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At least 181 records · Page 10

Atmosphere Assessment for MARS Science Laboratory Entry, Descent and Landing Operations

On August 6, 2012, the Mars Science Laboratory rover, Curiosity, successfully landed on the surface of Mars. The Entry, Descent and Landing (EDL) sequence was designed using atmospheric conditions estimated from mesoscale numerical models. The models, developed by two independent organizations (Oregon State University and the Southwest Research Institute), were validated against observations at Mars from three prior years. In the weeks and days before entry, the MSL "Council of Atmospheres" (CoA), a group of atmospheric scientists and modelers, instrument experts and EDL simulation engineers, evaluated the latest Mars data from orbiting assets including the Mars Reconnaissance Orbiter's Mars Color Imager (MARCI) and Mars Climate Sounder (MCS), as well as Mars Odyssey's Thermal Emission Imaging System (THEMIS). The observations were compared to the mesoscale models developed for EDL performance simulation to determine if a spacecraft parameter update was necessary prior to entry. This paper summarizes the daily atmosphere observations and comparison to the performance simulation atmosphere models. Options to modify the atmosphere model in the simulation to compensate for atmosphere effects are also presented. Finally, a summary of the CoA decisions and recommendations to the MSL project in the days leading up to EDL is provided.

Cianciolo, Alicia D.↗

Argument-Based Airworthiness Assurance of Small UAS

Presently, there are three avenues by which Unmanned Aircraft System (UAS) operations are authorized in the U.S. National Airspace System (NAS): obtaining either (i) a certificate of authorization (COA), or (ii) a special airworthiness certificate (SAC) in either the experimental, or the restricted category, or (iii) an exemption from an airworthiness certificate together with a civil COA. The first is meant primarily for public entities, such as NASA; the remaining two are the only available means for civil UAS operations. Recently, the Federal Aviation Administration (FAA) has also proposed a regulatory framework targeted for certain small UAS, specifically those weighing 55 pounds or less, although final rulemaking remains pending. We have previously shown how an assurance case can aggregate heterogeneous reasoning and safety evidence, with application to UAS safety. In this paper, we describe how assurance cases can serve as a common framework to justify overall system safety, unifying both operational aspects and airworthiness, in particular system design assurance. We also show how this approach can coexist with, and augment, existing safety analysis processes and best-practices, by transforming the artifacts they produce into structured assurance arguments. To illustrate the applicability and utility of our approach, we have been applying it for the design assurance of an unmanned rotorcraft system, intended for precision agriculture operations, as part of the NASA Unmanned Aircraft System (UAS) Integration in the National Airspace System (NAS) project.

Rotocraft↗

Highland Lakes Water Resources: Using NASA Earth Observations to Improve Detection Systems for Harmful Algal Events in the Highland Lakes in Central Texas

Beginning in 2019, harmful algal events in Austin, Texas, caused canine deaths in the Lady Bird Lake and Lake Travis reservoirs. These reservoirs are part of the larger Highland Lakes chain, managed by the Lower Colorado River Authority (LCRA) and the City of Austin Department of Watershed Protection (CoA DWP), which fulfill municipal, commercial, and agricultural water demands. Given the recent increase in favorable algal event conditions in central Texas, the LCRA and CoA DWP partnered with NASA DEVELOP to improve algal event early-warning systems through the application of remote sensing and machine learning. An Earth observation-based algal monitoring system will assist the responsible agencies in predicting algal conditions and communicating hazards to the public. The NASA DEVELOP team utilized Landsat 8 Operational Land Imager (OLI) and Sentinel-2 Multispectral Instrument (MSI) data to produce products including chlorophyll-a concentrations, cyanobacteria detections, turbidity, and water surface temperature. Chlorophyll-a concentrations were retrieved with a pre-trained machine learning model (mixture density network) and spectral indices, while the other products were derived from spectral indices. In situ field data were used to validate and quantify uncertainties for each product. The validations show strong correlations for chlorophyll-a and water surface temperature. Time series analyses of chlorophyll-a concentrations show peaks in the severe drought years (2015 and 2016). This project's resulting products enable monitoring of environmental proxies relevant to algal event presence in the Highland Lakes chain and will ultimately support water management, decision making, and risk communication.

Kaitlynn Hietpas↗

The Application of Remote Sensing and Machine Learning to Improve Early Warning Systems for Harmful Algal Events in the Highland Lake Chain, TX

Beginning in 2019, harmful algal events have caused canine deaths in both Lady Bird Lake and Lake Travis located near Austin, Texas. These two reservoirs are part of the larger Highland Lakes chain, managed by the City of Austin Department of Watershed Protection (COA DWP) and the Lower Colorado River Authority (LCRA), which fulfill municipal, commercial, and agricultural water demands. Given the recent increase in favorable environmental conditions for algal events in central Texas, NASA DEVELOP partnered with LCRA and COA DWP to improve monitoring and early detection of algal events, utilizing satellite remote sensing and machine learning. Spatially and temporally varied chlorophyll a concentrations, cyanobacteria detections, turbidity, and water surface temperature products are used as environmental proxies. Landsat 8 Operational Land Imager (OLI) and the Sentinel-2 MultiSpectral Instrument (MSI) data were used to provide a combined revisit time of up to ~3 days and < 30 m per pixel products. Chlorophyll a concentrations were estimated using a pre-trained Mixture Density Network, and cyanobacteria detection was accomplished using the Broad Wavelength Algae Index, which can differentiate algal blooms from algal proliferations (mats). In situ data were used to validate remotely sensed measurements and quantify uncertainties. Preliminary results show a good fit between the modeled output and in situ observations, suggesting that remote sensing data can be used to retrieve biogeochemical properties and/or inherent optical properties (IOPs) of water columns in these inland human-made lakes. Uncertainties were introduced from the sensitivity to atmospheric correction, inherent mismatch between satellite and sampling data, and a relatively lower signal-to-noise ratio over water. The resulting products enable near real-time monitoring of environmental proxies relevant to algal event presence in the Highland Lakes chain, and will ultimately support water management, decision making, and risk communication.

Shuyu Chang↗

Safety Case for Small Uncrewed Aircraft Systems (sUAS) Beyond Visual Line of Sight (BVLOS) Operations at NASA Langley Research Center

This Technical Memorandum (TM) is written to provide for dissemination of the methods and safety considerations for operations of small Uncrewed Aerial Systems (sUAS) Beyond Visual Line-of-Sight (BVLOS)at NASA Langley Research Center. It includes the Safety Case used to acquire a BVLOS Certificate of Authorization (COA) from the FAA and is being published to enable others to benefit from this work. The intended operations, subject to approval from the Federal Aviation Administration (FAA) and the National Aeronautics and Space Administration (NASA), will include a combination of Within Visual Line of Sight (WVLOS) and Beyond Visual Line of Sight (BVLOS) flights, comprising of at most five sUAS operating concurrently, with no more than three operating BVLOS. Flights will occur in a subset of the Langley Air Force Base (LAFB) Class D airspace (KLFI) at a maximum altitude of 400 ft AGL. Most operations within this subset will take place in the City Environment Range Testing for Autonomous Integrated Navigation (CERTAIN) Range. The CERTAIN Range includes airspace inside the borders of NASA Langley Research Center (LaRC). Additional airspace over the northern section of CERTAIN will be requested as part of the Certificate of Authorization (COA). NASA LaRC BVLOS operations on the CERTAIN Range can be broken down into five critical components needed to meet the 14 CFR § 91.113 see and avoid requirement: 1) procedural deconfliction with LAFB for UAS operations at or below 400 ft and manned aircraft at or above 900’ AGL; 2) ground equipment for detection of intruder aircraft and to support communications between crewmembers ; 3) sUAS vehicles with advanced onboard automation capable of autonomously maintaining safe separation; 4) BVLOS standardized operating procedures (SOPs); 5) and personnel to execute the flight operations in accordance with the SOPs and respond to airborne contingencies. The introduction of new ground equipment includes the use of the Remote Operations for Autonomous Missions (ROAM) UAS Operations Center, development and use of an Integrated Airspace Display (IAD), use of the L-STAR and GA-9120 radars, and the incorporation of standardized Vertiports. The ROAM Operations Center will be the central point for all BVLOS sUAS operations. All command and control (C2), voice communications and airspace awareness displays will reside inside ROAM. The IAD will provide raw data from ADS-B, FLARM, radar tracks and telemetered GPS vehicle positions for interpretation by an Airspace Monitor. The radars will search the class D airspace around the CERTAIN Range and serve as a backup to procedural deconfliction procedures coordinated with LAFB. In the event of a procedural deconfliction breakdown, radar detections of non-participating aircraft will be available so that the 91.113 see and avoid requirement can still be safely met. Finally, the incorporation of Vertiports will have video and network connectivity that enables large numbers of sUAS launches and recoveries from a single location. This is a continuation of the remote command and control of unpiloted aircraft component focused on evaluating unpiloted aircraft flight crew roles and responsibilities, control interfaces and the associated data links needed to operate a fleet of aircraft within a UAM Ecosystem. This work supports the development of future aviation operational concepts based on an Urban Air Mobility Maturity Level (UML) 4 environment (Patterson, 2020). It is assumed that future airspace will include hundreds of simultaneous aircraft operations within the airspace, therefore scalable operations are essential for enabling this future airspace to become a reality. Follow on work includes envisioned flights that expand operations beyond the CERTAIN range and lead to an effective Maritime Surveillance capability.

Matthew W Coldsnow↗

Off-Nominal Event Analysis in Autonomous Flights Based on Explainable Artificial Intelligence

A key objective in the Urban Air Mobility program at NASA is to intelligently perform an autonomous flight in a complex urban environment under all weather conditions with guaranteed levels of safety. To accomplish this, the mission manager (central decision-making module) of the vehicle needs to make informed decisions between various Courses of Action (CoA) based on its' interpretation of the inputs it receives. If an off-nominal event is detected either based on the amalgamation of sensor data or the use of machine learning models, the mission manager may greatly benefit from identification of the input features that most likely contributed to that specific event. Such an understanding is usually not possible to obtain from the classical machine learning models (deep learning) due to the inherent black box like structure. However, this understanding is achieved using eXplainable Artificial Intelligence (XAI) models that provide a human interpretable rationale for the predictions made. This work presents a game theory inspired XAI model for the off-nominal assessment of autonomous flights. The proposed approach based on Shapley values is model agnostic, provides local as well as global explanation and satisfies the four axioms (efficiency, symmetry, dummy, additivity) to achieve fair contribution. The versatility of the approach is first demonstrated on a simulated dataset in which the significance of each input to flight phase prediction is clearly identified. Subsequently, data from simulated flight trajectories are fed into the model which reveal the input features that most likely contributed to a rotor failure event thereby empowering the mission manager to take the appropriate CoA.

autonomy↗

Off-Nominal Event Analysis in Autonomous Flights Based on Explainable Artificial Intelligence

A key objective in the Urban Air Mobility program at NASA is to intelligently perform an autonomous flight in a complex urban environment under all weather conditions with guaranteed levels of safety. To accomplish this, the mission manager (central decision-making module) of the vehicle needs to make informed decisions between various Courses of Action (CoA) based on its' interpretation of the inputs it receives. If an off-nominal event is detected either based on the amalgamation of sensor data or the use of machine learning models, the mission manager may greatly benefit from identification of the input features that most likely contributed to that specific event. Such an understanding is usually not possible to obtain from the classical machine learning models (deep learning) due to the inherent black box like structure. However, this understanding is achieved using eXplainable Artificial Intelligence (XAI) models that provide a human interpretable rationale for the predictions made. This work presents a game theory inspired XAI model for the off-nominal assessment of autonomous flights. The proposed approach based on Shapley values is model agnostic, provides local as well as global explanation and satisfies the four axioms (efficiency, symmetry, dummy, additivity) to achieve fair contribution. The versatility of the approach is first demonstrated on a simulated dataset in which the significance of each input to flight phase prediction is clearly identified. Subsequently, data from simulated flight trajectories are fed into the model which reveal the input features that most likely contributed to a rotor failure event thereby empowering the mission manager to take the appropriate CoA.

autonomy↗

Synthetic pathway for biological carbon dioxide sequestration

This invention relates to methods for increasing carbon fixation and/or increasing biomass production in a plant, comprising: introducing into a plant, plant part, and/or plant cell heterologous polynucleotides encoding (1) a succinyl CoA synthetase, (2) a 2-oxoglutarate:ferredoxin oxidoreductase, (3) a 2-oxoglutarate carboxylase, (4) an oxalosuccinate reductase, or (5) an isocitrate lyase, or (6) a succinyl CoA synthetase and a 2-oxoglutarate:ferredoxin oxidoreductase, (7) a 2-oxoglutarate carboxylase and an oxalosuccinate reductase polypeptide, and/or (8) a 2-oxoglutarate carboxylase polypeptide, an oxalosuccinate reductase polypeptide and an isocitrate lyase polypeptide to produce a stably transformed plant, plant part, and/or plant cell, wherein said heterologous polynucleotides are from a bacterial and/or an archaeal species. Additionally, transformed plants, plant parts, and/or plant cells are provided as well as products produced from the transformed plants, plant parts, and/or plant cells.

09 BIOMASS FUELS↗

Engineered microbes for conversion of organic compounds to medium chain length alcohols and methods of use

This disclosure provides a genetically-modified bacterium from the genus Pseudomonas that comprises an exogenous nucleic acid encoding an enoyl-CoA reductase and an exogenous nucleic acid encoding an acyl-CoA reductase that produces medium chain length alcohols. The disclosure further provides methods for producing medium chain alcohols using such genetically-modified bacterium. This disclosure provides a renewable, bio-based production platform for valuable mcl-alcohols that have a wide range of industrial applications. Current production of mcl-alcohols typically occurs through the hydrogenation of plant oils and waxes. This process leads to issues of deforestation and is largely unsustainable. Utilizing waste lignin streams as the carbon source provides a more sustainable feedstock that can be generated from plant waste like corn stover. Along with this, the use of lignin avoids competition with food resources as traditional starch and sugar feedstocks.

Guss, Adam M.↗

Genetically modified cells that produce C6-C10 fatty acid derivatives

Genes encoding mutant 3-ketoacyl-CoA synthases are introduced into host cells. Certain of the mutants enhance the production of shorter-chain fatty acids and derivatives by the cell than do the wild-type (unmutated) enzymes. In other cases, the chain length is not significantly affected, but productivity is enhanced. In specific cases, both a shift toward lower chain length and higher productivity is seen. Cells producing the mutant 3-ketoacyl-CoA synthases are especially suitable for producing C6-C10 fatty acids and derivatives.

Liao, Hans H.↗

Recombinant microorganisms and uses therefor

Microorganisms are genetically engineered to produce 3-hydroxypropionate (3-HP). The microorganisms are carboxydotrophic acetogens. The microorganisms produce acetyl-coA using the Wood-Ljungdahl pathway for fixing CO/CO 2 . A β-alanine pyruvate aminotransferase from a microorganism that contains such an enzyme is introduced. Additionally, an acetyl-coA carboxylase may also be introduced. The production of 3-HP can be improved. This can be effected by improved promoters or higher copy number or enzymes that are catalytically more efficient.

Liew, Fungmin↗

Enhancing monolignol ferulate conjugate levels in poplar lignin via OsFMT1

The phenolic polymer lignin is one of the primary chemical constituents of the plant secondary cell wall. Due to the inherent plasticity of lignin biosynthesis, several phenolic monomers have been shown to be incorporated into the polymer, as long as the monomer can undergo radicalization so it can participate in coupling reactions. In this study, we significantly enhance the level of incorporation of monolignol ferulate conjugates into the lignin polymer to improve the digestibility of lignocellulosic biomass. Overexpression of a rice Feruloyl-CoA Monolignol Transferase (FMT), OsFMT1, in hybrid poplar (Populus alba x grandidentata) produced transgenic trees clearly displaying increased cell wall-bound ester-linked ferulate, p-hydroxybenzoate, and p-coumarate, all of which are in the lignin cell wall fraction, as shown by NMR and DFRC. We also demonstrate the use of a novel UV–Vis spectroscopic technique to rapidly screen plants for the presence of both ferulate and p-hydroxybenzoate esters. Lastly we show, via saccharification assays, that the OsFMT1 transgenic poplars have significantly improved processing efficiency compared to wild-type and Angelica sinensis-FMT-expressing poplars. The findings demonstrate that OsFMT1 has a broad substrate specificity and a higher catalytic efficiency compared to the previously published FMT from Angelica sinensis (AsFMT). Importantly, enhanced wood processability makes OsFMT1 a promising gene to optimize the composition of lignocellulosic biomass.

09 BIOMASS FUELS↗

Engineering Rhodosporidium toruloides for production of 3-hydroxypropionic acid from lignocellulosic hydrolysate

Microbial production of valuable bioproducts is a promising route towards green and sustainable manufacturing. The oleaginous yeast, Rhodosporidium toruloides, has emerged as an attractive host for the production of biofuels and bioproducts from lignocellulosic hydrolysates. 3-hydroxypropionic acid (3HP) is an attractive platform molecule that can be used to produce a wide range of commodity chemicals. This study focuses on establishing and optimizing the production of 3HP in R. toruloides. As R. toruloides naturally has a high metabolic flux towards malonyl-CoA, we exploited this pathway to produce 3HP. Upon finding the yeast capable of catabolizing 3HP, we then implemented functional genomics and metabolomic analysis to identify the catabolic pathways. Deletion of a putative malonate semialdehyde dehydrogenase gene encoding an oxidative 3HP pathway was found to significantly reduce 3HP degradation. We further explored monocarboxylate transporters to promote 3HP transport and identified a novel 3HP transporter in Aspergillus pseudoterreus by RNA-seq and proteomics. Combining these engineering efforts with media optimization in a fed-batch fermentation resulted in 45.4 g/L 3HP production. This represents one of the highest 3HP titers reported, and the highest titer produced from lignocellulosic hydrolysate to the best of our knowledge. This work demonstrates microbial production of 3HP in R. toruloides from lignocellulosic hydrolysate at high titers, and it represents a significant step toward enabling industrial production of 3HP in the future.

3-Hydroxypropionic acid↗

“Multiagent” Screening Improves Directed Enzyme Evolution by Identifying Epistatic Mutations

Enzyme evolution has enabled numerous advances in biotechnology and synthetic biology, yet still requires many iterative rounds of screening to identify optimal mutant sequences. This is due to the sparsity of the fitness landscape, which is caused by epistatic mutations that only offer improvements when combined with other mutations. We report an approach that incorporates diverse substrate analogues in the screening process, where multiple substrates act like multiple agents navigating the fitness landscape, identifying epistatic mutant residues without a need for testing the entire combinatorial search space. We initially validate this approach by engineering a malonyl-CoA synthetase and identify numerous epistatic mutations improving activity for several diverse substrates. The majority of these mutations would have been missed upon screening for a single substrate alone. We expect that this approach can accelerate a wide array of enzyme engineering programs.

60 APPLIED LIFE SCIENCES↗

Transcriptomic Analysis Reveals Novel Regulators of the Scots Pine Stilbene Pathway

Stilbenes accumulate in Scots pine heartwood where they have important roles in protecting wood from decaying fungi. They are also part of active defense responses, and their production is induced by different (a)biotic stressors. The specific transcriptional regulators as well as the enzyme responsible for activating the stilbene precursor cinnamate in the pathway are still unknown. UV-C radiation was the first discovered artificial stress activator of the pathway. Here, we describe a large-scale transcriptomic analysis of pine needles in response to UV-C and treatment with translational inhibitors, both activating the transcription of stilbene pathway genes. We used the data to identify putative candidates for the missing CoA ligase and for pathway regulators. We further showed that the pathway is transcriptionally activated by phosphatase inhibitor, ethylene and jasmonate treatments, as in grapevine, and that the stilbene synthase promoter retains its inducibility in some of the tested conditions in Arabidopsis, a species that normally does not synthesize stilbenes. Shared features between gymnosperm and angiosperm regulation and partially retained inducibility in Arabidopsis suggest that pathway regulation occurs not only via ancient stress-response pathway(s) but also via species-specific regulators. Understanding which genes control the biosynthesis of stilbenes in Scots pine aids breeding of more resistant trees.

4-Coumarate-CoA ligase↗

Insights into genetic determinants of volatile fatty acid catabolism in Cupriavidus necator H16

The soil bacterium Cupriavidus necator H16 is a promising host for upgrading waste-derived volatile fatty acids (VFAs) into renewable biochemicals. While bacterial VFA metabolic pathways are well understood, the C. necator genome encodes multiple enzymes for each catabolic step, and the degree of substrate specificity among these homologs is currently unknown. To gain insight into the catabolism of VFA substrates in C. necator, we performed transcriptomics on cells grown with acetate, propionate, butyrate, valerate, or hexanoate as the sole source of carbon and energy. These data revealed that C. necator upregulates multiple sets of genes putatively involved in substrate activation and β-oxidation in response to VFAs. To better understand this redundancy, we performed biochemical and genetic deletion studies of acyl-CoA synthetase enzymes upregulated during growth on VFA substrates. These results demonstrated the functional redundancy of the C. necator VFA catabolism and led to the identification of a gene cluster, H16_B1332-H16_B1337, that contains several genes that are important for the efficient catabolism of hexanoate. Constitutive expression of a second copy of these hexanoate catabolism genes did not improve growth of C. necator on hexanoate, suggesting that other factors (e.g., redox, transport, or toxicity) may be limiting for growth. Collectively, this work provides new insight into how C. necator uses metabolic regulation to effectively utilize VFA substrates and uncovers the important role of the gene cluster H16_B1332-H16_B1337 in the catabolism of hexanoate.

09 BIOMASS FUELS↗

Accumulation of Succinyl Coenzyme A Perturbs the Methicillin-Resistant Staphylococcus aureus (MRSA) Succinylome and Is Associated with Increased Susceptibility to Beta-Lactam Antibiotics

mecA -dependent methicillin resistance in MRSA is subject to regulation by numerous accessory factors involved in cell wall biosynthesis, nucleotide signaling, and central metabolism. Here, we report that mutations in the TCA cycle gene, sucC , increased susceptibility to β-lactam antibiotics and was accompanied by significant accumulation of succinyl-CoA, which in turn perturbed lysine succinylation in the proteome.

59 BASIC BIOLOGICAL SCIENCES↗

High–level β–carotene production from xylose by engineered Saccharomyces cerevisiae without overexpression of a truncated HMG1 (t HMG1 )

Beta-carotene is a natural pigment and health-promoting metabolite, and has been widely used in the nutraceutical, feed, and cosmetic industries. Here, we engineered a GRAS yeast Saccharomyces cerevisiae to produce beta-carotene from xylose, the sec- ond most abundant and inedible sugar component of lignocellulose biomass. Specifically, a beta-carotene biosynthetic pathway containing crtYB, crtI, and crtE from Xanthophyllomyces dendrorhous was introduced into a xylose-fermenting S. cerevisiae. Here, the resulting strain produced beta-carotene from xylose at a titer threefold higher than from glucose. Interestingly, overexpression of tHMG1, which has been reported as a critical genetic perturbation to enhance metabolic fluxes in the mevalonate pathway and beta-carotene production in yeast when glucose is used, did not further improve the production of beta-carotene from xylose. Through fermentation profiling, metabolites analysis, and transcriptional studies, we found the advantages of using xylose as a carbon source, instead of glucose, for beta-carotene production to be a more respiratory feature of xylose consumption, a larger cytosolic acetyl!CoA pool, and an upregulated expression level of rate-limiting genes in the beta-carotene!producing pathway, including ACS1 and HMG1. As a result, 772.8 mg/L of beta-carotene was ob- tained in a fed-batch bioreactor culture with xylose feeding. Considering the in- evitable large scale production of xylose when cellulosic biomass!based bioeconomy is implemented, our results suggest xylose utilization is a promising strategy for overproduction of carotenoids and other isoprenoids in engineered S. cerevisiae.

09 BIOMASS FUELS↗