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Evolved Gas Analyses of Sedimentary Rocks from the Glen Torridon Clay-Bearing Unit, Gale Crater, Mars: Results From the Mars Science Laboratory Sample Analysis at Mars Instrument Suite

Evolved gas analysis (EGA) data from the Sample Analysis at Mars (SAM) instrument suite indicated Fe-rich smectite, carbonate, oxidized organics, Fe/Mg sulfate, and chloride in sedimentary rocks from the Glen Torridon (GT) region of Gale crater that displayed phyllosilicate spectral signatures from orbit. SAM evolved H2O data indicated that the primary phyllosilicate in all GT samples was an Fe-rich dioctahedral smectite (e.g., nontronite) with lesser amounts of a phyllosilicate such as mixed layer talc-serpentine or greenalite-minnesotaite. CO(2) data supported the identification of siderite in several samples, and CO(2) and CO data was also consistent with trace oxidized organic compounds such as oxalate salts. SO(2) data indicated trace and/or amorphous Fe sulfates in all samples and one sample may contain Fe sulfides. SO(2) data points to significant Mg sulfates in two samples, and lesser amounts in several other samples. A lack of evolved O(2) indicated the absence of oxychlorine salts and Mn3+/Mn4+ oxides. The lack of, or very minor, evolved NO revealed absent or very trace nitrate/nitrite salts. HCl data suggested chloride salts in GT samples. Constraints from EGA data on mineralogy and chemistry indicated that the environmental history of GT involved alteration with fluids of variable redox potential, chemistry and pH under a range of fluid-to-rock ratio conditions. Several of the fluid episodes could have provided habitable environmental conditions and carbon would have been available to any past microbes though the lack of significant N could have been a limiting factor for microbial habitability in the GT region.

A. C. McAdam↗

Factors influencing the rate of non-enzymatic activation of carboxylic and amino acids by ATP

The nonenzymatic formation of adenylate anhydrides of carboxylic and amino acids is discussed as a necessary step in the origin of the genetic code and protein biosynthesis. Results of studies are presented which have shown the rate of activation to depend on the pKa of the carboxyl group, the pH of the medium, temperature, the divalent metal ion catalyst, salt concentration, and the nature of the amino acid. In particular, it was found that of the various amino acids investigated, phenylalanine had the greatest affinity for the adenine derivatives adenosine and ATP. Results thus indicate that selective affinities between amino acids and nucleotides were important during prebiotic chemical evolution, and may have played a major role in the origin of protein synthesis and genetic coding.

Mullins, D. W., Jr.↗

Very High Energy Outburst of Markarian 501 in May 2009

The very high energy (VHE; E great than 100 GeV) blazar Markarian 501 was observed between April 17 and May 5 (MJD 5493854956), 2009, as part of an extensive multi-wavelength campaign from radio to VHE. Strong VHE -ray activity was detected on May 1st with Whipple and VERITAS, when the flux (E greater than 400 GeV) increased to 10 times the pre-flare baseline flux (3.9 x 10(exp -11 ph cm(exp -2 S(exp -1), reaching five times the flux of the Crab Nebula. This coincided with a decrease in the optical polarization and a rotation of the polarization angle by 15deg. This VHE flare showed a fast flux variation with an increase of a factor approximately 4 in 25 min, and a falling time of approximately 50 min. We present the observations of the quiescent state previous to the flare and of the high state after the flare, focusing on the flux and spectral variability from Whipple, VERITAS, Fermi-LAT, RXTE, and Swift combined with optical and radio data.

BL Lacertae objects: individual: Mrk 501 / gamma r↗

Biosensors for EVA: Muscle Oxygen and pH During Walking, Running and Simulated Reduced Gravity

During lunar excursions in the EVA suit, real-time measurement of metabolic rate is required to manage consumables and guide activities to ensure safe return to the base. Metabolic rate, or oxygen consumption (VO2), is normally measured from pulmonary parameters but cannot be determined with standard techniques in the oxygen-rich environment of a spacesuit. Our group developed novel near infrared spectroscopic (NIRS) methods to calculate muscle oxygen saturation (SmO2), hematocrit, and pH, and we recently demonstrated that we can use our NIRS sensor to measure VO2 on the leg during cycling. Our NSBRI-funded project is looking to extend this methodology to examine activities which more appropriately represent EVA activities, such as walking and running and to better understand factors that determine the metabolic cost of exercise in both normal and lunar gravity. Our 4 year project specifically addresses risk: ExMC 4.18: Lack of adequate biomedical monitoring capability for Constellation EVA Suits and EPSP risk: Risk of compromised EVA performance and crew health due to inadequate EVA suit systems.

Lee, S. M. C.↗

Evidence Report: Risk of Renal Stone Formation

The formation of renal stones poses an in-flight health risk of high severity, not only because of the impact of renal colic on human performance but also because of complications that could potentially lead to crew evacuation, such as hematuria, infection, hydronephrosis, and sepsis. Evidence for risk factors comes from urine analyses of crewmembers, documenting changes to the urinary environment that are conducive to increased saturation of stone-forming salts, which are the driving force for nucleation and growth of a stone nidus. Further, renal stones have been documented in astronauts after return to Earth and in one cosmonaut during flight. Biochemical analysis of urine specimens has provided indication of hypercalciuria and hyperuricemia, reduced urine volumes, and increased urine saturation of calcium oxalate and calcium phosphate. A major contributor to the risk for renal stone formation is bone atrophy with increased turnover of the bone minerals. Dietary and fluid intakes also play major roles in the risk because of the influence on urine pH (more acidic) and on volume (decreased). Historically, specific assessments on urine samples from some Skylab crewmembers indicated that calcium excretion increased early in flight, notable by day 10 of flight, and almost exceeded the upper threshold for normal excretion (300mg/day in males). Other crewmember data documented reduced intake of fluid and reduced intake of potassium, phosphorus, magnesium, and citrate (an inhibitor of calcium stone formation) in the diet. Hence, data from both short-duration and long-duration missions indicate that space travel induces risk factors for renal stone formation that continue to persist after flight; this risk has been documented by reported kidney stones in crewmembers.

Sibonga, Jean D.↗

A Gamma-Ray Burst Trigger Toolkit

The detection rate of a gamma-ray burst detector can be increased by using a count rate trigger with many accumulation times DELTAt and energy bands DELTAE Because a burst's peak flux varies when averaged over different DELTAt and DELTAE the nominal sensitivity (the numerical value of the peak flux) of a trigger system is less important than how much fainter a burst could be at the detection threshold as DELTAt and DELTAE are changed. The relative sensitivity of different triggers can be quantified by referencing the detection threshold back to the peak flux for a fiducial value of DELTAt and DELTA E. This mapping between peak flux values for different sets of DELTAt and DELTAE varies from burst to burst. Quantitative estimates of the burst detection rate for a given detector and trigger system can be based on the observed rate at a measured peak flux value in this fiducial trigger. Predictions of a proposed trigger's burst detection rate depend on the assumed burst population, and these predictions can be wildly in error for triggers that differ significantly from previous missions. I base the fiducial rate on the BATSE observations: 550 bursts per sky above a peak flux of 0.3 ph per square centimeter per second averaged over DELTAt=1.024 sec and DELTAE=50-300 keV. Using a sample of 100 burst lightcurves I find that triggering on any value of DELTAt that is a multiple of 0.064 sec decreases the average threshold peak flux on the 1.024 sec timescale by a factor of 0.6. Extending DELTAE to lower energies includes the large flux of the X-ray background, increasing the background count rate. Consequently a low energy DELTAE is advantageous only for very soft bursts. Whether a large fraction of the population of bright bursts is soft is disputed; the new population of X-ray Flashes is soft but relatively faint.

Band, David L.↗

Continuous pH monitoring in a perfused bioreactor system using an optical pH sensor

Monitoring and regulating the pH of the solution in a bioprocess is one of the key steps in the success of bioreactor operation. An in-line optical pH sensor, based on the optical absorption properties of phenol red present in the medium, was developed and tested in this work for use in NASA space bioreactors based on a rotating wall-perfused vessel system supporting a baby hamster kidney (BHK-21) cell culture. The sensor was tested over three 30-day and one 124-day cell runs. The pH sensor initially was calibrated and then used during the entire cell culture interval. The pH reported by the sensor was compared to that measured by a fiber optically coupled Shimadzu spectrophotometer and a blood gas analyzer. The maximum standard error of prediction for all the four cell runs for development pH sensor against BGA was +/-0.06 pH unit and for the fiber optically coupled Shimadzu spectrophotometer against the blood gas analyzer was +/-0.05 pH unit. The pH sensor system performed well without need of recalibration for 124 days. Copyright 2002 Wiley Periodicals, Inc.

Bioreactors↗

Spectral Measurements of PMCs from SBUV/2 Instruments

The SBUV/2 (Solar Backscattered Ultraviolet, model 2) instrument is designed to monitor ozone stratospheric profile and total column ozone using measurements of the Earth's backscattered ultraviolet albedo. We have previously demonstrated that the normal radiance measurements from SBUV/2 instruments, which sample 12 discrete wavelengths between 252 and 340 nm during each scan, can be used to identify polar mesospheric clouds (PMCs). Some SBUV/2 instruments also periodically view the earth in continuous scan mode, covering the wavelength range 160-400 nm with 0.15 nm sampling. Analysis of these data show PMC occurrence rates similar to the normal discrete scan results, although the observation technique reduces the number of daily measurements by a factor of six. PMC observed by SBUV/2 instruments show a monotonic variation in the residual spectral albedo over the wavelength range 250 300 nm, with maximum enhancements of 10 15% at 250 nm. This result is consistent with microphysical model predictions from Jensen [1989. A numerical model of polar mesospheric cloud formation and evolution, Ph. D. Thesis, University of Colorado]. We find no evidence for a systematic localized increase in PMC residual albedo for wavelengths near 260 nm, in contrast to the recently reported results from the MSX UVISI instrument [Carbary J.F., et al., 2004. Evidence for bimodal particle distribution from the spectra of polar mesospheric clouds. Geophysics Research. Letters 31, L13108]. This result is observed for three different SBUV/2 instruments in both Northern and Southern Hemisphere data over a 13-year span. Our Mie scattering calculations show that the location and magnitude of the 260 nm hump feature is dependent upon the specific scattering angles appropriate to the MSX measurements. Although it explains the MSX spectrum, the bimodal size distribution proposed by Carbary et al. (2004), cannot explain the lack of scattering angle dependence of the SBUV/2 spectral shapes. The spectral signature of the SBUV/2 continuous scan PMC data is thus inconsistent with the bimodal particle size distribution suggested by Carbary et al. (2004).

DeLand, Matthew T.↗

Neurotrophic factor - Characterization and partial purification

Recent evidence suggests that neurotrophic activity is required for the normal proliferation and development of muscle cells. The present paper reports a study of the purification and characterization of a neurotrophic factor (NTF) from adult chicken ischiatic-peroneal nerves using two independent quantitative in vitro assay systems. The assays were performed by the measurement of the incorporation of tritiated thymidine or the sizes of single-cell clones by chick muscle cells grown in culture. The greatest amount of neutrotrophic activity is found to be extracted at a pH of 8; aqueous suspensions of the activity are stable to long-term storage at room temperature. The specific activity of the substance is doubled upon precipitation with ammonium sulfate or after gel filtration, and increase 4 to 5 fold after salt gradient elution from DEAE cellulose columns. The active fraction obtained after gel filtration and rechromatography on DEAE cellulose exhibits a 7 to 10-fold increase in specific activity. Electrophoresis of the most highly purified material yields a greatly concentrated band at around 80,000 daltons. Although NTF is purified almost 10-fold as indicated by the increase in specific activity, the maximum activity of the partially purified material is greatly reduced, possibly due to a requirement for a cofactor for the expression of maximum activity.

Popiela, H.↗

The Validity Chlorophyll-a Estimation by Sun Induced Fluorescence in Estuarine Waters: An Analysis of Long-term (2003-2011) Water Quality Data from Tampa Bay, Florida (USA)

Satellite observation of phytoplankton concentration or chlorophyll-a is an important characteristic, critically integral to monitoring coastal water quality. However, the optical properties of estuarine and coastal waters are highly variable and complex and pose a great challenge for accurate analysis. Constituents such as suspended solids and dissolved organic matter and the overlapping and uncorrelated absorptions in the blue region of the spectrum renders the blue-green ratio algorithms for estimating chlorophyll-a inaccurate. Measurement of sun-induced chlorophyll fluorescence, on the other hand, which utilizes the near infrared portion of the electromagnetic spectrum, may provide a better estimate of phytoplankton concentrations. While modelling and laboratory studies have illustrated both the utility and limitations of satellite baseline algorithms based on the sun induced chlorophyll fluorescence signal, few have examined the empirical validity of these algorithms using a comprehensive long term in situ data set. In an unprecedented analysis of a long term (2003-2011) in situ monitoring data from Tampa Bay, Florida (USA), we assess the validity of the FLH product from the Moderate Resolution Imaging Spectrometer (MODIS) against chlorophyll ]a and a suite of water quality parameters taken in a variety of conditions throughout a large optically complex estuarine system. A systematic analysis of sampling sites throughout the bay is undertaken to understand how the relationship between FLH and in situ chlorophyll-a responds to varying conditions within the estuary including water depth, distance from shore and structures and eight water quality parameters. From the 39 station for which data was derived, 22 stations showed significant correlations when the FLH product was matched with in situ chlorophyll-alpha data. The correlations (r2) for individual stations within Tampa Bay ranged between 0.67 (n=28, pless than 0.01) and-0.457 (n=12, p=.016), indicating that for some areas within the Bay, FLH can be a good predictor of chlorophyll-alpha concentration and hence a useful tool for the analysis of water quality. Overall, the results show a 106% increase in the validity of chlorophyll -a concentration estimates using FLH over the standard the blue-green OC3M algorithm. This analysis also illustrates that the correlations between FLH and in situ chlorophyll -a measurements increases with increasing water depth and distance of the monitoring sites from both the shore and structures. However, due to confounding factors related to the complexity of the estuarine system, a linear improvement in the FLH to chlorophyll ]a relationship was not clearly noted with increasing depth and distance from shore alone. Correlations of FLH with turbidity, nutrients (total nitrogen and total phosphorous) biological oxygen demand, salinity, sea surface temperature correlated positively with FLH concentrations, while dissolved oxygen and pH showed negative correlations. Principle component analyses are employed to further describe the relationships between the multivariate water quality parameters and the FLH product. The majority of sites with higher and very significant correlations (pless than 0.01) also showed high correlation values for nutrients, turbidity and biological oxygen demand. These sites were on average in greater than seven meters of water and over five kilometers from shore. A thorough understanding of the relationship between the MODIS FLH product and in situ water quality parameters will enhance our understanding of the accuracy MODIS fs global FLH algorithm and assist in optimizing its calibration for use in monitoring the quality of estuarine and coastal waters worldwide.

Moreno-Madrinan, Max Jacobo↗

Auxin-induced growth of Avena coleoptiles involves two mechanisms with different pH optima

Although rapid auxin-induced growth of coleoptile sections can persist for at least 18 hours, acid-induced growth lasts for a much shorter period of time. Three theories have been proposed to explain this difference in persistence. To distinguish between these theories, the pH dependence for auxin-induced growth of oat (Avena sativa L.) coleoptiles has been determined early and late in the elongation process. Coleoptile sections from which the outer epidermis was removed to facilitate buffer entry were incubated, with or without 10 micromolar indoleacetic acid, in 20 millimolar buffers at pH 4.5 to 7.0 to maintain a fixed wall pH. During the first 1 to 2 hours after addition of auxin, elongation occurs by acid-induced extension (i.e. the pH optimum is <5 and the elongation varies inversely with the solution pH). Auxin causes no additional elongation because the buffers prevent further changes in wall pH. After 60 to 90 minutes, a second mechanism of auxin-induced growth, whose pH optimum is 5.5 to 6.0, predominates. It is proposed that rapid growth responses to changes in auxin concentration are mediated by auxin-induced changes in wall pH, whereas the prolonged, steady-state growth rate is controlled by a second, auxin-mediated process whose pH optimum is less acidic.

NASA Program Space Biology↗

Effect of the Liquid-Vacuum Transition on the Relative Abundances of Amino and Fatty Acids Sought as Biosignatures on Icy Ocean Worlds

Interpreting measurements by robotic missions searching for signs of life in material erupted by cryovolcanic ocean worlds (e.g., Enceladus, Europa, Ceres) requires linking the composition of this material to that of its subsurface liquid source. Irrespective of the properties of –and processes along– the path of ejection, material erupted on airless worlds undergoes a transition from liquid to vacuum. Here, we investigate experimentally the effect of a direct transition from liquid to vacuum environments on the relative abundances of amino and carboxylic (fatty) acids, a metric used to distinguish between biological and abiotic sources for these compounds. Amino acids were dissolved in parent solutions prepared with 1 wt.% NaCl and pH 9–10 to match properties inferred from Enceladus plume grains. Compositional analysis of the dry solids resulting from the injection of this solution into vacuum indicates a < 50% change in amino acid abundances relative to glycine. Injection of two fatty acids, phenylacetic acid (soluble and undersaturated in the parent solution) and palmitic acid (insoluble and supersaturated) resulted in a change of less than a factor of 7 in their relative abundances. At this bulk scale (all grains aggregated together), proportions of amino and fatty acids were sufficiently conserved to allow distinction between end-member example biological and abiotic sources. We did not find strong correlations between the relative enrichment or depletion of amino or fatty acids with molecular properties such as molecular mass, hydrophobicity, functionality, or charge, other than high under-or supersaturation. Most of the organic residue was deposited along the experimental injection path, which suggests that cryovolcanic conduit walls on icy ocean worlds may be enriched in organic material relative to material having undergone the liquid-vacuum transition.

Amino acids↗

Investigation of Global Particulate Nitrate from the AeroCom Phase III Experiment

An assessment of global particulate nitrate and ammonium aerosol based on simulations from nine models participating in the Aerosol Comparisons between Observations and Models (AeroCom) phase III study is presented. A budget analysis was conducted to understand the typical magnitude, distribution, and diversity of the aerosols and their precursors among the models. To gain confidence regarding model performance, the model results were evaluated with various observations globally, including ground station measurements over North America, Europe, and east Asia for tracer concentrations and dry and wet depositions, as well as with aircraft measurements in the Northern Hemisphere mid-to-high latitudes for tracer vertical distributions. Given the unique chemical and physical features of the nitrate occurrence, we further investigated the similarity and differentiation among the models by examining (1) the pH-dependent NH3 wet deposition; (2) the nitrate formation via heterogeneous chemistry on the surface of dust and sea salt particles or thermodynamic equilibrium calculation including dust and sea salt ions; and (3) the nitrate coarse-mode fraction (i.e., coarse/total). It is found that HNO3, which is simulated explicitly based on full O3-HOx-NOx-aerosol chemistry by all models, differs by up to a factor of 9 among the models in its global tropospheric burden. This partially contributes to a large difference in NO3(-), whose atmospheric burden differs by up to a factor of 13. The atmospheric burdens of NH3 and NHC 4 differ by 17 and 4, respectively. Analyses at the process level show that the large diversity in atmospheric burdens of NO3(-), NH3, and NHC4(+) is also related to deposition processes. Wet deposition seems to be the dominant process in determining the diversity in NH3 and NHC 4 lifetimes. It is critical to correctly account for contributions of heterogeneous chemical production of nitrate on dust and sea salt, because this process overwhelmingly controls atmospheric nitrate production (typically greater than 80 %) and determines the coarse- and fine-mode distribution of nitrate aerosol.

Ammonium aerosol based on simulations↗

4,4,4-trifluoro-3-(indole-3-)butyric acid promotes root elongation in Lactuca sativa independent of ethylene synthesis and pH

We studied the mode of action of 4,4,4-trifluoro-3- (indole-3-) butyric acid (TFIBA), a recently described root growth stimulator, on primary root growth of Lactuca sativa L. seedlings. TFIBA (100 micromoles) promoted elongation of primary roots by 40% in 72 h but inhibited hypocotyl growth by 35%. TFIBA induced root growth was independent of pH. TFIBA did not affect ethylene production, but reduced the inhibitory effect of ethylene on root elongation. TFIBA promoted root growth even in the presence of the ethylene biosynthesis inhibitor L-alpha-(2-aminoethoxyvinyl)glycine. TFIBA and the ethylene-binding inhibitor silver thiosulphate (STS) had a similar effect on root elongation. The results indicate that TFIBA-stimulated root elongation was neither pH-dependent nor related to inhibition of ethylene synthesis, but was possibly related to ethylene action.

Non-NASA Center↗

Oligomerization of mononucleotides on montmorillonite: A potential approach to the prebiotic synthesis of RNA

The condensation of the 5'-phosphorimidazolide of adenosine (ImpA) on montmorillonite in a pH 8 aqueous solution yields oligomers containing up to 10 monomer units. The regiospecificity of 3',5'-phosphodiester bond formation is enhanced by addition of 10% diadenosine pyrophosphate (AppA) to the reaction mixture. A series of activated derivatives of 5'-AMP was prepared to investigate the effect of the leaving group on oligomer formation. The benzimidazole and p-dimethylamino-pyridine derivatives gave the best yields of oligomers. Factors important for oligomer formation is discussed.

Ferris, James P.↗

A New Method for Evaluating the Carbon Isotope Characteristics of Carbonate Formed Under Cryogenic Conditions Analogous to Mars

The two upcoming robotic missions to Mars, Phoenix and MSL, will both have the capability of measuring the carbon isotopic composition of CO2 in the martian atmosphere, as well as possible CO2 trapped in carbonate minerals in the Martian soil. Results from orbital and landed missions now clearly indicate that no large scale deposits of carbonate materials exist at the surface. However, some results from orbital remote sensing have been interpreted to indicate that carbonate minerals are present as fine particles interspersed at low concentrations (approx. 2%) in the martian dust. One likely mechanism for the production of these carbonates is during the freezing of transient water near the surface. Large deposits of near surface ice and photographic evidence for flowing water on the surface suggest that transient melting and refreezing of H2O is an active process on Mars. Any exposure of these fluids to the CO2 rich atmosphere should al-low the production of HCO3- solutions. Carbonates are likely precipitates from these solutions during freezing as extensive CO2 degassing, driven by the fluid s decreasing volume, drives CO2 out. This rapid CO2 degassing increases the pH of the solution and drives carbonate precipitation. It has been shown in previous studies that this rapid CO2 degassing also results in a kinetic isotopic fractionation where the CO2 gas has a much lighter isotopic composition causing a large isotope enrichment of C-13 in the precipitated carbonate. This kinetic isotope enrichment may be very common in the current martian environment, and may be a very important factor in understanding the very high deltaC-13 values of carbonates found in the martian meteorites. However, while previous studies have succeeded in generally quantifying the magnitude of this effect, detailed studies of the consistency of this effect, and the freezing rates needed to produce it are needed to understand any carbon isotope analyses from carbonate minerals in the martian soil or dust. This study demonstrates an innovative new method for measuring the isotopic composition of gas evolved from the freezing of carbonate solutions in real time, which allows for a much clearer view of the chemical processes involved. This method now sets the stage for detailed analysis of the chemical and isotopic mechanisms that produce cryogenic carbonates.

Niles, P. B.↗

Metabolic alkalosis during immobilization in monkeys (M. nemestrina)

The systemic and renal acid-base response of monkeys during ten weeks of immobilization was studied. By three weeks of immobilization, arterial pH and bicarbonate concentrations were elevated (chronic metabolic alkalosis). Net urinary acid excretion increased in immobilized animals. Urinary bicarbonate excretion decreased during the first three weeks of immobilization, and then returned to control levels. Sustained increases in urinary ammonium excretion were seen throughout the time duration of immobilization. Neither potassium depletion nor hypokalemia was observed. Most parameters returned promptly to the normal range during the first week of recovery. Factors tentatively associated with changes in acid-base status of monkeys include contraction of extracellular fluid volume, retention of bicarbonate, increased acid excretion, and possible participation of extrarenal buffers.

Young, D. R.↗

Solid Rocket Booster Hydraulic Pump Port Cap Joint Load Testing

The solid rocket booster uses hydraulic pumps fabricated from cast C355 aluminum alloy, with 17-4 PH stainless steel pump port caps. Corrosion-resistant steel, MS51830 CA204L self-locking screw thread inserts are installed into C355 pump housings, with A286 stainless steel fasteners installed into the insert to secure the pump port cap to the housing. In the past, pump port cap fasteners were installed to a torque of 33 Nm (300 in-lb). However, the structural analyses used a significantly higher nut factor than indicated during tests conducted by Boeing Space Systems. When the torque values were reassessed using Boeing's nut factor, the fastener preload had a factor of safety of less than 1, with potential for overloading the joint. This paper describes how behavior was determined for a preloaded joint with a steel bolt threaded into steel inserts in aluminum parts. Finite element models were compared with test results. For all initial bolt preloads, bolt loads increased as external applied loads increased. For higher initial bolt preloads, less load was transferred into the bolt, due to external applied loading. Lower torque limits were established for pump port cap fasteners and additional limits were placed on insert axial deformation under operating conditions after seating the insert with an initial preload.

Gamwell, W. R.↗