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At least 1,711 records · Page 95

Directed Shotgun Proteomics Guided by Saturated RNA-seq Identifies a Complete Expressed Prokaryotic Proteome

Prokaryotes, due to their moderate complexity, are particularly amenable to the comprehensive identification of the protein repertoire expressed under different conditions. We applied a generic strategy to identify a complete expressed prokaryotic proteome, which is based on the analysis of RNA and proteins extracted from matched samples. Saturated transcriptome profiling by RNA-seq provided an endpoint estimate of the protein-coding genes expressed under two conditions which mimic the interaction of Bartonella henselae with its mammalian host. Directed shotgun proteomics experiments were carried out on four subcellular fractions. By specifically targeting proteins which are short, basic, low abundant, and membrane localized, we could eliminate their initial underrepresentation compared to the estimated endpoint. A total of 1250 proteins were identified with an estimated false discovery rate below 1%. This represents 85% of all distinct annotated proteins and ~90% of the expressed protein-coding genes. Genes that were detected at the transcript but not protein level, were found to be highly enriched in several genomic islands. Furthermore, genes that lacked an ortholog and a functional annotation were not detected at the protein level; these may represent examples of overprediction in genome annotations. A dramatic membrane proteome reorganization was observed, including differential regulation of autotransporters, adhesins, and hemin binding proteins. Particularly noteworthy was the complete membrane proteome coverage, which included expression of all members of the VirB/D4 type IV secretion system, a key virulence factor.

Environmental Molecular Sciences Laboratory↗

Influence of Global Atmospheric Change on the Feeding Behavior and Growth Performance of a Mammalian Herbivore, Microtus ochrogaster

Global atmospheric change is influencing the quality of plants as a resource for herbivores. We investigated the impacts of elevated carbon dioxide (CO 2 ) and ozone (O 3 ) on the phytochemistry of two forbs, Solidago canadensis and Taraxacum officinale, and the subsequent feeding behavior and growth performance of weanling prairie voles (Microtus ochrogaster) feeding on those plants. Plants for the chemical analyses and feeding trials were harvested from the understory of control (ambient air), elevated CO 2 (560 µl CO 2 l -1 ), and elevated O 3 (ambient × 1.5) rings at the Aspen FACE (Free Air CO 2 Enrichment) site near Rhinelander, Wisconsin. We assigned individual voles to receive plants from only one FACE ring and recorded plant consumption and weanling body mass for seven days. Elevated CO 2 and O 3 altered the foliar chemistry of both forbs, but only female weanling voles on the O 3 diet showed negative responses to these changes. Elevated CO 2 increased the fiber fractions of both plant species, whereas O 3 fumigation elicited strong responses among many phytochemical components, most notably increasing the carbon-to-nitrogen ratio by 40% and decreasing N by 26%. Consumption did not differ between plant species or among fumigation treatments. Male voles were unaffected by the fumigation treatments, whereas female voles grew 36% less than controls when fed O 3 -grown plants. These results demonstrate that global atmospheric change has the potential to affect the performance of a mammalian herbivore through changes in plant chemistry.

59 BASIC BIOLOGICAL SCIENCES↗

Radioactive aerosols training

An introduction to radioactive aerosols will be presented in this class. There will be a specific focus on transuranic materials, typical to those used in operations at Los Alamos. The uptake of radioactive aerosols is affected by the physical characteristics of the airborne material, the behavior of the aerosol in air (or gas), and the interaction of the material with the worker. Aerosol behavior in the radiological workplace will influence filtration, release control, and dose considerations. This course will review the basics of aerosol technology, distinguishing between the classical (Newtonian) and molecular (quantized) regimes. Particles from about 1 to 10 microns (diameter) are described by classical physics, but particles from about 0.01 to 0.3 microns must be understood through modern molecular theory (i.e. Einstein, 1905). The class will discuss relevant LANL accidents that involved aerosol releases from the years 1971, 1994, 2000, 2003 and 2005.

61 RADIATION PROTECTION AND DOSIMETRY↗

Controlled synthesis and ordered assembly of Co 3 O 4 nanowires using genetically engineered bacterial flagella as biotemplates (Final Technical Report)

The major goals of the project are to gain a fundamental understanding on the influence of the displayed peptides and other processing parameters on the flagella assembly and the resulting biomimetic control over the synthesis and assembly of Co 3 O 4 nanoparticles (NPs) and nanowires (NWs). To pursue these goals, we designed two specific aims: Aim 1: Understanding the self-assembly of genetically engineered flagella and its outcome for site-specific nucleation and growth of Co 3 O 4 NPs and NWs. Aim 2: Understanding controllable alignment of 1D Co 3 O 4 -flagella NWs into highly ordered 2D and 3D structures.

59 BASIC BIOLOGICAL SCIENCES↗

Biokinetic and Dosimetric Models [Slides]

Presentation describes internal dosimetry as an intersection of disciplines involving Physiology, Anatomy, Physics, Mathematics and Computer Science, which some people call "The art of Internal Dosimetry". The basic components for internal dose calculations are: Biokinetic (metabolic) models describing the intake, distribution, retention and excretion of radionuclides in the body, Dosimetric models describing the interaction of radiation within the several body organs and tissues, and System of dose limitation.

61 RADIATION PROTECTION AND DOSIMETRY↗

Drought-Stress Tolerance in Wheat Seedlings Conferred by Phenazine-Producing Rhizobacteria

The specific role of phenazines produced by rhizosphere-colonizing Pseudomonas in mediating wheat seedling drought-stress tolerance and recovery from water deficit was investigated using Pseudomonas chlororaphis 30-84 and isogenic derivatives deficient or enhanced in phenazine production compared to wild type. Following a 7-day water deficit, seedlings that received no-inoculum or were colonized by the phenazine mutant wilted to collapse, whereas seedlings colonized by phenazine producers displayed less severe symptoms. After a 7-day recovery period, survival of seedlings colonized by phenazine-producing strains exceeded 80%, but was less than 60% for no-inoculum controls. A second 7-day water deficit reduced overall survival rates to less than 10% for no-inoculum control seedlings, whereas survival was ∼50% for seedlings colonized by phenazine-producers. The relative water content of seedlings colonized by phenazine-producers was 10–20% greater than for the no-inoculum controls at every stage of water deficit and recovery, resulting in higher recovery indices than observed for the no-inoculum controls. For 10-day water deficits causing the collapse of all seedlings, survival rates remained high for plants colonized by phenazine-producers, especially the enhanced phenazine producer (∼74%), relative to the no-inoculum control (∼25%). These observations indicate that seedlings colonized by the phenazine-producing strains suffered less from dehydration during water deficit and recovered better, potentially contributing to better resilience from a second drought/recovery cycle. Seedlings colonized by phenazine-producing strains invested more in root systems and produced 1.5 to 2 fold more root tips than seedlings colonized by the phenazine mutant or the no-inoculum controls when grown with or without water deficit. The results suggest that the presence of phenazine-producing bacteria in the rhizosphere provides wheat seedlings with a longer adjustment period resulting in greater drought-stress avoidance and resilience.

59 BASIC BIOLOGICAL SCIENCES↗

PcsR2 Is a LuxR-Type Regulator That Is Upregulated on Wheat Roots and Is Unique to Pseudomonas chlororaphis

LuxR solos are common in plant-associated bacteria and increasingly recognized for playing important roles in plant-microbe interkingdom signaling. Unlike the LuxR-type transcriptional regulators of prototype LuxR/LuxI quorum sensing systems, luxR solos do not have a LuxI-type autoinducer synthase gene associated with them. LuxR solos in plant-pathogenic bacteria are important for virulence and in plant endosymbionts contribute to symbiosis. In the present study, we characterized an atypical LuxR solo, PcsR2, in the biological control species Pseudomonas chlororaphis 30–84 that is highly conserved among sequenced P. chlororaphis strains. Unlike most LuxR solos in the plant-associated bacteria characterized to date, pcsR2 is not associated with a proline iminopeptidase gene and the protein has an atypical N-terminal binding domain. We created a pcsR2 deletion mutant and used quantitative RT-PCR to show that the expression of pcsR2 and genes in the operon immediately downstream was upregulated ~10-fold when the wild type strain was grown on wheat roots compared to planktonic culture. PcsR2 was involved in upregulation. Using a GFP transcriptional reporter, we found that expression of pcsR2 responded specifically to root-derived substrates as compared to leaf-derived substrates but not to endogenous AHLs. Compared to the wild type, the mutant was impaired in the ability to utilize root carbon and nitrogen sources in wheat root macerate and to colonize wheat roots. Phenazine production and most biofilm traits previously shown to be correlated with phenazine production also were diminished in the mutant. Gene expression of several of the proteins in the phenazine regulatory network including PhzR, Pip (phenazine inducing protein) and RpeA/RpeB were reduced in the mutant, and overexpression of these genes in trans restored phenazine production in the mutant to wild-type levels, indicating PcsR2 affects the activity of the these regulatory genes upstream of RpeA/RpeB via an undetermined mechanism. Our results indicate PcsR2 upregulates the expression of the adjacent operon in response to unknown wheat root-derived signals and belongs to a novel subfamily of LuxR-type transcriptional regulators found in sequenced P. chlororaphis strains.

59 BASIC BIOLOGICAL SCIENCES↗

Phage K gp102 Drives Temperature-Sensitive Antibacterial Activity on USA300 MRSA

There is widespread interest in using obligately lytic bacteriophages (“phages”) to treat human bacterial infections. Among Staphylococcus aureus infections, the USA300 lineage is a frequent cause of invasive disease. We observed that phage K, a model S. aureus myophage, exhibits temperature-sensitive growth on USA300 strains, with the wild-type phage providing poorer growth suppression in broth and forming smaller and fainter plaques at 37 °C vs. 30 °C. We isolated 65 mutants of phage K that had improved plaquing characteristics at 37 °C when compared to the parental phage. In all 65 mutants, this phenotype was attributable to loss-of-function (LoF) mutations in gp102, which encodes a protein of unknown function that has homologs only among the Herelleviridae (SPO1-like myophages infecting gram-positive bacteria). Additional experiments with representative mutants consistently showed that the temperature-sensitive plaque phenotype was specific to USA300 MRSA strains and that Gp102 disruption was correlated with improved suppression of bacterial growth in broth and improved antibacterial activity in a mouse model of upper respiratory tract infection. The same genotype and in vitro phenotypes could be replicated in close relatives of phage K. Gp102 disruption did not have a detectable effect on adsorption but did delay cell culture lysis relative to wild-type under permissive infection conditions, suggesting that gp102 conservation might be maintained by selective pressure for more rapid replication. Expression of gp102 on a plasmid was toxic to both an MSSA and a USA300 MRSA strain. Molecular modeling predicts a protein with two helix-turn-helix domains that displays some similarity to DNA-binding proteins such as transcription factors. While its function remains unclear, gp102 is a conserved gene that is important to the infection process of Kayvirus phages, and it appears that the manner in which USA300 strains defend against them at 37 °C can be overcome by gp102 LoF mutations.

59 BASIC BIOLOGICAL SCIENCES↗

Sikorsky interactive graphics surface design/manufacturing system

An interactive graphics system conceived to be used in the design, analysis, and manufacturing of aircraft components with free form surfaces was described. In addition to the basic surface definition and viewing capabilities inherent in such a system, numerous other features are present: surface editing, automated smoothing of control curves, variable milling patch boundary definitions, surface intersection definition and viewing, automatic creation of true offset surfaces, digitizer and drafting machine interfaces, and cutter path optimization. Documented costs and time savings of better than six to one are being realized with this system. The system was written in FORTRAN and GSP for use on IBM 2250 CRT's in conjunction with an IBM 370/158 computer.

Robbins, R.↗

Fast electrons in small solar flares

This review summarizes both the direct spacecraft observations of nonrelativistic solar electrons, and observations of the X-ray and radio emission generated by these particles at the sun and in the interplanetary medium. These observations bear on the basic astrophysical process of particle acceleration in tenuous plasmas. We find that in many small solar flares, the nearly 5-100 keV electrons accelerated during flash phase constitute the bulk of the total flare energy. Thus the basic flare mechanism in these flares essentially converts the available flare energy into fast electrons. These electrons may produce the other flare electromagnetic emissions through their interactions with the solar atmosphere. In large proton flares these electrons may provide the energy to eject material from the sun and to create a shock wave which could accelerate nuclei and electrons to much higher energies.

Lin, R. P.↗

Modeling Io volcanism: Maximum volcanic temperatures, depths of melting and magma composition

Interim results of thermal and structural modeling of volcanism on Io were presented. The final results of the modeling are summarized. The basic analysis is an evaluation of the magma trigger mechanism for initiating and maintaining eruptions. Secondary aspects include models of the mechanical mode of magma emplacement, interactions with a sulphur-rich upper crust, and more speculative implications for Io's volcanism.

Crumpler, L. S.↗

Temperature Humidity and Sea Level Pressure Increments Induced by 1DVAR Analysis of GPS Refractivity

The Global Positioning System (GPS) transmitted signals are affected by the atmosphere. Using the radio occultation technique, where a receiver is placed on a low-Earth-orbiting platform. it is possible to perform soundings. by measuring the bending angles of the rays. The information can be converted into atmospheric refractivity. We have developed a one dimensional variational (1DVAR) analysis that uses GPS/MET 1995 refractivity and 6-hour FVDAS (Finite Volume Data Assimilation System) forecasts as background information to constrain the retrievals. The analysis increments are defined as 1DVAR minus background temperature, humidity and sea level pressure. Before assimilating the 1DVAR profiles into the FVDAS. the increments need to be understood. First, some bias could be induced in the retrievals when confronted with actual biased data: second. bias in the back-round could create undesired bias in the retrievals. Anv bias in the analyses will ultimately change the climatology of the model the retrievals will be assimilated into. We relate the increments to the reduction of the difference between observed minus computed refractivity profiles. We also point out the difference in the mean increments using backgrounds which have assimilated either NESDIS TIROS Operational Vertical Sounder (TOVS) operational retrievals or Data Assimilation Office (DAO) TOVS interactive retrievals. The climatology of the model in terms of refractivity is significantly different and this impacts the GPS 1DVAR increments. This testifies that changing the basic load of assimilated data has an influence on the impact the GPS data may have in a DAS.

Poli, Paul↗

Composting on Mars or the Moon: I. Comparative evaluation of process design alternatives

As a candidate technology for treating solid wastes and recovering resources in bioregenerative Advanced Life Support, composting potentially offers such advantages as compactness, low mass, near ambient reactor temperatures and pressures, reliability, flexibility, simplicity, and forgiveness of operational error or neglect. Importantly, the interactions among the physical, chemical, and biological factors that govern composting system behavior are well understood. This article comparatively evaluates five Generic Systems that describe the basic alternatives to composting facility design and control. These are: 1) passive aeration; 2) passive aeration abetted by mechanical agitation; 3) forced aeration--O2 feedback control; 4) forced aeration--temperature feedback control; 5) forced aeration--integrated O2 and temperature feedback control. Each of the five has a distinctive pattern of behavior and process performance characteristics. Only Systems 4 and 5 are judged to be viable candidates for ALS on alien worlds, though which is better suited in this application is yet to be determined.

Review↗

Improved Oxygen-Beam Texturing of Glucose-Monitoring Optics

An improved method has been devised for using directed, hyperthermal beams of oxygen atoms and ions to impart desired textures to the tips of polymethylmethacrylate [PMMA] optical fibers to be used in monitoring the glucose content of blood. The improved method incorporates, but goes beyond, the method described in Texturing Blood-Glucose- Monitoring Optics Using Oxygen Beams (LEW-17642-1), NASA Tech Briefs, Vol. 29, No. 4 (April 2005), page 11a. The basic principle of operation of such a glucose-monitoring sensor is as follows: The textured surface of the optical fiber is coated with chemicals that interact with glucose in such a manner as to change the reflectance of the surface. Light is sent down the optical fiber and is reflected from, the textured surface. The resulting change in reflectance of the light is measured as an indication of the concentration of glucose. The required texture on the ends of the optical fibers is a landscape of microscopic cones or pillars having high aspect ratios (microscopic structures being taller than they are wide). The average distance between hills must be no more than about 5 mso that blood cells (which are wider) cannot enter the valleys between the hills, where they would interfere with optical sensing of glucose in the blood plasma. On the other hand, the plasma is required to enter the valleys, and high aspect ratio structures are needed to maximize the surface area in contact with the plasma, thereby making it possible to obtain a given level of optical glucose-measurement sensitivity with a relatively small volume of blood. There is an additional requirement that the hills be wide enough that a sufficient amount of light can propagate into them and, after reflection, can propagate out of them. The method described in the cited prior article produces a texture comprising cones and pillars that conform to the average-distance and aspect-ratio requirements. However, a significant fraction of the cones and pillars are so narrow that not enough light can propagate along them. The improved method makes it possible to form wider cones and pillars while still satisfying the average-distance and aspect-ratio requirements. In the improved method, as in the previously reported method, multiple optical fibers are first bundled together for simultaneous texturing of their distal tips. However, prior to texturing by exposure to an oxygen beam, the tips are first coated by vapor deposition of a thin, sparse layer of aluminum: The exposure to the aluminum vapor must be short enough (typically of the order of seconds) so that the aluminum nucleates into islands separated by uncoated areas. The coated tips are textured by exposure to a directed beam of hyperthermal (kinetic energy >1 eV) oxygen atoms and/or ions in a vacuum chamber, as in the previously reported method. The aluminum islands partially shield the underlying PMMA from oxidation and erosion by the beam, so that the cones or pillars remaining after texturing are wider than they would otherwise be. To some extent, the dimensions of the hills and the distances between them can be tailored through choice of the thickness of the aluminum coat and/or the oxygen-beam fluence. The figure illustrates an example of texturing of the tip of a PMMA optical fiber without and with prior aluminum coating.

Banks, Bruce A.↗

Coaxial Rotor Flow Phenomena in Forward Flight

Coaxial rotors are nding use in advanced rotorcraft concepts. Combined with lift offset rotor technology, they offer a solution to the problems of dynamic stall and re-verse ow that often limit single rotor edgewise forward ight speeds. Lower tip speed means reduced high-speed impulsive noise. The need for an anti-torque tail rotor is eliminated, a major boon during operation in conned areas. However, the operation of two counter-rotating ro-tors in close proximity generates many possibilities for aerodynamic interactions between rotor blades, blades and vortices, and between vortices. The parameter de-sign space is very large, and requires efcient computations as well as basic experiments to explore important physics to determine performance, loads, and acoustics. Computations are done on the classic HarringtonDingeldein rotor test case from the 1950s using the ROTUNS Navier Stokes code. Two regimes are explored: very low advance ratio as a perturbation from hover, and high advance ratio. Flow eld properties from RotUNS are used with 2-D OVERFLOW computations to capture blade crossing effects including those of higher subsonic Mach numbers. Bladeblade and bladevortex intersection events are captured using a MatLab-based predictor.

Flow Phenomena↗

A Summary of Normal Incidence Tube Tests of NASA'S Bio-inspired Broadband Acoustic Absorber

Motivated by the need to reduce noise pollution from aircraft engines, NASA has continued to design, manufacture and test recently patented structures that mimic the geometry and the broadband and low frequency acoustic absorption of assemblies of natural reeds. This report documents a screening test, which is a precursor to more controlled modelling experiments. The purpose of this experiment was to identify variables and prototype design features that might play a dominant role in the acoustic performance of these structures. To date, twenty-four prototypes, each with an overall shape of a cube (nominally 50 mm x 50 mm x 50 mm), were designed and additively manufactured from thermoplastics using a Fused Filament Fabrication technique. The prototypes were tested in the NASA Glenn and Langley Normal Incidence Tubes to experimentally determine the acoustic absorption as a function of frequency from 400 Hz to 3000 Hz. Results indicate that a variety of structures exhibit substantial acoustic absorption in that frequency range, with an average absorption coefficient greater than 0.6, especially in the frequency range of 400 Hz to 1000 Hz. Six basic prototype designs were chosen to inform future modeling research. Results of these experiments can be used to evaluate existing physics-based models of the interaction of sound waves with these structures or to develop new models. Validated physics-based models of these structures can help engineers optimize the designs for different commercial noise control applications.

Acoustics↗