs to model the individual HVAC components either from the measured data or based on the knowledge of the underlying physical phenomenon. Online model characterization is critical for improving the performance of real-time model-based fault detection and diagnosis (FDD) strategies. For HVAC control, models can be used to optimize the supervisory and local feedback control strategies to improve the energy consumption efficiency, or for providing ancillary services to the grid. It has been reported that, fans in HVAC systems of commercial buildings alone can provide substantial frequency regulation service, with little change in the indoor environment. In this paper, a real-time data-driven Air Handling Unit (AHU) fan model was developed based on recursive multi regression model. A generic nonlinear polynomial model has been studied to cover scenarios with different combinations of measurement variables, variable orders as well as different training and prediction horizons. Typical measurements including static pressure, mass flow rate, and damper positions are utilized as inputs to model the power consumption of the fan. The developed models have been validated both with simulation data from EnergyPlus-Dymola co-simulation model and with field measurement data for small to medium commercial buildings. The validation results show that the online model proposed can provide an effective prediction of the AHU fan power consumption.

Dong, Jin↗

DOE OSTI2018

Phenomena identification and ranking table study for thermal hydraulics for Advanced High Temperature Reactor

The Advanced High-Temperature Reactor (AHTR) is a conceptual design of a Fluoride salt-cooled High-temperature Reactor (FHR) utilizing 7 LiF-BeF 2 (66-34 mol%) as its primary coolant. To identify key phenomena that impose potential challenges on thermal hydraulics modeling and simulation of such a reactor system, a thermal hydraulics phenomena identification and ranking table (TH-PIRT) study was performed for the AHTR in a Department of Energy Nuclear Energy University Program (NEUP) integrated research project led by Georgia Institute of Technology. A panel of experts from regulators, industries, national laboratories, and academia was assembled for the study. In this paper, the TH-PIRTs identified by the panel for two events, namely, station blackout and simultaneous withdrawal of all control rods, are summarized and discussed in detail. In addition, the key phenomena that warrant further study and research for AHTR analysis are identified to support the validation of thermal hydraulics system-level analysis codes and computational fluid dynamics simulation tools, as well as future FHR reactor licensing.

42 ENGINEERING↗

DOE OSTI2015

Power requirements and environmental impact of a pedelec. A case study based on real-life applications

This paper describes the methodologies to appraise the power requests and environmental analysis of an electrically assisted bicycle under real driving conditions, also containing regulations and technical-science-related aspects. For this purpose, in this study, the on-road test program of an electrically assisted bicycle was executed in the urban area of Naples on different test tracks, so a general assessment about its driving behavior under several driving conditions was performed. The power requirements in different typical riding situations were estimated by a procedure based on the experimental kinematic parameters that characterize the driving dynamics collected during the real-life applications. An environmental analysis was also performed, with a methodology that takes into account the environmental assessment of a moped by measuring the experimental moped exhaust emissions of the regulated pollutants. Starting from the results acquired during the different test samples, besides, an assessment of the electric traction offered by this pedelec on the driving comfort was evaluated for different riding situations. - Highlights: • The power requirements of a pedelec in typical riding conditions were identified. • The estimated electricity consumption for battery recharging was defined. • An environmental valuation of the tested pedelec and of a moped was performed. • Emissions that could be saved utilizing a pedelec instead of a moped were derived.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2025

Predicting metal-binding proteins and structures through integration of evolutionary-scale and physics-based modeling

Metals are essential elements in all living organisms, binding to approximately 50% of proteins. They serve to stabilize proteins, catalyze reactions, regulate activities, and fulfill various physiological and pathological functions. While there have been many advancements in determining the structures of protein-metal complexes, numerous metal-binding proteins still need to be identified through computational methods and validated through experiments. Here, to address this need, we have developed the ESMBind workflow, which combines evolutionary scale modeling (ESM) for metal-binding prediction and physics-based protein-metal modeling. Our approach utilizes the ESM-2 and ESM-IF models to predict metal-binding probability at the residue level. In addition, we have designed a metal-placement method and energy minimization technique to generate detailed 3D structures of protein-metal complexes. Our workflow outperforms other models in terms of residue and 3D-level predictions. To demonstrate its effectiveness, we applied the workflow to 142 uncharacterized fungal pathogen proteins and predicted metal-binding proteins involved in fungal infection and virulence.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2015

Scarless genome editing and stable inducible expression vectors for Geobacter sulfurreducens

Metal reduction by members of the Geobacteraceae is encoded by multiple gene clusters, and the study of extracellular electron transfer often requires biofilm development on surfaces. Genetic tools that utilize polar antibiotic cassette insertions limit mutant construction and complementation. In addition, unstable plasmids create metabolic burdens that slow growth, and the presence of antibiotics such as kanamycin can interfere with the rate and extent of Geobacter biofilm growth. We report here genetic system improvements for the model anaerobic metal-reducing bacterium Geobacter sulfurreducens . A motile strain of G. sulfurreducens was constructed by precise removal of a transposon interrupting the fgrM flagellar regulator gene using SacB/sucrose counterselection, and Fe(III) citrate reduction was eliminated by deletion of the gene encoding the inner membrane cytochrome imcH . We also show that RK2-based plasmids were maintained in G. sulfurreducens for over 15 generations in the absence of antibiotic selection in contrast to unstable pBBR1 plasmids. Therefore, we engineered a series of new RK2 vectors containing native constitutive Geobacter promoters, and modified one of these promoters for VanR-dependent induction by the small aromatic carboxylic acid vanillate. Inducible plasmids fully complemented Δ imcH mutants for Fe(III) reduction, Mn(IV) oxide reduction, and growth on poised electrodes. A real-time, high-throughput Fe(III) citrate reduction assay is described that can screen numerous G. sulfurreducens strain constructs simultaneously and shows the sensitivity of imcH expression by the vanillate system. Lastly, these tools will enable more sophisticated genetic studies in G. sulfurreducens without polar insertion effects or need for multiple antibiotics.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Targeting hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase for lignin modification in Brachypodium distachyon

Abstract Background Hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase (HCT) is a central enzyme of the so-called “esters” pathway to monolignols. As originally envisioned, HCT functions twice in this pathway, to form coumaroyl shikimate and then, in the “reverse” direction, to convert caffeoyl shikimate to caffeoyl CoA. The discovery of a caffeoyl shikimate esterase (CSE) that forms caffeic acid directly from caffeoyl shikimate calls into question the need for the reverse HCT reaction in lignin biosynthesis. Loss of function of HCT gives severe growth phenotypes in several dicot plants, but less so in some monocots, questioning whether this enzyme, and therefore the shikimate shunt, plays the same role in both monocots and dicots. The model grass Brachypodium distachyon has two HCT genes, but lacks a classical CSE gene. This study was therefore conducted to evaluate the utility of HCT as a target for lignin modification in a species with an “incomplete” shikimate shunt. Results The kinetic properties of recombinant B. distachyon HCTs were compared with those from Arabidopsis thaliana , Medicago truncatula , and Panicum virgatum (switchgrass) for both the forward and reverse reactions. Along with two M. truncatula HCTs, B. distachyon HCT2 had the least kinetically unfavorable reverse HCT reaction, and this enzyme is induced when HCT1 is down-regulated. Down regulation of B. distachyon HCT1, or co-down-regulation of HCT1 and HCT2, by RNA interference led to reduced lignin levels, with only modest changes in lignin composition and molecular weight. Conclusions Down-regulation of HCT1, or co-down-regulation of both HCT genes, in B. distachyon results in less extensive changes in lignin content/composition and cell wall structure than observed following HCT down-regulation in dicots, with little negative impact on biomass yield. Nevertheless, HCT down-regulation leads to significant improvements in biomass saccharification efficiency, making this gene a preferred target for biotechnological improvement of grasses for bioprocessing.

09 BIOMASS FUELS↗

DOE OSTI2016

ChloroSeq, an optimized chloroplast RNA-Seq bioinformatic pipeline, reveals remodeling of the organellar transcriptome under heat stress

Although RNA-Seq has revolutionized transcript analysis, organellar transcriptomes are rarely assessed even when present in published datasets. Here, we describe the development and application of a rapid and convenient method, ChloroSeq, to delineate qualitative and quantitative features of chloroplast RNA metabolism from strand-specific RNA-Seq datasets, including processing, editing, splicing, and relative transcript abundance. The use of a single experiment to analyze systematically chloroplast transcript maturation and abundance is of particular interest due to frequent pleiotropic effects observed in mutants that affect chloroplast gene expression and/or photosynthesis. To illustrate its utility, ChloroSeq was applied to published RNA-Seq datasets derived from Arabidopsis thaliana grown under control and abiotic stress conditions, where the organellar transcriptome had not been examined. The most appreciable effects were found for heat stress, which induces a global reduction in splicing and editing efficiency, and leads to increased abundance of chloroplast transcripts, including genic, intergenic, and antisense transcripts. Moreover, by concomitantly analyzing nuclear transcripts that encode chloroplast gene expression regulators from the same libraries, we demonstrate the possibility of achieving a holistic understanding of the nucleus-organelle system. In conclusion, ChloroSeq thus represents a unique method for streamlining RNA-Seq data interpretation of the chloroplast transcriptome and its regulators.

RNA splicing↗

DOE OSTI2022

Hydropower Capital and O&M Costs: An Exploration of the FERC Form 1 Data

This report explores the potential for using responses from the Federal Energy Regulatory Commission’s (FERC’s) “Electric Utility Annual Report,” also known as FERC Form 1, as a cost database for conventional and pumped storage hydropower (PSH). The report outlines the process used to compile an easy-to-access database from the original FERC Form 1 data and discusses the historical cost/performance data. The steps for developing the database include downloading the annual FoxPro files from the FERC website (FERC, 2020), reading them into an Excel format, and reconciling naming issues across years, repeated entries, and shared assets, among others. The cleaned and compiled Form 1 database is available on Oak Ridge National Laboratory’s (ORNL’s) HydroSource website. For this report, the compiled Form 1 database was linked to two other hydropower databases, the National Inventory of Dams and the ORNL Existing Hydropower Assets, that provide additional information on the characteristics and other features of plants in the Form 1 data. Explorations of the plant characteristics, performance, capital costs, and operation and maintenance (O&M) cost data in the combined database were presented separately for conventional hydropower and PSH projects. Form 1 has several advantages as a hydropower cost database. First, the cost data is reported directly by plant owners. Second, Form 1 provides cost breakdowns of capital and operating costs for large conventional and PSH plants enabling more detailed tracking of hydropower costs compared with typical total cost estimates. Third, although the Form 1 data is not reported by all operational hydropower plants in the United States, the available data represents a wide range of plant characteristics and a sizable proportion of the hydropower fleet (61% of PSH plants and 22% of conventional hydropower plants by capacity). The lower proportion of the conventional hydropower fleet is because Form 1 reporting requirements apply only to private utilities that meet given size thresholds, leaving out federally owned facilities and many smaller hydropower plants. Overall, the Form 1 cost database represents a unique, publicly available database on hydropower asset capital and O&M costs. This report leads to a compiled database for the reporting years 1994 to 2020 that helps resolve issues with accessibility and use of the FERC Form 1 data by hydropower plant owners, project developers, technology developers, and regulators.

13 HYDRO ENERGY↗

DOE OSTI2024

T-Type Modular DC Circuit Breaker (T-Breaker) for Future DC Networks

The developed T-Type Modular DC Circuit Breaker (T-Breaker) technology offers an all-in-one solution to challenges in DC networks. This includes swift fault detection and protection, power transient stability, and power quality improvement, achieved through the utilization of wide bandgap (WBG) power semiconductors and energy storage devices. The T-Breaker not only facilitates rapid fault current detection and interruption but also implements fault current limiting through active insertion of storage devices or by operating WBG devices in the saturation region. Additionally, with the assistance of energy storage devices, potential overvoltage issues on power devices induced by control signal misalignment can be mitigated. The T-Breaker can be regulated to perform shunt current injection/absorption using the vertical arm and series voltage insertion via the horizontal arm, thereby enhancing DC system stability during voltage or load power fluctuation transients. The OSU team and Raytheon team actively worked together on designing, fabricating, assembling, and testing of two T-Breaker prototypes. The first prototype is rated at 1 kV, 500 A with half-bridge (unipolar) structure to validate the T-Breaker concept. The second prototype is rated at 20 kV, 50 A with full-bridge (bipolar) topology which can reach an efficiency of 99.977%, realize a power density of 60.2 MW/m3, and eliminate the 500-A fault current with a fault response time of around 20 µs. The prototypes show great feasibility of adopting this technology in multiple applications including electrified aircraft, super charging stations, data centers, etc.

24 POWER TRANSMISSION AND DISTRIBUTION↗

NASA NTRS2003

Magnetic Microspheres and Tissue Model Studies for Therapeutical Applications

Hyperthermia is a well known cancer therapy and consists of heating a tumor region to the elevated temperatures in the range of 40-45 C for an extended period of time (2-8 hours). This leads to thermal inactivation of cell regulatory and growth processes with resulting widespread necrosis, carbonization and coagulation. Moreover, heat boosts the tumor response to other treatments such as radiation, chemotherapy or immunotherapy. Of particular importance is careful control of generated heat in the treated region and keeping it localized. Higher heating, to about 56 C can lead to tissue thermo-ablation. With accurate temperature control, hyperthermia has the advantage of having minimal side effects. Several heating techniques are utilized for this purpose, such as whole body hyperthermia, radio-frequency (RF) hyperthermia, ultrasound technique, inductive microwave antenna hyperthermia, inductive needles (thermoseeds), and magnetic fluid hyperthermia (MFH).MFH offers many advantages as targeting capability by applying magnets. However, this technology still suffers significant inefficiencies due to lack of thermal control. This paper will provide a review of the topic and outline the ongoing work in this area. The main emphasis is in devising ways to overcome the technical difficulty in hyperthermia breast therapy of achieving a uniform therapeutic temperature over the required region of the body and holding it steady for an extended period (2-3 hours). The basic shortcomings of the presently utilized heating methods stem from the non-uniform thermal properties of the tissue and the point heating characteristics of the techniques without any thermal control. Our approach is to develop a novel class of magnetic fluids, which have inherent thermoregulating properties. We have identified a few magnetic alloys which can serve as suitable nano to micron-size particle material. The objective is to synthesize, characterize and evaluate the efficacy of Thermo Regulating Magnetic Fluids (TRMF) for hyperthermia therapy. The development of a tissue model and testing the fluid dynamics of particle motion, settling, distribution in the tissue matrix and heat generation will be discussed.

Ramachandran, N.↗

NASA NTRS2011

Automatic Facial Expression Recognition and Operator Functional State

The prevalence of human error in safety-critical occupations remains a major challenge to mission success despite increasing automation in control processes. Although various methods have been proposed to prevent incidences of human error, none of these have been developed to employ the detection and regulation of Operator Functional State (OFS), or the optimal condition of the operator while performing a task, in work environments due to drawbacks such as obtrusiveness and impracticality. A video-based system with the ability to infer an individual's emotional state from facial feature patterning mitigates some of the problems associated with other methods of detecting OFS, like obtrusiveness and impracticality in integration with the mission environment. This paper explores the utility of facial expression recognition as a technology for inferring OFS by first expounding on the intricacies of OFS and the scientific background behind emotion and its relationship with an individual's state. Then, descriptions of the feedback loop and the emotion protocols proposed for the facial recognition program are explained. A basic version of the facial expression recognition program uses Haar classifiers and OpenCV libraries to automatically locate key facial landmarks during a live video stream. Various methods of creating facial expression recognition software are reviewed to guide future extensions of the program. The paper concludes with an examination of the steps necessary in the research of emotion and recommendations for the creation of an automatic facial expression recognition program for use in real-time, safety-critical missions.

Blanson, Nina↗

NASA NTRS2012

Automatic Facial Expression Recognition and Operator Functional State

Blanson, Nina↗

DOE OSTI2017

Final Technical Report for GO17004 Regulatory Logic: Codes and Standards for the Hydrogen Economy

The objectives of this project are to: develop a robust supporting research and development program to provide critical hydrogen behavior data and a detailed understanding of hydrogen combustion and safety across a range of scenarios, needed to establish setback distances in building codes and minimize the overall data gaps in code development; support and facilitate the completion of technical specifications by the International Organization for Standardization (ISO) for gaseous hydrogen refueling (TS 20012) and standards for on-board liquid (ISO 13985) and gaseous or gaseous blend (ISO 15869) hydrogen storage by 2007; support and facilitate the effort, led by the NFPA, to complete the draft Hydrogen Technologies Code (NFPA 2) by 2008; with experimental data and input from Technology Validation Program element activities, support and facilitate the completion of standards for bulk hydrogen storage (e.g., NFPA 55) by 2008; facilitate the adoption of the most recently available model codes (e.g., from the International Code Council [ICC]) in key regions; complete preliminary research and development on hydrogen release scenarios to support the establishment of setback distances in building codes and provide a sound basis for model code development and adoption; support and facilitate the development of Global Technical Regulations (GTRs) by 2010 for hydrogen vehicle systems under the United Nations Economic Commission for Europe, World Forum for Harmonization of Vehicle Regulations and Working Party on Pollution and Energy Program (ECE-WP29/GRPE); and to Support and facilitate the completion by 2012 of necessary codes and standards needed for the early commercialization and market entry of hydrogen energy technologies.

08 HYDROGEN↗

DOE OSTI2018

ALK is required for NLRP3 inflammasome activation in macrophages

Highlights: • ALK promotes LPS-induced NLRP3 priming. • ALK promotes ATP-induced NLRP3 activation. • ALK promotes NLRP3-NEK7 complex formation. • ALK-mediated lipid peroxidation promotes NLRP3 activation. The NLRP3 inflammasome is a key mediator of host immune responses through the induction of pyroptosis and the release of cytokines. Although the pathologic role of inflammasome in infection and sterile inflammation is well known, the mechanism and regulation of NLRP3 inflammasome activation remains obscure. Here, we report that anaplastic lymphoma kinase (ALK) is a novel regulator of NLRP3 inflammasome activation in macrophages. Pharmacologic or genetic inhibition of ALK through targeted drugs (ceritinib and lorlatinib) or RNAi blocked extracellular ATP-induced NLRP3 inflammasome activation in macrophages. Mechanically, ALK-mediated NF-κB activation was required for the priming step of NLRP3 upregulation, whereas ALK-mediated lipid peroxidation contributed to the sensing step of NLRP3-NEK7 complex formation. These studies indicate that inhibition of ALK could be utilized to treat NLRP3-related inflammatory diseases.

60 APPLIED LIFE SCIENCES↗