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At least 1,567 records · Page 87

Genomic, Transcriptional, and Phenotypic Analysis of the Glucose Derepressed Clostridium beijerinckii Mutant Exhibiting Acid Crash Phenotype

Clostridium beijerinckii is a predominant solventogenic bacterium that is used for the ABE fermentation. Various C. beijerinckii mutants are constructed for desirable phenotypes. The C. beijerinckii mutant BA105 harboring a glucose derepression phenotype was previously isolated and demonstrated the enhanced amylolytic activity in the presence of glucose. Despite its potential use, BA105 is not further characterized and utilized. Therefore, the authors investigate fermentation phenotypes of BA105 in this study. Under the typical batch fermentation conditions, BA105 consistently exhibits acid crash phenotype resulting in limited glucose uptake and cell growth. However, when the culture pH is maintained above 5.5, BA105 exhibits the increased glucose uptake and butanol production than did the wild‐type. To further analyze BA105, the authors perform genome sequencing and RNA sequencing. Genome analysis identifies two SNPs unique to BA105, in the upstream region of AbrB regulator (Cbei_4885) and the ROK family glucokinase (Cbei_4895) which are involved in catabolite repression and regulation of sugar metabolism. Transcriptional analysis of BA105 reveals significant differential expression of the genes associated with the PTS sugar transport system and acid production. This study improves understanding of the acid crash phenomenon and provides the genetic basis underlying the catabolite derepression phenotype of C. beijericnkii .

Seo, Seung‐Oh↗

Highly multiplexed design of an allosteric transcription factor to sense new ligands

Allosteric transcription factors (aTF) regulate gene expression through conformational changes induced by small molecule binding. Although widely used as biosensors, aTFs have proven challenging to design for detecting new molecules because mutation of ligand-binding residues often disrupts allostery. Here, we develop Sensor-seq, a high-throughput platform to design and identify aTF biosensors that bind to non-native ligands. We screen a library of 17,737 variants of the aTF TtgR, a regulator of a multidrug exporter, against six non-native ligands of diverse chemical structures – four derivatives of the cancer therapeutic tamoxifen, the antimalarial drug quinine, and the opiate analog naltrexone – as well as two native flavonoid ligands, naringenin and phloretin. Sensor-seq identifies biosensors for each of these ligands with high dynamic range and diverse specificity profiles. The structure of a naltrexone-bound design shows shape-complementary methionine-aromatic interactions driving ligand specificity. To demonstrate practical utility, we develop cell-free detection systems for naltrexone and quinine. Sensor-seq enables rapid and scalable design of new biosensors, overcoming constraints of natural biosensors.

59 BASIC BIOLOGICAL SCIENCES↗

Testing and Analysis of Grid Forming Inverter Control for Achieving Resilient and Economic Operation of an Islanded Microgrid

This investigation examines the feasibility of operating a battery energy storage system (BESS) in parallel with synchronous generation by using grid forming (GFM) control in order to achieve frequency control objectives while mitigating increases to operating costs in the context of an islanded microgrid. The BESS GFM control system, which is based on conventional droop techniques, is modeled along with the overall microgrid using the Real Time Digital Simulator (RTDS) to allow for integration of genset controller hardware. A series of simulations are performed to test the voltage and frequency regulation capability of the BESS control system when the primary frequency regulating genset is tripped offline. The results of the simulations suggest that the GFM control scheme will successfully maintain frequency and voltage stability, which will enable operation without a back-up genset while not compromising the microgrid resiliency to contingencies.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Application of ERTS-1 data to the protection and management of New Jersey's coastal environment

The author has identified the following significant results. The principal thrust of this ERTS-1 experiment is to develop quasi-operational information products from analysis of ERTS-1 imagery and collateral aerial photography and to apply these products to the practical regulation, protection, and management of New Jersey's coastal environment. Incorporated into this goal is the development of procedures for the operational use of ERTS-1 data products within New Jersey's Department of Environmental Protection. Analysis and product preparation for operational needs has centered on four major coastal resource problem areas: detection of land use changes in the coastal zone; siting of ocean outfalls; monitoring of offshore waste disposal; and calculation of recession rates along the Atlantic Shore. The relative utility and estimated monetary benefits derived from ERTS and aircraft imagery for each problem area have been determined. Of equal importance is the development to a capability within the State to use and understand remote sensor-derived information, and the application of this information to meet the requirements of current and anticipated coastal zone legislation.

Yunghans, R. S.↗

Self-regulating proportionally controlled heating apparatus and technique

A self-regulating proportionally controlled heating apparatus and technique is provided wherein a single electrical resistance heating element having a temperature coefficient of resistance serves simultaneously as a heater and temperature sensor. The heating element is current-driven and the voltage drop across the heating element is monitored and a component extracted which is attributable to a change in actual temperature of the heating element from a desired reference temperature, so as to produce a resulting error signal. The error signal is utilized to control the level of the heater drive current and the actual heater temperature in a direction to reduce the noted temperature difference. The continuous nature of the process for deriving the error signal feedback information results in true proportional control of the heating element without the necessity for current-switching which may interfere with nearby sensitive circuits, and with no cyclical variation in the controlled temperature.

Strange, M. G.↗

Uncrewed Aerial Systems for Emergency Medical First Response: A Market Research Report

This report presents the findings from market research conducted for NASA’s Aerial Aid Convergent Aeronautics Solutions (CAS) exploration project, which aims to assess the current state of the market and technological readiness for Uncrewed Aerial Systems (UAS) for medical emergency first response. The research reveals a robust and rapidly growing market for UAS, with a notable emerging sector for Drones as First Responders (DFR). Despite this growth, DFR applications are currently limited by regulatory, technical, and other challenges, which restrict their use primarily to manned remote video surveillance, and therefore are primarily employed by police units. To our knowledge, there is no evidence of UAS being utilized by medical first responders for scene assessment. Limited evidence exists for closely related applications; however, these are mostly confined to pilot programs for the delivery of medical supplies or equipment. Although there has been discussion around fully autonomous DFR applications for medical purposes such as UAS ambulances or patient transport drones, these applications are generally not yet operational in practice. The technology for full autonomy, especially in guidance and control, has seen significant advancements, and recent Federal Aviation Administration (FAA)regulations are likely to accelerate adoption. Computer vision algorithms for fully autonomous medical emergency response scene surveillance are primed for advancement and deployment. A notable gap likely exists between advancements in computer vision research and what is being integrated in the commercial DFR sector. This gap is primarily due to challenges such as quality assurance for autonomous systems, the availability of application-specific training datasets for computer vision algorithms, regulatory constraints, and public perception and privacy concerns.

Joshua M Fody↗

Hydrogen Detection Strategies to Support H2@SCALE - The NREL Sensor Laboratory

Hydrogen represents a major pathway to decarbonize and stabilize the national and international energy industry and select manufacturing markets. To facilitate the development of hydrogen markets, the US Department of Energy initiated H2@Scale to bring together stakeholders to advance affordable hydrogen production, transport, storage, and utilization to increase revenue opportunities in multiple energy sectors. One major impediment to hydrogen implementation is cost. To expedite the use of hydrogen in energy and other markets, the United States announced in 2021 the Hydrogen Shot, which seeks to reduce the cost of clean hydrogen by 80% to $1 per 1 kilogram in 1 decade ("1 1 1"). As the cost of hydrogen drops, new applications will emerge that will require unique configurations of existing equipment and infrastructure, and eventually lead to advances in the generation and utilization of hydrogen. As the hydrogen economy expands, sensors and detection methods will need to adapt to changing infrastructure demands to address the primary targets of health & safety, emissions monitoring, and process control. The NREL Sensor Laboratory is playing a pivotal role in advancing the use of hydrogen sensors and detection methodologies in each of these categories to support DOE's mission for safe and efficient utilization in emerging markets. Health & safety monitors are required to ensure that operators and facilities can react to unintended hydrogen releases, either as GH2, LH2, or as a constituent of blends (e.g., natural gas or ammonia). Current detection methodologies focus on safety applications to detect near its lower flammable limit (4 vol %), and typically include point sensors in applications such as fixed or mobile detectors (e.g., personal gas monitors). Methodologies amenable for area detection include acoustic, emerging optical imaging methods, and flame detectors. Comparable detection strategies can be utilized for emissions monitoring and quantization, however few methods can simultaneously cover both low (emissions) and high (health & safety) levels. Deployment of emission level detectors will be required to 1) reduce product loss through small but potentially significant leaks from an environmental or cost perspective, 2) reduce downtime of high demand systems by early identification of eminent system failures (leaks through pump or compressor seals indicative of impending failure), and 3) address potential emission monitoring requirements that may be set by regulating bodies. The first two points should be adopted by industry to reduce the cost-of-goods-sold. The third main category for hydrogen detection relates to process control and may be advantageous for many existing applications. Two main applications are emerging. For example, the purity requirements for hydrogen that is dispensed from refueling systems for hydrogen fuel cell electric vehicles (FCEV) is rigorously regulated by the Standard SAE J2719, which prescribes maximum allowable levels of multiple impurities in the hydrogen fuel and must be verified by a regulatory body. Hydrogen contaminant detectors (HCD) integrated to the fueling station can assure this compliance. HCDs must be able operate in 100% H2 backgrounds and be able to distinguish between multiple contaminants at low ppm to low ppb levels. Secondly, as a strategy to decarbonize the natural gas grid, there are proposals to blend hydrogen with natural gas. This blending will affect transport applications (pipeline infrastructure), stationary combustion systems (turbines), and consumer and commercial appliances. In the short-term, hydrogen levels up to 20% are proposed. Variations in the hydrogen level can have dramatic impact on the combustion process and on the potential response of safety sensors. These mixtures may be regulated so that the concentration at a delivery point must be monitored with high precision. However, routine maintenance may introduce background gases such as ambient air (with water) or maintenance gases (introduced with welding processes or adhesive outgassing.) Therefore, the detection methodology must be robust enough to recover or respond to various contaminants. Several reviews can be found in literature addressing sensing and detection technologies, including their limitations and applications. However, for most applications, limitations can be alleviated by combining various detection techniques either through system integration or implementation of machine learning methods (artificial intelligence). In this presentation, we will discuss several applications, highlight their current approach for hydrogen detection, and suggest detection strategies to supplement their limitations.

ENERGY STORAGE,HYDROGEN↗

The SURE House (Solar Decathlon 2015)

Coastal towns and cities across the Northeastern US, with their high population density, aged utility infrastructure, and unique geography, are increasingly vulnerable to climate change related storm events. In October 2012 superstorm Sandy highlighted the fragility of our current coastal building types and made clear the need for a new model of design and construction which works to understand and mitigate these weaknesses. Dramatic changes in public policy, championed by both The Federal Emergency Management Agency (FEMA) and the National Flood Insurance Program (NFIP) are driving the rebuilding of these shore communities, often resulting in costly renovations, un-sustainable neighborhood configurations and in direct conflict with concurrent government policies such as The American with Disabilities Act (ADA). The SURE HOUSE demonstrates a series of new design solutions to these conflicting public policies and environmental imperatives. At Stevens Institute of Technology, the 2015 Solar Decathlon started with the challenge: Can we design a home for coastal New Jersey that dramatically reduces its energy use while protecting itself from the realities of a changing, more extreme climate? The SURE HOUSE merges the iconic 20th century shore home with 21st century building science. Utilizing innovative renewable energy technologies, a ‘Passive House’ level building envelope, and rugged glass-fiber-composite materials to flood-proof the home, the SURE HOUSE is a high-performance, net-zero-energy home, armored against extreme weather, designed for the contemporary lifestyle of the Jersey Shore and other vulnerable coastal communities. SUSTAINABLE At Stevens, we recognize that energy use in the home and workplace is directly connected to the growing problem of climate change. Reducing our energy consumption by designing higher performing, compact homes that are both functional, comfortable and desirable is the first critical step towards a modern, sustainable architecture for New Jersey and beyond. This is what informed the architectural design of the house. Configured about a compact form, thickly insulated and air-sealed walls eliminate thermal bridging and minimize energy losses while advanced glazing brings in free solar heat during the winter months. As a result of these passive design strategies, the SURE HOUSE has a greatly reduced carbon footprint requiring 91% less energy than a typical New Jersey home. Photovoltaic (PV) arrays on both the rooftop and operable shutters easily provide energy in excess of the home’s modest demands. The Stevens team considers a truly sustainable home in the era of climate change, one that prioritizes low energy use, and integrates right-sized renewable generation to supply the home’s needs. Low consumption, low production. RESILIENT In October of 2012, Hurricane Sandy wreaked havoc along the east coast of the US. In New Jersey alone there was an estimated 29.4 billion dollars in damages, 346,000 homes affected, and almost two and a half million people left without power, in some cases for over 10 days. Recovery from this storm and associated flooding is ongoing to this day, as many New Jersey homeowners grapple with the large costs of rebuilding and struggle to adapt to complicated new home building regulations. Damage from this storm to Hoboken, the home of the Stevens Institute of Technology’s campus, and to the New Jersey shore was extensive and many students on the SURE HOUSE team were directly affected by this historic event. The Stevens design team recognizes that in a world of more frequent and stronger storms, the ability to absorb and adapt to change is more important than ever. Successfully weathering the next storm and its aftermath is one of the primary goals in the design of the SURE HOUSE prototype. The SURE HOUSE introduces unique ‘dry flood-proofing’ methods to residential construction. Innovative wall and floor flood-proofing, utilizing durable composite sheathing materials adapted from the boating industry, were developed by the student team to render the SURE House’s building envelope flood proof up to the FEMA AE 6/7 Zone (+ 6/7 feet of water above sea-level). Designed and fabricated utilizing glass-fiber composite materials, the custom storm shutter system serves to protect the large glazed openings of the home from both air-borne debris impact and water infiltration during a storm event while also providing deterrence from the vandalism that often occurs in the aftermath of a calamitous event. During extended power outages, a ‘resilient’ solar array is capable of supplying critical amounts of energy and hot water to the home, without the use of battery storage or grid infrastructure. PV Description AC GRID-TIED PV SYSTEM The SURE HOUSE solar system consists of two distinct arrays, the grid-tied rooftop array which produces AC Powerand the shutter-mounted array which converts DC power generated by the modules directly into usable heat for the domestic hot water (DHW). The roof-mounted AC grid-tied array is comprised of three strings, two 11 module strings connected to an SMA SB 5000TL-US-223 inverter and a single 10 module string equipped and an SMA SB 3000TL-US-22 central inverter. All modules for the rooftop AC array are LG MonoX 2804 watt solar panels, chosen for their durability and efficiency. Inverter sizing and string size are optimized to the electrical characteristics of the chosen PV modules and the desired energy production, see Appendix 5B. The SURE HOUSE is projected to produce 12,353 kWh per year to meet 6,157 kWh per year of estimated consumption. Solar modules for the two AC grid-tied sub arrays are mounted on the main roof surface of the SURE HOUSE with a 10 degree tilt angle using a partially-ballasted polyethylene roof mounting system made by Renusol, which is particularly suited to the corrosive salt air of a coastal environment. This method was chosen for its installation simplicity and to limit detrimental roof penetrations. The 10 degree tilt optimizes the energy generated per roof area at a lower price, see Appendix 5A, and ensures that there is minimal wind uplift. DC SOLAR ELECTRIC DOMESTIC HOT WATER The SURE HOUSE’s approach to sustainable and resilient domestic hot water (DHW) consists of a unique, custom engineered DC solar electric hot water system. Employing the use of Advanced Energy’s DC electric PV heater (DCPVH)5 to heat domestic water well beyond the draw temperature, the SURE HOUSE is able to obtain a remarkably high solar fraction of 75% or more. The custom modified 80 gallon Vaughn DHW tank acts as a large solar ‘battery’ that stores heat energy in the form of hot water at almost 150 degrees, harvesting energy when the sun is out for use later in the day or overnight. The system, which uses electricity rather than heated fluid, is distinct from traditional ‘solar thermal’ systems by eliminating the dangers and associated maintenance issues of fluid based systems. By forgoing the use of external fluid loops, overheated solar collectors and pipes are not an issue for the SURE HOUSE system which never needs any yearly or seasonal flushing. In weather conditions where the integrated DCPVH unit cannot produce enough hot water via solar energy alone, the Vaughn hot water tank comes factory made with an AC powered 2.6 COP heat pump and a AC heating coil that serve as back-up heating elements. The DCPVH is powered by a DC array consisting of 10 custom-made Solbian by PVilion 180 watt, monocrystalline, flexible solar modules6. The Solbian modules are directly mounted to the top half of the operable storm shutters on the south façade of the SURE HOUSE. The SURE HOUSE team worked closely with PVillion, a PV system designer specializing in flexible architectural photovoltaics, to design an adhesive system that works with the glass-fiber storm shutter surface in a reliable manner over an extended period. Because this system operates in DC-only mode and never connects to the municipal power grid, the system can continue to create hot water safely and effectively even during grid disruption events. This hot water, and the energy stored within it, could be used for a range of activities including washing and cooking to more elaborate hydronic heating systems as desired. See Appendix 6 for system engineering, sizing and modeling.

14 SOLAR ENERGY↗

Evaluating the Feasibility of Transactive Approach for Voltage Management Using Inverters of a PV Plant

This article evaluates the feasibility of a double-auction-based Transactive Energy System (TES) for engaging the reactive power (Var) capability of inverters in a utility-scale pho¬tovoltaic (PV) plant for voltage management in distribution feed¬ers. In this approach, a PV plant owner (seller) provides Var supply curves in terms of price-quantity pairs consider¬ing the inverters’ P-Q capability, efficiency, opportunity cost for real power curtail¬ment, losses, etc. At the same time, the utility (buyer) proposes Var demand curves in price-quantity pairs exhibiting the marginal price of Var. Then, market clearing points are obtained from the intersection points of supply and demand curves which are then used to create Var dispatch sig-nals from the PV plant. To conduct a fea¬sibility study of this approach, an Australian distribution network with a utility-scale PV plant (capacity 3.275 MWP) is modeled in RSCAD and simu¬lated in real-time on MATLAB, RSCAD-RTDS co-simulation platform using real historical data. Consid¬ering a use case, namely, conservation voltage reduction (CVR), the operational feasibility along with cost-benefit analysis is demonstrated in software-in-the-loop (SIL) and hardware-in-the-loop (HIL) platform. Another use case, exploring the benefit from tap-change reduction of step-voltage-regulator, is investigated. These studies explore further use cases, e.g., PV hosting capacity, network reconfiguration, etc., to be incorporated in TES for experiencing substantial economic benefits.

Alam, Mollah Rezaul↗

Responses of vascular plant fine roots and associated microbial communities to whole-ecosystem warming and elevated CO 2 in northern peatlands

Warming and elevated CO 2 (eCO 2 ) are expected to facilitate vascular plant encroachment in peatlands. The rhizosphere, where microbial activity is fueled by root turnover and exudates, plays a crucial role in biogeochemical cycling, and will likely at least partially dictate the response of the belowground carbon cycle to climate changes. We leveraged the Spruce and Peatland Responses Under Changing Environments (SPRUCE) experiment, to explore the effects of a whole-ecosystem warming gradient (+0°C to 9°C) and eCO 2 on vascular plant fine roots and their associated microbes. We combined trait-based approaches with the profiling of fungal and prokaryote communities in plant roots and rhizospheres, through amplicon sequencing. Warming promoted self-reliance for resource uptake in trees and shrubs, while saprophytic fungi and putative chemoorganoheterotrophic bacteria utilizing plant-derived carbon substrates were favored in the root zone. Conversely, eCO 2 promoted associations between trees and ectomycorrhizal fungi. Trees mostly associated with short-distance exploration-type fungi that preferentially use labile soil N. Additionally, eCO 2 decreased the relative abundance of saprotrophs in tree roots. Our results indicate that plant fine-root trait variation is a crucial mechanism through which vascular plants in peatlands respond to climate change via their influence on microbial communities that regulate biogeochemical cycles.

59 BASIC BIOLOGICAL SCIENCES↗

SU-E-J-12: An Image-Guided Soft Robotic Patient Positioning System for Maskless Head-And-Neck Cancer Radiotherapy: A Proof-Of-Concept Study

Purpose: We propose a surface-image-guided soft robotic patient positioning system for maskless head-and-neck radiotherapy. The ultimate goal of this project is to utilize a soft robot to realize non-rigid patient positioning and real-time motion compensation. In this proof-of-concept study, we design a position-based visual servoing control system for an air-bladder-based soft robot and investigate its performance in controlling the flexion/extension cranial motion on a mannequin head phantom. Methods: The current system consists of Microsoft Kinect depth camera, an inflatable air bladder (IAB), pressured air source, pneumatic valve actuators, custom-built current regulators, and a National Instruments myRIO microcontroller. The performance of the designed system was evaluated on a mannequin head, with a ball joint fixed below its neck to simulate torso-induced head motion along flexion/extension direction. The IAB is placed beneath the mannequin head. The Kinect camera captures images of the mannequin head, extracts the face, and measures the position of the head relative to the camera. This distance is sent to the myRIO, which runs control algorithms and sends actuation commands to the valves, inflating and deflating the IAB to induce head motion. Results: For a step input, i.e. regulation of the head to a constant displacement, the maximum error was a 6% overshoot, which the system then reduces to 0% steady-state error. In this initial investigation, the settling time to reach the regulated position was approximately 8 seconds, with 2 seconds of delay between the command start of motion due to capacitance of the pneumatics, for a total of 10 seconds to regulate the error. Conclusion: The surface image-guided soft robotic patient positioning system can achieve accurate mannequin head flexion/extension motion. Given this promising initial Result, the extension of the current one-dimensional soft robot control to multiple IABs for non-rigid positioning control will be pursued.

60 APPLIED LIFE SCIENCES↗

Separate, separated, and together: the transcriptional program of the Clostridium acetobutylicum-Clostridium ljungdahlii syntrophy leading to interspecies cell fusion

ABSTRACT Syntrophic cocultures (hitherto assumed to be commensalistic) of Clostridium acetobutylicum and Clostridium ljungdahlii , whereby CO 2 and H 2 produced by the former feed the latter, result in interspecies cell fusion involving large-scale exchange of protein, RNA, and DNA between the two organisms. Although mammalian cell fusion is mechanistically dissected, the mechanism for such microbial-cell fusions is unknown. To start exploring this mechanism, we used RNA sequencing to identify genes differentially expressed in this coculture using two types of comparisons. One type compared coculture to the two monocultures, capturing the combined impact of interactions through soluble signals in the medium and through direct cell-to-cell interactions. The second type compared membrane-separated versus -unseparated cocultures, isolating the impact of interspecies physical contact. While we could not firmly identify specific genes that might drive cell fusion, consistent with our hypothesized model for this interspecies microbial cell fusion, we observed differential regulation of genes involved in C. ljungdahlii’s autotrophic Wood-Ljungdahl pathway metabolism and genes of the motility machinery. Unexpectedly, we also identified differential regulation of biosynthetic genes of several amino acids, and notably of arginine and histidine. We verified that they are produced by C. acetobutylicum and are metabolized by C. ljungdahlii to its growth advantage. These and other findings, and notably upregulation of C. acetobutylicum ribosomal-protein genes, paint a more complex syntrophic picture and suggest a mutualistic relationship, whereby beyond CO 2 and H 2 , C. acetobutylicum feeds C. ljungdahlii with growth-boosting amino acids, while benefiting from the H 2 utilization by C. ljungdahlii . IMPORTANCE The construction and study of synthetic microbial cocultures is a growing research area due to the untapped potential of defined multi-species industrial bioprocesses and the utility of defined cocultures for generating insight into complex, undefined, natural microbial consortia. Our previous work showed that coculturing C. acetobutylicum and C. ljungdahlii leads to a unique metabolic phenotype (production of isopropanol) and heterologous cell fusion events. Here, we used RNAseq to explore genes involved in and impacted by these fusions. First, we compared gene expression in coculture to each monoculture. Second, we utilized a transwell system to compare gene expression in mixed cocultures to cocultures with both species physically separated by a permeable membrane, isolating the impact of interspecies “touching” on the transcriptome. This study deepens our mechanistic understanding of the C. acetobutylicum-C. ljungdahlii coculture phenotype, laying the groundwork for reverse genetic studies of heterologous cell fusion in Clostridium cocultures.

Willis, Noah B. (ORCID:0009000689365955)↗

Mechanical coupling in the nitrogenase complex

The enzyme nitrogenase reduces dinitrogen to ammonia utilizing electrons, protons, and energy obtained from the hydrolysis of ATP. Mo-dependent nitrogenase is a symmetric dimer, with each half comprising an ATP-dependent reductase, termed the Fe Protein, and a catalytic protein, known as the MoFe protein, which hosts the electron transfer P-cluster and the active-site metal cofactor (FeMo-co). A series of synchronized events for the electron transfer have been characterized experimentally, in which electron delivery is coupled to nucleotide hydrolysis and regulated by an intricate allosteric network. We report a graph theory analysis of the mechanical coupling in the nitrogenase complex as a key step to understanding the dynamics of allosteric regulation of nitrogen reduction. This analysis shows that regions near the active sites undergo large-scale, large-amplitude correlated motions that enable communications within each half and between the two halves of the complex. Computational predictions of mechanically regions were validated against an analysis of the solution phase dynamics of the nitrogenase complex via hydrogen-deuterium exchange. These regions include the P-loops and the switch regions in the Fe proteins, the loop containing the residue β-188Ser adjacent to the P-cluster in the MoFe protein, and the residues near the protein-protein interface. In particular, it is found that: (i) within each Fe protein, the switch regions I and II are coupled to the [4Fe-4S] cluster; (ii) within each half of the complex, the switch regions I and II are coupled to the loop containing β-188Ser; (iii) between the two halves of the complex, the regions near the nucleotide binding pockets of the two Fe proteins (in particular the P-loops, located over 130 Å apart) are also mechanically coupled. Notably, we found that residues next to the P-cluster (in particular the loop containing β-188Ser) are important for communication between the two halves.

59 BASIC BIOLOGICAL SCIENCES↗

On the state estimation of structures with second order observers

The use of Linear Quadratic Regulator (LQR) control synthesis techniques implies the availability of full state feedback. For vibration control of structures, usually only a limited number of states are measured from which an observer model reconstructs the full state. It is shown that using second order observers is a viable technique for reconstructing the unmeasured states of structures under mildly restrictive conditions. Moreover, the computational advantages of the second order observer as compared to a first order observer indicate that significantly larger observer models may be utilized. Numerical examples are used to demonstrate the performance of second order observers. The implications of second order observers in the development of Controls-Structures Interaction (CSI) technology is discussed.

Belvin, W. Keith↗

To Decommission or Not To Decommission? That is the Question... Revisited

Major Highlights Comparing 1998 to 2014: Favoring Plant Shutdown: Electric utility deregulation has created greater competition; Markets have been disrupted by cheap natural gas; Despite high availability, cost of electric power from natural gas still below break-even cost of nuclear generation; Emerging ISO acceptance of fluctuating renewable energy on the grid. Favoring DECON: D and D technologies have matured; Reactor Vessel and Internals segmentation methods still being refined; Nuclear workforce is 16 years older and leaving the nuclear industry (Impact on D and D TBD); Avoiding risk of more restrictive regulation and higher LLW disposal costs. Favoring SAFSTOR: Full funding of NDT still major driver for deferring immediate dismantlement. SAFSTOR vs. near term D and D influenced by: Status of and confidence in decommissioning fund (NDT) and cost estimate; Local and state politics; Public pressure. Most early shutdown plants tend to have under-funded decommissioning funds and opt for SAFSTOR. SAFSTOR allows for fund growth through interest compounding, but must assume that cost increase is less than fund growth: May not be true across the board, Much uncertainty/risk in waste disposal costs, regulatory requirements, etc. when projected out several decades. Once initiated SAFSTOR decision is not easily changed to DECON: Plant infrastructure equipment and components not readily available, RCS chemical decontamination can be very challenging, Key/knowledgeable plant personnel are gone. Consideration for phased approach to DECON: Remove >95% of the radiological source term under today's known regulations and cost by full System Decontamination of the primary circuit systems, and removal of NSSS and decontamination of the Containment Building; Improves risk profile in those areas most vulnerable to excessive cost growth; Able to defer D and D of the remainder of the plant to allow for the NDT to grow with interest.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Lightweight Materials for Automotive Application: An Assessment of Material Production Data for Magnesium and Carbon Fiber

The use of lightweight materials in vehicle components, also known as “lightweighting,” can result in automobile weight reduction, which improves vehicle fuel economy and generally its environmental footprint. Materials often used for vehicle lightweighting include aluminum, magnesium, and polymers reinforced with either glass or carbon fiber. However, because alternative materials typically used for vehicle lightweighting require more energy to make on a per part basis than the material being replaced (often steel or iron), the fuel efficiency improvement induced by a weight reduction is partially offset by an increased energy for the vehicle material production. To adequately quantify this tradeoff, reliable and current values for life-cycle production energy are needed for both conventional and alternative materials. Our focus here is on the production of two such alternative materials: magnesium and carbon fibers. Both these materials are low density solids with good structural properties. These properties have enabled their use in applications where weight is an issue, not only for automobiles but also for aerospace applications. This report addresses the predominant production methods for these materials and includes a tabulation of available material and energy input data necessary to make them. The life cycle inventory (LCI) information presented herein represents a process chain analysis (PCA) approach to life cycle assessment (LCA) and is intended for evaluation as updated materials production data for magnesium and carbon fiber for inclusion into the Greenhouse gases, Regulated Emissions, and Energy use in Transportation model (GREET2_2012). The summary life-cycle metrics used to characterize the cradle-to-gate environmental performance of these materials are the cumulative energy demand (CED) and greenhouse gas emissions (GHG) per kilogram of material.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

The 2-micron plasmid as a nonselectable, stable, high copy number yeast vector

The endogenous 2-microns plasmid of Saccharomyces cerevisiae has been used extensively for the construction of yeast cloning and expression plasmids because it is a native yeast plasmid that is able to be maintained stably in cells at high copy number. Almost invariably, these plasmid constructs, containing some or all 2-microns sequences, exhibit copy number levels lower than 2-microns and are maintained stably only under selective conditions. We were interested in determining if there was a means by which 2-microns could be utilized for vector construction, without forfeiting either copy number or nonselective stability. We identified sites in the 2-microns plasmid that could be used for the insertion of genetic sequences without disrupting 2-microns coding elements and then assessed subsequent plasmid constructs for stability and copy number in vivo. We demonstrate the utility of a previously described 2-microns recombination chimera, pBH-2L, for the manipulation and transformation of 2-microns as a pure yeast plasmid vector. We show that the HpaI site near the STB element in the 2-microns plasmid can be utilized to clone yeast DNA of at least 3.9 kb with no loss of plasmid stability. Additionally, the copy number of these constructs is as high as levels reported for the endogenous 2-microns.

Non-NASA Center↗

Pore-Filling Induced Solid Electrolyte Failure of Ti-Doped Na 3 Zr 2 Si 2 PO 12 Characterized by Operando Synchrotron X-Ray Tomography

Solid-state batteries (SSBs), particularly those utilizing sodium metal, are emerging as a promising technology due to their potential for enhanced safety, higher energy density, and longer cycle life. NASICON (Na superionic conductor) materials, known for their robust crystalline structure and high ionic conductivity, are pivotal in the development of efficient sodium all-solid-state batteries. These materials exhibit high room-temperature ionic conductivity and electrochemical stability, making them ideal for various applications. Research has focused on improving NASICON's ionic conductivity and stability through doping, interface regulation, and composite anode design. Recent advancements include Ti-doped Na 3 Zr 2 Si 2 PO 12 (Ti-NZSP), which demonstrates improved surface stability, higher ionic conductivity, and increased critical current density. However, challenges such as Na dendrite formation and mechanical integrity under operational conditions persist. Advanced imaging techniques like operando synchrotron X-ray tomography have provided insights into failure mechanisms, revealing that pore-filling and dendrite growth are significant issues. Understanding these processes is essential for enhancing the performance and safety of Na-based SSBs. Here, this study underscores the need for continued research to address these challenges and develop reliable, high-performance solid-state electrolytes for future energy storage solutions.

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