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At least 1,567 records · Page 87

Genome, transcriptome and secretome analyses of the antagonistic, yeast-like fungus Aureobasidium pullulans to identify potential biocontrol genes

Aureobasidium pullulans is an extremotolerant, cosmopolitan yeast-like fungus that successfully colonises vastly different ecological niches. The species is widely used in biotechnology and successfully applied as a commercial biocontrol agent against postharvest diseases and fireblight. However, the exact mechanisms that are responsible for its antagonistic activity against diverse plant pathogens are not known at the molecular level. Thus, it is difficult to optimise and improve the biocontrol applications of this species. As a foundation for elucidating biocontrol mechanisms, we have de novo assembled a high-quality reference genome of a strongly antagonistic A. pullulans strain, performed dual RNA-seq experiments, and analysed proteins secreted during the interaction with the plant pathogen Fusarium oxysporum. Based on the genome annotation, potential biocontrol genes were predicted to encode secreted hydrolases or to be part of secondary metabolite clusters (e.g., NRPS-like, NRPS, T1PKS, terpene, and β-lactone clusters). Transcriptome and secretome analyses defined a subset of 79 A. pullulans genes (among the 10,925 annotated genes) that were transcriptionally upregulated or exclusively detected at the protein level during the competition with F. oxysporum. These potential biocontrol genes comprised predicted secreted hydrolases such as glycosylases, esterases, and proteases, as well as genes encoding enzymes, which are predicted to be involved in the synthesis of secondary metabolites. This study highlights the value of a sequential approach starting with genome mining and consecutive transcriptome and secretome analyses in order to identify a limited number of potential target genes for detailed, functional analyses.

transcriptome↗

SERDP PFAS Database

The PNNL-PFAS database was constructed to provide quantitative infrared spectra of per- and polyfluoroalkyl substances (PFAS) in the gas phase. These fluorine-rich, organic compounds have been used as non-flammable solvents, cleaning agents, and in the manufacture of a diverse variety of consumer products. Due to their widespread use and relative inertness, PFAS have accumulated in the natural environment, and exposure to these chemicals may be linked to harmful health effects. Development of methods for detecting and remediating PFAS contamination are thus active areas of research. This database supports PFAS investigations where gas-phase infrared signatures may be measured (e.g., during treatment by thermal decomposition). The database currently contains infrared spectra for 15 compounds; these were selected based on the potential of these compounds being thermal degradation products of the manufactured chemicals mentioned above. • Additional compounds will be added to the database in the future. • Additional information about the database may be obtained from [link to future paper about the PFAS database].

PFAS, infrared(IR)spectroscopy, DATABASE, FTIR, sp↗

High Temperature Stability of Aqueous Foams for Potential Application in Enhanced Geothermal System (EGS)

Enhanced Geothermal Systems (EGS) are typically constructed by injecting high-pressure water into the deep hot dry rocks (HDR) under carefully controlled conditions to create new or re-open existing fractures, which usually utilizes an immense quantity of water. Alternatively, aqueous foam-based fracturing fluids may offer potential advantages over conventional water-based fracturing fluid, including reduced consumption of water and environmental impact. Although foam-based fracturing has shown promising results in oil and gas industries, its applicability to EGS is unknown. Foams are complex mixtures of liquids with gaseous phases that are thermodynamically unstable. Their stability will decrease over time due to liquid drainage, bubble coarsening, and coalescence. Therefore, it is essential to stabilize foam fluids at high temperatures for EGS related applications such as fracking of HDRs.This paper is focused on investigating high-temperature stability of N2-in-water or N2 foams containing different surfactants and stabilizing agents. Foam stability was characterized by the half-life, which is defined as the time a foam decreases to 50% of its original height due to drainage. Surfactants with various concentrations dissolved in deionized water were investigated, including alfa olefin sulfonate (AOS), sodium dodecyl sulfonate (SDS), TergitolTM (NP – 40), and cetyltrimethylammonium chloride (CTAC). Also, the effects of other stabilizing agents, including guar gum, bentonite clay, crosslinker, graphene oxide (GO) dispersion on foam stability were tested. Initial results showed that foam stability decreased dramatically as temperature increased. On the other hand, foams became relatively more stable as pressure increased. Certain stabilizing agents, such as guar gum and crosslinker, can enhance foam stability at a wide range of temperatures. Our results indicate that it may be possible to obtain stable foams at high-temperature, high-pressure conditions with appropriate stabilizing agents.

Thakore, Virensinh↗

Structure- and Interaction-Based Design of Anti-SARS-CoV-2 Aptamers

Aptamer selection against novel infections is a complicated and time-consuming approach. Synergy can be achieved by using computational methods together with experimental procedures. In this study, we aim to develop a reliable methodology for a rational aptamer in silico et vitro design. The new approach combines multiple steps: (1) Molecular design, based on screening in a DNA aptamer library and directed mutagenesis to fit the protein tertiary structure; (2) 3D molecular modeling of the target; (3) Molecular docking of an aptamer with the protein; (4) Molecular dynamics (MD) simulations of the complexes; (5) Quantum-mechanical (QM) evaluation of the interactions between aptamer and target with further analysis; (6) Experimental verification at each cycle for structure and binding affinity by using small-angle X-ray scattering, cytometry, and fluorescence polarization. By using a new iterative design procedure, structure- and interaction-based drug design (SIBDD), a highly specific aptamer to the receptor-binding domain of the SARS-CoV-2 spike protein, was developed and validated. The SIBDD approach enhances speed of the high-affinity aptamers development from scratch, using a target protein structure. The method could be used to improve existing aptamers for stronger binding. This approach brings to an advanced level the development of novel affinity probes, functional nucleic acids. It offers a blueprint for the straightforward design of targeting molecules for new pathogen agents and emerging variants.

60 APPLIED LIFE SCIENCES↗

Transactive Energy Communications Interface Standards Landscape

Transactive energy represents distributed, optimal-seeking coordination approaches for managing the operation of large numbers of energy-related devices and systems. The concept assumes intelligent agents interact with other agents on behalf of their owners to balance the supply and demand of energy and coordinate other operation services in an electric system. They do this by exchanging value signals for services in near-term and forward time periods using market mechanisms. The technique is particularly applied to coordination of flexibility in operating distributed energy resources. The integration of large numbers of devices requires that they be able to connect and interoperate easily and reliably. Given the many technologies and solution providers integrating products, communications interfaces based on clear, unambiguous specifications with supporting tests require standardization and adoption by the community of system integrators. This report reviews the state of communications interface standards that show promise for transactive energy approaches to the coordination of distributed energy resources. It reviews promising standards to highlight challenges and gaps. It offers structured comparisons between standards and the features offered by their ecosystems of participants.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Proteomic Retrieval from Nucleic Acid Depleted Space-Flown Human Cells

Compared to experiments utilizing humans in microgravity, cell-based approaches to questions about subsystems of the human system afford multiple advantages, such as crew safety and the ability to achieve statistical significance. To maximize the science return from flight samples, an optimized method was developed to recover protein from samples depleted of nucleic acid. This technique allows multiple analyses on a single cellular sample and when applied to future cellular investigations could accelerate solutions to significant biomedical barriers to human space exploration. Cell cultures grown in American Fluoroseal bags were treated with an RNA stabilizing agent (RNAlater - Ambion), which enabled both RNA and immunoreactive protein analyses. RNA was purified using an RNAqueous(registered TradeMark) kit (Ambion) and the remaining RNA free supernatant was precipitated with 5% trichloroacetic acid. The precipitate was dissolved in SDS running buffer and tested for protein content using a bicinchoninic acid assay (1) (Sigma). Equal loads of protein were placed on SDS-PAGE gels and either stained with CyproOrange (Amersham) or transferred using Western Blotting techniques (2,3,4). Protein recovered from RNAlater-treated cells and stained with protein stain, was measured using Imagequant volume measurements for rectangles of equal size. BSA treated in this way gave quantitative data over the protein range used (Fig 1). Human renal cortical epithelial (HRCE) cells (5,6,7) grown onboard the International Space Station (ISS) during Increment 3 and in ground control cultures exhibited similar immunoreactivity profiles for antibodies to the Vitamin D receptor (VDR) (Fig 2), the beta isoform of protein kinase C (PKC ) (Fig 3), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Fig 4). Parallel immunohistochemical studies on formalin-fixed flight and ground control cultures also showed positive immunostaining for VDR and other biomarkers (Fig 5). These results are consistent with data from additional antigenic recovery experiments performed on human Mullerian tumor cells cultured in microgravity (8).

Hammond, D. K.↗

Team play with a powerful and independent agent: operational experiences and automation surprises on the Airbus A-320

Research and operational experience have shown that one of the major problems with pilot-automation interaction is a lack of mode awareness (i.e., the current and future status and behavior of the automation). As a result, pilots sometimes experience so-called automation surprises when the automation takes an unexpected action or fails to behave as anticipated. A lack of mode awareness and automation surprises can he viewed as symptoms of a mismatch between human and machine properties and capabilities. Changes in automation design can therefore he expected to affect the likelihood and nature of problems encountered by pilots. Previous studies have focused exclusively on early generation "glass cockpit" aircraft that were designed based on a similar automation philosophy. To find out whether similar difficulties with maintaining mode awareness are encountered on more advanced aircraft, a corpus of automation surprises was gathered from pilots of the Airbus A-320, an aircraft characterized by high levels of autonomy, authority, and complexity. To understand the underlying reasons for reported breakdowns in human-automation coordination, we also asked pilots about their monitoring strategies and their experiences with and attitude toward the unique design of flight controls on this aircraft.

NASA Discipline Space Human Factors↗

Durable polymer-aerogel based superhydrophobic coatings: a composite material

Provided are polymer-aerogel composite coatings, devices and articles including polymer-aerogel composite coatings, and methods for preparing the polymer-aerogel composite. The exemplary article can include a surface, wherein the surface includes at least one region and a polymer-aerogel composite coating disposed over the at least one region, wherein the polymer-aerogel composite coating has a water contact angle of at least about 140° and a contact angle hysteresis of less than about 1°. The polymer-aerogel composite coating can include a polymer and an ultra high water content catalyzed polysilicate aerogel, the polysilicate aerogel including a three dimensional network of silica particles having surface functional groups derivatized with a silylating agent and a plurality of pores.

Kissel, David J.↗

Real-Time and Online Monitoring of Glucose Contents by Using Molecular Imprinted Polymer-Based IDEs Sensor

A highly sensitive, selective, reversible, and reusable glucose sensor is developed by using molecularly imprinted polymer-based artificial receptors onto interdigital transducer. Sensor receptors were synthesized through bulk imprinting technology by using styrene as monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker, and AIBN as free radical initiator. Topography of the synthesized receptors was investigated by scanning electron microscopy (SEM). Fabricated sensor showed concentration-dependent linear and reversible response with lower limit of detection of 30 ppb and upper limit of detection ~ 500 ppm. Furthermore, newly fabricated sensor is highly selective towards its analyte of interest in the presence of other competing agents, and the regeneration of sensor response has been assessed with the percentage error of less than 2% under the period of 1 year at room temperature and pressure conditions. The reported sensor may have potential technological applications in the field of medical diagnostics, food, and pharmaceutical industry.

60 APPLIED LIFE SCIENCES↗

Molecular Technique to Reduce PCR Bias for Deeper Understanding of Microbial Diversity

Current planetary protection policies require that spacecraft targeted to sensitive solar system bodies be assembled and readied for launch in controlled cleanroom environments. A better understanding of the distribution and frequency at which high-risk contaminant microbes are encountered on spacecraft surfaces would significantly aid in assessing the threat of forward contamination. However, despite a growing understanding of the diverse microbial populations present in cleanrooms, less abundant microbial populations are probably not adequately taken into account due to technological limitations. This novel approach encompasses a wide spectrum of microbial species and will represent the true picture of spacecraft cleanroom-associated microbial diversity. All of the current microbial diversity assessment techniques are based on an initial PCR amplification step. However, a number of factors are known to bias PCR amplification and jeopardize the true representation of bacterial diversity. PCR amplification of a minor template appears to be suppressed by the amplification of a more abundant template. It is widely acknowledged among environmental molecular microbiologists that genetic biosignatures identified from an environment only represent the most dominant populations. The technological bottleneck overlooks the presence of the less abundant minority population and may underestimate their role in the ecosystem maintenance. DNA intercalating agents such as propidium monoazide (PMA) covalently bind with DNA molecules upon photolysis using visible light, and make it unavailable for DNA polymerase enzyme during polymerase chain reaction (PCR). Environmental DNA samples will be treated with suboptimum PMA concentration, enough to intercalate with 90 99% of the total DNA. The probability of PMA binding with DNA from abundant bacterial species will be much higher than binding with DNA from less abundant species. This will increase the relative DNA concentration of previously "shadowed" less abundant species available for PCR amplification. These PCR products obtained with and without PMA treatment will then be subjected to downstream diversity analyses such as sequencing and DNA microarray. It is expected that PMA-coupled PCR will amplify the "minority population" and help in understanding microbial diversity spectrum of an environmental sample at a much deeper level. This new protocol aims to overcome the major potential biases faced when analyzing microbial 16S rRNA gene diversity. This study will lead to a technological advancement and a commercial product that will aid microbial ecologists in understanding microbial diversity from various environmental niches. Implementation of this technique may lead to discoveries of novel microbes and their functions in sustenance of the ecosystem.

Vaishampayan, Parag A.↗

An Automated Approach to Map the History of Forest Disturbance from Insect Mortality and Harvest with Landsat Time-Series Data

Forests contain a majority of the aboveground carbon (C) found in ecosystems, and understanding biomass lost from disturbance is essential to improve our C-cycle knowledge. Our study region in the Wisconsin and Minnesota Laurentian Forest had a strong decline in Normalized Difference Vegetation Index (NDVI) from 1982 to 2007, observed with the National Ocean and Atmospheric Administration's (NOAA) series of Advanced Very High Resolution Radiometer (AVHRR). To understand the potential role of disturbances in the terrestrial C-cycle, we developed an algorithm to map forest disturbances from either harvest or insect outbreak for Landsat time-series stacks. We merged two image analysis approaches into one algorithm to monitor forest change that included: (1) multiple disturbance index thresholds to capture clear-cut harvest; and (2) a spectral trajectory-based image analysis with multiple confidence interval thresholds to map insect outbreak. We produced 20 maps and evaluated classification accuracy with air-photos and insect air-survey data to understand the performance of our algorithm. We achieved overall accuracies ranging from 65% to 75%, with an average accuracy of 72%. The producer's and user's accuracy ranged from a maximum of 32% to 70% for insect disturbance, 60% to 76% for insect mortality and 82% to 88% for harvested forest, which was the dominant disturbance agent. Forest disturbances accounted for 22% of total forested area (7349 km2). Our algorithm provides a basic approach to map disturbance history where large impacts to forest stands have occurred and highlights the limited spectral sensitivity of Landsat time-series to outbreaks of defoliating insects. We found that only harvest and insect mortality events can be mapped with adequate accuracy with a non-annual Landsat time-series. This limited our land cover understanding of NDVI decline drivers. We demonstrate that to capture more subtle disturbances with spectral trajectories, future observations must be temporally dense to distinguish between type and frequency in heterogeneous landscapes.

Forest↗

Chaotrope-Based Approach for Rapid In Vitro Assembly and Loading of Bacterial Microcompartment Shells

Bacterial microcompartments (BMCs) are proteinaceous organelles that self-assemble into selectively permeable shells that encapsulate enzymatic cargo. BMCs enhance catalytic pathways by reducing crosstalk among metabolites, preventing harmful intermediates from leaking into the cytosol and increasing reaction efficiency via enzyme colocalization. The intrinsic properties of BMCs make them attractive for biotechnological engineering. However, in vivo expression methods for shell synthesis have significant drawbacks that limit the potential design space for these nanocompartments. Here, we describe the development of an efficient and rapid method for the in vitro assembly of BMC shells from their protein building blocks. Our method enables large-scale construction of BMC shells by utilizing urea as a chaotropic agent to control self-assembly and provides an approach for encapsulation of both biotic and abiotic cargo under a broad range of reaction conditions. We demonstrate an enhanced level of control over the assembly of BMC shells in vitro and expand the design parameter space for engineering BMC systems with specialized and enhanced catalytic properties.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Developing A Dependable Multi-Agent Rover Swarm Using cFS

The future of space exploration lies in cooperative autonomous systems. Ensuring their high integrity remains a challenge. The Robust Software Engineering group at NASA Ames Research Center has been developing the Troupe project to explore the challenges with developing and assuring high integrity of cooperative autonomous robotic systems. In particular, Troupe aims to develop a swarm of autonomous rovers capable of mapping unknown terrain and assure their high integrity using the advanced V&V tools developed in the group. In this paper, we present the evolution of the design of Troupe. We focus on the lessons learned in developing and assuring the rover swarm using core Flight System (cFS). In particular, we discuss the benefits and challenges in applying model-based development to develop the rover swarm.

space systems↗

Developing A Dependable Multi-Agent Rover Swarm Using cFS

The future of space exploration lies in cooperative autonomous systems. Ensuring their high integrity remains a challenge. The Robust Software Engineering group at NASA Ames Research Center has been developing the Troupe project to explore the challenges with developing and assuring high integrity of cooperative autonomous robotic systems. In particular, Troupe aims to develop a swarm of autonomous rovers capable of mapping unknown terrain and assure their high integrity using the advanced V&V tools developed in the group. In this paper, we present the evolution of the design of Troupe. We focus on the lessons learned in developing and assuring the rover swarm using core Flight System (cFS). In particular, we discuss the benefits and challenges in applying model-based development to develop the rover swarm.

space systems↗

Distributed optimization for multi-commodity urban traffic control

A distributed method for concurrent traffic signal and routing control of traffic networks is proposed. The method is based on the multi-commodity store-and-forward model, in which the destinations are the commodities. The system benefits from the communication between vehicles and infrastructure, providing optimal signal timings to intersections and routes to vehicles on a link-by-link basis. Using the augmented Lagrangian to model the constraints into the objective, the baseline centralized problem is decomposed into a set of objective-coupled subproblems, one for each intersection, enabling the solution to be computed by a distributed- gradient projection algorithm. Further, the intersection agents only need to communicate and coordinate with neighboring intersections to ensure convergence to the optimal solution while tolerating suboptimal iterations that offer more flexibility, unlike other distributed approaches. Through microsimulation, we demonstrate the effectiveness of the proposed algorithm in traffic networks with time-varying demand. Computational analysis shows that the distributed problem is suitable for real-time applications. A robustness analysis show that the distributed formulation enables a graceful degradation of the system in case of failure.

Augmented Lagrangian↗

Distributional Deep Reinforcement Learning-Based Emergency Frequency Control

Emergency frequency control is one of the most critical approaches to maintain power system stability after major disturbances. With the increasing number of grid-connected renewable energy sources, existing model-based methods of frequency control are facing up with challenges of computational speed and scalability for large-scale systems. In this paper, the emergency frequency control problem is formulated as a Markov Decision Process (MDP) and solved through a novel Distributional Deep Reinforcement Learning (DDRL) method, namely the distributional soft actor critic (DSAC) method. Compared with other RL methods that only estimate the mean value, the proposed DSAC model estimates the distribution of value function over returns. This advancement can lead to more insights and knowledge for the agent, with the benefit of a much faster and more stable learning process, and the improved frequency control performance. Here, the simulation results on IEEE 39-bus and IEEE 118-bus systems demonstrate the effectiveness and robustness of proposed models, as well as the advantage compared to other state-of-the-art DRL algorithms.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Nanoparticle Injection Technology for Remediating Leaks of CO 2 Storage Formation (Final Report)

Leakage of a well can occur through the wellbore, the annulus between well tubing and steel casing, or on the outside of the casing. The overall goal of the project is to develop a new technology that can repair or reduce the leakage of wellbore. The new technology is based on simultaneously injecting nanoparticles electrochemically and removing some of the harmful ions in the system so the distressed cement materials can be densified and the corrosion risk of steel casing can be reduced. There were two objectives. One is the development of the injection technology for leakage repair and the other is the development of new numerical simulation models that can simulate and predict the performance of the new repair technology. An electro-migration testing unit and a in-house wellbore testing system were developed; healing agents with nanoparticles were selected, numerical models were developed to simulate the electro-migration technology, and the effectiveness of the new technology were evaluated.

42 ENGINEERING↗

Nanoparticle Injection Technology for Remediating Leaks of CO 2 Storage Formation (Final Technical Report)

Leakage of a well can occur through the wellbore, the annulus between well tubing and steel casing, or on the outside of the casing. The overall goal of the project is to develop a new technology that can repair or reduce the leakage of wellbore. The new technology is based on simultaneously injecting nanoparticles electrochemically and removing some of the harmful ions in the system so the distressed cement materials can be densified and the corrosion risk of steel casing can be reduced. There were two objectives. One is the development of the injection technology for leakage repair and the other is the development of new numerical simulation models that can simulate and predict the performance of the new repair technology. An electro-migration testing unit and an in-house wellbore testing system were developed; healing agents with nanoparticles were selected, numerical models were developed to simulate the electro-migration technology, and the effectiveness of the new technology was evaluated.

42 ENGINEERING↗