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At least 775 records · Page 43

Origin of replication discovery for environmentally isolated Pantoea strain enables expression of heterologous proteins, pathways and products

Leveraging predicted origin sequences from a previously characterized groundwater plasmidome, we constructed a barcoded plasmid library to screen for previously unknown origins. Testing this library against a panel of representative bacterial strains led to the identification of 3 previously unknown origins that replicate in gram-negative bacteria not previously associated with these origin sequences. Experimental validation confirmed that a plasmid bearing origin 6911 as the sole origin could replicate with a copy number of 9 (±2) in Pantoea sp. MT58, a fast growing and metal tolerant, environmentally important bacterium. Plasmids based on this new origin were used to express the reporter protein GFP, and non-native metabolite pathways for the natural product indigoidine and the terpenoid compound isoprenol. Functional previously unknown origins of replication in such non-model organisms can expand the toolkit for genetic manipulations of both model and less-studied bacteria.

molecular biology↗

Numerical comparisons of nonlinear convergence accelerators

As part of a continuing program of numerical tests of convergence accelerators, the iterated Aitken's Delta-squared method, Wynn's epsilon algorithm, Brezinski's theta algorithm, and Levin's u transform are compared on a broad range of test problems: linearly convergence alternating, monotone, and irregular-sign series, logarithmically convergent series, power method and Bernoulli method sequences, alternating and monotone asymptotic series, and some perturbation series arising in applications. In each category either the epsilon algorithm or the u transform gives the best results of the four methods tested. In some cases differences among methods are slight, and in others they are quite striking.

Smith, D. A.↗

Eclipsing binary stars as tests of gravity theories - The apsidal motion of AS Camelopardalis

AS Camelopardalis is an 8th-magnitude eclipsing binary that consists of two main-sequence (B8 V and a B9.5 V) components in an eccentric orbit (e = 0.17) with an orbital period of 3.43 days. Like the eccentric eclipsing system DI Herculis, and a few other systems, AS Cam is an important test case for studying relativistic apsidal motion. In these systems, the theoretical general relativistic apsidal motion is comparable to that expected from classical effects arising from tidal and rotational deformation of the stellar components. Accurate determinations of the orbital and stellar properties of AS Cam have been made by Hilditch (1972) and Khalliulin and Kozyreva (1983) that permit the theoretical relativistic and classical contributions to the apsidal motion to be determined reasonably well. All the published timings of primary and secondary minima have been gathered and supplemented with eclipse timings from 1899 to 1920 obtained from the Harvard plate collection. Least-squares solutions of the eclipse timings extending over an 80 yr interval yield a smaller than expected apsidal motion, in agreement with that found by Khalliulin and Kozyreva from a smaller set of data. The observed apsidal motion for AS Cam is about one-third that expected from the combined relativistic and classical effects. Thus, AS Cam joins DI Her in having an observed apsidal motion significantly less than that predicted from theory.

Maloney, Frank P.↗

Extending evolutionary forecasts across bacterial species

Improving evolutionary forecasting requires progressing from studying repeated evolution of a single genotype under identical conditions to formulating broad principles. These principles should enable predictions of how similar species will adapt to similar selective pressures. Evolve-and-resequence experiments with multiple species allow testing forecasts on different biological levels and elucidating the causes for failed predictions. Here, we show that forecasts for adaptation to static culture conditions can be extended to multiple species by testing previous predictions for Pseudomonas syringae and Pseudomonas savastanoi. In addition to sequence divergence, these species differ in their repertoire of biofilm regulatory genes and structural components. Consistent with predictions, both species repeatedly produced biofilm mutants with a wrinkly spreader phenotype. Predominantly, mutations occurred in the wsp operon, with less frequent promoter mutations near uncharacterized diguanylate cyclases. However, mutational patterns differed on the gene level, which was explained by a lack of conservation in relative fitness of mutants between more divergent species. The same mutation was the most frequent for both species suggesting that conserved mutation hotspots can increase parallel evolution. This study shows that evolutionary forecasts can be extended across species, but that differences in the genotype–phenotype–fitness map and mutational biases limit predictability on a detailed molecular level.

59 BASIC BIOLOGICAL SCIENCES↗

Long-Term and Low-Level Envelope C2V3 Stimulation by Highly Diverse Virus Isolates Leads to Frequent Development of Broad and Elite Antibody Neutralization in HIV-1-Infected Individuals

A minority of HIV-1-infected patients produce broadly neutralizing antibodies (bNAbs). Identification of viral and host correlates of bNAb production may help develop vaccines. We aimed to characterize the neutralizing response and viral and host-associated factors in Angola, which has one of the oldest, most dynamic, and most diverse HIV-1 epidemics in the world. Three hundred twenty-two HIV-1-infected adults from Angola were included in this retrospective study. Phylogenetic analysis of C2V3C3 env gene sequences was used for virus subtyping. Env-binding antibody reactivity was tested against polypeptides comprising the C2, V3, and C3 regions. Neutralizing-antibody responses were determined against a reference panel of tier 2 Env pseudoviruses in TZM-bl cells; neutralizing epitope specificities were predicted using ClustVis. All subtypes were found, along with untypeable strains and recombinant forms. Notably, 56% of the patients developed cross neutralizing, broadly neutralizing, or elite neutralizing responses. Broad and elite neutralization was associated with longer infection time, subtype C, lower CD4 + T cell counts, higher age, and higher titer of C2V3C3-specific antibodies relative to failure to develop bNAbs. Neutralizing antibodies targeted the V3-glycan supersite in most patients. V3 and C3 regions were significantly less variable in elite neutralizers than in weak neutralizers and nonneutralizers, suggesting an active role of V3C3-directed bNAbs in controlling HIV-1 replication and diversification. In conclusion, prolonged and low-level envelope V3C3 stimulation by highly diverse and ancestral HIV-1 isolates promotes the frequent elicitation of bNAbs. These results provide important clues for the development of an effective HIV-1 vaccine. Studies on neutralization by antibodies and their determinants in HIV-1-infected individuals have mostly been conducted in relatively recent epidemics caused by subtype B and C viruses. Results have suggested that elicitation of broadly neutralizing antibodies (bNAbs) is uncommon. The mechanisms underlying the elicitation of bNAbs are still largely unknown. We performed the first characterization of the plasma neutralizing response in a cohort of HIV-1-infected patients from Angola. Angola is characterized by an old and dynamic epidemic caused by highly diverse HIV-1 variants. Remarkably, more than half of the patients produced bNAbs, mostly targeting the V3-glycan supersite in HIV-1. This was associated with higher age, longer infection time, lower CD4 + T cell counts, subtype C infection, or higher titer of C2V3C3-specific antibodies relative to patients that did not develop bNAbs. These results may help develop the next generation of vaccine candidates for HIV-1.

59 BASIC BIOLOGICAL SCIENCES↗

Modeling Exhaust-Generated Aerodynamic Pressure Loads on Airfield Matting Repair Systems

Airfield matting systems are commonly used for rapid repair of damaged runways to facilitate continuity of critical operations. Under normal service conditions, matting repair systems are subject not only to wheel loads exerted by airfield traffic but also to aerodynamic pressure loads resulting from high-speed, turbulent exhaust plumes produced by jets during taxi and take-off. Matting systems employed by the U.S. Air Force have been tested with respect to wheel loads, but probabilities of failure because of pressure loads produced by jet exhaust have not yet been established. This study presents a numerical approach for preliminary estimation of worst-case matting anchor forces resulting from exhaust-generated pressure loads. Using a two-dimensional computational fluid dynamics model, system behavior is evaluated by means of a parametric study of six system variables, of which the most significant are: (1) distance between engine and matting, (2) depth of cavity openings at matting edges, and (3) engine exhaust velocity. The results demonstrate that matting systems are likely to experience net uplift in typical service scenarios, driven by the combined effects of flow separation and cavity pressurization. Worst-case anchor pull-out forces, computed according to a tributary-area approach, are estimated to fall in the range of 130 lb (581 N) to 979 lb (4,350 N), depending on assumed load-sharing behavior among anchors and the size of the repair site. Field testing of instrumented matting systems during jet taxi and take-off sequences is recommended as the best next step toward understanding system behavior.

Engineering↗

Tracking and Rejection of Biased Sinusoidal Signals Using Generalized Predictive Controller

Some novel applications require the tracking/rejection of biased sinusoidal reference/distur-bances. According to the internal model principle (IMP), a controller must embed the model of a biased sinusoidal signal to track references and also reject perturbations modeled through the aforementioned signal. However, the design of that kind of controller is not straightforward, especially when they are implemented in digital processors. This paper presents a controller, based on generalized predictive control (GPC), designed for tracking/rejection of biased sinusoidal signals. In general, GPC is based on the prediction of the plant responses through an augmented prediction model. The proposed approach develops an augmented model that predicts the future errors. The prediction model and the control law used in the proposed approach embed the discrete-time model of a biased sinusoidal signal. Thus, the proposed controller can track/reject biased sinusoidal references/disturbances. The predicted errors and the future inputs of the proposed augmented model are used to define the cost function that measures the control performance. An optimization technique was applied to obtain the solution of the cost function, which is the optimal sequence of future model inputs that allows defining the control law. Experimental tests prove that the proposed controller can asymptotically track and reject biased sinusoidal signals.

42 ENGINEERING↗

Land utilization and water resource inventories over extended test sites

In addition to the work on the corn blight this year, several other analysis tests were completed which resulted in significant findings. These aspects are discussed as follows: (1) field spectral measurements of soil conditions; (2) analysis of extended test site data; this discussion involves three different sets of data analysis sequences; (3) urban land use analysis, for studying water runoff potentials; and (4) thermal data quality study, as an expansion of our water resources studies involving temperature calibration techniques.

Hoffer, R. M.↗

Development and performance of power processor system for 2-gigahertz, 200-watt amplifier for communications technology satellite

The electrical and environmental requirements for a power processor system (PPS) designed to supply the appropriate voltages and currents to a 200-watt traveling wave tube (TWT) for a communication technology satellite is described. A block diagram of the PPS, the interface requirements between the PPS and spacecraft, the interface requirements between the PPS and 200-watt TWT, and the environmental requirements of the PPS are presented. Also included are discussions of protection circuits, interlocking sequences, and transient requirements. Predictions of the flight performance, based on ground test data, are provided.

Siegert, C. E.↗

A comparison of cloud motion winds from ATS 6 images with coinciding SMS 1 winds

A methodology is developed for accurate measurement of cloud motion winds from the geosynchronous ATS 6 image data. Attitude changes between consecutive images (as a function of scan-line number) are accounted for in wind computations through measurement of the earth-edge displacements between the successive infrared images. Also, an image matching procedure is used to remove obvious and distracting image distortions. The availability of SMS imagery coinciding with ATS 6 imagery makes SMS an excellent reference against which the quality of ATS 6 winds can be tested. The resulting winds inferred from cloud displacement measurements taken from a sequence of the corrected images are found to agree better than 2 m/sec rms with winds measured from coincident SMS 1 imagery.

Kuhlow, W. W.↗

On testing stuck-open faults in CMOS combinational circuits

Recently it has been found that a class of failure related to a particular technology (CMOS) cannot be modelled as the conventional stuck-at fault model. These failures change the combinational behavior of CMOS logic gates into a sequential one. Such a failure is modelled as a fault, called the Stuck-Open fault (SOP). The object of this paper is to develop a procedure to detect single SOPs in combinational circuits. It is shown, that in general, tests generated for stuck-at faults when applied in a particular sequence will detect all single SOP faults. In case of single redundancy in the network, the SOP fault on the redundant line cannot be detected. When there is reconvergent fan-out in the network, there is a one-one correspondence between the conditions for stuck-at fault and stuck-open fault detectability.

Chandramouli, R.↗

Scaling effects in the static large deflection response of graphite-epoxy composite beams

Scaling effects in the large deflection response of graphite-epoxy composite beams was investigated. Eight different scale model beams ranging from 1/6 to full-scale were subjected to an eccentric axial compressive load to promote large bending deformations and failures. Beams having laminate stacking sequences including unidirectional, angle ply, cross ply, and quasi-isotropic were tested to examine a wide variety of composite response and failure modes. The model beams were loaded under scaled test conditions until catastrophic failure. Data acquired included load, end displacement, and strain measurements, and qualitative failure measurements. The experimental data is compared to a large rotation beam analysis and a finite element model analysis. Results from the tests indicate that the beam response becomes nonlinear. Failure modes are consistent between scale models within a laminate family, however, a significant scale effect is observed in strength of the scaled beams.

Jackson, Karen E.↗

Implementation of a state of the art automated system for the production of cloud/water vapor motion winds from geostationary satellites

The thrust of the proposed effort under this contract is aimed at improving techniques to track water vapor data in sequences of imagery from geostationary satellites. In regards to this task, significant testing, evaluation, and progress was accomplished during this period. Sets of winds derived from Meteosat data were routinely produced during Atlantic hurricane events in the 1993 season. These wind sets were delivered via Internet in real time to the Hurricane Research Division in Miami for their evaluation in a track forecast model. For eighteen cases in which 72-hour forecasts were produced, thirteen resulted in track forecast improvements (some quite significant). In addition, quality-controlled Meteosat water vapor winds produced by NESDIS were validated against rawinsondes, yielding an 8 m/s RMS. This figure is comparable to upper-level cloud drift wind accuracies. Given the complementary horizontal coverage in cloud-free areas, we believe that water vapor vectors can supplement cloud-drift wind information to provide good full-disk coverage of the upper tropospheric flow. The impact of these winds on numerical analysis and forecasts will be tested in the next reporting period.

Velden, Christopher S.↗

A coronagraphic search for brown dwarfs around nearby stars

Brown dwarf companions have been searched for around stars within 10 pc of the Sun using the Johns-Hopkins University Adaptive Optics Coronagraph (AOC), a stellar coronagraph with an image stabilizer. The AOC covers the field around the target star with a minimum search radius of 1 sec .5 and a field of view of 1 arcmin sq. We have reached an unprecedented dynamic range of Delta m = 13 in our search for faint companions at I band. Comparison of our survey with other brown dwarf searches shows that the AOC technique is unique in its dynamic range while at the same time just as sensitive to brown dwarfs as the recent brown dwarf surveys. The present survey covered 24 target stars selected from the Gliese catalog. A total of 94 stars were detected in 16 fields. The low-latitude fields are completely dominated by background star contamination. Kolmogorov-Smirnov tests were carried out for a sample restricted to high latitudes and a sample with small angular separations. The high-latitude sample (b greater than or equal to 44 deg) appears to show spatial concentration toward target stars. The small separation sample (Delta Theta less than 20 sec) shows weaker dependence on Galactic coordinates than field stars. These statistical tests suggest that both the high-latitude sample and the small separation sample can include a substantial fraction of true companions. However, the nature of these putative companions is mysterious. They are too faint to be white dwarfs and too blue for brown dwarfs. Ignoring the signif icance of the statistical tests, we can reconcile most of the detections with distant main-sequence stars or white dwarfs except for a candidate next to GL 475. Given the small size of our sample, we conclude that considerably more targets need to be surveyed before a firm conclusion on the possibility of a new class of companions can be made.

Nakajima, T.↗

Bantam System Technology Project Ground System Requirements Document

The Low Cost Booster Project (LCBP), also known as Bantam, is an element of the Advanced Space Transportation Program focused on Low Cost Booster Technologies. During FY 99 flight demonstrations are planned to demonstrate the feasibility of producing a booster capable of inserting a 150 kg payload into low earth orbit. The ground support system is an element of the full launch system. The ground support system provides for integration of the payload with the launch vehicle, preparation of the vehicle for launch (including maintenance, integration and test of the vehicle flight software), monitor and control of the launch sequence, range safety during launch, and collection of telemetry during the flight up to payload release. The ground support system is intended to make the maximum possible use of Government Off-the-Shelf (GOTS) or Commercial Off-the-Shelf (COTS) hardware and software to obtain the best value in terms of development operations support and ultimate life cycle cost for the launch system.

Moon, J. M.↗

Implementation and Testing of Turbulence Models for the F18-HARV Simulation

This report presents three methods of implementing the Dryden power spectral density model for atmospheric turbulence. Included are the equations which define the three methods and computer source code written in Advanced Continuous Simulation Language to implement the equations. Time-history plots and sample statistics of simulated turbulence results from executing the code in a test program are also presented. Power spectral densities were computed for sample sequences of turbulence and are plotted for comparison with the Dryden spectra. The three model implementations were installed in a nonlinear six-degree-of-freedom simulation of the High Alpha Research Vehicle airplane. Aircraft simulation responses to turbulence generated with the three implementations are presented as plots.

Yeager, Jessie C.↗

Soft-Decision Decoding of Binary Linear Block Codes Based on an Iterative Search Algorithm

This correspondence presents a suboptimum soft-decision decoding scheme for binary linear block codes based on an iterative search algorithm. The scheme uses an algebraic decoder to iteratively generate a sequence of candidate codewords one at a time using a set of test error patterns that are constructed based on the reliability information of the received symbols. When a candidate codeword is generated, it is tested based on an optimality condition. If it satisfies the optimality condition, then it is the most likely (ML) codeword and the decoding stops. If it fails the optimality test, a search for the ML codeword is conducted in a region which contains the ML codeword. The search region is determined by the current candidate codeword and the reliability of the received symbols. The search is conducted through a purged trellis diagram for the given code using the Viterbi algorithm. If the search fails to find the ML codeword, a new candidate is generated using a new test error pattern, and the optimality test and search are renewed. The process of testing and search continues until either the MEL codeword is found or all the test error patterns are exhausted and the decoding process is terminated. Numerical results show that the proposed decoding scheme achieves either practically optimal performance or a performance only a fraction of a decibel away from the optimal maximum-likelihood decoding with a significant reduction in decoding complexity compared with the Viterbi decoding based on the full trellis diagram of the codes.

Lin, Shu↗

Pre-Test Analysis Predictions for the Shell Buckling Knockdown Factor Checkout Tests - TA01 and TA02

This report summarizes the pre-test analysis predictions for the SBKF-P2-CYL-TA01 and SBKF-P2-CYL-TA02 shell buckling tests conducted at the Marshall Space Flight Center (MSFC) in support of the Shell Buckling Knockdown Factor (SBKF) Project, NASA Engineering and Safety Center (NESC) Assessment. The test article (TA) is an 8-foot-diameter aluminum-lithium (Al-Li) orthogrid cylindrical shell with similar design features as that of the proposed Ares-I and Ares-V barrel structures. In support of the testing effort, detailed structural analyses were conducted and the results were used to monitor the behavior of the TA during the testing. A summary of predicted results for each of the five load sequences is presented herein.

Thornburgh, Robert P.↗