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At least 73 records · Page 4

Complete biosynthesis of salicylic acid from phenylalanine in plants

Salicylic acid (SA) is a pivotal phytohormone for plant responses to biotic and abiotic stresses. Plants have evolved two pathways to produce SA: the isochorismate synthase and phenylalanine ammonia lyase (PAL) pathways. Whereas the isochorismate synthase pathway has been fully identified, the PAL pathway remains incomplete. Here we report the full characterization of the PAL pathway for SA biosynthesis via functional analysis of rice (Oryza sativa) SA-DEFICIENT GENE 1 (OSD1) to OSD4. The cinnamoyl-coenzyme A (CoA) ligase OSD1 catalyses the conversion of trans-cinnamic acid to cinnamoyl-CoA, which is subsequently transformed to benzoyl-CoA via the β-oxidative pathway in peroxisomes. The resulting benzoyl-CoA is further converted to benzyl benzoate by the peroxisomal benzoyltransferase OSD2. Benzyl benzoate is subsequently hydroxylated to benzyl salicylate by the endoplasmic reticulum membrane-resident cytochrome P450 OSD3, which is ultimately hydrolysed to salicylic acid by the cytoplasmic carboxylesterase OSD4. Evolutionary analyses reveal that the PAL pathway was first assembled before the divergence of gymnosperms and has been conserved in most seed plants. Activation of the PAL pathway in rice significantly enhances salicylic acid levels and plant immunity. Completion of the PAL pathway provides critical insights into the primary salicylic acid biosynthetic pathway across plant species and offers a precise target for modulating crop immunity.

59 BASIC BIOLOGICAL SCIENCES↗

A 6 MW/m 2 High Heat Flux Testing Facility of Irradiated Materials Using Infrared Plasma-Arc Lamps

Assessing the effect of neutron irradiation of plasma-facing materials has been challenging due to both the technical and radiological challenges involved. In an effort to address the radiological challenges, a facility was developed to conduct high heat flux testing (HHFT) of inherently small samples of neutron-irradiated materials. A new line-focus reflector was designed and fabricated at Oak Ridge National Laboratory for a plasma-arc lamp (PAL) to attain a source heat flux of 12 MW/m 2 . The new reflector was fabricated with two ports for monitoring specimen condition during HHFT. At the same operational conditions for PAL, the absorbed heat flux in tungsten was increased from 1.39 MW/m 2 with the uniform irradiance reflector to 5.12 MW/m 2 for the line-focus reflector. This fourfold increase in the heat flux, at the same PAL electrode lifetimes, enabled cost-effective facility operation for a high number of cyclic high heat flux tests. Specifically, the test section is confined to a hemispherical dome, and specimens are bolted directly to a water-cooled copper alloy rod. Temperature measurement in the PAL facility was a main challenge due to a limited line of sight. For the first time in a PAL facility operating at high heat fluxes, the specimen surface temperature was directly measured during HHFT with a pyrometer. Finally, the HHFT data, which were obtained in this upgraded PAL facility, demonstrated the facility readiness for irradiated materials.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

DETAILED CHEMICAL ABUNDANCES OF FOUR STARS IN THE UNUSUAL GLOBULAR CLUSTER PALOMAR 1

Detailed chemical abundances for 21 elements are presented for four red giants in the anomalous outer halo globular cluster Palomar 1 (R{sub GC} = 17.2 kpc, Z = 3.6 kpc) using high-resolution (R = 36, 000) spectra from the High Dispersion Spectrograph on the Subaru Telescope. Pal 1 has long been considered unusual because of its low surface brightness, sparse red giant branch, young age, and its possible association with two extragalactic streams of stars. This paper shows that its chemistry further confirms its unusual nature. The mean metallicity of the four stars, [Fe/H] = -0.60 {+-} 0.01, is high for a globular cluster so far from the Galactic center, but is low for a typical open cluster. The [{alpha}/Fe] ratios, though in agreement with the Galactic stars within the 1{sigma} errors, agree best with the lower values in dwarf galaxies. No signs of the Na/O anticorrelation are detected in Pal 1, though Na appears to be marginally high in all four stars. Pal 1's neutron-capture elements are also unusual: its high [Ba/Y] ratio agrees best with dwarf galaxies, implying an excess of second-peak over first-peak s-process elements, while its [Eu/{alpha}] and [Ba/Eu] ratios show that Pal 1's contributions from the r-process must have differed in some way from normal Galactic stars. Therefore, Pal 1 is unusual chemically, as well in its other properties. Pal 1 shares some of its unusual abundance characteristics with the young clusters associated with the Sagittarius dwarf galaxy remnant and the intermediate-age LMC clusters, and could be chemically associated with the Canis Majoris overdensity; however, it does not seem to be similar to the Monoceros/Galactic Anticenter Stellar Stream.

79 ASTRONOMY AND ASTROPHYSICS↗

Down-regulation of kelch domain-containing F-box protein in Arabidopsis enhances the production of (poly)phenols and tolerance to ultraviolet radiation

Phenylpropanoid biosynthesis in plants engenders myriad phenolics with diverse biological functions. Phenylalanine ammonia-lyase (PAL) is the first committed enzyme in the pathway, directing primary metabolic flux into a phenylpropanoid branch. Previously, we demonstrated that the Arabidopsis Kelch-domain containing F-box proteins, AtKFB01, -20, and -50, function as the negative regulators controlling phenylpropanoid biosynthesis via mediating PAL’s ubiquitination and subsequent degradation. Here, we reveal that Arabidopsis KFB39, a close homolog of AtKFB50, also interacts physically with PAL isozymes and modulates PALs' stability and activity. Disturbing the expression of KFB39 reciprocally affects the accumulation/deposition of a set of phenylpropanoid end products, suggesting that KFB39 is an additional post-translational regulator responsible for the turnover of PAL and negatively controlling phenylpropanoid biosynthesis. Furthermore, we discover that exposure of Arabidopsis to UV-B radiation suppresses the expression of all four KFB genes while inducing the transcription of PAL isogenes; these data suggest that Arabidopsis consolidates both transcriptional and post-translational regulation mechanisms to maximize its responses to UV stress. Simultaneous down-regulation of all four identified KFBs significantly enhances the production of (poly)phenols and the plant’s tolerance to UV irradiation. This study offers a biotechnological approach for engineering the production of useful phenolic chemicals and for increasing a plant’s resistance to environmental stress.

59 BASIC BIOLOGICAL SCIENCES↗

Portable Airborne Laser System Measures Forest-Canopy Height

(PALS) is a combination of laser ranging, video imaging, positioning, and data-processing subsystems designed for measuring the heights of forest canopies along linear transects from tens to thousands of kilometers long. Unlike prior laser ranging systems designed to serve the same purpose, the PALS is not restricted to use aboard a single aircraft of a specific type: the PALS fits into two large suitcases that can be carried to any convenient location, and the PALS can be installed in almost any local aircraft for hire, thereby making it possible to sample remote forests at relatively low cost. The initial cost and the cost of repairing the PALS are also lower because the PALS hardware consists mostly of commercial off-the-shelf (COTS) units that can easily be replaced in the field. The COTS units include a laser ranging transceiver, a charge-coupled-device camera that images the laser-illuminated targets, a differential Global Positioning System (dGPS) receiver capable of operation within the Wide Area Augmentation System, a video titler, a video cassette recorder (VCR), and a laptop computer equipped with two serial ports. The VCR and computer are powered by batteries; the other units are powered at 12 VDC from the 28-VDC aircraft power system via a low-pass filter and a voltage converter. The dGPS receiver feeds location and time data, at an update rate of 0.5 Hz, to the video titler and the computer. The laser ranging transceiver, operating at a sampling rate of 2 kHz, feeds its serial range and amplitude data stream to the computer. The analog video signal from the CCD camera is fed into the video titler wherein the signal is annotated with position and time information. The titler then forwards the annotated signal to the VCR for recording on 8-mm tapes. The dGPS and laser range and amplitude serial data streams are processed by software that displays the laser trace and the dGPS information as they are fed into the computer, subsamples the laser range and amplitude data, interleaves the subsampled data with the dGPS information, and records the resulting interleaved data stream.

Nelson, Ross↗

Comparison of high-resolution airborne soil moisture retrievals to SMAP soil moisture during the SMAP validation experiment 2016 (SMAPVEX16)

The NASA's Soil Moisture Active Passive (SMAP) mission conducted a field experiment with its partners over two 40-km agricultural domains in Iowa and Manitoba in the summer of 2016 to address concerns observed in SMAP soil moisture (SM) retrievals over agricultural areas. The experiment featured airborne Passive Active L-band System (PALS) flights over each domain with intensive ground measurements and dense networks of SM monitoring stations. With two intensive observation periods separated in time (May 28–June 20 and July 14–August 16), the flights captured both early-season/low vegetation and later-season/high-vegetation conditions. The comparison of the PALS brightness temperature (TB) measurements to the SMAP TB observed over the sites resulted in root mean square difference (RMSD) of 2.8 K and 4.0 K for vertical and horizontal polarizations, respectively. The subsequent SM analysis rescaled the PALS TB with the SMAP TB to allow equitable comparisons between the SM retrievals from the two instruments. The PALS SM retrieval algorithm used the SM sampled by the ground teams during the overpass days for tuning, and was parameterized by a high-resolution vegetation water content product calibrated using vegetation samples collected during the experiment. The tuning process was not able to find a satisfactory result with a temporally constant set of parameters in the single channel algorithm for the two intensive observation periods of the experiment. This result indicated that the rapid change in the vegetation structure during the growth stages and likely variation in the surface roughness conditions were not compatible with rigid parameterization over the entire period. However, using seasonally variable parameters we found that it was possible to retrieve soil moisture with satisfactory accuracy. Comparative analysis with the SMAP SM product included aggregation of the PALS SM to the SMAP pixel-scale. The RMSD between the PALS SM and the aggregated manual field samples was <0.04 m3/m3 with Pearson correlation >0.85 for both sites. The comparison between different in situ sources indicated that the soil moisture network measurements were not the source of the large biases observed for SMAP over the sites reported in earlier studies. Therefore, the results suggested the rapidly growing vegetation and the early-season surface condition changes not captured by the SMAP algorithm caused the SMAP retrieval errors. In addition, the significant deviations of the vegetation water content used by the SMAP product from the calibrated vegetation water content obtained during the experiment compounds the problem.

Andreas Colliander↗

Identifying attributes of CO 2 leakage zones in shallow aquifers using a parametric level set method: Original Research Article: Identifying attributes of CO 2 leakage zones in shallow aquifers using a parametric level set method

Leakage through abandoned wells and geologic faults poses the greatest risk to CO 2 storage permanence. For shallow aquifers, secondary CO 2 plumes emanating from the leak zones may go undetected for a sustained period of time and has the greatest potential to cause large-scale and long-term environmental impacts. Identification of the attributes of leak zones, including their shape, location, and strength, is required for environmental risk assessment. This study applies a parametric level set (PaLS) method to characterize leak zones. Level set methods are appealing for tracking topological changes and recovering unknown shapes of objects. However, level set evolution using the conventional level set methods is computationally challenging. In PaLS, the level set function is approximated by using a weighted sum of basis functions and the level set evolution problem is replaced by an optimization problem. The efficacy of PaLS is demonstrated by reconstructing attributes of a CO 2 leak zone in a carbonate aquifer. Our results show that PaLS is a robust source identification method that can recover the approximate leak zone locations in the presence of measurement errors, model parameter uncertainty, and inaccurate guesses of source flux strengths. Accurate delineation of source geometry is achievable when a relatively dense observation network is used. The PaLS-based, source recovery framework introduced in this work is generic and can be adapted for any reactive transport model by simply switching the pre- and post-processing routines.

54 ENVIRONMENTAL SCIENCES↗

The intense slow positron beam facility at the PULSTAR reactor and applications in nano-materials study

An intense slow positron beam has been established at the PULSTAR nuclear research reactor of North Carolina State University. The slow positrons are generated by pair production in a tungsten moderator from gammarays produced in the reactor core and by neutron capture reactions in cadmium. The moderated positrons are electrostatically extracted and magnetically guided out of the region near the core. Subsequently, the positrons are used in two spectrometers that are capable of performing positron annihilation lifetime spectroscopy (PALS) and positron Doppler broadening spectroscopy (DBS) to probe the defect and free volume properties of materials. One of the spectrometers (e{sup +}-PALS) utilizes an rf buncher to produce a pulsed beam and has a timing resolution of 277 ps. The second spectrometer (Ps-PALS) uses a secondary electron timing technique and is dedicated to positronium lifetime measurements with an approximately 1 ns timing resolution. PALS measurements have been conducted in the e{sup +}-PALS spectrometer on a series of nano-materials including organic photovoltaic thin films, membranes for filtration, and polymeric fibers. These studies have resulted in understanding some critical issues related to the development of the examined nano-materials.

43 PARTICLE ACCELERATORS↗

TIDAL STREAM MORPHOLOGY AS AN INDICATOR OF DARK MATTER HALO GEOMETRY: THE CASE OF PALOMAR 5

This paper presents an example where the morphology of a single stellar stream can be used to rule out a specific galactic potential form without the need for velocity information. We investigate the globular cluster Palomar 5 (Pal 5), which is tidally disrupting into a cold, thin stream mapped over 22 deg on the sky with a typical width of 0.7 deg. We generate models of this stream by fixing Pal 5's present-day position, distance, and radial velocity via observations, while allowing its proper motion to vary. In a spherical dark matter halo we easily find models that fit the observed morphology. However, no plausible Pal 5 model could be found in the triaxial potential of Law and Majewski, which has been proposed to explain the properties of the Sagittarius stream. In this case, the long, thin, and curved morphology of the Pal 5 stream alone can be used to rule out such a potential configuration. Pal 5-like streams in this potential are either too straight, missing the curvature of the observations, or show an unusual morphology which we dub stream-fanning: a signature sensitive to the triaxiality of a potential. We conclude that the mere existence of other thin tidal streams must provide broad constraints on the orientation and shape of the dark matter halo they inhabit.

79 ASTRONOMY AND ASTROPHYSICS↗

Plasma-arc lamp high heat flux cycling exposure of neutron irradiated tungsten materials

Thick plate, unalloyed W was neutron irradiated in the High Flux Isotope Reactor (HFIR) at 550 °C to a fast fluence of 1.24 × 10 25 n m -2 E > 0.1 MeV (~0.24 dpa). Unirradiated and irradiated specimens of the material were high heat flux (HHF) tested in the Plasma Arc Lamp (PAL) facility. The PAL uses a high-power photon source to provide a broad and even heat distribution on the sample surface. To simulate on/off cycling of normal operating plasma, the samples were exposed to approximately 800 cycles at 4.73 MW m -2 absorbed heat flux (incident heat fluxes of 10.95 MW m -2 ). Additionally, after PAL exposure, slight changes were observed on the surfaces of the samples with SEM. The samples showed some annealing in the near surface polished region, but they were all below the damage threshold for cracking or other destructive features. The PAL has a large parameter space for future testing. The use of the HFIR and PAL to sequentially expose neutron irradiated samples to HHF will be a powerful tool for understanding materials behavior in a fusion-like environment.

36 MATERIALS SCIENCE↗

Chloroplast Dynamics and Photosynthetic Efficiency: Final Technical Report

This project investigated the mechanism by which chloroplasts position themselves to maximize solar energy utilization, to enhance gas exchange, to minimize environmental stress, and to promote efficient exchange of metabolites with other compartments within the plant cell. Chloroplasts move within leaf cells to optimize light levels, moving toward levels of light useful for photosynthesis while moving away from excess light. Plastids sometimes extend their reach by sending out projections (stromules) that can connect anchor chloroplasts in position within the cell or provide close contacts with plasma membrane, mitochondria, peroxisomes, endoplasmic reticulum, and the nucleus. The intracellular location of chloroplasts in relation to other organelles with which they share biosynthetic pathways, such as peroxisomes and mitochondria in photorespiration, affects metabolite flow. This work contributed to the knowledge of the mechanisms of organelle movement and anchoring in specific locations in plant cells and how proteins traffic within the cell. We identified two domains on 12 of the 13 Arabidopsis myosins that were similar to the vacuole-binding (V) domain characterized in yeast and to the DIL domain characterized in yeast and mouse as required for secretory vesicle or melanosome movement, respectively. Because all of the Arabidopsis regions with homology to the V domain contain the amino acid sequence PAL, we refer to this region as the Arabidopsis PAL domain. We have used the yeast Myo2p tail structural information to model the 12 myosin XI tail domains containing the homologous PAL and DIL domains. Eight YFP::DIL domain fusions labeled peroxisomes; none labeled mitochondria or chloroplasts. Six myosin XI Vacuole domains labeled mitochondria and seven labeled Golgi bodies. The Arabidopsis myosin XI-F PAL domain and the homologous myosin XI-F PAL domain from N. benthamiana labels chloroplasts and stromules in N. benthamiana leaves. Using an Arabidopsis line containing hotoconvertible GFP, we observed transfer of protein from one plastid to another and within a stromule from single plastids. We provided time-lapse movies demonstrating movement of both the photoconvertible GFP and standard GFP between plastids. We previously demonstrated the lack of a plastid network within plant cells. We provided protocols explaining how to use fluorescent protein technology to track plastids and stromules within plant cells. We demonstrated that standard GFP unexpectedly could be photoconverted to a red form under certain conditions, allowing the use of GFP lines for studies that require photoconversion.

59 BASIC BIOLOGICAL SCIENCES↗

Reduced phenylalanine ammonia-lyase and tyrosine ammonia-lyase activities and lignin synthesis in wheat grown under low pressure sodium lamps

Wheat (Triticum aestivum L. cv Fremont) grown in hydroponic culture under 24-hour continuous irradiation at 560 to 580 micromoles per square meter per second from either metalhalide (MH), high pressure sodium (HPS), or low pressure sodium (LPS) lamps reached maturity in 70 days. Grain yields were similar under all three lamps, although LPS-grown plants lodged at maturity. Phenylalanine ammonia-lyase (PAL) and a tyrosine ammonia lyase (TAL) with lesser activity were detected in all extracts of leaf, inflorescence, and stem. Ammonia-lyase activities increased with age of the plant, and plants grown under the LPS lamp displayed PAL and TAL activities lower than wheat cultured under MH and HPS radiation. Greenhouse solar-grown wheat had the highest PAL and TAL activities. Lignin content of LPS-grown wheat was also significantly reduced from that of plants grown under MH or HPS lamps or in the greenhouse, showing a correlation with the reduced PAL and TAL activities. Ratios of far red-absorbing phytochrome to total phytochrome were similar for all three lamps, but the data do not yet warrant a conclusion about specific wavelengths missing from the LPS lamps that might have induced PAL and TAL activities in plants under the other lamps.

Triticum/enzymology/growth & development/metabolis↗

Characterization of Space Shuttle Thermal Protection System (TPS) Materials for Return-to-Flight following the Shuttle Columbia Accident Investigation

During the Space Shuttle Columbia Accident Investigation, it was determined that a large chunk of polyurethane insulating foam (= 1.67 lbs) on the External Tank (ET) came loose during Columbia's ascent on 2-1-03. The foam piece struck some of the protective Reinforced Carbon-Carbon (RCC) panels on the leading edge of Columbia's left wing in the mid-wing area. This impact damaged Columbia to the extent that upon re-entry to Earth, superheGed air approaching 3,000 F caused the vehicle to break up, killing all seven astronauts on board. A paper after the Columbia Accident Investigation highlighted thermal analysis testing performed on External Tank TPS materials (1). These materials included BX-250 (now BX-265) rigid polyurethane foam and SLA-561 Super Lightweight Ablator (highly-filled silicone rubber). The large chunk of foam from Columbia originated fiom the left bipod ramp of the ET. The foam in this ramp area was hand-sprayed over the SLA material and various fittings, allowed to dry, and manually shaved into a ramp shape. In Return-to-Flight (RTF) efforts following Columbia, the decision was made to remove the foam in the bipod ramp areas. During RTF efforts, further thermal analysis testing was performed on BX-265 foam by DSC and DMA. Flat panels of foam about 2-in. thick were sprayed on ET tank material (aluminum alloys). The DSC testing showed that foam material very close to the metal substrate cured more slowly than bulk foam material. All of the foam used on the ET is considered fully cured about 21 days after it is sprayed. The RTF culminated in the successful launch of Space Shuttle Discovery on 7-26-05. Although the flight was a success, there was another serious incident of foam loss fiom the ET during Shuttle ascent. This time, a rather large chunk of BX-265 foam (= 0.9 lbs) came loose from the liquid hydrogen (LH2) PAL ramp, although the foam did not strike the Shuttle Orbiter containing the crew. DMA testing was performed on foam samples taken fiom a simulated PAL ramp panel. It was found that the smooth rind on the foam facing the cable tray did significantly affect the properties of foam at the PAL ramp surface. The smooth rind increased the storage modulus E' of the foam as much as 20- 40% over a temperature range of -145 to 95 C. Because of foam loss fiom the PAL ramp, future Shuttle flights were grounded indefinitely to allow further testing to better understand foam properties. The decision was also made to remove foam from the LH2 PAL, ramp. Other RTF efforts prior to the launch of Discovery included

Wingard, Doug↗

Physiologically-based pharmacokinetic model for Fentanyl in support of the development of Provisional Advisory Levels

Provisional Advisory Levels (PALs) are tiered exposure limits for toxic chemicals in air and drinking water that are developed to assist in emergency responses. Physiologically-based pharmacokinetic (PBPK) modeling can support this process by enabling extrapolations across doses, and exposure routes, thereby addressing gaps in the available toxicity data. Here, we describe the development of a PBPK model for Fentanyl – a synthetic opioid used clinically for pain management – to support the establishment of PALs. Starting from an existing model for intravenous Fentanyl, we first optimized distribution and clearance parameters using several additional IV datasets. We then calibrated the model using pharmacokinetic data for various formulations, and determined the absorbed fraction, F, and time taken for the absorbed amount to reach 90% of its final value, t90. For aerosolized pulmonary Fentanyl, F = 1 and t90 < 1 min indicating complete and rapid absorption. The F value ranged from 0.35 to 0.74 for oral and various transmucosal routes. Oral Fentanyl was absorbed the slowest (t90 ∼ 300 min); the absorption of intranasal Fentanyl was relatively rapid (t90 ∼ 20–40 min); and the various oral transmucosal routes had intermediate absorption rates (t90 ∼ 160–300 min). Based on these results, for inhalation exposures, we assumed that all of the Fentanyl inhaled from the air during each breath directly, and instantaneously enters the arterial circulation. We present model predictions of Fentanyl blood concentrations in oral and inhalation scenarios relevant for PAL development, and provide an analytical expression that can be used to extrapolate between oral and inhalation routes for the derivation of PALs. - Highlights: • We develop a Fentanyl PBPK model for relating external dose to internal levels. • We calibrate the model to oral and inhalation exposures using > 50 human datasets. • Model predictions are in good agreement with the available pharmacokinetic data. • The model can be used for extrapolating across routes, doses and exposure durations. • We illustrate how the model can be used for developing Provisional Advisory Levels.

60 APPLIED LIFE SCIENCES↗

HST/WFC3 OBSERVATIONS OF LOW-MASS GLOBULAR CLUSTERS AM 4 AND PALOMAR 13: PHYSICAL PROPERTIES AND IMPLICATIONS FOR MASS LOSS

We investigate the loss of low-mass stars in two of the faintest globular clusters known, AM 4 and Palomar 13 (Pal 13), using HST/WFC3 F606W and F814W photometry. To determine the physical properties of each cluster—age, mass, metallicity, extinction, and present day mass function (MF)—we use the maximum likelihood color-magnitude diagram (CMD) fitting program MATCH and the Dartmouth, Padova, and BaSTI stellar evolution models. For AM 4, the Dartmouth models provide the best match to the CMD and yield an age of >13 Gyr, metallicity log Z/Z {sub ☉} = –1.68 ± 0.08, a distance modulus (m – M) {sub V} = 17.47 ± 0.03, and reddening A{sub V} = 0.19 ± 0.02. For Pal 13 the Dartmouth models give an age of 13.4 ± 0.5 Gyr, log Z/Z {sub ☉} = –1.55 ± 0.06, (m – M) {sub V} = 17.17 ± 0.02, and A{sub V} = 0.43 ± 0.01. We find that the systematic uncertainties due to choice in assumed stellar model greatly exceed the random uncertainties, highlighting the importance of using multiple stellar models when analyzing stellar populations. Assuming a single-sloped power-law MF, we find that AM 4 and Pal 13 have spectral indices α = +0.68 ± 0.34 and α = –1.67 ± 0.25 (where a Salpeter MF has α = +1.35), respectively. Comparing our derived slopes with literature measurements of cluster integrated magnitude (M{sub V} ) and MF slope indicates that AM 4 is an outlier. Its MF slope is substantially steeper than clusters of comparable luminosity, while Pal 13 has an MF in line with the general trend. We discuss both primordial and dynamical origins for the unusual MF slope of AM 4 and tentatively favor the dynamical scenario. However, MF slopes of more low luminosity clusters are needed to verify this hypothesis.

79 ASTRONOMY AND ASTROPHYSICS↗

Arabidopsis myosin XI sub-domains homologous to the yeast myo2p organelle inheritance sub-domain target subcellular structures in plant cells

Myosin XI motor proteins transport plant organelles on the actin cytoskeleton. The Arabidopsis gene family that encodes myosin XI has 13 members, 12 of which have sub-domains within the tail region that are homologous to well-characterized cargo-binding domains in the yeast myosin V myo2p. Little is presently known about the cargo-binding domains of plant myosin XIs. Prior experiments in which most or all of the tail regions of myosin XIs have been fused to yellow fluorescent protein (YFP) and transiently expressed have often not resulted in fluorescent labeling of plant organelles. We identified 42 amino-acid regions within 12 Arabidopsis myosin XIs that are homologous to the yeast myo2p tail region known to be essential for vacuole and mitochondrial inheritance. A YFP fusion of the yeast region expressed in plants did not label tonoplasts or mitochondria. We investigated whether the homologous Arabidopsis regions, termed by us the “PAL” sub-domain, could associate with subcellular structures following transient expression of fusions with YFP in Nicotiana benthamiana. Seven YFP::PAL sub-domain fusions decorated Golgi and six were localized to mitochondria. In general, the myosin XI PAL sub-domains labeled organelles whose motility had previously been observed to be affected by mutagenesis or dominant negative assays with the respective myosins. Simultaneous transient expression of the PAL sub-domains of myosin XI-H, XI-I, and XI-K resulted in inhibition of movement of mitochondria and Golgi.

59 BASIC BIOLOGICAL SCIENCES↗

Manipulation of Micro Scale Particles in Optical Traps Using Programmable Spatial Light Modulation

1064 nm light, from an Nd:YAG laser, was polarized and incident upon a programmable parallel aligned liquid crystal spatial light modulator (PAL-SLM), where it was phase modulated according to the program controlling the PAL-SLM. Light reflected from the PAL-SLM was injected into a microscope and focused. At the focus, multiple optical traps were formed in which 9.975 m spheres were captured. The traps and the spheres were moved by changing the program of the PAL-SLM. The motion of ordered groups of micro particles was clearly demonstrated.

Seibel, Robin E.↗

Estimation of the Unsteady Aerodynamic Load on Space Shuttle External Tank Protuberances from a Component Wind Tunnel Test

At the wake of the Columbia (STS-107) accident it was decided to remove the Protuberance Aerodynamic Load (PAL) Ramp that was originally intended to protect various protuberances outside of the Space Shuttle External Tank from high buffet load induced by cross-flows at transonic speed. In order to establish the buffet load without the PAL ramp, a wind tunnel test was conducted where segments of the protuberances were instrumented with dynamic pressure transducers; and power-spectra of sectional lift and drag forces at various span-wise locations between two adjacent support brackets were measured under different cross flow angles, Mach number and other conditions. Additionally, frequency-dependent spatial correlations between the sectional forces were also established. The sectional forces were then adjusted by the correlation length to establish span-averaged spectra of normal and lateral forces that can be suitably "added" to various other unsteady forces encountered by the protuberance. This paper describes the methodology used for calculating the correlation-adjusted power spectrum of the buffet load. A second part of the paper describes wind-tunnel results on the difference in the buffet load on the protuberances with and without the PAL ramp. In general when the ramp height is the same as that of the protuberance height, such as that found on the liquid Oxygen part of the tank, the ramp is found to cause significant reduction of the unsteady aerodynamic load. However, on the liquid Hydrogen part of the tank, where the Oxygen feed-line is far larger in diameter than the height of the PAL ramp, little protection is found to be available to all but the Cable Tray.

Panda, J.↗