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At least 757 records · Page 42

Evolutionary flexibility and rigidity in the bacterial methylerythritol phosphate (MEP) pathway

Terpenoids are a diverse class of compounds with wide-ranging uses including as industrial solvents, pharmaceuticals, and fragrances. Efforts to produce terpenoids sustainably by engineering microbes for fermentation are ongoing, but industrial production still largely relies on nonrenewable sources. The methylerythritol phosphate (MEP) pathway generates terpenoid precursor molecules and includes the enzyme Dxs and two iron–sulfur cluster enzymes: IspG and IspH. IspG and IspH are rate limiting-enzymes of the MEP pathway but are challenging for metabolic engineering because they require iron–sulfur cluster biogenesis and an ongoing supply of reducing equivalents to function. Therefore, identifying novel alternatives to IspG and IspH has been an on-going effort to aid in metabolic engineering of terpenoid biosynthesis. We report here an analysis of the evolutionary diversity of terpenoid biosynthesis strategies as a resource for exploration of alternative terpenoid biosynthesis pathways. Using comparative genomics, we surveyed a database of 4,400 diverse bacterial species and found that some may have evolved alternatives to the first enzyme in the pathway, Dxs making it evolutionarily flexible. In contrast, we found that IspG and IspH are evolutionarily rigid because we could not identify any species that appear to have enzymatic routes that circumvent these enzymes. The ever-growing repository of sequenced bacterial genomes has great potential to provide metabolic engineers with alternative metabolic pathway solutions. With the current state of knowledge, we found that enzymes IspG and IspH are evolutionarily indispensable which informs both metabolic engineering efforts and our understanding of the evolution of terpenoid biosynthesis pathways.

59 BASIC BIOLOGICAL SCIENCES↗

Evolutionary Flexibility and Rigidity in the Bacterial Methylerythritol Phosphate (Mep) Pathway

Terpenoids are a diverse class of compounds with wide-ranging uses including as industrial solvents, pharmaceuticals, and fragrances. Efforts to produce terpenoids sustainably by engineering microbes for fermentation are ongoing, but industrial production still largely relies on nonrenewable sources. The methylerythritol phosphate (MEP) pathway generates terpenoid precursor molecules and includes the enzyme Dxs and two iron–sulfur cluster enzymes: IspG and IspH. IspG and IspH are rate limiting-enzymes of the MEP pathway but are challenging for metabolic engineering because they require iron–sulfur cluster biogenesis and an ongoing supply of reducing equivalents to function. Therefore, identifying novel alternatives to IspG and IspH has been an on-going effort to aid in metabolic engineering of terpenoid biosynthesis. We report here an analysis of the evolutionary diversity of terpenoid biosynthesis strategies as a resource for exploration of alternative terpenoid biosynthesis pathways. Using comparative genomics, we surveyed a database of 4,400 diverse bacterial species and found that some may have evolved alternatives to the first enzyme in the pathway, Dxs making it evolutionarily flexible. In contrast, we found that IspG and IspH are evolutionarily rigid because we could not identify any species that appear to have enzymatic routes that circumvent these enzymes. The ever-growing repository of sequenced bacterial genomes has great potential to provide metabolic engineers with alternative metabolic pathway solutions. With the current state of knowledge, we found that enzymes IspG and IspH are evolutionarily indispensable which informs both metabolic engineering efforts and our understanding of the evolution of terpenoid biosynthesis pathways.

Bailey Marshall↗

Application of the metabolic modeling pipeline in KBase to categorize reactions, predict essential genes, and predict pathways in an isolate genome

The DOE Systems Biology Knowledgebase (KBase) platform offers a range of powerful tools for the reconstruction, refinement, and analysis of genome-scale metabolic models built from microbial isolate genomes. In this chapter, we describe and demonstrate these tools in action with an analysis of isoprene production in the Bacillus subtilis DSM genome. Two different methods are applied to build initial metabolic models for the DSM genome, then the models are gapfilled in three different growth conditions. Next, flux balance analysis (FBA) and flux variability analysis (FVA) techniques are applied to both study the growth of these models in minimal media and classify reactions within each model based on essentiality and functionality. The models are applied with the FBA method to predict essential genes, which are then compared to an updated list of essential genes obtained for B. subtilis 168, a very similar strain to the DSM isolate. The models are also applied to simulate Biolog growth conditions, and these results are compared with Biolog data collected for B. subtilis 168. Finally, the DSM metabolic models are applied to explore the pathways and genes responsible for producing isoprene in this strain. These studies demonstrate the accuracy and utility of models generated from the KBase pipelines, as well as exploring the tools available for analyzing these models.

DOE knowledgebase↗

Sustainable Biofuels for Low-Carbon Maritime Transportation

The marine shipping sector heavily depends on fossil fuels and is one of the largest petroleum fuel consumers [1,2]. The annual global marine fuel consumption was estimated to be around 400 million metric tons in 2019 (2.5 billion barrels). Moreover, ocean shipping is one of the most significant contributors to sulfur oxides, nitrogen oxides, and particulate matter emissions. Global shipping contributes 13% of human-caused sulfur emissions and 2.6% of anthropogenic carbon dioxide emissions. As a major source of pollutant emissions, the marine industry faces several challenges related to emission regulations. The International Maritime Organization (IMO) has established a framework for reducing the carbon intensity of shipping: 40% reduction relative to 2008 levels by 2030 and 70% reduction by 2050. As the aviation sector, the maritime shipping sector is difficult to decarbonize through electrification. Biofuels offer the best opportunities for decarbonizing marine shipping in the near and medium-term. Advanced biofuels such as pyrolysis bio-oil offer the low-cost potential for meeting carbon reduction goals. For instance, the pyrolysis bio-oil exhibited promising marginal CO2 abatement costs at less than $100/tonne CO2-equivalent at a heavy fuel oil price greater than $1.10/gal [1]. As a potential biofuel option for low-carbon maritime shipping, this presentation focuses on a comparative techno-economic analysis (TEA) of bio-oils produced via a fast pyrolysis-based conversion pathway. The pathway converts a 50/50 blend of forest residues and clean pine to bio-oil via three process options: fast pyrolysis without vapor upgrading, and fast pyrolysis with vapor phase upgrading over ZSM-5 zeolite catalyst and Pt/TiO2 catalyst. The process configuration and operation variation led to different capital and operating costs, as well as the resulting raw bio-oil’s yield and quality, e.g., the water content, total acid number, and carboxylic acid number. The study also determined the minimum upgrading of bio-oils required to enable blending with very low sulfur fuel oil (VLSFO), with the associated costs reflected in TEA. This study shows that bio-oil could be a cost-effective fuel option for decarbonizing maritime shipping. Further research is required with respect to biofuel blending properties, such as compatibility with existing fuel system infrastructure and suitable engine performance.

BIOMASS FUELS↗

Techno-Economic Analysis of Integrated Gasification Fuel Cell Systems

This National Energy Technology Laboratory (NETL) study describes the results of a pathway study for coal-based integrated gasification and solid oxide fuel cell (IGFC) power systems with and without carbon capture and storage. Two pathway scenarios are considered, based on the operating pressure of the SOFC. Three gasification technologies, including a conventional technology, differing in the syngas methane content, which has a significant effect on the IGFC system performance, were considered. The performance and cost benefits were estimated for a series of projected gains made through the development of advances in the component technologies or improvements in plant costs and availability. The objective of the study is to provide targeted research and development guidance to the FECM SOFC Program and SOFC commercial developers.

30 DIRECT ENERGY CONVERSION↗

Mirostructure Characterization of Friction Consolidated Copper-Nickel using a Machine Learning Approach: Developing Process to Microstructure Associations

Friction consolidation (FC) is a solid phase processing approach where discrete material forms such as powders, chips, nuggets, etc. are densified via shear deformation. The precursors are placed in a billet container and brought in contact with a rotating tool that applying the desirable amount of normal force. Under the combined action of the rotation and normal pressure, the discrete precursor is consolidated through porosity reduction and shear deformation. FC is increasingly being studied as an attractive approach to manufacturing fully dense parts from powder forms owing to its ability to mix, alloy and consolidate difficult-to-process precursors in minimal number of process steps. Material consolidation and deformation in shear consolidation processes have been studied extensively previously for different material combinations previously. However, despite the extensive research in this area, understanding of the mechanistic processes in pore consolidation, deformation-induced mixing and material solubility during FC is still evolving. Material development using solid phase processing approaches such as FC is often performed based on research experience/education, which can be biased. Conventional analysis and simulation tools in this area tend to be successful only when material thermodynamic pathways and microstructural evolution sequences resulting from processing are clearly defined or known. They are not as effective for emerging advanced manufacturing technologies where material evolution pathways are not well established. The ability to predict optimal process parameters based on material chemistry and bulk properties is essential to accelerate materials design and processing, as are an understanding of the relevant structure-processing-property relationships. These structure-processing-property-performance relationships are at the core of materials science research. Microstructure characterization provides the link to these four core areas, often through visualizing material microstructure using imaging techniques. However, linking microstructure image data (i.e., micrographs) to variables of interest (e.g., processing parameters, material chemistry) in a reproducible, generalizable, and quantitative manner is a significant challenge. Typically, quantitatively linking image data to processing history relies on significant domain knowledge and manual or subject matter expert (SME)-heuristic based image analysis. Such an approach to image analysis has the potential to be biased, inefficient, and difficult to replicate.

36 MATERIALS SCIENCE↗

Comprehensive techno-economic and life cycle greenhouse gases analysis of green ammonia production utilizing PV and wind energy: Jordan as a case study

Ammonia (NH 3 ) has emerged as a critical player in the global energy transition due to its potential as a low-carbon fuel. Conventional ammonia production methods, primarily through steam methane reforming, are significant contributors to global CO 2 emissions. This study investigates the potential of green ammonia production in Jordan by leveraging the country's abundant solar photovoltaic and wind energy resources. A comprehensive techno-economic and life cycle greenhouse gas analysis was conducted to compare green ammonia production with conventional grey and blue ammonia pathways. Here, the study utilized spatial mapping to assess renewable energy capacity factors across Jordan and optimized the integration of hybrid PV/wind systems with proton exchange membrane (PEM) electrolyzers. The results indicate that the levelized cost of green ammonia (LCOA) in Jordan varies between 900 USD/kg NH 3 and 2500 USD/kg NH 3 which is significantly higher than grey ammonia (similar to 360 USD/kg NH 3 ). Nevertheless, the carbon intensity of green ammonia production (between 0.1 kg CO 2eq /kg NH 3 and 0.5 kg CO 2eq /kg NH 3 ) is much lower than the grey ammonia (1.8 kg CO 2eq /kg NH 3 ). Finally, the results indicate that considering future reductions in capital costs and advancement in renewable energy systems and PEM electrolyzers as well as the ability to sell the co-product O 2 can reduce the LCOA by up to 92 % and make it competitive with grey ammonia.

Green Ammonia↗

Nitric-oxide synthase is a mechanical signal transducer that modulates talin and vinculin expression

Mechanical stimuli can cause changes in muscle mass and structure which indicate that mechanisms exist for transducing mechanical stimuli into signals that influence gene expression. Myotendinous junctions show adaptations to modified muscle loading which suggest that these are transcriptionally distinct domains in muscle fibers that may experience local regulation of expression of structural proteins that are concentrated at these sites. Vinculin and talin are cytoskeletal proteins that are highly enriched at myotendinous junctions that we hypothesize to be subject to local transcriptional regulation. Our findings show that mechanical stimulation of muscle cells in vivo and in vitro causes an increase in the expression of vinculin and talin that is mediated by nitric oxide. Furthermore, nitric oxide-stimulated increases in vinculin and talin expression occur through a protein kinase G-dependent pathway and therefore differ from other mechanisms through which nitric oxide has been shown previously to modulate transcription. Analysis of vinculin mRNA distribution in mechanically stimulated muscle fibers shows that the mRNA is highly concentrated at myotendinous junctions, which supports the hypothesis that myotendinous junctions are distinct domains in which the expression of cytoskeletal proteins is modulated by mechanical stimuli through a nitric oxide and protein kinase G-dependent pathway.

NASA Discipline Musculoskeletal↗

Overview of Integrated Pathway Analyses to Meet the Hydrogen Energy Earthshot Goal

This slide deck was presented at the 2021 DOE Hydrogen Shot Summit on Aug. 31, 2021. The presentation describes two ongoing NETL system analysis efforts: "Comparison of Commercial, State-of-the-Art, Fossil-Based Hydrogen Production Technologies" (nearing completion), and "Hydrogen Energy Earthshot Initiative Screening Analysis" (which has recently started). The former study will detail costs and greenhouse gas emissions profiles of current commercial technologies (e.g., reforming and gasification routes). The latter will focus on our efforts to identify pathways that may meet the goal of the DOE Hydrogen Energy Earthshot Initiative (i.e., $1/kg H2 by 2030) as well as low carbon emissions.

Lewis, Eric↗

Rewiring yeast metabolism for producing 2,3-butanediol and two downstream applications: Techno-economic analysis and life cycle assessment of methyl ethyl ketone (MEK) and agricultural biostimulant production

Rising concerns for sustainability and global climate change have driven the development of sustainable pro- duction pathways for biofuels and chemicals from lignocellulosic biomass via integrated biological and chemical processes. We constructed an engineered Saccharomyces cerevisiae capable of producing 2,3-butanediol (2,3-BDO) from glucose without accumulating ethanol and glycerol, which hinder downstream processing of 2,3-BDO, through extensive metabolic reprogramming. Specifically, we introduced heterologous 2,3-BDO biosynthetic enzymes and deleted the major isozymes of ethanol and glycerol biosynthetic enzymes. In addition, we introduced an NAD + regenerating Pyruvate-Malate (PM) cycle and enhanced the NAD + regenerating capability of the PM cycle to resolve the redox imbalance from the deletion of ethanol and glycerol production pathways. The resulting engineered yeast produced 109.9 g/L of 2,3-BDO with a productivity of 1.0 g/L/h and a yield of 0.36 g/ g glucose in a fed-batch fermentation. We also conducted techno-economic analysis (TEA) and life cycle assessment (LCA) of the production of methyl ethyl ketone (MEK) through catalytic dehydration of 2,3-BDO. A TEA based on the experimental results indicated that the minimum product selling price (MPSP) was estimated to be $1.90/kg. Regarding cradle-to-grave LCA, 100-year global warming potential (GWP100) and fossil energy consumption (FEC) were found to be 0.37 kg CO 2 eq/kg and 3.1 MJ/kg, respectively. These results demonstrated the feasibility of cost-competitive and sustainable bio-based MEK production via yeast fermentation. In addition, we explored the possibility of using the fermentation broth containing 2,3-BDO as a biostimulant inducing drought tolerance in plants. As a result, the yeast 2,3-BDO fermentation broth can induce drought tolerance in Arabidopsis thaliana without a complicated purification process.

2,3-butanediol (2,3-BDO)↗

Design and full core fuel performance assessment of high burnup cores for 4-loop PWRs

Increasing the fuel discharge burnup of current light water reactors (LWRs) promises reductions in fuel cycle and/or operations costs. By assuming a constant core power density, the economic gain is enabled by better fuel utilization and/or an increased capacity factor. In this effort to investigate greater than 62 MWd/kgU maximum rod average burnup for 110+ kW/l core power density, two core designs have been developed for a standard 17x17, 193 fuel assemblies pressurized water reactor (PWR). The levelized unit cost methodology is employed to evaluate fuel cycle, operation and maintenance, and capital cost impacts and to examine the economic viability of both core design pathways. Core design and optimization are performed using the commercial STUDSVIK code package. Fuel performance analysis is realized in full core configuration via auditing FRAPCON4.1, FAST1.2, and the high-fidelity code BISON. To provide a realistic assessment, the core design process takes into consideration best practices in current PWR core design. It features acceptable performance in terms of various core design constraints on maximum allowable peaking and boron concentration. Gadolinia (Gd2O3) is used as a burnable poison with a maximum of 9 wt% concentration while feeding 89 or 77 fuel assemblies in a 3-batch refueling scheme. Full core fuel performance simulation, which allows for characterization of relevant fuel temperatures, plenum pressures, stresses, and strains, is performed with respect to two bounding burnup levels. Such performance is potentially licensable for the 18-month high burnup core (<68 MWd/kgU peak pin), while it is more challenging for the 24-month high burnup core design pathway (<75 MWd/kgU peak pin). Maximum rod plenum pressure is identified as the most limiting fuel performance parameter. Here, while the scope of the present study focuses on the steady-state plus overpower conditions, the acceptability of the new discharge burnup has to be further assessed by considering uncertainties and impacts under accident scenarios in the future.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Description, Usage, and Validation of the MVL-15 Modified Vortex Lattice Analysis Capability

MVL-15 is the most recent version of the Modified Vortex-Lattice (MVL) code developed within the Aerodynamics Systems Analysis Branch (ASAB) at NASA LaRC. The term "modified" refers to the primary modification of the core vortex-lattice methodology: inclusion of viscous aerodynamics tables that are linked to the linear solution via iterative processes. The inclusion of the viscous aerodynamics inherently converts the MVL-15 from a purely analytic linearized method to a semi-empirical blend which retains the rapid execution speed of the linearized method while empirically characterizing the section aerodynamics at all spanwise lattice points. The modification provides a means to assess non-linear effects on lift that occur at angles of attack near stall, and provides a means to determine the drag associated with the application of design strategies for lift augmentation such as the use of flaps or blowing. The MVL-15 code is applicable to the analyses of aircraft aerodynamics during cruise, but it is most advantageously applied to the analysis of aircraft operating in various high-lift configurations. The MVL methodology has been previously conceived and implemented; the initial concept version was delivered to the ASAB in 2001 (van Dam, C.), subsequently revised (Gelhausen, P. and Ozoroski, T. 2002 / AVID Inc., Gelhausen, P., and Roberts, M. 2004), and then overhauled (Ozoroski, T., Hahn, A. 2008). The latest version, MVL-15 has been refined to provide analysis transparency and enhanced to meet the analysis requirements of the Environmentally Responsible Aviation (ERA) Project. Each revision has been implemented with reasonable success. Separate applications of the methodology are in use, including a similar in-house capability, developed by Olson, E. that is tailored for structural and acoustics analyses. A central premise of the methodology is that viscous aerodynamic data can be associated with analytic inviscid aerodynamic results at each spanwise wing section, thereby providing a pathway to map viscous data to the inviscid results. However, a number of factors can sidetrack the analysis consistency during various stages of this process. For example, it should be expected that the final airplane lift curve and drag polar results depend strongly on the geometry and aerodynamics of the airfoil section; however, flap deflections and flap chord extensions change the local reference geometry of the input airfoil, the airplane wing, the tabulated non-dimensional viscous aerodynamics, and the spanwise links between the linear and the viscous aerodynamics. These changes also affect the bound circulation and therefore, calculation and integration of the induced angle of attack and induced drag. MVL-15 is configured to ensure these types of challenges are properly addressed. This report is a comprehensive manual describing the theory, use, and validation of the MVL-15 analysis tool. Section 3 summarizes theoretical, procedural, and characteristic features of MVL-15, and includes a list of the files required to setup, execute, and summarize an analysis. Section 4, Section 5, Section 6, and Section 7 combine to comprise the User's Guide portions of this report. The MVL-15 input and output files are described in Section 4 and Section 5, respectively; the descriptions are supplemented with example files and information about the file formats, parameter definitions, and typical parameter values. Section 6 describes the Wing Geometry Setup Utility and the 2d-Variants Utility files that simplify and assist setting up a consistent set of MVL-15 geometry and aerodynamics input parameters and input files. Section 7 describes the use of the 3d-Results Presentation Utility file that can be used to automatically create summary tables and charts from the MVL-15 output files. Section 8 documents the Validation Results of an extensive and varied validation test matrix, including results of an airplane analysis representative of the ERA Program. A start-to-finish example of the airplane analysis procedure is described in Section 7.

Ozoroski, Thomas A.↗

A multilayer network analysis of Alzheimer's disease pathogenesis: Roles for p‐tau, synaptic peptides, and physical activity

INTRODUCTION: In the aging brain, cognitive abilities emerge from the coordination of complex pathways arising from a balance between protective lifestyle and environmental factors and accumulation of neuropathologies. METHODS: As part of the Rush Memory and Aging Project (n = 440), we measured accelerometer-based actigraphy, cognitive performance, and after brain autopsy, selected reaction monitoring mass spectrometry. Multilevel network analysis was used to examine the relationships among the molecular machinery of vesicular neurotransmission, Alzheimer's disease (AD) neuropathology, cognition, and late-life physical activity. RESULTS: Synaptic peptides involved in neuronal secretory function were the most influential contributors to the multilayer network, reflecting the complex interdependencies among AD pathology, synaptic processes, and late-life cognition. Older adults with lower physical activity evidenced stronger adverse relationships among phosphorylated tau peptides, markers of synaptic integrity, and tangle pathology. DISCUSSION: Network-based approaches simultaneously model interdependent biological processes and advance understanding of the role of physical activity in age-associated cognitive impairment. Highlights: Network-based approaches simultaneously model interdependent biological processes. Secretory synaptic peptides were influential contributors to the multilayer network. Older adults with lower physical activity had adverse relationships among pathology. There was interdependence among phosphorylated tau, synaptic integrity, and tangles. Network methods elucidate the role of physical activity in cognitive impairment.

60 APPLIED LIFE SCIENCES↗

Activation of c-Jun N-terminal kinase and apoptosis in endothelial cells mediated by endogenous generation of hydrogen peroxide

Reactive oxygen species have been implicated in the activation of signal transduction pathways. However, extracellular addition of oxidants such as hydrogen peroxide (H2O2) often requires concentrations that cannot be readily achieved under physiological conditions to activate biological responses such as apoptosis. Explanations for this discrepancy have included increased metabolism of H2O2 in the extracellular environment and compartmentalization within the cell. We have addressed this issue experimentally by examining the induction of apoptosis of endothelial cells induced by exogenous addition of H2O2 and by a redox cycling agent, 2,3-dimethoxy-1,4-naphthoquinone, that generates H2O2 in cells. Here we show that low nanomolar steady-state concentrations (0.1-0.5 nmol x min(-1) x 10(6) cells) of H2O2 generated intracellularly activate c-Jun N terminal kinase and initiate apoptosis in endothelial cells. A comparison with bolus hydrogen peroxide suggests that the low rate of intracellular formation of this reactive oxygen species results in a similar profile of activation for both c-Jun N terminal kinase and the initiation of apoptosis. However, a detailed analysis reveals important differences in both the duration and profile for activation of these signaling pathways.

Non-NASA Center↗

Application of the Metabolic Modeling Pipeline in KBase to Categorize Reactions, Predict Essential Genes, and Predict Pathways in an Isolate Genome

The DOE Systems Biology Knowledgebase (KBase) platform offers a range of powerful tools for the reconstruction, refinement, and analysis of genome-scale metabolic models built from microbial isolate genomes. In this chapter, we describe and demonstrate these tools in action with an analysis of isoprene production in the Bacillus subtilis DSM genome. Two different methods are applied to build initial metabolic models for the DSM genome, then the models are gapfilled in three different growth conditions. Next, flux balance analysis (FBA) and flux variability analysis (FVA) techniques are applied to both study the growth of these models in minimal media and classify reactions within each model based on essentiality and functionality. The models are applied with the FBA method to predict essential genes, which are then compared to an updated list of essential genes obtained for B. subtilis 168, a very similar strain to the DSM isolate. The models are also applied to simulate Biolog growth conditions, and these results are compared with Biolog data collected for B. subtilis 168. Finally, the DSM metabolic models are applied to explore the pathways and genes responsible for producing isoprene in this strain. These studies demonstrate the accuracy and utility of models generated from the KBase pipelines, as well as exploring the tools available for analyzing these models.

Allen, Benjamin↗

A Concept of Operations (ConOps) of an In-time Aviation Safety Management System (IASMS) for Advanced Air Mobility (AAM)

The growth of new emerging operations involving Advanced Air Mobility (AAM) necessitates developing a perspective for an In-time Aviation Safety Management System (IASMS). This perspective advances from the National Academies report on IASMS and its recommendation for developing a Concept of Operations (ConOps) for IASMS. A ConOps has been developed for In-time System-Wide Safety Assurance (ISSA) from which the IASMS ConOps pivots to provide a robust scope commensurate with the broad vision defined by the National Academies. The IASMS ConOps focuses on emerging operations and spans innovations in Unmanned Aircraft System (UAS) and an increasingly complex ecosystem comprised of a widening mix of vehicles and technologies, Urban Air Mobility (UAM) with industry-federated services, traditional operations, as well as new supersonic aircraft and space launch systems.The challenge for the IASMS ConOps is to be broad to encompass innovations in the coming years and decades while agile to ensure levels of safety compatible with operational and certification requirements of the National Airspace System (NAS). The IASMS ConOps interweaves increasing complexity of operational safety capabilities and unlocking UAS Maturity Levels (UMLs). The relationships between increased complexity of automation and automated systems, fewer operators who are not as traditionally higher skilled, more complex operational environments, and aviation operations management with mixed aircraft and equipage pose a multi-dimensional space for IASMS capabilities essential for safety assurance and risk management. Instantiating IASMS capabilities and how they would be integral to AAM operations and increasing maturity of UAM could be accomplished through a series of Safety Demonstrators. These Safety Demonstrators could provide increased understanding and insight into use of controls for risk mitigation, means of compliance for certification, and operational experience with safety services such as in relation to contingency management. The IASMS capabilities can be viewed as initially residing with the vehicle, airspace, and Supplemental Data Service Provider (SDSP). For example, vehicle capabilities include communications including the command and control link, Remote Identification (ID), conflict advisory/alerting, and UAS system monitoring. These capabilities monitor and assess data such as battery health, aircraft state, and human performance. Complexity of ISAMS capabilities depends on a number of factors. These factors are intendedonly as a notional categorization with the purpose being to reflect the complexity of the AAM ecosystem that would drive up the complexity of ISSA capabilities including systems, sensors, models, standards, and controls. Factors could include the Vehicle Flight Management, Environment, Airspace, and Contingency Management. Each of these factors can be comprised of multiple sub-factors that contribute to increasing complexity. For example, Airspace at a lower level of complexity could be dedicated to UTM operations that are unmonitored, and at a higher level of complexity could involve mixed UTM and ATM operations. The IASMS concept includes safety services that provide data and information to different participants in AAM. The roles and responsibilities of participants can be defined using the Responsible-Accountable-Consulted-Informed (RACI) analysis. For example, for the safety service involving the Remote ID, the Operator would be accountable for providing the data, the Vehicle would be responsible for transmitting it, and the USS, SDSP, Vertiports, FIMS (FAA), and Public Entities such as safety services would be informed by receiving the data. The IASMS ConOps identifies the capabilities needed for risk mitigation and safety assurance in the increasingly complex national airspace. The ConOps serves as a pathway for engaging with industry to gain operational experience including through the Safety Demonstrator series, the RACI analysis, and operational complexity factors. The ConOps serves to integrate these different perspectives to build a cohesive and cogent approach to an AAM safety management system.

In-Time Aviation Safety Management System↗

Transporters and ascorbate–glutathione metabolism for differential cadmium accumulation and tolerance in two contrasting willow genotypes

Salix matsudana Koidz is a low cadmium (Cd)-accumulating willow, whereas its cultivated variety, Salix matsudana var. matsudana f. umbraculifera Rehd., is a high Cd-accumulating and tolerant willow (HCW). The physiological and molecular mechanisms underlying differential Cd accumulation and tolerance in the two Salix species are poorly understood. In this study, we confirmed that the differential Cd translocation capacity from roots to the shoots leads to the differential Cd accumulation in their aboveground parts between these two willow genotypes. Cadmium accumulation happens preferentially in the transport pathway, and Cd is mainly located in the vacuolar, cell wall and intercellular space in HCW bark by cadmium location analysis at tissue and subcellular levels. Comparative transcriptome analysis revealed that higher expressions of several metal transporter genes (ATP-binding cassette transporters, K + transporters/channels, yellow stripe-like proteins, zinc-regulated transporter/iron-regulated transporter-like proteins, etc.) are involved in root uptake and translocation capacity in HCW; meanwhile, ascorbate–glutathione metabolic pathways play essential roles in Cd detoxification and higher tolerance of the Cd-accumulator HCW. These results lay the foundation for further understanding the molecular mechanisms of Cd accumulation in woody plants and provide new insights into molecular-assisted-screening woody plant varieties for phytoremediation.

59 BASIC BIOLOGICAL SCIENCES↗

iGCE and MitoFlyght Spaceflight Missions: Unraveling Oxidative Stress Responses in Space

Thriving In DEep Space (TIDES) initiative aims to comprehensively understand how hostile environments such as the Moon and Mars affect human physiology. Here we present two NASA-selected spaceflight experiments under the TIDES portfolio, iGCE (Integrated Gravity Continuum Experiment) and MitoFlyght (Mitochondrial Investigation of Oxidative Stress in Flies). We hypothesize that exposure to spaceflight conditions induces oxidative stress responses that negatively impact physiology. The iGCE mission employs two well-established spaceflight models, Drosophila melanogaster and C.elegans, using Redwire’s Multi-use Variable-g Platform (MVP) hardware to assess changes in cardiac, muscle, and nervous systems across five different gravities: Hypergravity (2g), Earth (1g), Mars (0.37g), Moon (0.16g), and microgravity (ug). This mission focuses on uncovering alterations in protein homeostasis, autophagy, and mitochondrial function conserved across species. In the MitoFlyght mission to the ISS, Drosophila will be housed in the Vented Fly Box (VFB). This mission evaluates the oxidative stress response and autophagic pathway in muscle, heart, and nervous system. Additionally, we will (a) use the genetic mutant, Tor7/P to test whether increased autophagy is beneficial or a maladaptive response to the spaceflight stressors, and (b) utilize fly lines with tissue-specific expression (neuronal, muscle, and cardiac) of an antioxidant gene, SOD2 (superoxide dismutase) as a potential countermeasure. Data from these missions will be compared with previous LEO-based datasets to identify shared signatures. Furthermore, cross-species analysis of the transcriptomic data from other invertebrate and vertebrate spaceflight studies will help determine evolutionarily conserved pathways perturbed by space stressors. Overall, both these missions aim to provide crucial insights into the mechanisms underlying oxidative stress responses, synaptic changes, and heart and muscle deficits, facilitating the identification of diagnostic and therapeutic targets to mitigate the adverse health effects of long-duration space habitation. Ultimately, this research will enhance our ability to thrive in deep space and inform future missions.

Janani Iyer↗