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At least 793 records · Page 44

Implementation of real-time digital signal processing systems

Special purpose hardware implementation of DFT Computers and digital filters is considered in the light of newly introduced algorithms and IC devices. Recent work by Winograd on high-speed convolution techniques for computing short length DFT's, has motivated the development of more efficient algorithms, compared to the FFT, for evaluating the transform of longer sequences. Among these, prime factor algorithms appear suitable for special purpose hardware implementations. Architectural considerations in designing DFT computers based on these algorithms are discussed. With the availability of monolithic multiplier-accumulators, a direct implementation of IIR and FIR filters, using random access memories in place of shift registers, appears attractive. The memory addressing scheme involved in such implementations is discussed. A simple counter set-up to address the data memory in the realization of FIR filters is also described. The combination of a set of simple filters (weighting network) and a DFT computer is shown to realize a bank of uniform bandpass filters. The usefulness of this concept in arriving at a modular design for a million channel spectrum analyzer, based on microprocessors, is discussed.

Narasimha, M.↗

A study of parameter identification

A set of definitions for deterministic parameter identification ability were proposed. Deterministic parameter identificability properties are presented based on four system characteristics: direct parameter recoverability, properties of the system transfer function, properties of output distinguishability, and uniqueness properties of a quadratic cost functional. Stochastic parameter identifiability was defined in terms of the existence of an estimation sequence for the unknown parameters which is consistent in probability. Stochastic parameter identifiability properties are presented based on the following characteristics: convergence properties of the maximum likelihood estimate, properties of the joint probability density functions of the observations, and properties of the information matrix.

Herget, C. J.↗

The Local ISM and its Interaction with the Winds of Nearby Late-type Stars

We present new Goddard High-Resolution Spectrograph (GHRS) observations of the Ly-alpha and Mg II absorption lines seen toward the nearby stars 61 Cyg A and 40 Eri A. We use these data to measure interstellar properties along these lines of sight and to search for evidence of circumstellar hydrogen walls, which are produced by collisions between the stellar winds and the Local InterStellar Medium (LISM). We were able to model the Ly-alpha lines of both stars without hydrogen-wall absorption components, but for 61 Cyg A the fit required a stellar Ly-alpha, line profile with an improbably deep self-reversal, and for 40 Eri A the fit required a very low deuterium-to-hydrogen ratio that is inconsistent with previous GHRS measurements. Since these problems could be rectified simply by including stellar hydrogen-wall components with reasonable attributes, our preferred fits to the data include these components. We have explored several ways in which the hydrogen-wall properties measured here and in previous work can be used to study stellar winds and the LISM. We argue that the existence of a hydrogen wall around 40 Eri A and a low H I column density along that line of sight imply that either the interstellar density must decrease toward 40 Eri A or the hydrogen ionization fraction (chi) must increase. We find that hydrogen-wall temperatures are larger for stars with faster velocities through the LISM. The observed temperature-velocity relation is consistent with the predictions of hydromagnetic shock jump conditions. More precise comparison of the data and the jump conditions suggests crude upper limits for both chi and the ratio of magnetic to thermal pressure in the LISM (alpha): chi less than 0.6 and alpha less than 2. The latter upper limit corresponds to a limit on the LISM magnetic field of B less than 5 micro G. These results imply that the plasma Mach number of the interstellar wind flowing into the heliosphere is M(sub A) greater than 1.3, which indicates that the collision is supersonic and that there should therefore be a bow shock outside the heliopause in the upwind direction. Finally, we estimate stellar wind pressures (P sub wind) from the measured hydrogen-wall column densities. These estimates represent the first empirical measurements of wind properties for late-type main-sequence stars. The wind pressures appear to be correlated with stellar X-ray surface fluxes, F(x), in a manner consistent with the relation P(wind) varies as F(x)(exp -1/2), a relation that is also consistent with the variations of P(sub wind) and F(sub x) observed during the solar activity cycle. If this relation can in fact be generalized to solar-like stars, as is suggested by our data, then it is possible to estimate stellar wind properties simply by measuring stellar X-rays. One implication of this is that stellar wind pressures and mass-loss rates are then predicted to increase with time, since F(sub x) is known to decrease with stellar age.

Wood, Brian E.↗

Results from Testing Crew-Controlled Surface Telerobotics on the International Space Station

During Summer 2013, the Intelligent Robotics Group at NASA Ames Research Center conducted a series of tests to examine how astronauts in the International Space Station (ISS) can remotely operate a planetary rover. The tests simulated portions of a proposed lunar mission, in which an astronaut in lunar orbit would remotely operate a planetary rover to deploy a radio telescope on the lunar far side. Over the course of Expedition 36, three ISS astronauts remotely operated the NASA "K10" planetary rover in an analogue lunar terrain located at the NASA Ames Research Center in California. The astronauts used a "Space Station Computer" (crew laptop), a combination of supervisory control (command sequencing) and manual control (discrete commanding), and Ku-band data communications to command and monitor K10 for 11 hours. In this paper, we present and analyze test results, summarize user feedback, and describe directions for future research.

robotics↗

End-User Perspectives on Using Quantitative Real-Time PCR and Genomic Sequencing in the Field

Quantitative real-time PCR and genomic sequencing have become mainstays for performing molecular detection of biological threat agents in the field. There are notional assessments of the benefits, disadvantages, and challenges that each of these technologies offers according to findings in the literature. However, direct comparison between these two technologies in the context of field-forward operations is lacking. Most market surveys, whether published in print form or provided online, are directed to product manufacturers who can address their respective specifications and operations. One method for comparing these technologies is surveying end-users who are best suited for discussing operational capabilities, as they have hands-on experience with state-of-the-art molecular detection platforms and protocols. These end-users include operators in military defense and first response, as well as various research scientists in the public sector such as government and service laboratories, private sector, and civil society such as academia and nonprofit organizations performing method development and executing these protocols in the field. Our objective was to initiate a survey specific to end-users and their feedback. We developed a questionnaire that asked respondents to (1) determine what technologies they currently use, (2) identify the settings where the technologies are used, whether lab-based or field-forward, and (3) rate the technologies according to a set list of criteria. Of particular interest are assessments of sensitivity, specificity, reproducibility, scalability, portability, and discovery power. This article summarizes the findings from the end-user perspective, highlighting technical and operational challenges.

59 BASIC BIOLOGICAL SCIENCES↗

Defining the Minimal Set of Microbial Genes Required for Valorization of Lignin Biomass (Final Report)

Project Goals: Lignin is the second most abundant biopolymer on earth and represents a critically underutilized biomass resource for hydrocarbon feedstocks. Despite substantial effort, there is still no efficient process to convert lignin to useable carbon-based platform chemicals and materials. The goal of this project is identify a minimal set of microbial enzymes necessary for lignin breakdown and sufficient for the synthesis of valuable chemical intermediates from lignin isolated as a byproduct of lignocellulosic ethanol production. These genes will be then used to engineer functional whole cell biocatalysts for tunable lignin metabolism. To date, although a number of enzymes have been associated with lignin degradation, most have been tested in isolation (as individual enzymes) and on drastically different substrates -- often dyes that are not related to lignin. In contrast, lignin utilization in nature likely occurs by microbial consortia with multiple enzymes acting synergistically. We propose to examine two separate stages of lignin breakdown carried out by the microbes that do it best: (1) early breakdown of native polymeric lignin into soluble fragments by a set of sequenced wood-rotting fungal species, and (2) downstream metabolism of these soluble lignin fragments to useful chemical intermediates by a panel of sequenced soil saprophytes. Our approach involves testing sets of genes that will be assayed combinatorially in the context of a heterologous expression host. The resulting engineered strains will be systematically assayed using soluble lignin fragments, synthetic defined polymeric lignin, and finally lignin directly sourced from lignocellulosic processing streams. In addition to resulting in a functional whole cell biocatalyst for lignin utilization, we anticipate that this approach will allow us to address key unanswered questions about lignin metabolism in nature, including: (1) Why does the Trametes versicolor genome contain 25 different class II peroxidases? (2) What is the role of laccases in lignin metabolism? Why do some aggressive lignin degraders have many laccases (e.g. >7 in T. versicolor) while others have none (e.g. P. chrysosporium)? (3) How is peroxide provided in a controlled manner to drive peroxidase activity without causing the enzyme inhibition that is so often observed in vitro? (4) What strategies do microbial lignin degraders use to avoid the problem of repolymerization during active lignin degradation? and (5) Can microbial lignin metabolism be diverted for high level production of defined aromatics? A final critical question is whether combining key minimal sets of enzymes from a wide range of organisms will result in engineered strains capable of highly efficient, streamlined pathways for lignin utilization that can be tuned for a specific carbon output. This effort will leverage DOE investments in microbial genome sequencing, and secure a critical channel for lignin biomass utilization that will also help to render lignocellulosic a viable feedstock for the production of renewable liquid biofuels.

59 BASIC BIOLOGICAL SCIENCES↗

SFA-VirOmics

PNNL's Soil Microbiome Science Focus Area (SFA) is focused on understanding the basic biology underpinning how interactions among various soil microbial community members, across trophic levels, lead to the emergence of community functions. Moisture, in particular, drives microbial interactions and influences everything from cell function to substrate fate within soils. The group predicts this results in repeatable, predictable phenotypes. The sum of these phenotypes comprises the “soil metaphenome”. Understanding how the soil metaphenome shifts in response to moisture will provide a basis for modeling and predicting these shifts in reaction network responses. Visit the PNNL Soil Microbiome Science Focus Area Program homepage for more information. PNNL’s Soil Microbiome SFA Virome Dataset Annotation page is an extension to the PNNL Soil Microbiome SFA repository on DataHub allows for exploring and downloading integrated experimental omics dataset annotations, associated experimental metadata, pre- and post-processed viral data files, and other associated materials directly related to experimental viral project data.

59 BASIC BIOLOGICAL SCIENCES↗

Light-regulated root gravitropism: a role for, and characterization of, a calcium/calmodulin-dependent protein kinase homolog

Roots of many species grow downward (orthogravitropism) only when illuminated. Previous work suggests that this is a calcium-regulated response and that both calmodulin and calcium/calmodulin-dependent kinases participate in transducing gravity and light stimuli. A genomic sequence has been obtained for a calcium/calmodulin-dependent kinase homolog (MCK1) expressed in root caps, the site of perception for both light and gravity. This homolog consists of 7265 base pairs and contains 11 exons and 10 introns. Since MCK1 is expressed constitutively in both light and dark, it is unlikely that the light directly affects MCK1 expression, though the activity of the protein may be affected by light. In cultivars showing light-regulated gravitropism, we hypothesize that MCK1, or a homolog, functions in establishing the auxin asymmetry necessary for orthogravitropism.

NASA Discipline Plant Biology↗

INTEGRITY - Integrated Human Exploration Mission Simulation Facility

It is proposed to develop a high-fidelity ground facility to carry out long-duration human exploration mission simulations. These would not be merely computer simulations - they would in fact comprise a series of actual missions that just happen to stay on earth. These missions would include all elements of an actual mission, using actual technologies that would be used for the real mission. These missions would also include such elements as extravehicular activities, robotic systems, telepresence and teleoperation, surface drilling technology-all using a simulated planetary landscape. A sequence of missions would be defined that get progressively longer and more robust, perhaps a series of five or six missions over a span of 10 to 15 years ranging in duration from 180 days up to 1000 days. This high-fidelity ground facility would operate hand-in-hand with a host of other terrestrial analog sites such as the Antarctic, Haughton Crater, and the Arizona desert. Of course, all of these analog mission simulations will be conducted here on earth in 1-g, and NASA will still need the Shuttle and ISS to carry out all the microgravity and hypogravity science experiments and technology validations. The proposed missions would have sufficient definition such that definitive requirements could be derived from them to serve as direction for all the program elements of the mission. Additionally, specific milestones would be established for the "launch" date of each mission so that R&D programs would have both good requirements and solid milestones from which to .build their implementation plans. Mission aspects that could not be directly incorporated into the ground facility would be simulated via software. New management techniques would be developed for evaluation in this ground test facility program. These new techniques would have embedded metrics which would allow them to be continuously evaluated and adjusted so that by the time the sequence of missions is completed, the best management techniques will have been developed, implemented, and validated. A trained cadre of managers experienced with a large, complex program would then be available.

Henninger, Donald L.↗

Direct interaction of DNA repair protein tyrosyl DNA phosphodiesterase 1 and the DNA ligase III catalytic domain is regulated by phosphorylation of its flexible N-terminus

Tyrosyl DNA phosphodiesterase 1 (TDP1) and DNA Ligase IIIα (LigIIIα) are key enzymes in single-strand break (SSB) repair. TDP1 removes 3'-tyrosine residues remaining after degradation of DNA topoisomerase (TOP) 1 cleavage complexes trapped by either DNA lesions or TOP1 inhibitors. It is not known how TDP1 is linked to subsequent processing and LigIIIα-catalyzed joining of the SSB. Here we define a direct interaction between the TDP1 catalytic domain and the LigIII DNA-binding domain (DBD) regulated by conformational changes in the unstructured TDP1 N-terminal region induced by phosphorylation and/or alterations in amino acid sequence. Full-length and N-terminally truncated TDP1 are more effective at correcting SSB repair defects in TDP1 null cells compared with full-length TDP1 with amino acid substitutions of an N-terminal serine residue phosphorylated in response to DNA damage. TDP1 forms a stable complex with LigIII 170-755 , as well as full-length LigIIIα alone or in complex with the DNA repair scaffold protein XRCC1. Small-angle X-ray scattering and negative stain electron microscopy combined with mapping of the interacting regions identified a TDP1/LigIIIα compact dimer of heterodimers in which the two LigIII catalytic cores are positioned in the center, whereas the two TDP1 molecules are located at the edges of the core complex flanked by highly flexible regions that can interact with other repair proteins and SSBs. As TDP1and LigIIIα together repair adducts caused by TOP1 cancer chemotherapy inhibitors, the defined interaction architecture and regulation of this enzyme complex provide insights into a key repair pathway in nonmalignant and cancer cells.

59 BASIC BIOLOGICAL SCIENCES↗

Catalytic Difunctionalization of Cyclic Dienes: Direct Entry to Novel ROMP Monomers

We developed a catalytic platform to convert simple hydrocarbon feedstocks into valuable, tunable materials by leveraging nickel-catalyzed difunctionalization of cyclic dienes to access a novel class of cyclic alkene monomers. These monomers undergo ring-opening metathesis polymerization (ROMP) to yield sequence-controlled polymers with defined stereochemistry. Through mechanistic studies and catalyst optimization, we established a scalable, gram-level synthesis for selective diarylation, and expanded the reaction scope to include arylalkylation through rationally tuning the organoboron coupling partner. The resulting polymers were systematically studied to understand how steric, electronic, and stereochemical features influence polymerization behavior and bulk material properties. Functionalized derivatives bearing sulfonated groups were explored as proton-exchange membranes, and chemical recycling pathways were developed to recover monomers from the final materials. This work bridges small-molecule catalysis and macromolecular design, enabling access to tunable, recyclable polymers from abundant hydrocarbon starting materials.

36 MATERIALS SCIENCE↗

Space experiment development process

Described is a process for developing space experiments utilizing the Space Shuttle. The role of the Principal Investigator is described as well as the Principal Investigator's relation with the project development team. Described also is the sequence of events from an early definition phase through the steps of hardware development. The major interactions between the hardware development program and the Shuttle integration and safety activities are also shown. The presentation is directed to people with limited Shuttle experiment experience. The objective is to summarize the development process, discuss the roles of major participants, and list some lessons learned. Two points should be made at the outset. First, no two projects are the same so the process varies from case to case. Second, the emphasis here is on Code EN/Microgravity Science and Applications Division (MSAD).

Depauw, James F.↗

In search of the elusive flow field

A specific set of problems must be overcome before an optical flow field can be derived from an image sequence. These problems are discussed and an outline of a scheme for overcoming these obstacles is presented. A two-dimensional motion sensor is presented which is capable of measuring edge velocity in twelve separate directions at any point in the image. However, because of the aperture problem, the sensors can only determine the velocity component in the direction orthogonal to the edge. A second stage which uses a cosine-weighted voting scheme is used to derive the true flow field from the edge velocities. The whole system is based on a parallel network of connections and weights and is potentially a very fast method of obtaining the two-dimensional flow field.

Perrone, John A.↗

Inner Magnetospheric Electric Fields Derived from IMAGE EUV

The local and global patterns of plasmaspheric plasma transport reflect the influence of electric fields imposed by all sources in the inner magnetosphere. Image sequences of thermal plasma G:istribution obtained from the IMAGE Mission Extreme Ultraviolet Imager can be used to derive plasma motions and, using a magnetic field model, the corresponding electric fields. These motions and fields directly reflect the dynamic coupling of injected plasmasheet plasma and the ionosphere, in addition to solar wind and atmospheric drivers. What is being learned about the morphology of inner magnetospheric electric fields during storm and quite conditions from this new empirical tool will be presented and discussed.

Gallagher, D. L.↗

Bayesian Approach to Estimation of Water Table Elevations Using Historical Rasters as Prior Information 2019 - 20430

In cases of complex but only partially known geology and a lack of spatial control in observation well locations, water table elevation estimation is very challenging. In some cases, auxiliary information, such as observations of the movement of tracers, operation of injection and extraction wells, and calibration of groundwater models against the historical elevation data, can be combined with expert judgement to estimate flow directions in areas of sparse data and to aid in the production of more reliable contour maps (and associated rasters) than could be produced by relying on sparse well elevation data alone. Given a historical sequence of these raster maps, the question arises how to automate, to the extent possible, the process of producing new raster maps to reflect data from previous times, the current data and the operation of expert judgement. One solution is to adopt a Bayesian point of view and to regard the historical well elevation data, auxiliary information and historical raster maps as prior information. The well elevations for water table wells, as well as those for injection/extraction wells and the data associated with other relevant variables, can be viewed as predictors for the raster surface. From this prior information, we can, conditional on the values of these predictors for a new time period, compute an expected value map and a standard deviation map for the new raster. These then can be taken to specify a prior predictive distribution for the pixels in the new raster map. Then we condition the pixels, corresponding to water level observation wells within the raster, on the observed values in those wells (which in general will differ from the regression estimate) for the new time period. Given the smoothness of the water table surface, we then smooth the surface of deviations from the mean surface, based on the variograms of the historical rasters, and add this smoothed surface to the regression mean surface. The error structure of the produced raster map is defined by the regression error structure and the error due to smoothing based on the estimated variograms. This methodology has been developed and is being further refined for groundwater monitoring and remediation at LANL. It is a very flexible method that can also be applied with a variety of other predictors applied to model the water level wells in the area of interest over the historical record. The smoothness of the spatial process and its possible evolution over time can then be estimated from the residuals from this regression. This can be augmented by expert hydrogeological opinion based on site topography and hydrogeology. (authors)

54 ENVIRONMENTAL SCIENCES↗

Crystal structures of pinoresinol-lariciresinol and phenylcoumaran benzylic ether reductases and their relationship to isoflavone reductases

Despite the importance of plant lignans and isoflavonoids in human health protection (e.g. for both treatment and prevention of onset of various cancers) as well as in plant biology (e.g. in defense functions and in heartwood development), systematic studies on the enzymes involved in their biosynthesis have only recently begun. In this investigation, three NADPH-dependent aromatic alcohol reductases were comprehensively studied, namely pinoresinol-lariciresinol reductase (PLR), phenylcoumaran benzylic ether reductase (PCBER), and isoflavone reductase (IFR), which are involved in central steps to the various important bioactive lignans and isoflavonoids. Of particular interest was in determining how differing regio- and enantiospecificities are achieved with the different enzymes, despite each apparently going through similar enone intermediates. Initially, the three-dimensional x-ray crystal structures of both PLR_Tp1 and PCBER_Pt1 were solved and refined to 2.5 and 2.2 A resolutions, respectively. Not only do they share high gene sequence similarity, but their structures are similar, having a continuous alpha/beta NADPH-binding domain and a smaller substrate-binding domain. IFR (whose crystal structure is not yet obtained) was also compared (modeled) with PLR and PCBER and was deduced to have the same overall basic structure. The basis for the distinct enantio-specific and regio-specific reactions of PCBER, PLR, and IFR, as well as the reaction mechanism and participating residues involved (as identified by site-directed mutagenesis), are discussed.

NASA Discipline Plant Biology↗

Orthogonal glycolytic pathway enables directed evolution of noncanonical cofactor oxidase

Abstract Noncanonical cofactor biomimetics (NCBs) such as nicotinamide mononucleotide (NMN + ) provide enhanced scalability for biomanufacturing. However, engineering enzymes to accept NCBs is difficult. Here, we establish a growth selection platform to evolve enzymes to utilize NMN + -based reducing power. This is based on an orthogonal, NMN + -dependent glycolytic pathway in Escherichia coli which can be coupled to any reciprocal enzyme to recycle the ensuing reduced NMN + . With a throughput of >10 6 variants per iteration, the growth selection discovers a Lactobacillus pentosus NADH oxidase variant with ~10-fold increase in NMNH catalytic efficiency and enhanced activity for other NCBs. Molecular modeling and experimental validation suggest that instead of directly contacting NCBs, the mutations optimize the enzyme’s global conformational dynamics to resemble the WT with the native cofactor bound. Restoring the enzyme’s access to catalytically competent conformation states via deep navigation of protein sequence space with high-throughput evolution provides a universal route to engineer NCB-dependent enzymes.

59 BASIC BIOLOGICAL SCIENCES↗

Identifying candidate structured RNAs in CRISPR operons

Noncoding RNAs with secondary structures play important roles in CRISPR-Cas systems. Many of these structures likely remain undiscovered. We used a large-scale comparative genomics approach to predict 156 novel candidate structured RNAs from 36,111 CRISPR-Cas systems. A number of these were found to overlap with coding genes, including palindromic candidates that overlapped with a variety of Cas genes in type I and III systems. Among these 156 candidates, we identified 46 new models of CRISPR direct repeats and 1 tracrRNA. This tracrRNA model occasionally overlapped with predicted cas9 coding regions, emphasizing the importance of expanding our search windows for novel structure RNAs in coding regions. We also demonstrated that the antirepeat sequence in this tracrRNA model can be used to accurately assign thousands of predicted CRISPR arrays to type II-C systems. This study highlights the importance of unbiased identification of candidate structured RNAs across CRISPR-Cas systems.

59 BASIC BIOLOGICAL SCIENCES↗