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At least 73 records · Page 4
All sourcesNASA NTRSDOE OSTI
NASA NTRS2001

Optimization of an oligonucleotide microchip for microbial identification studies: a non-equilibrium dissociation approach

The utility of a high-density oligonucleotide microarray (microchip) for identifying strains of five closely related bacilli (Bacillus anthracis, Bacillus cereus, Bacillus mycoides, Bacillus medusa and Bacillus subtilis) was demonstrated using an approach that compares the non-equilibrium dissociation rates ('melting curves') of all probe-target duplexes simultaneously. For this study, a hierarchical set of 30 oligonucleotide probes targeting the 16S ribosomal RNA of these bacilli at multiple levels of specificity (approximate taxonomic ranks of domain, kingdom, order, genus and species) was designed and immobilized in a high-density matrix of gel pads on a glass slide. Reproducible melting curves for probes with different levels of specificity were obtained using an optimized salt concentration. Clear discrimination between perfect match (PM) and mismatch (MM) duplexes was achieved. By normalizing the signals to an internal standard (a universal probe), a more than twofold discrimination (> 2.4x) was achieved between PM and 1-MM duplexes at the dissociation temperature at which 50% of the probe-target duplexes remained intact. This provided excellent differentiation among representatives of different Bacillus species, both individually and in mixtures of two or three. The overall pattern of hybridization derived from this hierarchical probe set also provided a clear 'chip fingerprint' for each of these closely related Bacillus species.

Non-NASA Center↗
NASA NTRS1999

A model of EcoRII restriction endonuclease action: the active complex is most likely formed by one protein subunit and one DNA recognition site

To elucidate the mechanism of interaction of restriction endonuclease EcoRII with DNA, we studied by native gel electrophoresis the binding of this endonuclease to a set of synthetic DNA-duplexes containing the modified or canonical recognition sequence 5'-d(CCA/TGG)-3'. All binding substrate or substrate analogues tested could be divided into two major groups: (i) duplexes that, at the interaction with endonuclease EcoRII, form two types of stable complexes on native gel in the absence of Mg2+ cofactor; (ii) duplexes that form only one type of complex, observed both in the presence and absence of Mg2+. Unlike the latter, duplexes under the first group can be hydrolyzed by endonuclease. Data obtained suggest that the active complex is most likely formed by one protein subunit and one DNA recognition sequence. A model of EcoRII endonuclease action is presented.

Oligodeoxyribonucleotides/chemistry/metabolism↗
NASA NTRS1981

Flight test results for the F-8 digital fly-by-wire aircraft control sensor analytic redundancy management technique

This paper reviews the formulation and flight test results of an algorithm to detect and isolate the first failure of any one of twelve duplex control sensor signals being monitored. The technique uses like-signal differences for fault detection while relying upon analytic redundancy relationships among unlike quantities to isolate the faulty sensor. The fault isolation logic utilizes the modified sequential probability ratio test, which explicitly accommodates the inevitable irreducible low frequency errors present in the analytic redundancy residuals. In addition, the algorithm uses sensor output selftest, which takes advantage of the duplex sensor structure by immediately removing a highly erratic sensor from control calculations and analytic redundancy relationships while awaiting a definitive fault isolation decision via analytic redundancy. This study represents a proof of concept demonstration of a methodology that can be applied to duplex or higher flight control sensor configurations and, in addition, can monitor the health of one simplex signal per analytic redundancy relationship.

Deckert, J. C.↗
NASA NTRS1983

Analytic redundance management for flight control sensors

The formulation and flight test results of an algorithm to detect and isolate the first failure of any one of 12 duplex control sensor signals (24 in all) being monitored are reviewed. The technique uses like signal differences for fault detection while relying upon analytic redundancy relationships among unlike quantities to isolate the faulty sensor. The fault isolation logic utilizes the modified sequential probability ratio test, which explicitly accommodates the inevitable, irreducible low frequency errors present in the analytic redundancy residuals. In addition, the algorithm used sensor output selftest, which takes advantage of the duplex sensor structure by immediately removing a hard failed sensor from control calculations and analytic redundancy relationships while awaiting a definitive fault isolation decision via analytic redundancy. This study represents a proof of concept demonstration of a methodology that is applied to duplex or higher flight control sensor configurations and, in addition, can monitor the health of one simplex signal per analytic redundancy relationship.

Deckert, J. C.↗
NASA NTRS1993

Comparative analysis of different configurations of PLC-based safety systems from reliability point of view

The study of a comparative analysis of distinct multiplex and fault-tolerant configurations for a PLC-based safety system from a reliability point of view is presented. It considers simplex, duplex and fault-tolerant triple redundancy configurations. The standby unit in case of a duplex configuration has a failure rate which is k times the failure rate of the standby unit, the value of k varying from 0 to 1. For distinct values of MTTR and MTTF of the main unit, MTBF and availability for these configurations are calculated. The effect of duplexing only the PLC module or only the sensors and the actuators module, on the MTBF of the configuration, is also presented. The results are summarized and merits and demerits of various configurations under distinct environments are discussed.

Tapia, Moiez A.↗
NASA NTRS1998

Origin of the catalytic activity of bovine seminal ribonuclease against double-stranded RNA

Bovine seminal ribonuclease (RNase) binds, melts, and (in the case of RNA) catalyzes the hydrolysis of double-stranded nucleic acid 30-fold better under physiological conditions than its pancreatic homologue, the well-known RNase A. Reported here are site-directed mutagenesis experiments that identify the sequence determinants of this enhanced catalytic activity. These experiments have been guided in part by experimental reconstructions of ancestral RNases from extinct organisms that were intermediates in the evolution of the RNase superfamily. It is shown that the enhanced interactions between bovine seminal RNase and double-stranded nucleic acid do not arise from the increased number of basic residues carried by the seminal enzyme. Rather, a combination of a dimeric structure and the introduction of two glycine residues at positions 38 and 111 on the periphery of the active site confers the full catalytic activity of bovine seminal RNase against duplex RNA. A structural model is presented to explain these data, the use of evolutionary reconstructions to guide protein engineering experiments is discussed, and a new variant of RNase A, A(Q28L K31C S32C D38G E111G), which contains all of the elements identified in these experiments as being important for duplex activity, is prepared. This is the most powerful catalyst within this subfamily yet observed, some 46-fold more active against duplex RNA than RNase A.

Non-NASA Center↗
NASA NTRS1994

Crystal structure of a four-stranded intercalated DNA: d(C4)

The crystal structure of d(C4) solved at 2.3-A resolution reveals a four-stranded molecule composed of two interdigitated or intercalated duplexes. The duplexes are held together by hemiprotonated cytosine-cytosine base pairs and are parallel stranded, but the two duplexes point in opposite directions. The molecule has a slow right-handed twist of 12.4 degrees between covalently linked cytosine base pairs, and the base stacking distance is 3.1 A. This is in general agreement with the NMR studies. A biological role for DNA in this conformation is suggested.

Non-NASA Center↗
NASA NTRS2011

Atomic Simulation of Complex DNA DSBs and the Interactions with the Ku70/80 Heterodimer

DNA double strand breaks (DSBs) induced by ionizing radiation (IR) usually contain modified bases such as 8-oxo-7,8-dihydroguanine (8-oxoG) and thymine glycol, apurinic/apyrimidinic (AP) sites, 2-deoxyribonolactone, or single-strand breaks (SSBs). The presence of such lesions in close proximity to the DSB terminus makes the DNA nicks more difficult to repair and rejoin than endogenously induced simple DSBs, and as such a major determinant of the biological effects of high linear energy transfer (LET) radiation as encountered in space travel. In this study we conducted molecular dynamics simulations on a series of DNA duplexes with various complex lesions of 8-oxoG and AP sites, in an effort to investigate the effects of such lesions to the structural integrity and stability of DNA after insulted by IR. We also simulated the interaction of such complex DSBs with the Ku70/80 heterodimer, the first protein in mammalian cells to embark the non-homologous end joining (NHEJ) DNA repair pathway. The results indicate, compared to DNA with simple DSBs, the complex lesions can enhance the hydrogen bonds opening rate at the DNA terminus, and increase the mobility of the whole duplex, thus they present more deleterious effects to the genome integrity if not captured and repaired promptly in cells. Simulations also demonstrate the binding of Ku drastically reduces structural disruption and flexibility caused by the complex lesions, and the interactions of Ku with complex DSBs have a different potential energy landscape from the bound structure with simple DSB. In all complex DSBs systems, the binding of DSB terminus with Ku70 is softened while the binding of the middle duplex with Ku80 is tightened. This energy shift may help the Ku protein to secure at the DSB terminus for a longer time, so that other end processing factors or repair pathways can proceed at the lesions before NHEJ repair process starts. These atomic simulations may provide valuable new insight into the selective action of repair proteins on damaged DNA.

Hu, Shaowen↗
NASA NTRS2011

Atomistic Simulations of Complex DNA DSBs and the Interactions with Ku70/80 Heterodimer

Compared to DNA with simple DSBs, the complex lesions can enhance the hydrogen bonds opening rate at the DNA terminus, and increase the mobility of the whole duplex. Binding of Ku drastically reduces the structural disruption and flexibility caused by the complex lesions. In all complex DSBs systems, the binding of DSB terminus with Ku70 is softened while the binding of the middle duplex with Ku80 is tightened. Binding of Ku promotes the rigidity of DNA duplexes, due to the clamp structure of the inner surface of the rings of Ku70/80.

Hu, Shaowen↗
NASA NTRS1971

Effect of initial hydrogen content of a titanium alloy on susceptibility to hot-salt stress-corrosion

The Ti-8Al-1Mo-1V alloy was tested in four conditions: mill annealed (70 ppM H), duplex annealed (70 ppM H), vacuum annealed to an intermediate (36 ppM) and a low (9 ppM H) hydrogen level. Material annealed at 650 C (duplex condition) exhibited resistance to hot-salt stress corrosion superior to that exhibited by material in the mill-annealed condition. Reduction of the alloy hydrogen content from 70 to as low as 9 ppM did not influence resistance to hot-salt stress corrosion embrittlement or cracking.

Gray, H. R.↗
NASA NTRS1971

A fault-tolerant information processing concept for space vehicles.

A distributed fault-tolerant information processing system is proposed, comprising a central multiprocessor, dedicated local processors, and multiplexed input-output buses connecting them together. The processors in the multiprocessor are duplicated for error detection, which is felt to be less expensive than using coded redundancy of comparable effectiveness. Error recovery is made possible by a triplicated scratchpad memory in each processor. The main multiprocessor memory uses replicated memory for error detection and correction. Local processors use any of three conventional redundancy techniques: voting, duplex pairs with backup, and duplex pairs in independent subsystems.

Hopkins, A. L., Jr.↗
NASA NTRS1972

Effect of initial hydrogen content of a titanium alloy on susceptibility to hot salt stress corrosion.

The influence of the initial hydrogen content of a titanium alloy on subsequent resistance to hot salt stress corrosion embrittlement and cracking was investigated. A Ti-8Al-1Mo-1V alloy was tested in four conditions: mill annealed (70 ppm H), duplex annealed (70 ppm H), vacuum annealed to an intermediate (36 ppm H) and a low (9 ppm H) hydrogen level. Material annealed at 650 C (duplex condition) exhibited resistance to hot salt stress corrosion superior to that exhibited by material in the mill annealed condition. Reduction of the alloy hydrogen content from 70 to as low as 9 ppm did not influence resistance to hot salt stress corrosion embrittlement or cracking.

Gray, H. R.↗
NASA NTRS1974

Coatings for directional eutectics

Coating compositions were evaluated for oxidation protection of directionally solidified composite alloy NiTaC-13. These coatings included three NiCrAlY compositions (30-5-1, 25-10-1 and 20-15-1), two FeCrAlY compositions (30-5-1 and 25-10-1), a CoCrAlY composition (25-10-1), and one duplex coating, Ni-35Cr + Al. Duplicate pin samples of each composition were evaluated using two cyclic furnace oxidation tests of 100 hours at 871 C and 500 hours at 1093 C. The two best coatings were Ni-20Cr-15Al-lY and Ni-35Cr + Al. The two preferred coatings were deposited on pins and were evaluated in detail in .05 Mach cyclic burner rig oxidation to 1093 C. The NiCrAlY coating was protective after 830 hours of cycling, while the duplex coating withstood 630 hours. Test bars were coated and cycled for up to 500 hours. Tensile tests indicated no effect of coatings on strength. In 871 C air stress rupture, a degradation was observed for coated relative to bare material. The cycled NiCrAlY coating offered excellent protection with properties superior to the bare cycled NiTaC-13 in 1093 C air stress rupture.

Jackson, M. R.↗
NASA NTRS1974

High-temperature strength stability of three forms of chemically vapor deposited tungsten

Three types of CVD tungsten (fluoride-produced, chloride-produced, and a layered composite of the two-termed duplex) were evaluated to determine their high-temperature strength and microstructural stability following 5000-hr exposure to temperatures of 1540 and 1700 C. At the highest temperatures investigated (1540 and 1700 C), the tensile strengths of the two basic materials were essentially equal. At lower temperatures, chloride tungsten possessed lower yield strength but higher ultimate strength than fluoride tungsten while the behavior of the duplex material was generally intermediate. Apparent anomalies in high-temperature elongation behavior are explained on the basis of grain boundary cavity formation and recrystallization. The grain size of fluoride tungsten changed only slightly following 5000-h treatment at 1700 C. In contrast, chloride tungsten possessed both poor resistance to grain growth and an accompanying relatively high ductile-brittle transition temperature.

Bryant, W. A.↗
NASA NTRS1977

Real-time testing of titanium sheet and extrusion coupon specimens subjected to Mach 2.7 supersonic cruise aircraft wing stresses and temperatures

The accuracy of three accelerated flight-by-flight test methods for material selection, and fatigue substantiation of supersonic cruise aircraft structure was studied. The real time stresses and temperatures applied to the specimens were representative of the service conditions in the lower surface of a Mach 2.7 supersonic cruise aircraft wing root structure. Each real time flight lasted about 65 minutes, including about one hour at (500 F) in the cruise condition. Center notched coupon specimens from six titanium materials were tested: mill-annealed, duplex-annealed, and triplex-annealed Ti-8Al-1Mo-1V sheets; mill-annealed Ti-8Al-1Mo-1V extrusion; mill-annealed Ti-6Al-4V sheet; and solution-treated and aged Ti-6Al-4V extrusion. For duplex-annealed Ti-8Al-1Mo-1V sheet, specimens with single spotweld were also tested. The test results were studied in conjunction with other related data from the literature for: material selection, structural fabrication, fatigue resistance of supersonic cruise aircraft structure, and fatigue test acceleration procedures for supersonic cruise aircraft.

Lunde, T.↗
NASA NTRS1981

F-8 digital fly-by-wire aircraft analytic redundancy management flight test experience

The formulation and flight test results of an algorithm to detect and isolate the first failure of any one of twelve duplex control sensors being monitored are described. The technique uses like sensor output differences for fault detection while relying upon analytic redundancy relationships among unlike quantities to isolate the faulty sensor. The fault isolation logic utilizes the modified sequential probability ratio test, which explicitly accommodates the inevitable irreducible low frequency errors present in the analytic redundancy residuals. In addition, the algorithm uses sensor output selftest, which takes advantage of the duplex sensor structure by immediately removing a highly erratic sensor from control calculations and analytic redundancy relationships while awaiting a definitive fault isolation decision via analytic redundancy.

Deckert, J. C.↗
NASA NTRS1981

A new diffusion-inhibited oxidation-resistant coating for superalloys

A concept for enhanced protection of superalloys consists of adding an oxidation- and diffusion-resistant cermet layer between the superalloy and the outer oxidation-resistant metallic alloy coating. Such a duplex coating was compared with a physical-vapor-deposited (PVD) NiCrAlY coating in cyclic oxidation at 1150 C. The substrate alloy was MA 754 - an oxide-dispersion-strengthened superalloy that is difficult to coat. The duplex coating, applied by plasma spraying, outperformed the PVD coating on the basis of weight change and both macroscopic and metallographic observations.

Gedwill, M. A.↗
NASA NTRS1982

Evaluation of present thermal barrier coatings for potential service in electric utility gas turbines

The resistance of present-day thermal barrier coatings to combustion gases found in electric utility turbines was assessed. The plasma sprayed coatings, both duplex and graded types, were primarily zirconia-based, although a calcium silicate was also evaluated. Both atmospheric burner rig tests and high pressure tests (135 psig) showed that several present-day thermal barrier coatings have a high potential for service in gas turbines burning the relatively clean GT No. 2 fuel. However, coating improvements are needed for use in turbines burning lower grade fuel such as residual oil. The duplex ZrO2.8Y2O3/NiCrA1Y coating was ranked highest and selected for near-term field testing, with Ca2SiO4/NiCrA1Y ranked second. Graded coatings show potential for corrosive turbine operating conditions and warrant further development. The coating degradation mechanisms for each coating system subjected to the various environmental conditions are also described.

Bratton, R. J.↗