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At least 91 records · Page 5

Correlating transcription and protein expression profiles of immune biomarkers following lipopolysaccharide exposure in lung epithelial cells

Universal and early recognition of pathogens occurs through recognition of evolutionarily conserved pathogen associated molecular patterns (PAMPs) by innate immune receptors and the consequent secretion of cytokines and chemokines. The intrinsic complexity of innate immune signaling and associated signal transduction challenges our ability to obtain physiologically relevant, reproducible and accurate data from experimental systems. One of the reasons for the discrepancy in observed data is the choice of measurement strategy. Immune signaling is regulated by the interplay between pathogen-derived molecules with host cells resulting in cellular expression changes. However, these cellular processes are often studied by the independent assessment of either the transcriptome or the proteome. Correlation between transcription and protein analysis is lacking in a variety of studies. In order to methodically evaluate the correlation between transcription and protein expression profiles associated with innate immune signaling, we measured cytokine and chemokine levels following exposure of human cells to the PAMP lipopolysaccharide (LPS) from the Gram-negative pathogen Pseudomonas aeruginosa . Expression of 84 messenger RNA (mRNA) transcripts and 69 proteins, including 35 overlapping targets, were measured in human lung epithelial cells. We evaluated 50 biological replicates to determine reproducibility of outcomes. Following pairwise normalization, 16 mRNA transcripts and 6 proteins were significantly upregulated following LPS exposure, while only five (CCL2, CSF3, CXCL5, CXCL8/IL8, and IL6) were upregulated in both transcriptomic and proteomic analysis. This lack of correlation between transcription and protein expression data may contribute to the discrepancy in the immune profiles reported in various studies. The use of multiomic assessments to achieve a systems-level understanding of immune signaling processes can result in the identification of host biomarker profiles for a variety of infectious diseases and facilitate countermeasure design and development.

59 BASIC BIOLOGICAL SCIENCES↗

Lunar Impact Flash Locations from NASA's Lunar Impact Monitoring Program

Meteoroids are small, natural bodies traveling through space, fragments from comets, asteroids, and impact debris from planets. Unlike the Earth, which has an atmosphere that slows, ablates, and disintegrates most meteoroids before they reach the ground, the Moon has little-to-no atmosphere to prevent meteoroids from impacting the lunar surface. Upon impact, the meteoroid's kinetic energy is partitioned into crater excavation, seismic wave production, and the generation of a debris plume. A flash of light associated with the plume is detectable by instruments on Earth. Following the initial observation of a probable Taurid impact flash on the Moon in November 2005,1 the NASA Meteoroid Environment Office (MEO) began a routine monitoring program to observe the Moon for meteoroid impact flashes in early 2006, resulting in the observation of over 330 impacts to date. The main objective of the MEO is to characterize the meteoroid environment for application to spacecraft engineering and operations. The Lunar Impact Monitoring Program provides information about the meteoroid flux in near-Earth space in a size range-tens of grams to a few kilograms-difficult to measure with statistical significance by other means. A bright impact flash detected by the program in March 2013 brought into focus the importance of determining the impact flash location. Prior to this time, the location was estimated to the nearest half-degree by visually comparing the impact imagery to maps of the Moon. Better accuracy was not needed because meteoroid flux calculations did not require high-accuracy impact locations. But such a bright event was thought to have produced a fresh crater detectable from lunar orbit by the NASA spacecraft Lunar Reconnaissance Orbiter (LRO). The idea of linking the observation of an impact flash with its crater was an appealing one, as it would validate NASA photometric calculations and crater scaling laws developed from hypervelocity gun testing. This idea was dependent upon LRO finding a fresh impact crater associated with one of the impact flashes recorded by Earth-based instruments, either the bright event of March 2013 or any other in the database of impact observations. To find the crater, LRO needed an accurate area to search. This Technical Memorandum (TM) describes the geolocation technique developed to accurately determine the impact flash location, and by association, the location of the crater, thought to lie directly beneath the brightest portion of the flash. The workflow and software tools used to geolocate the impact flashes are described in detail, along with sources of error and uncertainty and a case study applying the workflow to the bright impact flash in March 2013. Following the successful geolocation of the March 2013 flash, the technique was applied to all impact flashes detected by the MEO between November 7, 2005, and January 3, 2014.

Moser, D. E.↗

NEAr Transporter (NEAT) Domains: Unique Surface Displayed Heme Chaperones That Enable Gram-Positive Bacteria to Capture Heme-Iron From Hemoglobin

Iron is an important micronutrient that is required by bacteria to proliferate and to cause disease. Many bacterial pathogens forage iron from human hemoglobin (Hb) during infections, which contains this metal within heme (iron–protoporphyrin IX). Several clinically important pathogenic species within the Firmicutes phylum scavenge heme using surface-displayed or secreted NEAr Transporter (NEAT) domains. In this review, we discuss how these versatile proteins function in the Staphylococcus aureus Iron-regulated surface determinant system that scavenges heme-iron from Hb. S. aureus NEAT domains function as either Hb receptors or as heme-binding chaperones. In vitro studies have shown that heme-binding NEAT domains can rapidly exchange heme amongst one another via transiently forming transfer complexes, leading to the interesting hypothesis that they may form a protein-wire within the peptidoglycan layer through which heme flows from the microbial surface to the membrane. In Hb receptors, recent studies have revealed how dedicated heme- and Hb-binding NEAT domains function synergistically to extract Hb’s heme molecules, and how receptor binding to the Hb-haptoglobin complex may block its clearance by macrophages, prolonging microbial access to Hb’s iron. The functions of NEAT domains in other Gram-positive bacteria are also reviewed.

59 BASIC BIOLOGICAL SCIENCES↗

Qualification of Processing Batches for Tank Closure Cesium Removal Through In-Tank Batch Contact Testing - 20470

The Tank Closure Cesium Removal (TCCR) process at the Savannah River Site (SRS) is currently processing waste from Tank 10H, generating decontaminated salt solution that is sent to Tank 11H. Transfers out of Tank 11H are then disposed of on site as a grout wasteform in the SRS Saltstone Processing Facility. This is a full-scale demonstration of the use of Crystalline Silicotitanate (CST) for the decontamination of aqueous nuclear waste supernate. CST primarily removes cesium from the high sodium alkaline feed, but also adsorbs strontium, actinides, and some other trace metals. The TCCR unit is an at-tank set of four ion exchange columns, with two typically being operated in series during waste processing. Tank 10H is serving a dual function as both the salt dissolution tank as well as the feed tank for the TCCR system. Prior to operation of TCCR, Tank 10H must undergo dissolution campaigns, dissolving the salt cake to form an aqueous salt solution (supernate). After each dissolution campaign, the supernate created must be qualified prior to processing through the TCCR system. Qualification includes detailed characterization of the supernate as well as in-tank batch contact tests to determine the equilibrium loading of Cs-137 on the CST ion exchange media (IONSIV{sup TM} R9120-Ba). In support of the in-tank batch contact testing, Savannah River National Laboratory (SRNL) developed a CST sample holder, also referred to as a 'tea-bag', to hold a measured amount (∼0.1 grams) of pretreated CST between stainless steel screens. The tea-bag fits within a standard stainless steel sample vial that has been modified to allow the flow of supernate to the CST. This sample holder is then immersed in the waste tank allowing free contact between the CST beads and the surrounding liquid, at an effectively infinite liquid-to-solid phase ratio. A pair of tea-bags are deployed in the tank for a period of 10 days for each batch, after which the CST is recovered, digested, and analyzed to determine the loading of cesium isotopes on the CST. This result then supports the TCCR column thermal loading limits for processing as part of the TCCR safety basis. This paper will discuss the design and testing of the CST sample holder, as well as results from the first few batches of waste qualified and subsequently processed through the TCCR system. In addition, results from analyses of Tank 11H samples, showing decontamination of the product will be included. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Actinide abundances in ordinary chondrites

Measurements of actinide and light REE (LREE) abundances and of phosphate abundances in equilibrated ordinary chondrites were obtained and were used to define the Pu abundance in the solar system and to determine the degree of variation of actinide and LREE abundances. The results were also used to compare directly the Pu/U ratio with the earlier obtained ratio determined indirectly, as (Pu/Nd)x(Nd/U), assuming that Pu behaves chemically as a LREE. The data, combined with high-accuracy isotope-dilution data from the literature, show that the degree of gram-scale variability of the Th, U, and LREE abundances for equilibrated ordinary chondrites is a factor of 2-3 for absolute abundances and up to 50 percent for relative abundances. The observed variations are interpreted as reflecting the differences in the compositions and/or proportions of solar nebula components accreted to ordinary chondrite parent bodies.

Hagee, B.↗

Effects of Inlet Icing on Performance of Axial-flow Turbojet Engine in Natural Icing Conditions

A flight investigation in natural icing conditions was conducted to determine the effect of inlet ice formations on the performance of axial-flow turbojet engines. The results are presented for icing conditions ranging from a liquid-water content of 0.1 to 0.9 gram per cubic meter and water-droplet size from 10 to 27 microns at ambient-air temperature from 13 to 26 degrees F. The data show time histories of jet thrust, air flow, tail-pipe temperature, compressor efficiency, and icing parameters for each icing encounter. The effect of inlet-guide-vane icing was isolated and shown to account for approximately one-half the total reduction in performance caused by inlet icing.

COMPRESSORS - AXIAL FLOW↗

The Use of Large Lithium-Drifted, Germanium Diodes for Gamma-Ray Spectral Measurements at Balloon Altitudes

A series of balloon-borne experiments has been initiated at the Oak Ridge National Laboratory (ORNL) in cooperation with the Space Sciences Laboratory of the NASA G. C. Marshall Space Flight Center to determine the feasibility of using large (greater than 20 cc) lithium-drifted, germanium diodes to investigate the spectrum of atmospheric gamma rays (60 keV < E < 1.5 MeV) at altitudes corresponding to a few grams per cm (sup 2) residual atmosphere. The results of these measurements will also provide a basis for designing an appropriate shield for a multi-diode, highly directional gamma-ray spectrometer for use in astrophysical measurements. Two flights were accomplished during 1967 using the same two diodes and basic instrument package. Both flights were launched at the NCAR Scientific Balloon Flight Base, Palestine, Texas, and attained a float altitude of 117, 000 ft. The measured atmospheric gamma-ray spectrum shows at least one distinct line at 511 keV (annihilation radiation) superimposed on a continuous distribution of gamma rays attributable to both energy-degraded gamma rays and brems-strahlung photons. A second diode, encased in a passive shield of plastic and lead, shows the expected lines resulting from both neutron inelastic scattering and capture in the germanium. Data acquisition was accomplished on an event-by-event basis through the use of 512-channel, on-board ADC and word-generator circuits and a ground-based, on-line telemetry decoder. The decoder makes it possible to store the data in the memory of a modified pulse-height analyzer simultaneously with storage on magnetic tape. This provides a real-time visual observation of the data as it is accumulated and greatly facilitates preflight calibrations.

Chapman, G. T.↗

Miniature Free-Flying Magnetometer Utilizing System-On-A-Chip Technology

Four Free-Flying Magnetometers (FFMs), developed at the Jet Propulsion Laboratory (JPL) for the Enstrophy mission, were successfully deployed from the payload of a sounding rocket launched from Poker Flats, Alaska on February 11, 1999. The FFMs functioned successfully by synchronously measuring the vector magnetic field at 4 points separate from the payload and at relative distances up to 3 km, and communicated their data, in bursts, to the ground. This is the first time synchronized in-situ multipoint measurements of the Earth's magnetic field utilizing miniature spin-stabilized "sensorcraft" have been performed. The data they provided have enabled, for the first time, the direct measure of field-aligned current density and are enabling new science by determining the fine-scale structure of the currents in the Earth's ionosphere involved in the production of aurora. These proof-of-concept "hockey puck" (80 mm diameter, 38 mm height, 250 gram mass) FFMs were built using off-the-shelf commercial, industrial, and military grade surface-mount electronic components. Radiation-hard electronics was not required for the Enstrophy mission's short sub-orbital flight. The successful design, implementation, and flight demonstration of this 1st generation FFM design has provided a solid base for further development of a 2nd generation FFM design for planetary science applications. A reliable ultra-miniature radiation-hard 2nd-generation FFM utilizing System-On-A-Chip (SOAC) technologies is proposed. This design would be targeted for long-term planetary missions to investigate magnetospheric field configurations in regions having small-scale structure and to separate spatial and temporal variations. A fleet of short-lived (expendable) FFMs would be deployed into a targeted region to gather multiprobe vector magnetic field data. The FFMs would be ejected from a parent spacecraft at a speed of a few m/sec and would cover spatial volumes of order tens of kilometers for times of order one hour. The parent spacecraft would carry a sufficient number of FFMs for multiple deployments. Additional information is contained in the original extended abstract.

Eyre, F. B.↗

Universal Approach to Fabricating Graphene-Supported Single-Atom Catalysts from Doped ZnO Solid Solutions

Single-atom catalysts (SACs) have attracted widespread interest for many catalytic applications because of their distinguishing properties. However, general and scalable synthesis of efficient SACs remains significantly challenging, which limits their applications. Here we report an efficient and universal approach to fabricating a series of high-content metal atoms anchored into hollow nitrogen-doped graphene frameworks (M-N-Grs; M represents Fe, Co, Ni, Cu, etc.) at gram-scale. The highly compatible doped ZnO templates, acting as the dispersants of targeted metal heteroatoms, can react with the incoming gaseous organic ligands to form doped metal-organic framework thin shells, whose composition determines the heteroatom species and contents in M-N-Grs. We achieved over 1.2 atom % (5.85 wt %) metal loading content, superior oxygen reduction activity over commercial Pt/C catalyst, and a very high diffusion-limiting current (6.82 mA cm -2 ). Both experimental analyses and theoretical calculations reveal the oxygen reduction activity sequence of M-N-Grs. Additionally, the superior performance in Fe-N-Gr is mainly attributed to its unique electron structure, rich exposed active sites, and robust hollow framework: This synthesis strategy will stimulate the rapid development of SACs for diverse energy-related fields.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A non-Gaussian statistical model for surface elevation of nonlinear random wave fields

Probability density function of the surface elevation of a nonlinear random wave field is obtained. The wave model is based on the Stokes expansion carried to the third order for both deep water waves and waves in finite depth. The amplitude and phase of the first-order component of the Stokes wave are assumed to be Rayleigh and uniformly distributed and slowly varying, respectively. The probability density function for the deep water case was found to depend on two parameters: the root-mean-square surface elevation and the significant slope. For water of finite depth, an additional parameter, the nondimensional depth, is also required. An important difference between the present result and the Gram-Charlier representation is that the present probability density functions are always nonnegative. It is also found that the 'constant' term in the Stokes expansion, usually neglected in deterministic studies, plays an important role in determining the details of the density function. The results compare well with laboratory and field experiment data.

Huang, N. E.↗

Fitness factors impacting survival of a subsurface bacterium in contaminated groundwater

Many factors contribute to the ability of a microbial species to persist when encountering complexly contaminated environments, including time of exposure, the nature and concentration of contaminants, availability of nutritional resources, and possession of a combination of appropriate molecular mechanisms needed for survival. Herein we sought to identify genes that are most important for survival of Gram-negative Enterobacteriaceae in contaminated groundwater environments containing high concentrations of nitrate and metals using the metal-tolerant Oak Ridge Reservation isolate, Pantoea sp. MT58 (MT58). Survival fitness experiments in which a randomly barcoded transposon insertion (RB-TnSeq) library of MT58 was exposed directly to contaminated Oak Ridge Reservation groundwater samples from across a nitrate and mixed metal contamination plume were used to identify genes important for survival with increasing exposure times and concentrations of contaminants, and availability of a carbon source. Genes involved in controlling and using carbon, encoding transcriptional regulators, and related to Gram-negative outer membrane processes were among those found to be important for survival in contaminated Oak Ridge Reservation groundwater. A comparative genomics analysis of 75 Pantoea genus strains allowed us to further separate the survival determinants into core and non-core genes in the Pantoea pangenome, revealing insights into the survival of subsurface microorganisms during contaminant plume intrusion.

59 BASIC BIOLOGICAL SCIENCES↗

Characterization of Interactions between CTX-M-15 and Clavulanic Acid, Desfuroylceftiofur, Ceftiofur, Ampicillin, and Nitrocefin

Cefotaximase-Munich (CTX-M) extended-spectrum beta-lactamases (ESBLs) are commonly associated with Gram-negative, hospital-acquired infections worldwide. Several beta-lactamase inhibitors, such as clavulanate, are used to inhibit the activity of these enzymes. To understand the mechanism of CTX-M-15 activity, we have determined the crystal structures of CTX-M-15 in complex with two specific classes of beta-lactam compounds, desfuroylceftiofur (DFC) and ampicillin, and an inhibitor, clavulanic acid. The crystal structures revealed that Ser70 and five other residues (Lys73, Tyr105, Glu166, Ser130, and Ser237) participate in catalysis and binding of those compounds. Based on analysis of steady-state kinetics, thermodynamic data, and molecular docking to both wild-type and S70A mutant structures, we determined that CTX-M-15 has a similar affinity for all beta-lactam compounds (ceftiofur, nitrocefin, DFC, and ampicillin), but with lower affinity for clavulanic acid. A catalytic mechanism for tested β-lactams and two-step inhibition mechanism of clavulanic acid were proposed. CTX-M-15 showed a higher activity toward DFC and nitrocefin, but significantly lower activity toward ampicillin and ceftiofur. The interaction between CTX-M-15 and both ampicillin and ceftiofur displayed a higher entropic but lower enthalpic effect, compared with DFC and nitrocefin. DFC, a metabolite of ceftiofur, displayed lower entropy and higher enthalpy than ceftiofur. This finding suggests that compounds containing amine moiety (e.g., ampicillin) and the furfural moiety (e.g., ceftiofur) could hinder the hydrolytic activity of CTX-M-15.

59 BASIC BIOLOGICAL SCIENCES↗

Natural icing of an axial-flow turbojet engine in flight for a single icing condition

An investigation has been conducted in natural icing conditions to determine the effect of ice formations on the performance of an axial-flow turbojet engine. One flight was made in an icing condition in which the liquid-water content varied from 0.077 to 0.490 gram per cubic meter and the average droplet size varied from 5.4 to 12.1 microns. During a period of 60 minutes in icing, at an engine speed of 11,000 rpm, the tail-pipe temperature increased from 865 degrees to 965 degrees F. and the jet thrust decreased from 1950 to 1700 pounds. Near the end of the icing period, the engine was satisfactorily accelerated to take-off power.

Acker, Loren W↗

Experimental studies of circumstellar, interstellar and interplanetary refractory grain analogs

Experimental techniques for studying the processes which occur during the life cycle of a refractory grain are reviewed. Consideration is given to the construction of a differentially-pumped beam-mass spectrometer which is expected to determine the relative stability of small clusters in the Fe-Mg-Si-O and Ca-Ti-Al-O systems as a function of temperature and pressure. Also, a smoke generator has been constructed to produce gram quantities of amorphous grains for use in thermal annealing and hydrous alteration studies of smokes containing Fe-Mg-Al-Ti-Ca-Si-O and Cl. Topics of the studies include the properties of grains produced in a flow condensation apparatus, the sorption efficiency of Si2O3 smokes for AR gas, the processing of ices containing refractory precursors such as SiH4 or Fe(CO)5 plus water, and the residue from 1 MeV bombardment and warming of an amorphous Fe-SiO smoke.

Nuth, J. A.↗

Compositional variations among whole-rock fragments of the L6 chondrite Bruderheim

Twenty about 100-mg random fragments of the Bruderheim chondrite were analyzed by INAA to determine extents of heterogeneity and to check earlier results that gave anomalous LREE concentrations. Considerable compositional heterogeneity was observed in both concentrations (maximum/minimum of 1.7 for Ir, 1.8 for Na, 6.8 for La, 7.4 for Co) and concentration ratios (greater than 3 x for Ni/Co and La/Eu). Compositions could be modeled quantitatively in terms of end-member components of constant composition representing the minerals olivine + orthopyroxene, clinopyroxene, plagioclase, kamacite, taenite, troilite, chromite, and phosphate. The model yields the proportions of minerals in each sample as well as the meteorite as a whole; accurate determination of the end members depends on high intrasample precision for many elements. No evidence of compositional heterogeneity of end members for trace elements was found. It is shown that these analyses, which used only about 2 grams of the meteorite, are equivalent in accuracy to a single sample of several tens of grams, and that knowledge of the extent of heterogeneity is essential to knowing the quality of sampling as well as for comparing results from one analysis with another and of one meteorite with another.

Haas, Johnson R.↗

Molecular simulation of lignin-related aromatic compound permeation through gram-negative bacterial outer membranes

Lignin, an abundant aromatic heteropolymer in secondary plant cell walls, is the single largest source of renewable aromatics in the biosphere. Leveraging this resource for renewable bioproducts through targeted microbial action depends on lignin fragment uptake by microbial hosts and subsequent enzymatic action to obtain the desired product. Recent computational work has emphasized that bacterial inner membranes are permeable to many aromatic compounds expected from lignin depolymerization processes. In this study, we expand on these findings through simulations for 42 lignin-related compounds across a gram-negative bacterial outer membrane model. Unbiased simulation trajectories indicate that spontaneous crossing for the full outer membrane is relatively rare at molecular simulation timescales, primarily due to preferential membrane partitioning and slow diffusion within the lipopolysaccharide layer within the outer membrane. Membrane partitioning and permeability coefficients were determined through replica exchange umbrella sampling simulations to overcome sampling limitations. We find that the glycosylated lipopolysaccharides found in the outer membrane increase the permeation barrier to many lignin-related compounds, particularly the most hydrophobic compounds. However, the effect is relatively modest; at industrially relevant concentrations, uncharged lignin-related compounds will readily diffuse across the outer membrane without the need for specific porins. Together, our results provide insight into the permeability of the bacterial outer membrane for assessing lignin fragment uptake and the future production of renewable bioproducts.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

A Study of Large Droplet Ice Accretion in the NASA Lewis IRT at Near-Freezing Conditions

Results of experiments designed to determine the effects of large droplet ice accretion on a NACA 23012 wing section are presented. Using primarily an icing condition with a median volumetric diameter droplet size of 160 micron and a liquid water content of 0.82 grams per cubic meter, the effects of various air temperatures, angles of attack, and de-icer boot cycle interval times on ice accretion were studied. Measurements of aerodynamic performance penalties due to the ice accretions were made. Results were also compared with similar tests conducted with a Twin Otter wing section in Part 1 of this study. The form of the ice from the large droplet cloud varied as a function of air total temperature; particularly at the near-freezing temperatures of 28 F to 34 F. Changing boot cycle interval time did not prevent formation of an ice ridge. The most detrimental aerodynamic effects occurred at an air total temperature of 28 F.

Addy, Harold E., Jr.↗

OSIRIS-REx Off-Nominal Re-entry Breakup Analysis

The Origins, Spectral Interpretation, Resource Identification, Security, Regolith Explorer (OSIRIS-REx) is a NASA asteroid sampling mission that launched on September 8, 2016. Its objectives are to study the asteroid Bennu for up to 505 days and obtain at least 60 grams of pristine regolith. The sample return to Earth is planned for September 2023. For contingency planning and risk assessment of a potential off-nominal Earth-return trajectory, a re-entry breakup analysis was performed to determine the response of the spacecraft to the environment and predict the breakup sequence and timeline, debris survival, and debris impact conditions. The failure scenario assumed a failure to separate between the bus and the sample return capsule (SRC), resulting in the combined bus+SRC configuration for the re-entry vehicle. Furthermore, consistent overburns or underburns were considered to produce three sets of initial conditions for the analysis consisting of nominal, steep, and shallow entry flight path angles. The results were compared to the breakup analysis performed for a similar vehicle, the Stardust spacecraft, which returned samples from the comet Wild 2 in 2006. This paper describes the OSIRIS-REx spacecraft and presents the results of the re-entry breakup analysis.

reentry breakup analysis↗