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At least 1,063 records · Page 59

A role for myosin VI in retinal pigment epithelium phagocytosis

Highlights: • Quashing myosin VI activity reduces rates of phagosome trafficking in RPE cells. • Trafficked phagosomes in RPE cells undergo two modes of motion. • The more randomly directed mode of phagosome trafficking is myosin VI dependent. • The more directed mode of phagosome trafficking is myosin VI independent. The retinal pigment epithelium (RPE) is a monolayer of pigmented cells adjacent to the choroid coat, the vascular layer of the eye. Among other functions, these cells are responsible for the phagocytosis of rod and cone cell waste shed from the photoreceptor outer segments. We describe here studies to understand the involvement of the motor protein myosin VI in the trafficking of internalized microspheres by a human retinal pigment epithelium primary cell line (ARPE-19). We perturbed the myosin VI-actin interaction by overexpressing a dominant negative myosin VI construct in these cells. We used single particle tracking to characterize the trajectories of internalized fluorescent microspheres. Analysis of the speed of the microspheres' motions revealed that the magnitude of the average speed over short time scales is reduced in the presence of the dominant negative motor. Analysis of the mean-squared displacement of these trajectories demonstrated that trafficking of phagosomes involves two modes of motion, a rapid, more randomly directed motion that is myosin VI dependent and a slower, more directed motion that is independent of the motor. From these data, we posit that this trafficking process involves an interplay between myosin VI motors, which are involved in motion through the actin periphery of the cell, and microtubule motors.

60 APPLIED LIFE SCIENCES↗

A polymer, random walk model for the size-distribution of large DNA fragments after high linear energy transfer radiation

DNA double-strand breaks (DSBs) produced by densely ionizing radiation are not located randomly in the genome: recent data indicate DSB clustering along chromosomes. Stochastic DSB clustering at large scales, from > 100 Mbp down to < 0.01 Mbp, is modeled using computer simulations and analytic equations. A random-walk, coarse-grained polymer model for chromatin is combined with a simple track structure model in Monte Carlo software called DNAbreak and is applied to data on alpha-particle irradiation of V-79 cells. The chromatin model neglects molecular details but systematically incorporates an increase in average spatial separation between two DNA loci as the number of base-pairs between the loci increases. Fragment-size distributions obtained using DNAbreak match data on large fragments about as well as distributions previously obtained with a less mechanistic approach. Dose-response relations, linear at small doses of high linear energy transfer (LET) radiation, are obtained. They are found to be non-linear when the dose becomes so large that there is a significant probability of overlapping or close juxtaposition, along one chromosome, for different DSB clusters from different tracks. The non-linearity is more evident for large fragments than for small. The DNAbreak results furnish an example of the RLC (randomly located clusters) analytic formalism, which generalizes the broken-stick fragment-size distribution of the random-breakage model that is often applied to low-LET data.

DNA/radiation effects↗

High-Throughput Screening of Catalytic H 2 Production

Hydrogenases, ferredoxins, and ferredoxin-NADP + reductases (FNR) are redox proteins that mediate electron metabolism in vivo, and are also potential components for biological H 2 production technologies. A high-throughput H 2 production assay device (H 2 PAD) is presented that enables simultaneous evaluation of 96 individual H 2 production reactions to identify components that improve performance. Using a CCD camera and image analysis software, H 2 PAD senses the chemo-optical response of Pd/WO 3 thin films to the H 2 produced. H 2 PAD-enabled discovery of hydrogenase and FNR mutants that enhance biological H 2 production is reported. From a library of 10 080 randomly mutated Clostridium pasteurianum [FeFe] hydrogenases, we found a mutant with nearly 3-fold higher H 2 production specific activity. From a library of 400 semi-randomly mutated Oryza sativa FNR, the top hit enabled a 60 % increase in NADPH-driven H 2 production rates. H 2 PAD can also facilitate elucidation of fundamental biochemical mechanisms within these systems.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Correlated Displacement of Dynamic Elastic Dipoles Produces Nonclassical Electrostriction in Zr-Doped Ceria

By combining experimental data with density functional theory-based ab initio molecular dynamics modeling, this work provides evidence that nonclassical electrostriction in isovalent Zr-doped ceria is due to the correlated anharmonic motion of dynamic elastic dipoles associated with multiple [ZrO 8 ]-local bonding units with a high Zr concentration (Zr 0.1 Ce 0.9 O 2 ). Introduction of 0.5 mol % trivalent or divalent codopants (Sc, Yb, La, or Ca) reduces the longitudinal electrostriction strain coefficient by more than a factor of 10, produces a 3-fold decrease in the relative dielectric permittivity, and increases the elastic modulus. Since these changes depend neither on the radius nor on the valency of the codopant, we conclude that the responsible species are charge-compensating oxygen vacancies (VO). For trivalent dopants (Do 0.005 Zr 0.1 Ce 0.895 O 1.9975 ), oxygen vacancies are present at a concentration ratio 1:40 with respect to Zr, giving, for random distribution, a characteristic interaction distance of ≤2.3 unit cells (1.2 nm). Oxygen vacancies participate in [ZrO 7 -V O ] local bonding units, disrupting the correlated dynamic displacements of the connected [ZrO 8 ]-local bonding units. Finally, such correlated motion of dynamic elastic dipoles may also explain the exponential increase in the longitudinal electrostriction strain coefficient with an increase in Zr concentration to <0.2 mole fraction and must be taken into account for further development of nonclassical electrostrictors based on Zr-doped ceria.

36 MATERIALS SCIENCE↗

Experimental and Theoretical Study of Azimuth Angle and Polarization Dependences of Sum-Frequency-Generation Vibrational Spectral Features of Uniaxially Aligned Cellulose Crystals

Comprehensive interpretation of vibrational sum-frequency-generation (SFG) spectroscopic features of SFG-active nanodomains interspersed in amorphous bulk requires the knowledge of nonlinear susceptibility, χ ijk (2) , of the SFG-active phase as a function of its spatial arrangement in the bulk as well as the polarizations of the probe lights. Our work reports the full analysis of the azimuth angle and polarization dependence of SFG signals from a control sample consisting of uniaxially aligned cellulose Iβ crystals. The χ ijk (2) terms of cellulose were estimated from quantum mechanics calculations using time-dependent density functional theory (TD-DFT), and a simple structural model was built with truncated glucose dimers. The theoretical azimuth angle and polarization dependences of characteristic CH/CH 2 and OH stretch modes of cellulose were compared with the experimentally observed trends. These comparisons revealed that the relative polarity of crystallites within the SFG coherence length, the random quasi phase-matching of polycrystalline domains, and the preferential packing of crystallites in the bulk play important roles governing the spectral features. Compared to that of small molecules, the difference between chiral and achiral responses in SFG spectra is more difficult to confirm because of the inhomogeneous distribution of crystallites in the bulk sample.

59 BASIC BIOLOGICAL SCIENCES↗

Atomic-level structural correlations across the morphotropic phase boundary of a ferroelectric solid solution: xBiMg 1/2 Ti 1/2 O 3 -(1$-$x)PbTiO 3

Revelation of unequivocal structural information at the atomic level for complex systems is uniquely important for deeper and generic understanding of the structure property connections and a key challenge in materials science. Here in this paper we report an experimental study of the local structure by applying total elastic scattering and Raman scattering analyses to an important non-relaxor ferroelectric solid solution exhibiting the so-called composition-induced morphotropic phase boundary (MPB), where concomitant enhancement of physical properties have been detected. The powerful combination of static and dynamic structural probes enabled us to derive direct correspondence between the atomic-level structural correlations and reported properties. The atomic pair distribution functions obtained from the neutron total scattering experiments were analysed through big-box atom-modelling implementing reverse Monte Carlo method, from which distributions of magnitudes and directions of off-centred cationic displacements were extracted. We found that an enhanced randomness of the displacement-directions for all ferroelectrically active cations combined with a strong dynamical coupling between the A- and B-site cations of the perovskite structure, can explain the abrupt amplification of piezoelectric response of the system near MPB. Finally, altogether this provides a more fundamental basis in inferring structure-property connections in similar systems including important implications in designing novel and bespoke materials.

36 MATERIALS SCIENCE↗

Investigating the effect of zirconium oxide microstructure on corrosion performance: a comparison between neutron, proton, and nonirradiated oxides - 2016-0069

Scanning precession electron diffraction in the transmission electron microscope has been used to simultaneously map the phase, orientation, and grain morphology of oxides formed on Zircaloy-2 after three and six cycles in a boiling water reactor in unprecedented detail. For comparison, a region of a preoxidized autoclave-formed oxide was also proton-irradiated at the Dalton Cumbrian Facility. The proton irradiation was observed to cause additional stabilization of the tetragonal phase that was attributed to the stabilizing effect of irradiation- induced defects in the oxide. In the reactor-formed oxides, no extra stabilization of the tetragonal grains was observed under neutron irradiation, as indicated by the similar tetragonal phase fraction and transformation twin-boundary distributions between the nonirradiated and reactor-formed oxides. It is suggested that the damage rate is too low in the newly formed oxide to cause significant stabilization of the tetragonal phase. This technique also reveals that the oxide formed under reactor conditions has a more heterogeneous microstructure, and the growth of well-oriented columnar monoclinic grains is significantly reduced compared with a nonirradiated oxide. High-angle annular dark-field scanning transmission electron microscopy also revealed the development of extensive networks of intergranular porosity and eventually grain de-cohesion in the reactor-formed oxides. These results suggest that the tetragonal-monoclinic transformation is not responsible for the accelerated corrosion exhibited under reactor conditions. It is proposed that the usual out-of-reactor oxide growth and nucleation processes are significantly modified under reactor conditions, resulting in a more heterogeneous and randomly oriented oxide microstructure with reduced columnar grain growth. It is suggested that this disordered oxide microstructure allows for the formation of extensive intergranular porosity that could lead to accelerated in-reactor corrosion. (authors)

36 MATERIALS SCIENCE↗

SUPERNOVA DRIVING. I. THE ORIGIN OF MOLECULAR CLOUD TURBULENCE

Turbulence is ubiquitous in molecular clouds (MCs), but its origin is still unclear because MCs are usually assumed to live longer than the turbulence dissipation time. Interstellar medium (ISM) turbulence is likely driven by supernova (SN) explosions, but it has never been demonstrated that SN explosions can establish and maintain a turbulent cascade inside MCs consistent with the observations. In this work, we carry out a simulation of SN-driven turbulence in a volume of (250 pc){sup 3}, specifically designed to test if SN driving alone can be responsible for the observed turbulence inside MCs. We find that SN driving establishes a velocity scaling consistent with the usual scaling laws of supersonic turbulence, suggesting that previous idealized simulations of MC turbulence, driven with a random, large-scale volume force, were correctly adopted as appropriate models for MC turbulence, despite the artificial driving. We also find that the same scaling laws extend to the interiors of MCs, and that the velocity–size relation of the MCs selected from our simulation is consistent with that of MCs from the Outer-Galaxy Survey, the largest MC sample available. The mass–size relation and the mass and size probability distributions also compare successfully with those of the Outer Galaxy Survey. Finally, we show that MC turbulence is super-Alfvénic with respect to both the mean and rms magnetic-field strength. We conclude that MC structure and dynamics are the natural result of SN-driven turbulence.

79 ASTRONOMY AND ASTROPHYSICS↗

Bayesian Cross-matching of High Proper-motion Stars in Gaia DR2 and Photometric Metallicities for ∼1.7 million K and M Dwarfs

We present a Bayesian method to cross-match 5,827,988 high proper-motion Gaia sources (μ > 40 mas yr{sup −1}) to various photometric surveys: Two Micron All Sky Survey, AllWISE data release from the Wide-field Infrared Explorer (WISE) mission, Galaxy Evolution Explorer, Radial Velocity Experiment, Sloan Digital Sky Survey, and Panoramic Survey Telescope and Rapid Response System (Pan-STARRS). To efficiently associate these objects across catalogs, we develop a technique that compares the multidimensional distribution of all sources in the vicinity of each Gaia star to a reference distribution of random field stars obtained by extracting all sources in a region on the sky displaced 2′. This offset preserves the local field stellar density and magnitude distribution, allowing us to characterize the frequency of chance alignments. The resulting catalog with Bayesian probabilities >95% has a marginally higher match rate than current internal Gaia data release 2 (DR2) matches for most catalogs. However, a significant improvement is found with Pan-STARRS, where ∼99.8% of the sample within the Pan-STARRS footprint is recovered, as compared to a low ∼20.8% in Gaia DR2. Using these results, we train a Gaussian process regressor to calibrate two photometric metallicity relationships. For dwarfs of 3500 < T{sub eff} < 5280 K, we use metallicity values of 4378 stars from the Apache Point Observatory Galactic Evolution Experiment and Hejazi et al. to calibrate the relationship, producing results with a 1σ precision of 0.12 dex and few systematic errors. We then indirectly infer the metallicity of 4018 stars with 2850 < T{sub eff} < 3500 K, which are wide companions of primaries whose metallicities are estimated with our first regressor, to produce a relationship with a 1σ precision of 0.21 dex and significant systematic errors. Additional work is needed to better remove unresolved binaries from this second sample to reduce these systematic errors.

47 OTHER INSTRUMENTATION↗

Analytical techniques for the study of some parameters of multispectral scanner systems for remote sensing

The author has identified the following significant results. The concept of average mutual information in the received spectral random process about the spectral scene was developed. Techniques amenable to implementation on a digital computer were also developed to make the required average mutual information calculations. These techniques required identification of models for the spectral response process of scenes. Stochastic modeling techniques were adapted for use. These techniques were demonstrated on empirical data from wheat and vegetation scenes.

Wiswell, E. R.↗

Fractals and cosmological large-scale structure

Observations of galaxy-galaxy and cluster-cluster correlations as well as other large-scale structure can be fit with a 'limited' fractal with dimension D of about 1.2. This is not a 'pure' fractal out to the horizon: the distribution shifts from power law to random behavior at some large scale. If the observed patterns and structures are formed through an aggregation growth process, the fractal dimension D can serve as an interesting constraint on the properties of the stochastic motion responsible for limiting the fractal structure. In particular, it is found that the observed fractal should have grown from two-dimensional sheetlike objects such as pancakes, domain walls, or string wakes. This result is generic and does not depend on the details of the growth process.

Luo, Xiaochun↗

A statistical model for interpreting computerized dynamic posturography data

Computerized dynamic posturography (CDP) is widely used for assessment of altered balance control. CDP trials are quantified using the equilibrium score (ES), which ranges from zero to 100, as a decreasing function of peak sway angle. The problem of how best to model and analyze ESs from a controlled study is considered. The ES often exhibits a skewed distribution in repeated trials, which can lead to incorrect inference when applying standard regression or analysis of variance models. Furthermore, CDP trials are terminated when a patient loses balance. In these situations, the ES is not observable, but is assigned the lowest possible score--zero. As a result, the response variable has a mixed discrete-continuous distribution, further compromising inference obtained by standard statistical methods. Here, we develop alternative methodology for analyzing ESs under a stochastic model extending the ES to a continuous latent random variable that always exists, but is unobserved in the event of a fall. Loss of balance occurs conditionally, with probability depending on the realized latent ES. After fitting the model by a form of quasi-maximum-likelihood, one may perform statistical inference to assess the effects of explanatory variables. An example is provided, using data from the NIH/NIA Baltimore Longitudinal Study on Aging.

NASA Discipline Neuroscience↗

Induction of a bystander mutagenic effect of alpha particles in mammalian cells

Ever since the discovery of X-rays was made by Rontgen more than a hundred years ago, it has always been accepted that the deleterious effects of ionizing radiation such as mutation and carcinogenesis are attributable mainly to direct damage to DNA. Although evidence based on microdosimetric estimation in support of a bystander effect appears to be consistent, direct proof of such extranuclear/extracellular effects are limited. Using a precision charged particle microbeam, we show here that irradiation of 20% of randomly selected A(L) cells with 20 alpha particles each results in a mutant fraction that is 3-fold higher than expected, assuming no bystander modulation effect. Furthermore, analysis by multiplex PCR shows that the types of mutants induced are significantly different from those of spontaneous origin. Pretreatment of cells with the radical scavenger DMSO had no effect on the mutagenic incidence. In contrast, cells pretreated with a 40 microM dose of lindane, which inhibits cell-cell communication, significantly decreased the mutant yield. The doses of DMSO and lindane used in these experiments are nontoxic and nonmutagenic. We further examined the mutagenic yield when 5-10% of randomly selected cells were irradiated with 20 alpha particles each. Results showed, likewise, a higher mutant yield than expected assuming no bystander effects. Our studies provide clear evidence that irradiated cells can induce a bystander mutagenic response in neighboring cells not directly traversed by alpha particles and that cell-cell communication process play a critical role in mediating the bystander phenomenon.

Non-NASA Center↗

The Terrestrial Organism and Biogeochemistry Spatial Sampling Design for the National Ecological Observatory Network

The National Ecological Observatory Network (NEON) seeks to facilitate ecological prediction at a continental scale by measuring processes that drive change and responses at sites across the United States for thirty years. The spatial distribution of observations of terrestrial organisms and soil within NEON sites is determined according to a “design‐based” sample design that relies on the randomization of sampling locations. Development of the sample design was guided by high‐level NEON objectives and the multitude of data products that will be subjected to numerous analytical approaches to address the causes and consequences of ecological change. A requirement framework permeates the NEON design, ensuring traceability from each facet of the design to the high‐level requirements that make the NEON mission statement actionable. Requirements were developed for the terrestrial sample design to guide the key components of the design: Randomizing the sample locations ensures the unbiased collection of data, is appropriate for organisms and soil, and provides data suitable for a variety of analyses. Stratification increases efficiency and allows sampling to focus on those parts of the landscape measured by other NEON observation platforms. Attention to the sample size and spatial plot allocation ensures that data products will be sufficient to inform questions asked of the data and the NEON objectives. Establishing a framework with the capacity for re‐evaluate and design iteration allows for adaption to unexpected challenges and optimization of the sample design based on early data returns. The utility of the NEON sampling design is highlighted by its application across terrestrial systems. The data generated from this unique design will be used to quantify patterns in: the abundance and diversity of small mammals, breeding birds, insects, and soil microbes; vegetation structure, biomass, productivity, and diversity; and soil biogeochemistry.

National Ecological Observatory Network↗

Shaping the spectrum of random-phase radar waveforms

The various technologies presented herein relate to generation of a desired waveform profile in the form of a spectrum of apparently random noise (e.g., white noise or colored noise), but with precise spectral characteristics. Hence, a waveform profile that could be readily determined (e.g., by a spoofing system) is effectively obscured. Obscuration is achieved by dividing the waveform into a series of chips, each with an assigned frequency, wherein the sequence of chips are subsequently randomized. Randomization can be a function of the application of a key to the chip sequence. During processing of the echo pulse, a copy of the randomized transmitted pulse is recovered or regenerated against which the received echo is correlated. Hence, with the echo energy range-compressed in this manner, it is possible to generate a radar image with precise impulse response.

47 OTHER INSTRUMENTATION↗

Computational experiments to characterize bubble formation and movement in waste glass foam layer

Computer simulation experiments are a valuable complement to physical experimentation for systems where there is limited access to quantities of interest. The motivation for this work is to characterize the fluid dynamic and heat transfer processes that affect melt rate of waste slurry during vitrification, which can limit the throughput rate of the melter. Heat transfer to the cold cap from the molten glass below this layer is an important factor determining melt rate. In this study, computational fluid dynamics is used to elucidate the behavior of gas-filled cavities that form underneath the cold cap in a laboratory-scale waste glass melter. The cold cap consists of a reacting feed layer above a foam layer. The cavities are a result of coalesced gases from collapsing primary foam and air from forced convection bubblers. Bubbles in the foam layer move horizontally with the circulation pattern of the melt and escape around the edges of the cold cap to the plenum above. The effects of varying thermophysical properties and foam parameters based on the bounds of high-level waste glass formulations were explored. Using the DAKOTA toolkit, a Latin hypercube space-filling design was used to randomly distribute samples in bins to achieve good coverage over the parameter space with minimal points. Using the results from the set of simulations, a Gaussian process model with a linear trend was used to create response surfaces for several key quantities of interest, including temperatures, temperature gradients, velocities, and foam extent beneath the cold cap.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Phase Calibration of Microphones by Measurement in the Free-field

Over the past several years, significant effort has been expended at NASA Langley developing new Micro-Electro-Mechanical System (MEMS)-based microphone directional array instrumentation for high-frequency aeroacoustic measurements in wind tunnels. This new type of array construction solves two challenges which have limited the widespread use of large channel-count arrays, namely by providing a lower cost-per-channel and a simpler method for mounting microphones in wind tunnels and in field-deployable arrays. The current generation of array instrumentation is capable of extracting accurate noise source location and directivity on a variety of airframe components using sophisticated data reduction algorithms [1-2]. Commercially-available MEMS microphones are condenser-type devices and have some desirable characteristics when compared with conventional condenser-type microphones. The most important advantages of MEMS microphones are their size, price, and power consumption. However, the commercially-available units suffer from certain important shortcomings. Based on experiments with array prototypes, it was found that both the bandwidth and the sound pressure limit of the microphones should be increased significantly to improve the performance and flexibility of the microphone array [3]. It was also desired to modify the packaging to eliminate unwanted Helmholtz resonance s exhibited by the commercial devices. Thus, new requirements were defined as follows: Frequency response: 100 Hz to 100 KHz (+/-3dB) Upper sound pressure limit: Design 1: 130 dB SPL (THD less than 5%) Design 2: 150-160 dB SPL (THD less than 5%) Packaging: 3.73 x 6.13 x 1.3 mm can with laser-etched lid. In collaboration with Novusonic Acoustic Innovation, NASA modified a Knowles SiSonic MEMS design to meet these new requirements. Coupled with the design of the enhanced MEMS microphones was the development of a new calibration method for simultaneously obtaining the sensitivity and phase response of the devices over their entire broadband frequency range. Traditionally, electrostatic actuators (EA) have been used to characterize air-condenser microphones; however, MEMS microphones are not adaptable to the EA method due to their construction and very small diaphragm size [4]. Hence a substitution based, free-field method was developed to calibrate these microphones at frequencies up to 80 kHz. The technique relied on the use of a random, ultrasonic broadband centrifugal sound source located in a small anechoic chamber. The free-field sensitivity (voltage per unit sound pressure) was obtained using the procedure outlined in reference 4. Phase calibrations of the MEMS microphones were derived from cross spectral phase comparisons between the reference and test substitution microphones and an adjacent and invariant grazing-incidence 1/8-inch standard microphone. The free-field calibration procedure along with representative sensitivity and phase responses for the new high-frequency MEMS microphones are presented here.

Shams, Qamar A.↗

Diets high in resistant starch increase plasma levels of trimethylamine-N-oxide, a gut microbiome metabolite associated with CVD risk

Production of trimethylamine- N -oxide (TMAO), a biomarker of CVD risk, is dependent on intestinal microbiota, but little is known of dietary conditions promoting changes in gut microbial communities. Resistant starches (RS) alter the human microbiota. We sought to determine whether diets varying in RS and carbohydrate (CHO) content affect plasma TMAO levels. We also assessed postprandial glucose and insulin responses and plasma lipid changes to diets high and low in RS. In a cross-over trial, fifty-two men and women consumed a 2-week baseline diet (41 percentage of energy (%E) CHO, 40 % fat, 19 % protein), followed by 2-week high- and low-RS diets separated by 2-week washouts. RS diets were assigned at random within the context of higher (51–53 %E) v . lower CHO (39–40 %E) intake. Measurements were obtained in the fasting state and, for glucose and insulin, during a meal test matching the composition of the assigned diet. With lower CHO intake, plasma TMAO, carnitine, betaine and γ -butyrobetaine concentrations were higher after the high- v . low-RS diet ( P <0·01 each). These metabolites were not differentially affected by high v . low RS when CHO intake was high. Although the high-RS meal reduced postprandial insulin and glucose responses when CHO intake was low ( P <0·01 each), RS did not affect fasting lipids, lipoproteins, glucose or insulin irrespective of dietary CHO content. In conclusion, a lower-CHO diet high in RS was associated with higher plasma TMAO levels. These findings, together with the absence of change in fasting lipids, suggest that short-term high-RS diets do not improve markers of cardiometabolic health.

59 BASIC BIOLOGICAL SCIENCES↗