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Congruence of Clusters Defined By Whole Genome Sequencing and MALDI-TOF for Bacteria Isolated From Cleanrooms

Introduction: Oligotrophic conditions can render cleanrooms inhospitable to microbes. Despite these constraints, fungi and bacteria are frequently isolated from surfaces in astromaterials cleanrooms at the Johnson Space Center. Bacillus species are of particular concern because endospores belonging to this genus are resilient and can affect astromaterials. Current monitoring programs rely on 16S rRNA sequencing and the VITEK2 Compact system. These methods have limited power to resolve Bacillus species. Matrix-assisted laser desorption - time of flight mass spectrometry (MALDI-TOF MS), provides a rapid, low cost, method of identifying bacterial isolates and has a higher resolution than 16S rRNA sequencing, particularly for Bacillus species; however, few studies have compared this method to the industry gold standard, whole genome sequencing (WGS). Methods: Based on 16S rRNA classification, we selected 14 isolates for analysis with MALDI-TOF and WGS. Mass spectra were generated with MALDI-TOF MS and processed with custom scripts to identify clusters of closely related isolates and calculate a matrix of pairwise cosine similarity scores. Hybrid Illumina and Nanopore sequencing were used to generate draft genomes. Pairwise similarity scores were calculated from these genomes based on the average amino acid identity (AAI) predicted from single copy core genes. Congruence of clustering between these methods, was assessed by calculating adjusted Rand and Wallace coefficients. Results: Clusters of species generated from MALDI-TOF MS showed good agreement of phylotypes generated with WGS. Pairs of strains that were > 94% similar to each other in terms of predicted amino acid sequences consistently showed cosine similarities of mass spectra > 0.65 and, of the 9 clusters identified with WGS, 8 were identical with MALDI-TOF. This corresponds to an adjusted Rand index of 0.95 and a 95% confidence interval of 0.80 – 1.00 for adjusted Wallace coefficients. The only discordance was for a pair of isolates that were classified as Paenibacillus species. This pair showed relatively high similarity (0.84) in terms of MALDI-TOF MS but only 85% similarity in terms of AAI. Conclusion: This study shows that MALDI-TOF and WGS exhibit a similar ability to delineate Bacillus species isolated from cleanrooms and taxonomic units described by these two methods are consistent with one another. Since MALDI-TOF MS is low in cost and high in throughput, this approach appears to be an ideal option for routine microbial monitoring and identifying Bacillus species.

Farnaz Mazhari↗

NASA NTRS2016

A New Load Residual Threshold Definition for the Evaluation of Wind Tunnel Strain-Gage Balance Data

A new definition of a threshold for the detection of load residual outliers of wind tunnel strain-gage balance data was developed. The new threshold is defined as the product between the inverse of the absolute value of the primary gage sensitivity and an empirical limit of the electrical outputs of a strain{gage. The empirical limit of the outputs is either 2.5 microV/V for balance calibration or check load residuals. A reduced limit of 0.5 microV/V is recommended for the evaluation of differences between repeat load points because, by design, the calculation of these differences removes errors in the residuals that are associated with the regression analysis of the data itself. The definition of the new threshold and different methods for the determination of the primary gage sensitivity are discussed. In addition, calibration data of a six-component force balance and a five-component semi-span balance are used to illustrate the application of the proposed new threshold definition to different types of strain{gage balances. During the discussion of the force balance example it is also explained how the estimated maximum expected output of a balance gage can be used to better understand results of the application of the new threshold definition.

load residuals↗

NASA NTRS1993

Analysis of matrix cracking and local delamination in (0/theta/-theta)s graphite epoxy laminates under tensile load

Three-dimensional element analyses of (0/theta/-theta)s graphite epoxy laminates, where theta = 15, 20, 25, 30, and 45 deg, subjected to axial tensile load, were performed. The interlaminar stresses in the theta/-theta interface were calculated with and without a matrix crack in the central -theta plies. The interlaminar normal stress changes from a small compressive stress when no matrix crack is present to a high tensile stress at the intersection of the matrix crack and the free edge. The analysis of local delamination from the -theta matrix crack indicates a high strain energy release rate and a localized Mode I component near the free edge, within one-ply distance from the matrix crack. To examine the stress state causing the matrix cracking, the maximum principal normal stress in a plane perpendicular to the fiber direction in the -theta ply was calculated in an uncracked laminate. The corresponding shear stress parallel to the fiber was also calculated. The principal normal stress at the laminate edge increased through the ply thickness and reached a very high tensile value at the theta/-theta interface indicating that the crack in the -theta ply may initiate at the theta/-theta interface.

Salpekar, S. A.↗

NASA NTRS1991

Analysis of matrix cracking and local delamination in (0/theta/-theta)sub s graphite epoxy laminates under tension load

Several 3D finite element analyses of (0/theta/-theta)sub s graphite epoxy laminates, where theta=15, 20, 25, 30, and 45 deg, subjected to axial tension load were performed. The interlaminar stresses in the theta/-theta interface were calculated with and without a matrix crack in the central -theta plies. The interlaminar normal stress changes from a small compressive stress when no matrix crack is present to a high tensile stress at the intersection of the matrix crack and free edge. The analysis of local delamination from the -theta matrix crack indicates a high strain energy release rate and a localized mode I component near the free edge, within one ply distance from the matrix crack. In order to examine the stress state causing the matrix cracking the maximum principal normal stress in a plane perpendicular to the fiber direction in the -theta ply was calculated in an uncracked laminate. The corresponding shear stress parallel to the fiber was also calculated. The principal normal stress at the laminate edge increases through the ply thickness and reached a very high tensile value at the theta/-theta interface indicating that the crack in the -theta ply may initiate at the theta/-theta interface. Crack profiles on the laminate edge in the -theta ply were constructed from the principal stress directions. The cracks were found to be more curved for layups with smaller theta angles, which is consistent with experimental observations in the literature.

Salpekar, S. A.↗

NASA NTRS1991

Analysis of matrix cracking and local delamination in (O/theta/-theta)sub s graphite epoxy laminates under tension load

Several 3D finite element analyses of (O/theta/-theta)sub s graphite epoxy laminates, where theta = 15, 20, 25, 30, and 45 deg, subjected to axial tension load were performed. The interlaminar stresses in the theta/-theta interface were calculated with and without a matrix crack in the central -theta plies. The interlaminar normal stress changes from a small compressive stress when no matrix crack is present to a high tensile stress at the intersection of the matrix crack and free edge. The analysis of local delamination from the -theta matrix crack indicates a high strain energy release rate and a localized mode I component near the free edge, within one ply distance from the matrix crack. In order to examine the stress state causing the matrix cracking the maximum principal normal stress in a plane perpendicular to the fiber direction in the -theta ply was calculated in an uncracked laminate. The corresponding shear stress parallel to the fiber was also calculated. The principal normal stress at the laminate edge increases through the ply thickness and reached a very high tensile value at the theta/-theta interface indicating that the crack in the -theta ply may initiate at the theta/-theta interface. Crack profiles on the laminate edge in the -theta ply were constructed from the principal stress directions. The cracks were found to be more curved for layups with smaller theta angles, which is consistent with experimental observations in the literature.

Salpekar, S. A.↗

NASA NTRS1976

Symmetric stiffness matrix for incompressible hyperelastic materials

Symmetric structure matrices are derived for solving plane strain and axisymmetric problems involving incompressible hyperelastic materials. An infinite hollow cylinder subjected to internal pressure is considered as an example. Displacement and hydrostatic pressure profiles are calculated using the Newton-Raphson iteration technique. The results are in good agreement with the exact curves.

Takamatsu, T.↗

NASA NTRS1992

Pancakelike domes on Venus

The shape of seven large domes on the plains of Venus, with volumes between 100 and 1000 cu km, is compared with that of an axisymmetric gravity current spreading over a rigid horizontal surface. Both the altimetric profiles and the horizontal projection of the line of intersection of domes on the SAR images agree well with the theoretical similarity solution for a newtonian fluid, but not with the shape calculated for a rigid-plastic rheology, nor with that for a static model with a strong skin. As a viscous current spreads, it generates an isotropic strain rate tensor whose magnitude is independent of radius. Such a flow can account for the randomly oriented cracks that are uniformly distributed on the surface of the domes. The stress induced by the flow in the plains material below is obtained, and is probably large enough to produce the short radial cracks in the surface of the plains beyond the domes. The viscosity of the domes can be estimated from their thermal time constants if spreading is possible only when the fluid is hot, and lies between 10(exp 14) and 10(exp 17) Pa s. Laboratory experiments show that such viscosities correspond to temperatures of 610 - 690 C in dry rhyolitic magmas. These temperatures agree with laboratory measurements of the solidus temperature of wet rhyolite. These results show that the development of the domes can be understood using simple fluid dynamical ideas, and that the magmas involved can be produced by wet melting at depths below 10 km, followed by eruption and degassing.

Mckenzie, Dan↗

NASA NTRS1995

Real time acousto-ultrasonic NDE technique for monitoring damage in ceramic composites under dynamic loads

Research effort was directed towards developing a near real-time, acousto-ultrasonic (AU), nondestructive evaluation (NDE) tool to study the failure mechanisms of ceramic composites. Progression of damage is monitored in real-time by observing the changes in the received AU signal during the actual test. During the real-time AU test, the AU signals are generated and received by the AU transducers attached to the specimen while it is being subjected to increasing quasi-static loads or cyclic loads (10 Hz, R = 1.0). The received AU signals for 64 successive pulses were gated in the time domain (T = 40.96 micro sec) and then averaged every second over ten load cycles and stored in a computer file during fatigue tests. These averaged gated signals are representative of the damage state of the specimen at that point of its fatigue life. This is also the first major attempt in the development and application of real-time AU for continuously monitoring damage accumulation during fatigue without interrupting the test. The present work has verified the capability of the AU technique to assess the damage state in silicon carbide/calcium aluminosilicate (SiC/CAS) and silicon carbide/ magnesium aluminosilicate (SiC/MAS) ceramic composites. Continuous monitoring of damage initiation and progression under quasi-static ramp loading in tension to failure of unidirectional and cross-ply SiC/CAS and quasi-isotropic SiC/MAS ceramic composite specimens at room temperature was accomplished using near real-time AU parameters. The AU technique was shown to be able to detect the stress levels for the onset and saturation of matrix cracks, respectively. The critical cracking stress level is used as a design stress for brittle matrix composites operating at elevated temperatures. The AU technique has found that the critical cracking stress level is 10-15% below the level presently obtained for design purposes from analytical models. An acousto-ultrasonic stress-strain response (AUSSR) model for unidirectional and cross-ply ceramic composites was formulated. The AUSSR model predicts the strain response to increasing stress levels using real-time AU data and classical laminated plate theory. The Weibull parameters of the AUSSR model are used to calculate the design stress for thermo-structural applications. Real-time AU together with the AUSSR model was used to study the failure mechanisms of SiC/CAS ceramic composites under static and fatigue loading. An S-N curve was generated for a cross-ply SiC/CAS ceramic composite material. The AU results are corroborated and complemented by other NDE techniques, namely, in-situ optical microscope video recordings and edge replication.

Tiwari, Anil↗

NASA NTRS2011

20 plus Years of Computational Fluid Dynamics for the Space Shuttle

This slide presentation reviews the use of computational fluid dynamics in performing analysis of the space shuttle with particular reference to the return to flight analysis and other shuttle problems. Slides show a comparison of pressure coefficient with the shuttle ascent configuration between the wind tunnel test and the computed values. the evolution of the grid system for the space shuttle launch vehicle (SSLv) from the early 80's to one in 2004, the grid configuration of the bipod ramp redesign from the original design to the current configuration, charts with the computations showing solid rocket booster surface pressures from wind tunnel data, calculated over two grid systems (i.e., the original 14 grid system, and the enhanced 113 grid system), and the computed flight orbiter wing loads are compared with strain gage data on STS-50 during flight. The loss of STS-107 initiated an unprecedented review of all external environments. The current SSLV grid system of 600+ grids, 1.8 Million surface points and 95+ million volume points is shown. The inflight entry analyses is shown, and the use of Overset CFD as a key part to many external tank redesign and debris assessments is discussed. The work that still remains to be accomplished for future shuttle flights is discussed.

Gomez, Reynaldo J., III↗

NASA NTRS2015

In-Flight Aeroelastic Stability of the Thermal Protection System on the NASA HIAD, Part II: Nonlinear Theory and Extended Aerodynamics

Conical shell theory and a supersonic potential flow aerodynamic theory are used to study the nonlinear pressure buckling and aeroelastic limit cycle behavior of the thermal protection system for NASA's Hypersonic Inflatable Aerodynamic Decelerator. The structural model of the thermal protection system consists of an orthotropic conical shell of the Donnell type, resting on several circumferential elastic supports. Classical Piston Theory is used initially for the aerodynamic pressure, but was found to be insufficient at low supersonic Mach numbers. Transform methods are applied to the convected wave equation for potential flow, and a time-dependent aerodynamic pressure correction factor is obtained. The Lagrangian of the shell system is formulated in terms of the generalized coordinates for all displacements and the Rayleigh-Ritz method is used to derive the governing differential-algebraic equations of motion. Aeroelastic limit cycle oscillations and buckling deformations are calculated in the time domain using a Runge-Kutta method in MATLAB. Three conical shell geometries were considered in the present analysis: a 3-meter diameter 70 deg. cone, a 3.7-meter 70 deg. cone, and a 6-meter diameter 70 deg. cone. The 6-meter configuration was loaded statically and the results were compared with an experimental load test of a 6-meter HIAD. Though agreement between theoretical and experimental strains was poor, the circumferential wrinkling phenomena observed during the experiments was captured by the theory and axial deformations were qualitatively similar in shape. With Piston Theory aerodynamics, the nonlinear flutter dynamic pressures of the 3-meter configuration were in agreement with the values calculated using linear theory, and the limit cycle amplitudes were generally on the order of the shell thickness. The effect of axial tension was studied for this configuration, and increasing tension was found to decrease the limit cycle amplitudes when the circumferential elastic supports were neglected, but resulted in more complex behavior when the supports were included. The nominal flutter dynamic pressure of the 3.7-meter configuration was significantly lower than that of the 3-meter, and it was found that two sets of natural modes coalesce to flutter modes near the same dynamic pressure. This resulted in a significant drop in the limit cycle frequencies at higher dynamic pressures, where the flutter mode with the lower frequency becomes more critical. Pre-buckling pressure loads and the aerodynamic pressure correction factor were studied for all geometries, and these effects resulted in significantly lower flutter boundaries compared with Piston Theory alone. The maximum dynamic pressure predicted by aerodynamic simulations of a proposed 3.7-meter HIAD vehicle was still lower than any of the calculated flutter dynamic pressures, suggesting that aeroelastic effects for this vehicle are of little concern.

Goldman, Benjamin D.↗

NASA NTRS1976

Thermal design and analysis of a hydrogen-burning wind tunnel model of an airframe-integrated scramjet

An aerodynamic model of a hydrogen burning, airframe integrated scramjet engine has been designed, fabricated, and instrumented. This model is to be tested in an electric arc heated wind tunnel at an altitude of 35.39 km (116,094 ft.) but with an inlet Mach number of 6 simulating precompression on an aircraft undersurface. The scramjet model is constructed from oxygen free, high conductivity copper and is a heat sink design except for water cooling in some critical locations. The model is instrumented for pressure, surface temperature, heat transfer rate, and thrust measurements. Calculated flow properties, heat transfer rates, and surface temperature distributions along the various engine components are included for the conditions stated above. For some components, estimates of thermal strain are presented which indicate significant reductions in plastic strain by selective cooling of the model. These results show that the 100 thermal cycle life of the engine was met with minimum distortion while staying within the 2669 N (600 lbf) engine weight limitation and while cooling the engine only in critical locations.

Guy, R. W.↗

NASA NTRS1979

Development of procedures for calculating stiffness and damping of elastomers in engineering applications. Part 5: Elastomer performance limits and the design and test of an elastomer damper

Tests were performed on elastomer specimens of the material polybutadiene to determine the performance limitations imposed by strain, temperature, and frequency. Three specimens were tested: a shear specimen, a compression specimen, and a second compression specimen in which thermocouples were embedded in the elastomer buttons. Stiffness and damping were determined from all tests, and internal temperatures were recorded for the instrumented compression specimen. Measured results are presented together with comparisons between predictions of a thermo-viscoelastic analysis and the measured results. Dampers of polybutadiene and Viton were designed, built, and tested. Vibration measurements were made and sensitivity of vibration to change in unbalance was also determined. Values for log decrement were extracted from the synchronous response curves. Comparisons were made between measured sensitivity to unbalance and log decrement and predicted values for these quantities.

Tecza, J. A.↗

NASA NTRS1987

Interlaminar shear fracture toughness and fatigue thresholds for composite materials

Static and cyclic end notched flexure tests were conducted on a graphite epoxy, a glass epoxy, and graphite thermoplastic to determine their interlaminar shear fracture toughness and fatigue thresholds for delamination in terms of limiting values of the mode II strain energy release rate, G-II, for delamination growth. The influence of precracking and data reduction schemes are discussed. Finite element analysis indicated that the beam theory calculation for G-II with the transverse shear contribution included was reasonably accurate over the entire range of crack lengths. Cyclic loading significantly reduced the critical G-II for delamination. A threshold value of the maximum cyclic G-II below which no delamination occurred after one million cycles was identified for each material. Also, residual static toughness tests were conducted on glass epoxy specimens that had undergone one million cycles without delamination. A linear mixed-mode delamination criteria was used to characterize the static toughness of several composite materials; however, a total G threshold criterion appears to characterize the fatigue delamination durability of composite materials with a wide range of static toughness.

Obrien, T. Kevin↗

NASA NTRS1989

Interlaminar shear fracture toughness and fatigue thresholds for composite materials

O'Brien, T. Kevin↗

NASA NTRS1991

Elasto-plastic coupled temperature-displacement finite element analysis of two-dimensional rolling-sliding contact with a translating heat source

The present paper describes a transient translating elastoplastic thermomechanical finite element model to study two-dimensional frictional rolling contact. Frictional two-dimensional contact is simulated by repeatedly translating a nonuniform thermomechanical distribution across the surface of an elastoplastic half space. The half space is represented by a two-dimensional finite element mesh with appropriate boundaries. Calculations are for an elastic-perfectly plastic material and the selected thermophysical properties are assumed to be temperature independent. The paper presents temperature variations, stress and plastic strain distributions and deformations. Residual tensile stresses are observed. The magnitude and depth of these stresses depends on (1) the temperature gradients and (2) the magnitudes of the normal and tangential tractions.

Rubin, C. A.↗

NASA NTRS1993

Composite micromechanical modeling using the boundary element method

The use of the boundary element method for analyzing composite micromechanical behavior is demonstrated. Stress-strain, heat conduction, and thermal expansion analyses are conducted using the boundary element computer code BEST-CMS, and the results obtained are compared to experimental observations, analytical calculations, and finite element analyses. For each of the analysis types, the boundary element results agree reasonably well with the results from the other methodologies, with explainable discrepancies. Overall, the boundary element method shows promise in providing an alternative method to analyze composite micromechanical behavior.

Goldberg, Robert K.↗

NASA NTRS1948

Effect of Centrifugal Force on the Elastic Curve of a Vibrating Cantilever Beam

A study was made to determine the effect of rotation on the dynamic-stress distribution in vibrating cantilever beams. The results of a mathematical analysis are presented together with experimental results obtained by means of stroboscopic photographs and strain gages. The theoretical analysis was confined to uniform cantilever beams; the experimental work was extended to include a tapered cantilever beam to simulate an aircraft propeller blade. Calculations were made on nondimensional basis for second and third mode vibration; the experiments were conducted on beams of various lengths, materials, and cross sections for second-mode vibration. From this investigation it was concluded that high vibratory-stress positions are unaffected by the addition of centrifugal force. Nonrotating vibration surveys of blades therefore are valuable in predicting high vibratory-stress locations under operating conditions.

Simpkinson, Scott H↗

NASA NTRS1995

Characterization of High Ge Content SiGe Heterostructures and Graded Alloy Layers Using Spectroscopic Ellipsometry

Si(x)Ge(1-x)heterostructures on Si substrates have been widely studied due to the maturity of Si technology. However, work on Si(x)Ge)1-x) heterostructures on Ge substrates has not received much attention. A Si(x)Ge(1-x) layer on a Si substrate is under compressive strain while Si(x)Ge(1-x) on Ge is under tensile strain; thus the critical points will behave differently. In order to accurately characterize high Ge content Si(x)Ge(1-x) layers the energy shift algorithm used to calculate alloy compositions, has been modified. These results have been used along with variable angle spectroscopic ellipsometry (VASE) measurements to characterize Si(x)Ge(1-x)/Ge superlattices grown on Ge substrates. The results agree closely with high resolution x-ray diffraction measurements made on the same samples. The modified energy shift algorithm also allows the VASE analysis to be upgraded in order to characterize linearly graded layers. In this work VASE has been used to characterize graded Si(x)Ge(1-x) layers in terms of the total thickness, and the start and end alloy composition. Results are presented for a 1 micrometer Si(x)Ge(1-x) layer linearly graded in the range 0.5 less than or equal to x less than or equal to 1.0.

Heyd, A. R.↗