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At least 1,603 records · Page 89

Comprehensive Cradle to Grave Life Cycle Analysis of On-Road Vehicles in the United States based on GREET

To properly compare and contrast the environmental performance of one vehicle technology against another, it is necessary to consider their production, operation, and end-of-life fates. Since 1995, Argonne’s GREET® life cycle analysis model (Greenhouse gases, Regulated Emissions, and Energy use in Technologies) has been annually updated to model and refine the latest developments in fuels and materials production, as well as vehicle operational and composition characteristics. Updated cradle-to-grave life cycle analysis results from the model’s latest release are described for a wide variety of fuel and powertrain options for U.S. light-duty and medium/heavy-duty vehicles. Light-duty vehicles include a passenger car, sports utility vehicle (SUV), and pick-up truck, while medium/heavy-duty vehicles include a Class 6 pickup-and-delivery truck, Class 8 day-cab (regional) truck, and Class 8 sleeper-cab (long-haul) truck. Powertrain coverage includes internal combustion (spark ignition and compression ignition) engines, hybrid electric, plug-in hybrid, full battery electric, and fuel cell vehicles powered by conventional and low carbon energy sources. The results offer insights into the current state of these technologies, as well as a projection of the likely environmental implications of future fuel and vehicle advancements through a time-series evaluation of life cycle greenhouse gas emissions.

Kelly, Jarod C.↗

Light-inducible protein degradation in E. coli with the LOVdeg tag

Molecular tools for optogenetic control allow for spatial and temporal regulation of cell behavior. In particular, light-controlled protein degradation is a valuable mechanism of regulation because it can be highly modular, used in tandem with other control mechanisms, and maintain functionality throughout growth phases. Here, we engineered LOVdeg, a tag that can be appended to a protein of interest for inducible degradation in Escherichia coli using blue light. We demonstrate the modularity of LOVdeg by using it to tag a range of proteins, including the LacI repressor, CRISPRa activator, and the AcrB efflux pump. Additionally, we demonstrate the utility of pairing the LOVdeg tag with existing optogenetic tools to enhance performance by developing a combined EL222 and LOVdeg system. Finally, we use the LOVdeg tag in a metabolic engineering application to demonstrate post-translational control of metabolism. Together, our results highlight the modularity and functionality of the LOVdeg tag system and introduce a powerful new tool for bacterial optogenetics.

59 BASIC BIOLOGICAL SCIENCES↗

Light-inducible protein degradation in E. coli with the LOVdeg tag

Molecular tools for optogenetic control allow for spatial and temporal regulation of cell behavior. In particular, light-controlled protein degradation is a valuable mechanism of regulation because it can be highly modular, used in tandem with other control mechanisms, and maintain functionality throughout growth phases. Here, we engineered LOVdeg, a tag that can be appended to a protein of interest for inducible degradation in Escherichia coli using blue light. We demonstrate the modularity of LOVdeg by using it to tag a range of proteins, including the LacI repressor, CRISPRa activator, and the AcrB efflux pump. Additionally, we demonstrate the utility of pairing the LOVdeg tag with existing optogenetic tools to enhance performance by developing a combined EL222 and LOVdeg system. Finally, we use the LOVdeg tag in a metabolic engineering application to demonstrate post-translational control of metabolism. Together, our results highlight the modularity and functionality of the LOVdeg tag system and introduce a powerful new tool for bacterial optogenetics.

Tague, Nathan (ORCID:0000000281146700)↗

Coupling device with improved thermal interface

The primary object of the present invention is to provide a simple, reliable, and lightweight coupling that will also have an efficient thermal interface. A further object of the invention is to provide a coupling that is capable of blind mating with little or no insertion forces. Another object of the invention is to provide a coupling that acts as a thermal regulator to maintain a constant temperature on one side of the coupling. Another object of the invention is to increase the available surface area of a coupling thus providing a larger area for the conduction of heat across the thermal interface. Another object of the invention is to provide a fluidic coupling that has no fluid passing across the interface, thus reducing the likelihood of leaks and contamination. The foregoing objects are achieved by utilizing, as in the prior art, a hot area (at an elevated temperature as compared to a cold area) with a need to remove excess heat from the hot area to a cold area. In this device, the thermal interface will occur not on a planar horizontal surface, but along a non-planar vertical surface, which will reduce the reaction forces and increase the thermal conductivity of the device. One non-planar surface is a surface on a cold pin extending from the cold area and the other non-planar surface is a surface on a hot pin extending from the hot area. The cold pin is fixed and does not move while the hot pin is a flexible member and its movement towards the cold pin will bring the two non-planar surfaces together forming the thermal interface. The actuating member for the device is a shape-memory actuation wire which is attached through an aperture to the hot pin and through another aperture to an actuation wire retainer. By properly programming the actuation wire, heat from the hot area will cause the actuation wire to bend the hot wire. Heat from the hot area will cause the actuation wire to bend the hot pin towards the cold pin forming the coupling and the desired thermal interface. The shape-memory actuation wire is made of a shape-memory-effect alloy such as Nitinol.

Milam, Malcolm Bruce↗

Valsalva maneuver: Insights into baroreflex modulation of human sympathetic activity

Valsalva's maneuver, voluntary forced expiration against a closed glottis, is a well-characterized research tool, used to assess the integrity of human autonomic cardiovascular control. Valsalva straining provokes a stereotyped succession of alternating positive and negative arterial pressure and heart rate changes mediated in part by arterial baroreceptors. Arterial pressure changes result primarily from fluctuating levels of venous return to the heart and changes of sympathetic nerve activity. Muscle sympathetic activity was measured directly in nine volunteers to explore quantitatively the relation between arterial pressure and human sympathetic outflow during pressure transients provoked by controlled graded Valsalva maneuvers. Our results underscore several properties of sympathetic regulation during Valsalva straining. First, muscle sympathetic nerve activity changes as a mirror image of changes in arterial pressure. Second, the magnitude of sympathetic augmentation during Valsalva straining predicts phase 4 arterial pressure elevations. Third, post-Valsalva sympathetic inhibition persists beyond the return of arterial and right atrial pressures to baseline levels which reflects an alteration of the normal relation between arterial pressure and muscle sympathetic activity. Therefore, Valsalva straining may have some utility for investigating changes of reflex control of sympathetic activity after space flight; however, measurement of beat-to-beat arterial pressure is essential for this use. The utility of this technique in microgravity can not be determined from these data. Further investigations are necessary to determine whether these relations are affected by the expansion of intrathoracic blood volume associated with microgravity.

Smith, Michael L.↗

Mission Risk Reduction Regulatory Change Management

NASA Headquarters Environmental Management Division supports NASA's mission to pioneer the future in space exploration, scientific discovery, and aeronautics research by integrating environmental considerations into programs and projects early-on, thereby proactively reducing NASA's exposure to institutional, programmatic and operational risk. As part of this effort, NASA established the Principal Center for Regulatory Risk Analysis and Communication (RRAC PC) as a resource for detecting, analyzing, and communicating environmental regulatory risks to the NASA stakeholder community. The RRAC PC focuses on detecting emerging environmental regulations and other operational change drivers that may pose risks to NASA programs and facilities, and effectively communicating the potential risks. For example, regulatory change may restrict how and where certain activities or operations may be conducted. Regulatory change can also directly affect the ability to use certain materials by mandating a production phase-out or restricting usage applications of certain materials. Regulatory change can result in significant adverse impacts to NASA programs and facilities due to NASA's stringent performance requirements for materials and components related to human-rated space vehicles. Even if a regulation does not directly affect NASA operations, U.S. and international regulations can pose program risks indirectly through requirements levied on manufacturers and vendors of components and materials. For example, manufacturers can change their formulations to comply with new regulatory requirements. Such changes can require time-consuming and costly requalification certification for use in human spaceflight programs. The RRAC PC has implemented a system for proactively managing regulatory change to minimize potential adverse impacts to NASA programs and facilities. This presentation highlights the process utilized by the RRACPC to communicate regulatory change and the associated potential risks within NASA, as well as the process for communicating and cooperating with other government agencies and industry partners, both domestic and international, to ensure mission success.

Scroggins, Sharon↗

The Histone Modification Domain of Paf1 Complex Subunit Rtf1 Directly Stimulates H2B Ubiquitylation through an Interaction with Rad6

The five-subunit yeast Paf1 Complex (Paf1C) regulates all stages of transcription and is critical for the monoubiquitylation of histone H2B (H2Bub), a modification that broadly influences chromatin structure and eukaryotic transcription. Here we show that the histone modification domain (HMD) of Paf1C subunit Rtf1 directly interacts with the ubiquitin conjugase Rad6 and stimulates H2Bub independently of transcription. We present the crystal structure of the Rtf1 HMD and use site-specific, in vivo crosslinking to identify a conserved Rad6 interaction surface. Utilizing ChIP-exo analysis, we define the localization patterns of the H2Bub machinery at high resolution and demonstrate the importance of Paf1C in targeting the Rtf1 HMD, and thereby H2Bub, to its appropriate genomic locations. Finally, we observe HMD-dependent stimulation of H2Bub in a transcription-free, reconstituted in vitro system. Taken together, our results argue for an active role for Paf1C in promoting H2Bub and ensuring its proper localization in vivo.

59 BASIC BIOLOGICAL SCIENCES↗

Single cell RNA sequencing reveals shifts in cell maturity and function of endogenous and infiltrating cell types in response to acute intervertebral disc injury

Intervertebral disc (IVD) degeneration contributes to disabling back pain. Degeneration can be initiated by injury and progressively leads to an irreversible loss of cells and function. IVD function restoration through cell replacement therapies have had limited success due to knowledge gaps in the critical cell populations important for repair. Here, in this study, we used single cell RNA sequencing to identify the transcriptional changes of IVD resident and infiltrating cell populations from Control and Injured coccygeal IVDs extracted from 12-week-old female C57BL/6J mice 7 days post injury. Clustering, gene ontology, and pseudotime trajectory analyses determined transcriptomic divergences with injury, flow cytometry identified they types of infiltrating immune cells, and immunofluorescence was utilized to define mesenchymal stem cell (MSC) localization. We identified 11 distinct clusters that included IVD, immune, vascular cells, and MSCs. Differential gene expression analysis determined that Outer Annulus Fibrosus, Neutrophils, Saa2-High MSCs, Macrophages, and Krt18 + Nucleus Pulposus (NP) cells were the major drivers of transcriptomic differences between Control and Injured cells. Gene ontology revealed that the most upregulated biological pathways were angiogenesis and T cell-related while wound healing and ECM regulation were downregulated. Pseudotime trajectory analyses revealed that IVD injury directed cells towards increased differentiation in all clusters, except for Krt18 + NP cells which remained in a less mature cell state. Saa2-High and Grem1-High MSCs populations shifted towards more differentiated IVD cells profiles with injury and localized distinctly within the IVD. This study revealed novel MSC populations with the potential to be leveraged for future IVD repair studies.

Cartilage↗

Revealing Fast Structural Dynamics in pH-Responsive Peptides with Time-Resolved X-ray Scattering

Many biomaterials can adapt to changes in local biological environment (such as pH, temperature or ionic composition) in order to regulate function or deliver a payload. Such adaptation to environmental perturbation is typically a hierarchical process that begins with a response on a local structural level and then propagates to supramolecular and macromolecular scales. Understanding fast structural dynamics that occur upon the perturbation is important for rational design of functional biomaterials. However, few nanosecond time-resolved methods can probe both intra- and inter-molecular scales simultaneously with a high structural resolution. Here we utilize time-resolved x-ray scattering to probe nanosecond to microsecond structural dynamics of polypeptides undergoing protonation via a pH jump initiated by photoexcitation of a photoacid. Our results provide insight into the protonation-induced hierarchical changes in packing of peptide chains, formation of helical structure and associated collapse of the peptide chain.

Rimmerman, Dolev↗

Resin-assisted enrichment of thiols as a general strategy for proteomic profiling of cysteine-based reversible modifications

Reversible modifications on cysteine thiols play a significant role in redox signaling and regulation. A number of reversible redox modifications, including disulfide formation, S-nitrosylation, and S-glutathionylation, have been recognized for their significance in various physiological and pathological processes. Here we describe in detail a resin-assisted thiol-affinity enrichment protocol for both biochemical and proteomics applications. This protocol serves as a general approach for specific isolation of thiol-containing proteins or peptides derived from reversible redox-modified proteins. This approach utilizes thiol-affinity resins to directly capture thiol-containing proteins or peptides through a disulfide exchange reaction followed by on-resin protein digestion and on-resin multiplexed isobaric labeling to facilitate LC-MS/MS based quantitative site-specific analysis of redox modifications. The overall approach requires a much simpler workflow with increased specificity compared to the commonly used biotin switch technique. By coupling different selective reduction strategies, the resin-assisted approach provides the researcher with a useful tool capable of enriching different types of reversible modifications on protein thiols. Procedures for selective enrichment and analyses of S-nitrosylation and total reversible cysteine oxidation are presented to demonstrate the utility of this general strategy.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A Data-Driven Voltage Control Strategy for Distribution Grids With Distributed Energy Resources

Traditionally, distribution system control approaches have been model-based. The deployment of advanced metering infrastructure has provided electric utilities with the capability of data-driven control with real-time measurements. The shift from model-based to data-driven control represents a significant advancement in the management of distribution systems, offering a more adaptive approach to system control because of the ability to dynamically adapt to changing conditions without the need for system modeling. Here, in this paper, a behavioral data-driven control method is developed to provide voltage regulation to an actual distribution system by controlling the legacy devices and distributed energy resource (DER) assets. The studied distribution system has a load tap changer and three capacitor banks as the legacy devices and photovoltaic systems as the DERs. The performance of the proposed control algorithm is validated using a laboratory test bed setup considering multiple scenarios. The results show that the proposed control achieved 99% voltage regulation.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Tariff Setting in the District Heating Sector: Recommendations Based on International Experience

Ukraine’s district heating (DH) system is an essential resource for the country and has the potential to provide efficient, local, and renewable energy to a large share of the population. However, inefficient regulation and management have caused the sector to experience a vicious cycle of financial, operational, and technological problems. Overcoming these challenges and building a sustainable DH system requires comprehensive reforms, which could attract significant investment, improve the quality of services, and increase the efficiency of DH. One of the key elements of a comprehensive reform program should be a comprehensive reform of the tariff system. The purpose of this report is to provide analysis of European models for tariff regulation and recommendations for reforms to Ukraine’s DH tariff system. The report presents several case studies from European countries, followed by analysis of the current tariff system in Ukraine and barriers to effective regulation. Based on these findings, the report proposes actions Ukraine could take to improve the tariff regulatory system and promote a sustainable DH sector.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Developing a Prototype Methodology to Rank CO2-EOR Wells and Assess Their Reuse Potential for Geologic Carbon Sequestration

This study presents a prototype methodology for evaluating the reuse potential of Class II CO₂-enhanced oil recovery (CO₂-EOR) wells as Class VI wells for geologic carbon sequestration. The approach focuses on assessing wellbore construction materials—casing, cement, tubing, and packers—based on U.S. Environmental Protection Agency (EPA) Class VI well conversion guidelines. Utilizing Python scripts and JSON representations, the methodology automates checks on digitized Texas Railroad Commission (TRRC) data to rank wells based on regulatory and integrity criteria. Key factors include casing integrity, cementing techniques, tubing compatibility, and packer selection. Due to limitations in digitized data, manual verification is required for certain sections. Future enhancements include incorporating non-digitized data via web scraping and machine learning. This research provides a practical framework for well owners and regulators, supporting informed decision-making for sustainable CO₂ storage.

carbon sequestration↗

Quasi-Linear Circuit

This work involved developing spacequalifiable switch mode DC/DC power supplies that improve performance with fewer components, and result in elimination of digital components and reduction in magnetics. This design is for missions where systems may be operating under extreme conditions, especially at elevated temperature levels from 200 to 300 degC. Prior art for radiation-tolerant DC/DC converters has been accomplished utilizing classical magnetic-based switch mode converter topologies; however, this requires specific shielding and component de-rating to meet the high-reliability specifications. It requires complex measurement and feedback components, and will not enable automatic re-optimization for larger changes in voltage supply or electrical loading condition. The innovation is a switch mode DC/DC power supply that eliminates the need for processors and most magnetics. It can provide a well-regulated voltage supply with a gain of 1:100 step-up to 8:1 step down, tolerating an up to 30% fluctuation of the voltage supply parameters. The circuit incorporates a ceramic core transformer in a manner that enables it to provide a well-regulated voltage output without use of any processor components or magnetic transformers. The circuit adjusts its internal parameters to re-optimize its performance for changes in supply voltage, environmental conditions, or electrical loading at the output

Bradley, William↗

Role of Storage and Demand Response, Greening the Grid

Greening the Grid provides technical assistance to energy system planners, regulators, and grid operators to overcome challenges associated with integrating variable renewable energy into the grid. This document, part of a Greening the Grid toolkit, examines storage and demand response as means to match renewable energy supply with demand.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Dynamic proteomic profiling of a unicellular cyanobacterium Cyanothece ATCC51142 across light-dark diurnal cycles

Unicellular cyanobacteria of the genus Cyanothece are recognized for their ability to execute nitrogen (N2)-fixation in the dark and photosynthesis in the light. Systems-wide dynamic proteomic profiling with mass spectrometry (MS) analysis reveals fundamental insights into the control and regulation of these functions. To expand upon the current knowledge of protein expression patterns in Cyanothece ATCC51142, we performed quantitative proteomic analysis using partial ("unsaturated") metabolic labeling and high mass accuracy LC-MS analysis. This dynamic proteomic profiling identified 721 actively synthesized proteins with significant temporal changes in expression throughout the light-dark cycles, of which 425 proteins matched with previously characterized cycling transcripts. The remaining 296 proteins contained a cluster of proteins uniquely involved in DNA replication and repair, protein degradation, tRNA synthesis and modification, transport and binding, and regulatory functions. Analysis of protein functions revealed that the expression of nitrogenase in the dark is mediated by higher respiration and glycogen metabolism. We have also shown that Cyanothece ATCC51142 utilizes alternative pathways for carbon (C) and nitrogen (N) acquisition, particularly, aspartic acid and glutamate as substrates of C and N, respectively. Utilization of phosphoketolase (PHK) pathway for the conversion of xylulose-5P to pyruvate and acetyl-P likely constitutes an alternative strategy to compensate higher ATP and NADPH demand. In conclusion, this study provides a deeper insight into how Cyanothece ATCC51142 modulates cellular functions to accommodate photosynthesis and N2-fixation within the single cell.

59 BASIC BIOLOGICAL SCIENCES↗

Case study on the utility of hepatic global gene expression profiling in the risk assessment of the carcinogen furan

Furan is a chemical hepatocarcinogen in mice and rats. Its previously postulated cancer mode of action (MOA) is chronic cytotoxicity followed by sustained regenerative proliferation; however, its molecular basis is unknown. To this end, we conducted toxicogenomic analysis of B3C6F1 mouse livers following three week exposures to non-carcinogenic (0, 1, 2 mg/kg bw) or carcinogenic (4 and 8 mg/kg bw) doses of furan. We saw enrichment for pathways responsible for cytotoxicity: stress-activated protein kinase (SAPK) and death receptor (DR5 and TNF-alpha) signaling, and proliferation: extracellular signal-regulated kinases (ERKs) and TNF-alpha. We also noted the involvement of NF-kappaB and c-Jun in response to furan, which are genes that are known to be required for liver regeneration. Furan metabolism by CYP2E1 produces cis-2-butene-1,4-dial (BDA), which is required for ensuing cytotoxicity and oxidative stress. NRF2 is a master regulator of gene expression during oxidative stress and we suggest that chronic NFR2 activity and chronic inflammation may represent critical transition events between the adaptive (regeneration) and adverse (cancer) outcomes. Another objective of this study was to demonstrate the applicability of toxicogenomics data in quantitative risk assessment. We modeled benchmark doses for our transcriptional data and previously published cancer data, and observed consistency between the two. Margin of exposure values for both transcriptional and cancer endpoints were also similar. In conclusion, using furan as a case study we have demonstrated the value of toxicogenomics data in elucidating dose-dependent MOA transitions and in quantitative risk assessment. - Highlights: • Global gene expression changes in furan-exposed mouse livers were analyzed. • A molecular mode of action for furan-induced hepatocarcinogenesis is proposed. • Key pathways include NRF2, SAPK, ERK and death receptor signaling. • Important roles for TNF-alpha, c-Jun, and NF-κB in tumorigenesis are proposed. • BMD and MoE values from transcriptional and apical data are compared.

60 APPLIED LIFE SCIENCES↗