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At least 433 records · Page 24

Binding profiles for 961 Drosophila and C. elegans transcription factors reveal tissue-specific regulatory relationships

A catalog of transcription factor (TF) binding sites in the genome is critical for deciphering regulatory relationships. Here, we present the culmination of the efforts of the modENCODE (model organism Encyclopedia of DNA Elements) and modERN (model organism Encyclopedia of Regulatory Networks) consortia to systematically assay TF binding events in vivo in two major model organisms,Drosophila melanogaster(fly) andCaenorhabditis elegans(worm). These data sets comprise 605 TFs identifying 3.6 M sites in the fly and 356 TFs identifying 0.9 M sites in the worm, and represent the majority of the regulatory space in each genome. We demonstrate that TFs associate with chromatin in clusters termed “metapeaks,” that larger metapeaks have characteristics of high-occupancy target (HOT) regions, and that the importance of consensus sequence motifs bound by TFs depends on metapeak size and complexity. Combining ChIP-seq data with single-cell RNA-seq data in a machine-learning model identifies TFs with a prominent role in promoting target gene expression in specific cell types, even differentiating between parent–daughter cells during embryogenesis. These data are a rich resource for the community that should fuel and guide future investigations into TF function. To facilitate data accessibility and utility, all strains expressing green fluorescent protein (GFP)-tagged TFs are available at the stock centers for each organism. The chromatin immunoprecipitation sequencing data are available through the ENCODE Data Coordinating Center, GEO, and through a direct interface that provides rapid access to processed data sets and summary analyses, as well as widgets to probe the cell-type-specific TF–target relationships.

Biochemistry & Molecular Biology↗

Silicon photonic chip for 16-channel wavelength division (de-)multiplexing in the O-band

We experimentally demonstrate a silicon photonic chip-scale 16-channel wavelength division multiplexer (WDM) operating in the O-band. The silicon photonic chip consists of a common-input bus waveguide integrated with a sequence of 16 spectral add-drop filters implemented by 4-port contra-directional Bragg couplers and resonant cladding modulated perturbations. The combination of these features reduces the spectral bandwidth of the filters and improves the crosstalk. An apodization of the cladding modulated perturbations between the bus and the add/drop waveguides is used to optimize the strength of the coupling coefficient in the propagation direction to reduce the intra-channel crosstalk on adjacent channels. The fabricated chip was validated experimentally with a measured intra-channel crosstalk of ∼−18.9 dB for a channel spacing of 2.6 nm. The multiplexer/demultiplexer chip was also experimentally tested with a 10 Gbps data waveform. The resulting eye-pattern indicates that this approach is suitable for datacenter WDM-based interconnects in the O-band with large aggregate bandwidths.

Davis, Jordan A. (ORCID:0000000343483295)↗

Projection system for display of parallax and perspective

A projection system for the display of parallax and perspective of a real image from a hologram is presented. A reference beam was projected in a sequence of several projections at selected angles of perspective through the hologram, this sequence being rapidly performed. The resulting angular spaced images emitting from the hologram were directed onto a mirror which was coordinately tilted to reflect all of the resulting images to register onto a screen where they appeared as a single three dimensional image.

Kurtz, R. L.↗

Mechanically fastened joints in woven fabric composites

Strength analysis for composite bolted joints involves the mating of a stress analysis with an appropriate mode specific failure criterion for each of the primary failure modes. The stress analysis and failure criteria are independent of each other and can be manipulated separately in order to optimize the strength analysis package formed by their coupling. Material properties tests were conducted on rubber toughened graphite-epoxy material to measure the basic strength and stiffness in the warp and fill directions and in shear. Test matrices are summarized for investigations of laminate configuration, stacking sequence, fastener diameter, edge distance, fastener half spacing, laminate thickness, and fastener torque. A three dimensional finite element analysis computer program was written and failure criteria for net tension, shearout, and bearing were determined.

Wilson, D. W.↗

Extreme Be stars in the Magellanic Clouds

In the course of a spectroscopic investigation of the O-type stars in the Magellanic Clouds, a population of luminous Be stars was serendipitously encountered. They are here referred to as extreme Be stars, following the terminology used by Schild (1966) for a population of Be stars in the region of h and Chi Persei which lie significantly above the main sequence. As members of the Clouds, their luminosities and intrinsic colors can be determined directly. A comparison of these new high-luminosity Be stars with known H-beta-emission-line supergiants in the Clouds is made in both the optical and UV spectral regions. The Galactic counterparts of these Be stars are also discussed. Although these Be stars are an average of several magnitudes brighter than those in the Galaxy, this may in part be a selection effect.

Garmany, C. D.↗

Architectures and algorithms for digital image processing; Proceedings of the Meeting, Cannes, France, December 5, 6, 1985

The conference presents papers on the architectures, algorithms, and applications of image processing. Particular attention is given to a very large scale integration system for image reconstruction from projections, a prebuffer algorithm for instant display of volume data, and an adaptive image sequence filtering scheme based on motion detection. Papers are also presented on a simple, direct practical method of sensing local motion and analyzing local optical flow, image matching techniques, and an automated biological dosimetry system.

Duff, Michael J. B.↗

Advanced Stirling Convertor Durability Testing: Plans and Interim Results

The U.S. Department of Energy (DOE), Lockheed Martin Corporation (LM), and NASA Glenn Research Center (GRC) have been developing the Advanced Stirling Radioisotope Generator (ASRG) for use as a power system for space science missions. In support of this program, GRC has been involved in testing Stirling convertors, including the Advanced Stirling Convertor (ASC), for use in the ASRG. This testing includes electromagnetic interference/compatibility (EMI/EMC), structural dynamics, advanced materials, organics, and unattended extended operation. The purpose of the durability tests is to experimentally demonstrate the margins in the ASC design. Due to the high value of the hardware, previous ASC tests focused on establishing baseline performance of the convertors within the nominal operating conditions. The durability tests present the first planned extension of the operating conditions into regions beyond those intended to meet the product spec, where the possibility exists of lateral contact, overstroke, or over-temperature events. These tests are not intended to cause damage that would shorten the life of the convertors, so they can transition into extended operation at the conclusion of the tests. This paper describes the four tests included in the durability test sequence: 1) start/stop cycling, 2) exposure to constant acceleration in the lateral and axial directions, 3) random vibration at increased piston amplitude to induce contact events, and 4) overstroke testing to simulate potential failures during processing or during the mission life where contact events could occur. The paper also summarizes the analysis and simulation used to predict the results of each of these tests.

Meer, David W.↗

Advanced Stirling Convertor Durability Testing: Plans and Interim Results

The U.S. Department of Energy (DOE), Lockheed Martin Corporation (LM), and NASA Glenn Research Center (GRC) have been developing the Advanced Stirling Radioisotope Generator (ASRG) for use as a power system for space science missions. In support of this program, NASA?s Glenn Research Center (GRC) has been involved in testing Stirling convertors, including the Advanced Stirling Convertor (ASC), for use in the ASRG. This testing includes electromagnetic interference/compatibility (EMI/EMC), structural dynamics, advanced materials, organics, and unattended extended operation. The purpose of the durability tests is to experimentally demonstrate the margins in the ASC design. Due to the high value of the hardware, previous ASC tests focused on establishing baseline performance of the convertors within the nominal operating conditions. The durability tests present the first planned extension of the operating conditions into regions beyond those intended to meet the product spec, where the possibility exists of lateral contact, overstroke, or over-temperature events. These tests are not intended to cause damage that would shorten the life of the convertors, so they can transition into extended operation at the conclusion of the tests. This paper describes the four tests included in the durability test sequence: 1) start/stop cycling, 2) exposure to constant acceleration in the lateral and axial directions, 3) random vibration at increased piston amplitude to induce contact events, and 4) overstroke testing to simulate potential failures during processing or during the mission life where contact events could occur. The paper also summarizes the analysis and simulation used to predict the results of each of these tests.

Meer, Dave↗

Field Exploration and Life Detection Sampling Through Planetary Analogue Sampling (FELDSPAR).

Exploration missions to Mars rely on rovers to perform analyses over small sampling areas; however, landing sites for these missions are selected based on large-scale, low-resolution remote data. The use of Earth analogue environments to estimate the multi-scale spatial distributions of key signatures of habitability can help ensure mission science goals are met. A main goal of FELDSPAR is to conduct field operations analogous to Mars sample return in its science, operations, and technology from landing site selection, to in-field sampling location selection, remote or stand-off analysis, in situ analysis, and home laboratory analysis. Lava fields and volcanic regions are relevant analogues to Martian landscapes due to desiccation, low nutrient availability, and temperature extremes. Operationally, many Icelandic lava fields are remote enough to require that field expeditions address several sampling constraints that are experienced in robotic exploration, including in situ and sample return missions. The Fimmvruhls lava field was formed by a basaltic effusive eruption associated with the 2010 Eyjafjallajkull eruption. Mlifellssandur is a recently deglaciated plain to the north of the Myrdalsjkull glacier. Holuhraun was formed by a 2014 fissure eruptions just north of the large Vatnajkull glacier. Dyngjusandur is an alluvial plain apparently kept barren by repeated mechanical weathering. Informed by our 2013 expedition, we collected samples in nested triangular grids every decade from the 10 cm scale to the 1 km scale (as permitted by the size of the site). Satellite imagery is available for older sites, and for Mlifellssandur, Holuhraun, and Dyngjusandur we obtained overhead imagery at 1 m to 200 m elevation. PanCam-style photographs were taken in the field by sampling personnel. In-field reflectance spectroscopy was also obtained with an ASD spectrometer in Dyngjusandur. All sites chosen were 'homogeneous' in apparent color, morphology, moisture, grain size, and reflectance spectra at all scales greater than 10 cm. Field lab assays were conducted to monitor microbial habitation, including ATP quantification, qPCR for fungal, bacterial, and archaeal DNA, and direct cell imaging using fluorescence microscopy. Home laboratory analyses include Raman spectroscopy and community sequencing. ATP appeared to be significantly more sensitive to small changes in sampling location than qPCR or fluorescence microscopy. Bacterial and archaeal DNA content were more consistent at the smaller scales, but similarly variable across more distant sites. Conversely, cell counts and fungal DNA content have significant local variation but appear relatively homogeneous over scales of 1 km. ATP, bacterial DNA, and archaeal DNA content were relatively well correlated at many spatial scales. While we have observed spatial variation at various scales and are beginning to observe how that variation fluctuates over time as biodiversity recovers after an eruption, we do not yet fully understand what parameters lead to the observed spatial variation. Home laboratory analyses will help us further understand the elemental and structural composition of the basaltic matrices, but further field analyses are vital for the understanding how temperature, moisture, incident radiation, and so forth influence the habitability of a microclimate.

Field↗

Identification of the prooxidant site of human ceruloplasmin: a model for oxidative damage by copper bound to protein surfaces

Free transition metal ions oxidize lipids and lipoproteins in vitro; however, recent evidence suggests that free metal ion-independent mechanisms are more likely in vivo. We have shown previously that human ceruloplasmin (Cp), a serum protein containing seven Cu atoms, induces low density lipoprotein oxidation in vitro and that the activity depends on the presence of a single, chelatable Cu atom. We here use biochemical and molecular approaches to determine the site responsible for Cp prooxidant activity. Experiments with the His-specific reagent diethylpyrocarbonate (DEPC) showed that one or more His residues was specifically required. Quantitative [14C]DEPC binding studies indicated the importance of a single His residue because only one was exposed upon removal of the prooxidant Cu. Plasmin digestion of [14C]DEPC-treated Cp (and N-terminal sequence analysis of the fragments) showed that the critical His was in a 17-kDa region containing four His residues in the second major sequence homology domain of Cp. A full length human Cp cDNA was modified by site-directed mutagenesis to give His-to-Ala substitutions at each of the four positions and was transfected into COS-7 cells, and low density lipoprotein oxidation was measured. The prooxidant site was localized to a region containing His426 because CpH426A almost completely lacked prooxidant activity whereas the other mutants expressed normal activity. These observations support the hypothesis that Cu bound at specific sites on protein surfaces can cause oxidative damage to macromolecules in their environment. Cp may serve as a model protein for understanding mechanisms of oxidant damage by copper-containing (or -binding) proteins such as Cu, Zn superoxide dismutase, and amyloid precursor protein.

NASA Discipline Regulatory Physiology↗

A genomic perspective on fungal diversity and evolution

Originating from aquatic unicellular ancestors, over the course of ~1 billion years, the fungi have evolved to occupy nearly all aerobic environments on the planet, diversified into millions of different ‘species’ and have developed complex multicellular structures. Their relatively small, simple genomes have facilitated massive-scale sequencing and allowed us to explore genome evolution across an ancient eukaryotic kingdom. With thousands of genomes from diverse lineages now available, this Review will discuss insights into fungal biology and evolution gleaned with genomics and other multi-omics approaches. Using published genomes available through GenBank and the Joint Genome Institute’s MycoCosm platform, we generated kingdom-wide phylogenies and used them to highlight how fungal genomes have changed over time. With this phylogeny as a guide, we also discuss major evolutionary transitions that occurred across the fungal kingdom. Although progress has been made, these efforts are hampered by biases in genome representation and limited characterization of gene functions. Here, in this study, we discuss these challenges and possible future directions to address them, including initiatives to characterize conserved genes of unknown function and scale up sequencing towards 10,000 annotated fungal genomes.

Mondo, Stephen J. [USDOE Joint Genome Institute (J↗

Nanopores and nucleic acids: prospects for ultrarapid sequencing

DNA and RNA molecules can be detected as they are driven through a nanopore by an applied electric field at rates ranging from several hundred microseconds to a few milliseconds per molecule. The nanopore can rapidly discriminate between pyrimidine and purine segments along a single-stranded nucleic acid molecule. Nanopore detection and characterization of single molecules represents a new method for directly reading information encoded in linear polymers. If single-nucleotide resolution can be achieved, it is possible that nucleic acid sequences can be determined at rates exceeding a thousand bases per second.

Review↗

Dissecting the dominant hot spring microbial populations based on community-wide sampling at single-cell genomic resolution

With advances in DNA sequencing and miniaturized molecular biology workflows, rapid and affordable sequencing of single-cell genomes has become a reality. Compared to 16S rRNA gene surveys and shotgun metagenomics, large-scale application of single-cell genomics to whole microbial communities provides an integrated snapshot of community composition and function, directly links mobile elements to their hosts, and enables analysis of population heterogeneity of the dominant community members. To that end, we sequenced nearly 500 single-cell genomes from a low diversity hot spring sediment sample from Dewar Creek, British Columbia, and compared this approach to 16S rRNA gene amplicon and shotgun metagenomics applied to the same sample. We found that the broad taxonomic profiles were similar across the three sequencing approaches, though several lineages were missing from the 16S rRNA gene amplicon dataset, likely the result of primer mismatches. At the functional level, we detected a large array of mobile genetic elements present in the single-cell genomes but absent from the corresponding same species metagenome-assembled genomes. Moreover, we performed a single-cell population genomic analysis of the three most abundant community members, revealing differences in population structure based on mutation and recombination profiles. While the average pairwise nucleotide identities were similar across the dominant species-level lineages, we observed differences in the extent of recombination between these dominant populations. Most intriguingly, the creek's Hydrogenobacter sp. population appeared to be so recombinogenic that it more closely resembled a sexual species than a clonally evolving microbe. Together, this work demonstrates that a randomized single-cell approach can be useful for the exploration of previously uncultivated microbes from community composition to population structure.

59 BASIC BIOLOGICAL SCIENCES↗

An Archaea 5S rRNA analog is stably expressed in Escherichia coli

Mini-genes for 5S-like rRNA were constructed. These genes had a sequence which largely resembles that of the naturally occurring 5S rRNA of a bacterium, Halococcus morrhuae, which phylogenetically belongs to the Archaea. Plasmids carrying the mini-genes were transformed into Escherichia coli (Ec). Ribosomal incorporation was not a prerequisite for stable accumulation of the RNA product. However, only those constructs with a well-base-paired helix I accumulated RNA product. This result strongly implies that this aspect of the structure is likely to be an important condition for stabilizing 5S rRNA-like products. The results are consistent with our current understanding of 5S rRNA processing in Ec. When used in conjunction with rRNA probe technology, the resulting chimeric RNA may be useful as a monitoring tool for genetically engineered microorganisms or naturally occurring organisms that are released into the environment.

Non-NASA Center↗

Whole Genome Sequencing Links Mycobacterium bovis From Cattle, Cheese and Humans in Baja California, Mexico

Mycobacterium bovis causes tuberculosis (TB) in cattle, which in turn can transmit the pathogen to humans. Tuberculosis in dairy cattle is of particular concern where the consumption of raw milk and dairy products is customary. Baja California (BCA), Mexico, presents high prevalence of TB in both cattle and humans, making it important to investigate the molecular epidemiology of the disease in the region. A long-term study was undertaken to fully characterize the diversity of M. bovis genotypes circulating in dairy cattle, cheese and humans in BCA by whole-genome sequencing (WGS). During a 2-year period, 412 granulomatous tissue samples were collected from local abattoirs and 314 cheese samples were purchased from local stores and vendors in BCA and sent to the laboratory for mycobacterial culture, histology, direct PCR and WGS. For tissue samples M. bovis was recovered from 86.8%, direct PCR detected 90% and histology confirmed 85.9% as mycobacteriosis-compatible. For cheese, M. bovis was recovered from 2.5% and direct PCR detected 6% of the samples. There was good agreement between diagnostic tests. Subsequently, a total of 345 whole-genome SNP sequences were obtained. Phylogenetic analysis grouped these isolates into 10 major clades. SNP analysis revealed putative transmission clusters where the pairwise SNP distance between isolates from different dairies was ≤3 SNP. Also, human and/or cheese isolates were within 8.45 (range 0–17) and 5.8 SNP (range 0–15), respectively, from cattle isolates. Finally, a comparison between the genotypes obtained in this study and those reported previously suggests that the genetic diversity of M. bovis in BCA is well-characterized, and can be used to determine if BCA is the likely source of M. bovis in humans and cattle in routine epidemiologic investigations and future studies. In conclusion, WGS provided evidence of ongoing local transmission of M. bovis among the dairies in this high-TB burden region of BCA, as well as show close relationships between isolates recovered from humans, cheese, and cattle. This confirms the need for a coordinated One Health approach in addressing the elimination of TB in animals and humans. Overall, the study contributes to the knowledge of the molecular epidemiology of M. bovis in BCA, providing insight into the pathogen's dynamics in a high prevalence setting.

59 BASIC BIOLOGICAL SCIENCES↗

Hydrodynamics of binary coalescence. 2: Polytropes with gamma = 5/3

We present a new numerical study of the equilibrium and stability properties of close binary systems. We use the smoothed-particle hydrodynamics (SPH) technique both to construct accurate equilibrium configurations in three dimensions and to follow their hydrodynamic evolution. We adopt a simple polytropic equation of state p = K(sub rho)(exp gamma) with gamma = 5/3 and K = constant within each star, applicable to low-mass degenerate dwarfs as well as low-mass main-sequence stars. For degenerate configurations, we set the two polytropic constants equal, K = K prime, independent of the mass ratio. For main-sequence stars, we adjust K and K prime so as to obtain a simple mass-radius relation of the form R/R prime = M/M prime, where R prime and M prime are the radius and mass of the secondary. Along a sequence of binary equilibrium configurations for two identical stars, we demonstrate the existence of both secular and dynamical instabilities, confirming directly the results of recent analytic work. We use the SPH method to calculate the nonlinear development of the dynamical instability and to determine the final fate of the system. We find that the two stars merge together into a single, rapidly rotating object in just a few orbital periods. Equilibrium sequences are also constructed for systems containing two nonidentical stars. These sequences terminate at a Roche limit, which we can determine very accurately using SPH. For two low-mass main-sequence stars with mass ratio q approximately less than 0.4 we find that the (synchronized) Roche limit configuration is secularly unstable. We discuss the implications of our results for the evolution of double white-dwarf systems and W Ursae Majoris binaries.

Rasio, Frederic A.↗

ASCA Observations of the T Tauri Star SU Aurigae and the Surrounding L1517 Dark Cloud

We present the results of a approximately equals 40 ks pointed Advanced Satellite for Cosmology and Astrophysics (ASCA) observation of the L1517 star-forming region, centered on the X-ray-bright T Tauri star SU Aurigae. This star has the highest X-ray luminosity of any classical T Tauri star in the Taurus-Auriga region, and its optical spectra show evidence for both mass inflow and outflow. Strong X-ray emission was detected from SU Aur (L(sub x) = 10(exp 30.9) ergs s(exp -1)) as well as weaker emission from five other pre-main-sequence stars. Although no large-amplitude flares were detected, the X-ray emission of SU Aur showed clear variability in the form of a slow decline in count rate during the 1.3 day observation. We provide the first direct comparison of the coronal differential emission measure (DEM) distribution of a classical T Tauri star with that of a young main-sequence star of similar spectral type. The DEM distributions of SU Aur (G2; age 3 Myr) and the young solar-like star EK Draconis (GO V; age 70 Myr) are qualitatively similar, with both showing a bimodal temperature distribution characterized by a cool plasma component peaking at approximately 8-9 MK and a hot component peaking at approximately 20-21 MK. However, there is a striking difference in the relative proportion of plasma at high temperatures in the two stars, with hot plasma (>20 MK) accounting for approximately equals 80% of the volume emission measure of SU Aur, compared to only approximately equals 40% in EK Dra. These results provide new insight into the changes that will occur in the corona of a T Tauri star as it descends onto the main sequence. A sharp decline in the fraction of coronal plasma at flarelike temperatures will occur during the late-T Tauri and post-T Tauri phases, and other recent X-ray studies have shown that this decline will continue after the young solar-like star reaches the main sequence.

Skinner, Stephen L.↗

ForceGen: End-to-end de novo protein generation based on nonlinear mechanical unfolding responses using a language diffusion model

Through evolution, nature has presented a set of remarkable protein materials, including elastins, silks, keratins and collagens with superior mechanical performances that play crucial roles in mechanobiology. However, going beyond natural designs to discover proteins that meet specified mechanical properties remains challenging. Here, we report a generative model that predicts protein designs to meet complex nonlinear mechanical property-design objectives. Our model leverages deep knowledge on protein sequences from a pretrained protein language model and maps mechanical unfolding responses to create proteins. Via full-atom molecular simulations for direct validation, we demonstrate that the designed proteins are de novo, and fulfill the targeted mechanical properties, including unfolding energy and mechanical strength, as well as the detailed unfolding force-separation curves. Our model offers rapid pathways to explore the enormous mechanobiological protein sequence space unconstrained by biological synthesis, using mechanical features as the target to enable the discovery of protein materials with superior mechanical properties.

59 BASIC BIOLOGICAL SCIENCES↗