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At least 127 records · Page 7

Buckling Design and Imperfection Sensitivity of Sandwich Composite Launch-Vehicle Shell Structures

Composite materials are increasingly being considered and used for launch-vehicle structures. For shell structures, such as interstages, skirts, and shrouds, honeycomb-core sandwich composites are often selected for their structural efficiency. Therefore, it is becoming increasingly important to understand the structural response, including buckling, of sandwich composite shell structures. Additionally, small geometric imperfections can significantly influence the buckling response, including considerably reducing the buckling load, of shell structures. Thus, both the response of the theoretically perfect structure and the buckling imperfection sensitivity must be considered during the design of such structures. To address the latter, empirically derived design factors, called buckling knockdown factors (KDFs), were developed by NASA in the 1960s to account for this buckling imperfection sensitivity during design. However, most of the test-article designs used in the development of these recommendations are not relevant to modern launch-vehicle constructions and material systems, and in particular, no composite test articles were considered. Herein, a two-part study on composite sandwich shells to (1) examine the relationship between the buckling knockdown factor and the areal mass of optimized designs, and (2) to interrogate the imperfection sensitivity of those optimized designs is presented. Four structures from recent NASA launch-vehicle development activities are considered. First, designs optimized for both strength and stability were generated for each of these structures using design optimization software and a range of buckling knockdown factors; it was found that the designed areal masses varied by between 6.1% and 19.6% over knockdown factors ranging from 0.6 to 0.9. Next, the buckling imperfection sensitivity of the optimized designs is explored using nonlinear finite-element analysis and the as-measured shape of a large-scale composite cylindrical shell. When compared with the current buckling design recommendations, the results suggest that the current recommendations are overly conservative and that the development of new recommendations could reduce the acreage areal mass of many composite sandwich shell designs by between 4% and 19%, depending on the structure.

Schultz, Marc R.↗

Improved field assessments of chemicals and explosives using high-resolution gamma-ray spectroscopy with a tagged neutron interrogation system

A fi eld-portable tagged neutron interrogation system is being developed for detection of explosives and chemical warfare agents by prompt gamma-ray neutron activation analysis. The system combines an associated particle imaging deuterium-tritium neutron generator with a high-purity germanium gamma-ray detector. Fine spatial resolution neutron and alpha detectors enable precise mapping of characteristic gamma-ray emission distributions onto high-clarity neutron radiographic images. A narrow coincidence gate associated with each tagged neutron ensures a large signal-to-background ratio, thus signifi cantly improving sensitivity to the chemical compositions of the items being assessed. Recent experimental results recorded from test objects involving mock explosives and other chemicals are presented.

46 - INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AN↗

Improved field assessments of chemicals and explosives using high-resolution gamma-ray spectroscopy with a tagged neutron interrogation system

A fi eld-portable tagged neutron interrogation system is being developed for detection of explosives and chemical warfare agents by prompt gamma-ray neutron activation analysis. The system combines an associated particle imaging deuterium-tritium neutron generator with a high-purity germanium gamma-ray detector. Fine spatial resolution neutron and alpha detectors enable precise mapping of characteristic gamma-ray emission distributions onto high-clarity neutron radiographic images. A narrow coincidence gate associated with each tagged neutron ensures a large signal-to-background ratio, thus signifi cantly improving sensitivity to the chemical compositions of the items being assessed. Recent experimental results recorded from test objects involving mock explosives and other chemicals are presented.

46 - INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AN↗

Building a COTS archive for satellite data

The goal of the NOAA/NESDIS Active Archive was to provide a method of access to an online archive of satellite data. The archive had to manage and store the data, let users interrogate the archive, and allow users to retrieve data from the archive. Practical issues of the system design such as implementation time, cost and operational support were examined in addition to the technical issues. There was a fixed window of opportunity to create an operational system, along with budget and staffing constraints. Therefore, the technical solution had to be designed and implemented subject to constraint imposed by the practical issues. The NOAA/NESDIS Active Archive came online in July of 1994, meeting all of its original objectives.

Singer, Ken↗

Structural health monitoring apparatus and methodology

Disclosed is an apparatus and methodology for structural health monitoring (SHM) in which smart devices interrogate structural components to predict failure, expedite needed repairs, and thus increase the useful life of those components. Piezoelectric wafer active sensors (PWAS) are applied to or integrated with structural components and various data collected there from provide the ability to detect and locate cracking, corrosion, and disbanding through use of pitch-catch, pulse-echo, electro/mechanical impedance, and phased array technology. Stand alone hardware and an associated software program are provided that allow selection of multiple types of SHM investigations as well as multiple types of data analysis to perform a wholesome investigation of a structure.

Giurgiutiu, Victor↗

Short-Term Stable Isotope Probing of Proteins Reveals Taxa Incorporating Inorganic Carbon in a Hot Spring Microbial Mat

The upper green layer of the chlorophototrophic microbial mats associated with the alkaline siliceous hot springs of Yellowstone National Park consists of oxygenic cyanobacteria ( Synechococcus spp.), anoxygenic Roseiflexus spp., and several other anoxygenic chlorophototrophs. Synechococcus spp. are believed to be the main fixers of inorganic carbon (C i ), but some evidence indicates that Roseiflexus spp. also contribute to inorganic carbon fixation during low-light, anoxic morning periods. Contributions of other phototrophic taxa have not been investigated. In order to follow the pathway of C i incorporation into different taxa, mat samples were incubated with [ 13 C]bicarbonate for 3 h during the early-morning, low-light anoxic period. Extracted proteins were treated with trypsin and analyzed by mass spectrometry, leading to peptide identifications and peptide isotopic profile signatures containing evidence of 13 C label incorporation. A total of 25,483 peptides, corresponding to 7,221 proteins, were identified from spectral features and associated with mat taxa by comparison to metagenomic assembly sequences. A total of 1,417 peptides, derived from 720 proteins, were detectably labeled with 13 C. Most 13 C-labeled peptides were derived from proteins of Synechococcus spp. and Roseiflexus spp. Chaperones and proteins of carbohydrate metabolism were most abundantly labeled. Proteins associated with photosynthesis, C i fixation, and N 2 fixation were also labeled in Synechococcus spp. Importantly, most proteins of the 3-hydroxypropionate bi-cycle for C i fixation in Roseiflexus spp. were labeled, establishing that members of this taxocene contribute to C i fixation. Other taxa showed much lower [ 13 C]bicarbonate incorporation. IMPORTANCE Yellowstone hot spring mats have been studied as natural models for understanding microbial community ecology and as modern analogs of stromatolites, the earliest community fossils on Earth. Stable-isotope probing of proteins (Pro-SIP) permitted short-term interrogation of the taxa that are involved in the important process of light-driven C i fixation in this highly active community and will be useful in linking other metabolic processes to mat taxa. Here, evidence is presented that Roseiflexus spp., which use the 3-hydroxypropionate bi-cycle, are active in C i fixation. Because this pathway imparts a lower degree of selection of isotopically heavy C i than does the Calvin-Benson-Bassham cycle, the results suggest a mechanism to explain why the natural abundance of 13 C in mat biomass is greater than expected if only the latter pathway were involved. Understanding how mat community members influence the 13 C/ 12 C ratios of mat biomass will help geochemists interpret the 13 C/ 12 C ratios of organic carbon in the fossil record.

59 BASIC BIOLOGICAL SCIENCES↗

Observation of an Intermediate to H 2 Binding in a Metal–Organic Framework

Coordinatively unsaturated metal sites within certain zeolites and metal–organic frameworks can strongly adsorb a wide array of substrates. While many classical examples involve electron-poor metal cations that interact with adsorbates largely through physical interactions, unsaturated electron-rich metal centers housed within porous frameworks can often chemisorb guests amenable to redox activity or covalent bond formation. Despite the promise that materials bearing such sites hold in addressing myriad challenges in gas separations and storage, very few studies have directly interrogated mechanisms of chemisorption at open metal sites within porous frameworks. Here, we show that nondissociative chemisorption of H 2 at the trigonal pyramidal Cu + sites in the metal–organic framework Cu I -MFU- 4 l occurs via the intermediacy of a metastable physisorbed precursor species. In situ powder neutron diffraction experiments enable crystallographic characterization of this intermediate, the first time that this has been accomplished for any material. Evidence for a precursor intermediate is also afforded from temperature-programmed desorption and density functional theory calculations. Here, the activation barrier separating the precursor species from the chemisorbed state is shown to correlate with a change in the Cu + coordination environment that enhances p-backbonding with H 2 . Ultimately, these findings demonstrate that adsorption at framework metal sites does not always follow a concerted pathway and underscore the importance of probing kinetics in the design of next-generation adsorbents.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Marine Shallow Cloud Adjustments to the Presence of Shortwave-Absorbing Aerosols: Advancing Understanding Through a Combined Analysis of Lasic Datasets and Process Modeling (Final Report)

This is a final report for this award, whose purpose was to support the analysis of data emanating from the LASIC (Layered Atlantic Smoke Interaction with Clouds) campaign. (Layered Atlantic Smoke Interactions with Clouds) is a strategy to improve our understanding of aged carbonaceous aerosol, its seasonal evolution, and the mechanisms by which clouds adjust to the presence of the aerosol. The observational strategy centers on deploying the AMF1 cloud, aerosol, and atmospheric profiling instrumentation to Ascension Island, located within the trade-wind shallow cumulus regime (14.50W, 80S) 3000 km offshore of continental Africa. The location is within the latitude zone of the maximum outflow of aerosol, with the deepening boundary layer known to entrain free-tropospheric smoke. The primary activities for LASIC are: 1) to improve current knowledge on aged biomass burning aerosol and its radiative properties as a function of the seasonal cycle; 2) to use surface-based remote sensing to sensitively interrogate the atmosphere for the relative vertical location of aerosol and clouds; 3) to improve our understanding of the cloud adjustments to the presence of shortwave-absorbing aerosol within the vertical column, both through aerosol-radiation and through aerosol-cloud interactions; 4) to aid low cloud parameterization efforts for climate models. This award serves to further these activities and includes a sub-award to Dr. Pablo Saide at UCLA for activities 3) and 4).

54 ENVIRONMENTAL SCIENCES↗

ARIEL: Autonomous Excavation Site Selection for Europa Lander Mission Concept

This paper presents ARIEL (Autonomous Ranking and Interrogation of Excavation Location), an autonomy system for selecting an excavation site on-board for NASA’s Europa Lander Mission Concept. Historically, excavation site selection has been performed by a lengthy ground-in-the-loop (GITL) process involving manual inspections, assessments, and decision making in past missions. However, as Europa Lander would have approximately 20 days of lifetime after the landing, many surface activities, including excavation site selection, must be autonomously performed on-board. This paper describes the overall system of ARIEL as well as its two major algorithmic components: vision-based candidate selection and smart interrogation, which estimates the physical properties of the icy surface through physical contact with the robotic arm’s endeffector. Preliminary results are presented using images from Earth analogue sites. The Europa Lander mission returned to the formulation phases in early 2019 while ARIEL was at an early stage of development. Described in this paper is a snapshot of ARIEL as of the project suspension. This paper also describes the remaining challenges to be solved, should the mission resume in the future.

Ono, Hiro↗

Enhancers that direct gene expression to central nervous system vascular endothelial cells in vivo

CNS vascular endothelial cells (ECs) exhibit a distinctive gene expression program that is foundational for the blood-brain barrier (BBB). Previous research identified candidate cis-regulatory elements (CREs) that were hypothesized to control this program. In this work, transgenic mice and recombinant adeno-associated virus (rAAV) vectors have been used to interrogate these candidate CREs in vivo. These experiments show that an 850 bp genomic DNA segment ∼60 kb 5′ of Slc2a1 possesses enhancer activity that is (1) specific for BBB+ CNS ECs and (2) both necessary and sufficient for BBB+ EC gene expression. A screen of >8,000 genomic DNA segments from CNS EC-specific CRE candidates reveals several hundred with enhancer activity. Transcription factors ERG and LEF1 are shown to occupy sites in brain ECs that are highly enriched in candidate and experimentally validated CREs, lending strong support to a model in which canonical Wnt signaling activates the BBB program via LEF1.

CUT&RUN↗

Environmental Contributions to Proton Sharing in Protein Low-Barrier Hydrogen Bonds

Hydrogen bonds (H-bonds) are central to biomolecular structure and dynamics. Although H-bonds are typically characterized by well-defined proton positions, proton delocalization has been proposed to play a role in facilitating enzyme catalysis and allostery in some systems. Experimentally locating protons is difficult, hampering the study of proton mobility in H-bonds. We used neutron crystallography, atomic resolution X-ray bond length analysis, and large quantum mechanics/molecular mechanics-Born–Oppenheimer molecular dynamics (QM/MM-BOMD) simulations to comprehensively characterize the shared proton/deuteron in a Glu–Asp low-barrier hydrogen bond (LBHB) in the bacterial protein YajL that is a conventional H-bond in the homologous disease-associated human protein DJ-1. X-ray bond length analysis of protiated and perdeuterated DJ-1 and YajL shows no significant effect of deuteron substitution on these carboxylic acid-carboxylate H-bonds but does reveal an effect at the active site glutamic acid near a cysteine thiolate. Residues in an H-bonded network that might favor LBHB formation in YajL were interrogated by the mutation of homologous residues in DJ-1. A distal DJ-1 substitution increases proton delocalization in the Glu–Asp H-bond, demonstrating that mutations within extended H-bond networks can modulate proton transfer barriers in carboxylic acid-carboxylate H-bonds. In addition, proton mobility in the H-bond is correlated with dimer-spanning motions in the QM/MM-BOMD simulations of YajL and DJ-1. Our results show that proton delocalization can be tuned using combined bioinformatic, structural, and computational information, opening the possibility of using engineered proton delocalization as a probe of H-bonding environments and as a tool to test hypotheses about LBHB function.

Lin, Jiusheng [University of Nebraska, Lincoln, NE↗

Assessment of enzyme active site positioning and tests of catalytic mechanisms through X-ray–derived conformational ensembles

How enzymes achieve their enormous rate enhancements remains a central question in biology, and our understanding to date has impacted drug development, influenced enzyme design, and deepened our appreciation of evolutionary processes. While enzymes position catalytic and reactant groups in active sites, physics requires that atoms undergo constant motion. Numerous proposals have invoked positioning or motions as central for enzyme function, but a scarcity of experimental data has limited our understanding of positioning and motion, their relative importance, and their changes through the enzyme’s reaction cycle. To examine positioning and motions and test catalytic proposals, we collected “room temperature” X-ray crystallography data for Pseudomonas putida ketosteroid isomerase (KSI), and we obtained conformational ensembles for this and a homologous KSI from multiple PDB crystal structures. Here, ensemble analyses indicated limited change through KSI’s reaction cycle. Active site positioning was on the 1- to 1.5-Å scale, and was not exceptional compared to noncatalytic groups. The KSI ensembles provided evidence against catalytic proposals invoking oxyanion hole geometric discrimination between the ground state and transition state or highly precise general base positioning. Instead, increasing or decreasing positioning of KSI’s general base reduced catalysis, suggesting optimized Ångstrom-scale conformational heterogeneity that allows KSI to efficiently catalyze multiple reaction steps. Ensemble analyses of surrounding groups for WT and mutant KSIs provided insights into the forces and interactions that allow and limit active-site motions. Most generally, this ensemble perspective extends traditional structure–function relationships, providing the basis for a new era of “ensemble–function” interrogation of enzymes.

59 BASIC BIOLOGICAL SCIENCES↗

Improved bacterial recombineering by parallelized protein discovery

Exploiting bacteriophage-derived homologous recombination processes has offered precise, multiplex editing of microbial genomes and the construction of billions of customized genetic variants in a single day. The techniques that enable this, multiplex automated genome engineering (MAGE) and directed evolution with random genomic mutations (DIvERGE), are however, currently limited to a handful of microorganisms for which single-stranded DNA-annealing proteins (SSAPs) that promote efficient recombineering have been identified. Thus, to enable genome-scale engineering in new hosts, efficient SSAPs must first be found. Here we present a high-throughput method for SSAP discovery that we call “serial enrichment for efficient recombineering” (SEER). By performing SEER in Escherichia coli to screen hundreds of putative SSAPs, we identify highly active variants PapRecT and CspRecT. CspRecT increases the efficiency of single-locus editing to as high as 50% and improves multiplex editing by 5- to 10-fold in E. coli , while PapRecT enables efficient recombineering in Pseudomonas aeruginosa , a concerning human pathogen. CspRecT and PapRecT are also active in other, clinically and biotechnologically relevant enterobacteria. We envision that the deployment of SEER in new species will pave the way toward pooled interrogation of genotype-to-phenotype relationships in previously intractable bacteria.

60 APPLIED LIFE SCIENCES↗

Nonpigmented PsbR is involved in the integrity of excitation landscape in higher plant photosystem II, a case study in Arabidopsis thaliana and a mutant

PsbR is a nonpigmented 10 kDa protein in Photosystem II (PSII) in algae and plants. A recent structural study clarified its enigmatic structural location in a Photosystem II megacomplex that has baffled the community for more than four decades. Our current study interrogates whether absence of PsbR affects the overall dynamics of excitation energy migration within light harvesting complexes (LHC) and PSII super assemblies using highly-active PSII membrane particles, so-called BBY particles, isolated from a PsbR deletion mutant (ΔPsbR) of Arabidopsis thaliana. A femto-second (fs)-time-resolved transient absorption experimentation recorded at 77 K with selective excitation of Chl b which is exclusively present in LHCs enabled us to resolve the temporal differences in LHC→LHC and LHC→PSII excitation energy transfer steps. By applying specific target spectro-kinetic models to the transient absorption datasets, we demonstrated that the time constants of Chl a LHC → Chl a LHC excitation transfer significantly elongates in the ΔPsbR LHC-PSII particles, suggestive of the decreased aggregation level of photosynthetic proteins in the mutant. These findings highlight excitation energy transfer integrity in LHC-PSII assembly is not only determined by the pigmented light-harvesting complexes, but also synergistically by the nonpigmented PSII components. Furthermore, the disturbed integrity in dynamics of excitation energy transfer pathway within LHC-PSII supercomplex is discussed in the context of the altered LHC-PSII megacomplexes type I and II architectures which result from the absence of the PsbR protein in higher plant PSII.

Chlorophyll↗

A Safe, Self-Calibrating, Wireless System for Measuring Volume of Any Fuel at Non-Horizontal Orientation

A system for wirelessly measuring the volume of fluid in tanks at non-horizontal orientation is predicated upon two technologies developed at Langley Research Center. The first is a magnetic field response recorder that powers and interrogates magnetic field response sensors [ Magnetic Field Response Measurement Acquisition System, (LAR-16908), NASA Tech Briefs, Vol. 30, No. 6 (June 2006), page 28]. Magnetic field response sensors are a class of sensors that are powered via oscillating magnetic fields and when electrically active respond with their own magnetic fields whose attributes are dependent upon the magnitude of the physical quantity being measured. The response recorder facilitates the use of the second technology, which is a magnetic field response fluid-level sensor ["Wireless Fluid- Level Sensors for Harsh Environments," (LAR-17155), NASA Tech Briefs, Vol. 33, No. 4 (April 2009), page 30]. The method for powering and interrogating the sensors allows them to be completely encased in materials (Fig. 1) that are chemically resilient to the fluid being measured, thereby facilitating measurement of substances (e.g., acids, petroleum, cryogenic, caustic, and the like) that would normally destroy electronic circuitry. When the sensors are encapsulated, no fluid (or fluid vapor) is exposed to any electrical component of the measurement system. There is no direct electrical line from the vehicle or plant power into a fuel container. The means of interrogating and powering the sensors can be completely physically and electrically isolated from the fuel and vapors by placing the sensor on the other side of an electrically non-conductive bulkhead (Fig. 2). These features prevent the interrogation system and its electrical components from becoming an ignition source.

Woodward, Stanley E.↗

Regulatory response to a hybrid ancestral nitrogenase in Azotobacter vinelandii

Biological nitrogen fixation, the microbial reduction of atmospheric nitrogen to bioavailable ammonia, represents both a major limitation on biological productivity and a highly desirable engineering target for synthetic biology. However, the engineering of nitrogen fixation requires an integrated understanding of how the gene regulatory dynamics of host diazotrophs respond across sequence-function space of its central catalytic metalloenzyme, nitrogenase. Here, we interrogate this relationship by analyzing the transcriptome of Azotobacter vinelandii engineered with a phylogenetically inferred ancestral nitrogenase protein variant. The engineered strain exhibits reduced cellular nitrogenase activity but recovers wild-type growth rates following an extended lag period. We find that expression of genes within the immediate nitrogen fixation network is resilient to the introduced nitrogenase sequence-level perturbations. Rather the sustained physiological compatibility with the ancestral nitrogenase variant is accompanied by reduced expression of genes that support trace metal and electron resource allocation to nitrogenase. Our results spotlight gene expression changes in cellular processes adjacent to nitrogen fixation as productive engineering considerations to improve compatibility between remodeled nitrogenase proteins and engineered host diazotrophs.

59 BASIC BIOLOGICAL SCIENCES↗

A Magnetic Field Response Recorder: A New Tool for Measurement Acquisition

A magnetic field response recorder was developed to facilitate a measurement acquisition method that uses magnetic fields to power and to interrogate all sensors. Sensors are designed as electrically passive inductive-capacitive or passive inductive-capacitive-resistive circuits that produce magnetic field responses when electrically activated by oscillating magnetic fields. When electrically activated, the sensor's magnetic field response attributes (frequency, amplitude and bandwidth) correspond to the one or more physical states that each sensor measures. The response recorder makes it possible to simultaneously measure two unrelated physical properties using this class of sensors. The recorder is programmable allowing it to analyze one or more response attributes simultaneously. A single sensor design will be used to demonstrate that the acquisition method and the sensor example can be used to for all phases of a component's life from manufacturing to damage that can destroy it.

Woodard, Stanley E.↗

Methods of Using a Magnetic Field Response Sensor Within Closed, Electrically Conductive Containers

Magnetic field response sensors are a class of sensors that are powered via oscillating magnetic fields, and when electrically active, respond with their own magnetic fields with attributes dependent upon the magnitude of the physical quantity being measured. A magnetic field response recorder powers and interrogates the magnetic sensors [see Magnetic-Field-Response Measurement- Acquisition System, NASA Tech Briefs Vol. 30, No, 6 (June 2006, page 28)]. Electrically conductive containers have low transmissivity for radio frequency (RF) energy and thus present problems for magnetic field response sensors. It is necessary in some applications to have a magnetic field response sensor s capacitor placed in these containers. Proximity to conductive surfaces alters the inductance and capacitance of the sensors. As the sensor gets closer to a conductive surface, the electric field and magnetic field energy of the sensor is reduced due to eddy currents being induced in the conductive surface. Therefore, the capacitors and inductors cannot be affixed to a conductive surface or embedded in a conductive material. It is necessary to have a fixed separation away from the conductive material. The minimum distance for separation is determined by the desired sensor response signal to noise ratio. Although the inductance is less than what it would be if it were not in proximity to the conductive surface, the inductance is fixed. As long as the inductance is fixed, all variations of the magnetic field response are due to capacitance changes. Numerous variations of inductor mounting can be utilized, such as providing a housing that provides separation from the conductive material as well as protection from impact damage. The sensor can be on the same flexible substrate with a narrow throat portion of the sensor between the inductor and the capacitor, Figure 1. The throat is of sufficient length to allow the capacitor to be appropriately placed within the container and the inductor placed outside the container. The throat is fed through the orifice in the container wall (e.g., fuel tank opening) and connects to the inductor and capacitor via electrical leads to form a closed circuit, Figure 2. Another embodiment is to have the inductor and capacitor fabricated as separate units. In this embodiment, the inductor is mounted external to the container, and the capacitor is mounted internal to the container, Figure 1. Electrical leads are fed through the orifice to connect the inductor and capacitor, Figure 2. When a container holding multiple sensors is made of a conductive material, an antenna can be placed internal to the container. An internal antenna allows all components of the sensors to reside inside the container. The antenna must be separated from the container wall s conductive surface. Additionally, the inductors must be maintained in a fixed position relative to and separated from the container

Woodward, Stanley E.↗