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Digital Droplet PCR and Mesocosm-Based Methods to Evaluate Biocontainment Strategies in a Native Soil Ecosystem

Genetically modified industrial production microbes and their associated bioproducts have emerged as an integral component of a sustainable bioeconomy. However, the rapid development of these innovative technologies raises biosecurity concerns, namely, the risk of environmental escape. Thus, the realization of a bioeconomy hinges not only on the development and deployment of microbial production hosts, but also on the development of secure biosystems and biocontainment designs. Current laboratory-based biocontainment testing systems do not accurately reflect the complexities found in natural environments, necessitating an environmentally relevant analysis pipeline that allows for the detection of rare escapees within a complex soil microbiome and differentiation between closely related strains. To this end, we have developed an approach that utilizes soil mesocosms and integrated digital droplet PCR (ddPCR) system to evaluate the efficacy of novel biocontainment strategies. We demonstrate the utility of this approach by modeling contamination with industrial microbial chasses versus their biocontained counterparts. Here we demonstrate the broad utility of this system by highlighting findings from strains of Saccharomyces cerevisiae that are contained with an inducible toxin anti-toxin system, strains of Synechocystis sp. PCC 6803 contained via gene knockout or toxin anti-toxin system, and strains of Escherichia coli that are contained via genomic recoding. We also show that ddPCR can be used to detect gene copies from E. coli equal to those counted by traditional spot plating assays. The resultant data demonstrates that this system has broad utility across diverse microbial chassis and biocontainment strategies and enables researchers to track the fate of our contaminating microbe with high sensitivity in the soil. The findings presented here support the use of this mesocosm-based approach to assess the environmental impact of industrial microbes and to validate biocontainment strategies.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH

Structural analysis model validation using modal test data

The design of a structure which can survive a prescribed dynamic environment is generally based on the use of an analytical model of the structure. The modal test is employed to verify the analytical model. This paper is concerned with the involved validation process, taking into account modal test results from realistic complex spacecraft structural systems as examples to demonstrate the proposed validation procedure. Attention is given to a load analysis model definition, a pretest analysis and test/analysis model, and the correlation parameters. Coordinate transformation is considered along with mode identification, frequency orthogonality, effective mass, mode shape, local kinetic energy, modal force, strain energy, and modal forces by acceleration measurements. Two examples are used to illustrate the procedure.

Chen, J.-C.

NASA Applications for Computational Electromagnetic Analysis

Computational Electromagnetic Software is used by NASA to analyze the compatibility of systems too large or too complex for testing. Recent advances in software packages and computer capabilities have made it possible to determine the effects of a transmitter inside a launch vehicle fairing, better analyze the environment threats, and perform on-orbit replacements with assured electromagnetic compatibility.

Lewis, Catherine C.

THz Instruments for Space Exploration

THz heterodyne spectrometers, capable of providing spectral resolution of >106 and detection sensitivity in the parts-per-billion range, provide a unique capability for space exploration. The exact operating frequency and technology for the instrument is driven largely by the science that is being investigated along with pragmatic concerns for mass and power requirements. These instruments are examples of highly complex systems that involve design, integration and testing of diverse technologies such as THz cryogenic detectors, optical elements, and microwave and submillimeter-wave components. This paper will review some of the advanced microwave and submillimeter-wave technologies that are being developed to create the next generation of THz instruments for space exploration.

Mehdi, Imran

Exploring the Use of Large Immersive Display Systems in the Nuclear Industry

Large immersive display systems play a critical role in the nuclear industry by enabling advanced training, design reviews, scientific visualization, and safety simulations. These systems, such as CAVE and powerwalls, allow engineers and operators to interact with virtual nuclear environments in real-time, providing a deeper understanding of complex systems. Their ability to simulate real-world scenarios for testing and optimization in a safe, controlled environment ensures that operators and engineers can refine process, enhance safety protocols, and troubleshoot complex challenges. This article outlines key lessons learned from deploying these systems, practical insights into their applications, and considerations for future improvements, aiming to guide their broader adoption and effective use in the nuclear industry.

99 - GENERAL AND MISCELLANEOUS

Spacecraft structural system identification by modal test

A structural parameter estimation procedure using the measured natural frequencies and kinetic energy distribution as observers is proposed. The theoretical derivation of the estimation procedure is described and its constraints and limitations are explained. This procedure is applied to a large complex spacecraft structural system to identify the inertia matrix using modal test results. The inertia matrix is chosen after the stiffness matrix has been updated by the static test results.

Chen, J.-C.

UTM Technical Capability Level 4 Testing

NASA's UTM project is conducting research on a traffic management concept for small unmanned aircraft systems (UAS) flying in low altitude, uncontrolled airspace. The project started in 2015 and is developing prototype UTM systems of successively complex technical capability levels (TCL) that are tested in the field. To date TCL levels 1-3 have been tested and TCL 4 will be tested in the summer of 2019. Project results are transferred to the FAA and industry to advance the adoption, implementation, and design standards of future UTM systems.

Johnson, Ronald D.

AdaStress

This is a tutorial on AdaStress, a tool for finding and analyzing the likeliest failures in a simulated system under test. The presentation outlines the adaptive stress testing framework, provides a demonstration of use, and showcases several examples of failure detection in a complex real-world system.

Reinforcement learning

Spinoff 2010

Topics covered include: Burnishing Techniques Strengthen Hip Implants; Signal Processing Methods Monitor Cranial Pressure; Ultraviolet-Blocking Lenses Protect, Enhance Vision; Hyperspectral Systems Increase Imaging Capabilities; Programs Model the Future of Air Traffic Management; Tail Rotor Airfoils Stabilize Helicopters, Reduce Noise; Personal Aircraft Point to the Future of Transportation; Ducted Fan Designs Lead to Potential New Vehicles; Winglets Save Billions of Dollars in Fuel Costs; Sensor Systems Collect Critical Aerodynamics Data; Coatings Extend Life of Engines and Infrastructure; Radiometers Optimize Local Weather Prediction; Energy-Efficient Systems Eliminate Icing Danger for UAVs; Rocket-Powered Parachutes Rescue Entire Planes; Technologies Advance UAVs for Science, Military; Inflatable Antennas Support Emergency Communication; Smart Sensors Assess Structural Health; Hand-Held Devices Detect Explosives and Chemical Agents; Terahertz Tools Advance Imaging for Security, Industry; LED Systems Target Plant Growth; Aerogels Insulate Against Extreme Temperatures; Image Sensors Enhance Camera Technologies; Lightweight Material Patches Allow for Quick Repairs; Nanomaterials Transform Hairstyling Tools; Do-It-Yourself Additives Recharge Auto Air Conditioning; Systems Analyze Water Quality in Real Time; Compact Radiometers Expand Climate Knowledge; Energy Servers Deliver Clean, Affordable Power; Solutions Remediate Contaminated Groundwater; Bacteria Provide Cleanup of Oil Spills, Wastewater; Reflective Coatings Protect People and Animals; Innovative Techniques Simplify Vibration Analysis; Modeling Tools Predict Flow in Fluid Dynamics; Verification Tools Secure Online Shopping, Banking; Toolsets Maintain Health of Complex Systems; Framework Resources Multiply Computing Power; Tools Automate Spacecraft Testing, Operation; GPS Software Packages Deliver Positioning Solutions; Solid-State Recorders Enhance Scientific Data Collection; Computer Models Simulate Fine Particle Dispersion; Composite Sandwich Technologies Lighten Components; Cameras Reveal Elements in the Short Wave Infrared; Deformable Mirrors Correct Optical Distortions; Stitching Techniques Advance Optics Manufacturing; Compact, Robust Chips Integrate Optical Functions; Fuel Cell Stations Automate Processes, Catalyst Testing; Onboard Systems Record Unique Videos of Space Missions; Space Research Results Purify Semiconductor Materials; and Toolkits Control Motion of Complex Robotics.

Source record

An Architecture for Real-Time Interpretation and Visualization of Structural Sensor Data in a Laboratory Environment

A visualization system is being developed out of the need to monitor, interpret, and make decisions based on the information from several thousand sensors during experimental testing to facilitate development and validation of structural health monitoring algorithms. As an added benefit the system will enable complete real-time sensor assessment of complex test specimens. Complex structural specimens are routinely tested that have hundreds or thousands of sensors. During a test, it is impossible for a single researcher to effectively monitor all the sensors and subsequently interesting phenomena occur that are not recognized until post-test analysis. The ability to detect and alert the researcher to these unexpected phenomena as the test progresses will significantly enhance the understanding and utilization of complex test articles. Utilization is increased by the ability to halt a test when the health monitoring algorithm response is not satisfactory or when an unexpected phenomenon occurs, enabling focused investigation potentially through the installation of additional sensors. Often if the test continues, structural changes make it impossible to reproduce the conditions that exhibited the phenomena. The prohibitive time and costs associated with fabrication, sensoring, and subsequent testing of additional test articles generally makes it impossible to further investigate the phenomena. A scalable architecture is described to address the complex computational demands of structural health monitoring algorithm development and laboratory experimental test monitoring. The researcher monitors the test using a photographic quality 3D graphical model with actual sensor locations identified. In addition, researchers can quickly activate plots displaying time or load versus selected sensor response along with the expected values and predefined limits. The architecture has several key features. First, distributed dissimilar computers may be seamlessly integrated into the information flow. Second, virtual sensors may be defined that are complex functions of existing sensors or other virtual sensors. Virtual sensors represent a calculated value not directly measured by particular physical instrument. They can be used, for example, to represent the maximum difference in a range of sensors or the calculated buckling load based on the current strains. Third, the architecture enables autonomous response to preconceived events, where by the system can be configured to suspend or abort a test if a failure is detected in the load introduction system. Fourth, the architecture is designed to allow cooperative monitoring and control of the test progression from multiple stations both remote and local to the test system. To illustrate the architecture, a preliminary implementation is described monitoring the Stitched Composite Wing recently tested at LaRC.

Doggett, William

Evaluation of modal testing methods

Modal tests are playing an increasingly important role in structural dynamics efforts which are in need of analytical model verification or trouble shootings. In the meantime, the existing modal testing methods are undergoing great changes as well as new methods are being created. Although devoted advocates of each method can be found to argue the relative advantages and disadvantages, the general superiority, if any, of one or the other is not yet evident. The Galileo spacecraft, a realistic, complex structural system, will be used as a test article for performing modal tests by various methods. The results will be used to evaluate the relative merits of the various modal testing methods.

Chen, J.-C.

Development of an Expert System for Representing Procedural Knowledge

A high level of automation is of paramount importance in most space operations. It is critical for unmanned missions and greatly increases the effectiveness of manned missions. However, although many functions can be automated by using advanced engineering techniques, others require complex reasoning, sensing, and manipulatory capabilities that go beyond this technology. Automation of fault diagnosis and malfunction handling is a case in point. The military have long been interested in this problem, and have developed automatic test equipment to aid in the maintenance of complex military hardware. These systems are all based on conventional software and engineering techniques. However, the effectiveness of such test equipment is severely limited. The equipment is inflexible and unresponsive to the skill level of the technicians using it. The diagnostic procedures cannot be matched to the exigencies of the current situation nor can they cope with reconfiguration or modification of the items under test. The diagnosis cannot be guided by useful advice from technicians and, when a fault cannot be isolated, no explanation is given as to the cause of failure. Because these systems perform a prescribed sequence of tests, they cannot utilize knowledge of a particular situation to focus attention on more likely trouble spots. Consequently, real-time performance is highly unsatisfactory. Furthermore, the cost of developing test software is substantial and time to maturation is excessive. Significant advances in artificial intelligence (AI) have recently led to the development of powerful and flexible reasoning systems, known as expert or knowledge-based systems. We have devised a powerful and theoretically sound scheme for representing and reasoning about procedural knowledge.

Georgeff, Michael P.

Investigating lab-scaled offshore wind aerodynamic testing failure and developing solutions for early anomaly detections

As offshore wind systems become more complex, the risk of human error or equipment malfunction increases during experimental testing. This study investigates a lab-scale incident involving a 1 : 50 scale 5 MW wind turbine, where a generator failure led to rotor overspeed and a blade–tower strike. To improve early fault detection, we propose a data-driven method based on multivariate long short-term memory (LSTM) models. High-frequency measurements are projected onto principal components, and anomalies are identified using reconstruction error and its time derivative. Two models are trained on different healthy datasets and tested using single- and multi-principal component (1PC and MPC) variations. Results show that combining both error and error derivative improves detection accuracy. The 1PC model detects faults faster, has a higher recall rate, and achieves a 43 % improvement in anomaly detection accuracy, while the MPC model yields higher precision. This approach provides a simple and effective tool for early anomaly detection in lab-scale experiments, helping to reduce the risk of future failures during the testing of new technologies.

17 WIND ENERGY

Practical Applications of Cosmic Ray Science: Spacecraft, Aircraft, Ground Based Computation and Control Systems and Human Health and Safety

In this paper we review the discovery of cosmic ray effects on the performance and reliability of microelectronic systems as well as on human health and safety, as well as the development of the engineering and health science tools used to evaluate and mitigate cosmic ray effects in earth surface, atmospheric flight, and space flight environments. Three twentieth century technological developments, 1) high altitude commercial and military aircraft; 2) manned and unmanned spacecraft; and 3) increasingly complex and sensitive solid state micro-electronics systems, have driven an ongoing evolution of basic cosmic ray science into a set of practical engineering tools (e.g. ground based test methods as well as high energy particle transport and reaction codes) needed to design, test, and verify the safety and reliability of modern complex electronic systems as well as effects on human health and safety. The effects of primary cosmic ray particles, and secondary particle showers produced by nuclear reactions with spacecraft materials, can determine the design and verification processes (as well as the total dollar cost) for manned and unmanned spacecraft avionics systems. Similar considerations apply to commercial and military aircraft operating at high latitudes and altitudes near the atmospheric Pfotzer maximum. Even ground based computational and controls systems can be negatively affected by secondary particle showers at the Earth's surface, especially if the net target area of the sensitive electronic system components is large. Accumulation of both primary cosmic ray and secondary cosmic ray induced particle shower radiation dose is an important health and safety consideration for commercial or military air crews operating at high altitude/latitude and is also one of the most important factors presently limiting manned space flight operations beyond low-Earth orbit (LEO).

Atwell, William

Preliminary Application of Formal Verification to An Autonomy Architecture for Unmanned Aircraft

There is a desire to design autonomous systems in such a way that capabilities can be easily added or re- combined to produce new behaviors while preserving their safety properties. ICAROUS, a prototype software architecture for building safety-centric autonomous unmanned aircraft applications, is designed to support this type of extensibility and re-configurability. In ICAROUS, core capabilities are implemented as individual soft- ware services, so that enabling access to new capabilities simply requires adding new services. To make use of these capabilities, ICAROUS includes a specialized service that provides a general framework for config- uring the relative priorities, conditions, and rules that govern how different modules should be engaged and disengaged during flight. The inherent complexity of coordinating multiple modules under changing conditions makes it difficult to determine whether a particular configuration could have erroneous behaviors in certain circumstances. A robust set of integration tests can help discover errors, but testing can only realistically cover a relatively small proportion of total system behaviors. Developing good tests and interpreting the results to pinpoint the cause of errors when they arise can also be very time-consuming. To supplement testing, formal methods can be used to model and analyze complex systems, achieving better coverage and simplifying the process of finding, understanding, and fixing errors. To demonstrate these benefits, this paper explores the ap- plication of formal methods to ICAROUS. In particular, the Spin model checker is used to specify requirements for and model portions of the system, then verify whether the model satisfies the requirements and find and fix errors when it does not.

Formal Methods

Preliminary Application of Formal Verification to An Autonomy Architecture for Unmanned Aircraft

There is a desire to design autonomous systems in such a way that capabilities can be easily added or re-combined to produce new behaviors while preserving their safety properties. ICAROUS, a prototype software architecture for building safety-centric autonomous unmanned aircraft applications, is designed to support this type of extensibility and re-configurability. In ICAROUS, core capabilities are implemented as individual soft- ware services, so that enabling access to new capabilities simply requires adding new services. To make use of these capabilities, ICAROUS includes a specialized service that provides a general framework for config- uring the relative priorities, conditions, and rules that govern how different modules should be engaged and disengaged during flight. The inherent complexity of coordinating multiple modules under changing conditions makes it difficult to determine whether a particular configuration could have erroneous behaviors in certain circumstances. A robust set of integration tests can help discover errors, but testing can only realistically cover a relatively small proportion of total system behaviors. Developing good tests and interpreting the results to pinpoint the cause of errors when they arise can also be very time-consuming. To supplement testing, formal methods can be used to model and analyze complex systems, achieving better coverage and simplifying the process of finding, understanding, and fixing errors. To demonstrate these benefits, this paper explores the ap- plication of formal methods to ICAROUS. In particular, the Spin model checker is used to specify requirements for and model portions of the system, then verify whether the model satisfies the requirements and find and fix errors when it does not.

Formal Methods

Models and Algorithms for Equilibrium Analysis of Mixed-Material Nucleic Acid Systems

Dynamic programming algorithms within the NUPACK software suite enable analysis of equilibrium base-pairing properties for complex and test tube ensembles containing arbitrary numbers of interacting nucleic acid strands. Currently, calculations are limited to single-material systems that are either all-RNA or all-DNA. Here, to enable analysis of mixed-material systems that are critical for modern applications in vitro, in situ, and in vivo, we develop physical models and dynamic programming algorithms that allow the material of the system to be specified at nucleotide resolution. Free energy parameter sets are constructed for both RNA/DNA and RNA/2'OMe-RNA mixed-material systems by combining available empirical mixed-material parameters with single-material parameter sets to enable treatment of the full complex and test tube ensembles. New dynamic programming recursions account for the material of each nucleotide throughout the recursive process. For a complex with N nucleotides, the mixed-material dynamic programming algorithms maintain the O(N 3 ) time complexity of the single-material algorithms, enabling efficient calculation of diverse physical quantities over complex and test tube ensembles (e.g., complex partition function, equilibrium complex concentrations, equilibrium base-pairing probabilities, minimum free energy secondary structure(s), and Boltzmann-sampled secondary structures) at a cost increase of roughly 2.0-3.5×. The results of existing single-material algorithms are exactly reproduced when applying the new mixed-material algorithms to single-material systems. Accuracy is significantly enhanced using mixed-material models and algorithms to predict RNA/DNA and RNA/2'OMe-RNA duplex melting temperatures from the experimental literature as well as RNA/DNA melt profiles from new experiments. In conclusion, mixed-material analyses can be performed online using the NUPACK web app (www.nupack.org) or locally using the NUPACK Python module.

2′OMe-RNA

Experiences with Testing the Largest Ground System NASA Has Ever Built

In the 1980s, the National Aeronautics and Space Administration (NASA) embarked upon a major Earth-focused program called Mission to Planet Earth. The Goddard Space Flight Center (GSFC) was selected to manage and develop a key component - the Earth Observing System (EOS). The EOS consisted of four major missions designed to monitor the Earth. The missions included 4 spacecraft. Terra (launched December 1999), Aqua (launched May 2002), ICESat (Ice, Cloud, and Land Elevation Satellite, launched January 2003), and Aura (scheduled for launch January 2004). The purpose of these missions was to provide support for NASA s long-term research effort for determining how human-induced and natural changes affect our global environment. The EOS Data and Information System (EOSDIS), a globally distributed, large-scale scientific system, was built to support EOS. Its primary function is to capture, collect, process, and distribute the most voluminous set of remotely sensed scientific data to date estimated to be 350 Gbytes per day. The EOSDIS is composed of a diverse set of elements with functional capabilities that require the implementation of a complex set of computers, high-speed networks, mission-unique equipment, and associated Information Technology (IT) software along with mission-specific software. All missions are constrained by schedule, budget, and staffing resources, and rigorous testing has been shown to be critical to the success of each mission. This paper addresses the challenges associated with the planning, test definition. resource scheduling, execution, and discrepancy reporting involved in the mission readiness testing of a ground system on the scale of EOSDIS. The size and complexity of the mission systems supporting the Aqua flight operations, for example, combined with the limited resources available, prompted the project to challenge the prevailing testing culture. The resulting success of the Aqua Mission Readiness Testing (MRT) program was due in no small measure to re-structuring the traditional programmatic and technical approach to a more efficient and robust program. Programmatically, it meant gaining the endorsement, commitment, and cooperation of the numerous subsystem element managers and other stakeholder organizations. Technically, it required an MRT program that was agile, could rapidly adapt to requirements changes, and was flexible in its overall approach. Furthermore, this paper addresses the following questions: 1. What are the key ingredients (e.g., test tools, organization) needed to conduct a successful MRT program? 2. What distinguishes EOS MRT from the traditional system testing approach? 3. Where should the focus of testing be since it is infeasible to test every element or subsystem? 4. How can MRT be applied effectively to other systems or missions? To provide answers to these questions, this paper relies heavily on real-life, hands-on experiences ("lessons learned") gained during mission readiness testing of the Terra ground system and, most recently, the Aqua and ICESat missions. Moreover, this paper explores how lessons learned were turned into lessons applied for the upcoming Aura mission. Although derived from the EOS missions, MRT techniques and strategies can be applied to enhance the testing of other missions.

Lehtonen, Ken