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At least 613 records · Page 34

Elemental profiling and genome-wide association studies reveal genomic variants modulating ionomic composition in Populus trichocarpa leaves

The ionome represents elemental composition in plant tissues and can be an indicator of nutrient status as well as overall plant performance. Thus, identifying genetic determinants governing elemental uptake and storage is an important goal for breeding and engineering biomass feedstocks with improved performance. In this study, we coupled high-throughput ionome characterization of leaf tissues with high-resolution genome-wide association studies (GWAS) to uncover genetic loci that modulate ionomic composition in leaves of poplar ( Populus trichocarpa ). Significant agreement was observed across the three ionomic profiling platforms tested: inductively coupled plasma-mass spectrometry (ICP-MS), neutron activation analysis (NAA) and laser-induced breakdown spectroscopy (LIBS). Relative quantification of 20 elements using ICP-MS across a population of 584 genotypes, revealed larger variation in micro-nutrients and trace elements content than for macro-nutrients across genotypes. The GWAS performed using a set of high-density (>8.2 million) single nucleotide polymorphisms, identified over 600 loci significantly associated with variations in these mineral elements, pointing to numerous uncharacterized candidate genes. A significant enrichment for genes related to ion homeostasis and transport was observed, including several members of the cation-proton antiporters (CPA) family and MATE efflux transporters, previously reported to be critical for plant growth and fitness in other species. Our results also included a polymorphic copy of the high-affinity molybdenum transporter MOT1 found directly associated to molybdenum content. For the first time in a perennial plant, our results provide evidence of genetic control of mineral content in a model tree species.

59 BASIC BIOLOGICAL SCIENCES↗

Dynamics of cell proliferation in the adult dentate gyrus of two inbred strains of mice

The output potential of proliferating populations in either the developing or the adult nervous system is critically dependent on the length of the cell cycle (T(c)) and the size of the proliferating population. We developed a new approach for analyzing the cell cycle, the 'Saturate and Survive Method' (SSM), that also reveals the dynamic behaviors in the proliferative population and estimates of the size of the proliferating population. We used this method to analyze the proliferating population of the adult dentate gyrus in 60 day old mice of two inbred strains, C57BL/6J and BALB/cByJ. The results show that the number of cells labeled by exposure to BUdR changes dramatically with time as a function of the number of proliferating cells in the population, the length of the S-phase, cell division, the length of the cell cycle, dilution of the S-phase label, and cell death. The major difference between C57BL/6J and BALB/cByJ mice is the size of the proliferating population, which differs by a factor of two; the lengths of the cell cycle and the S-phase and the probability that a newly produced cell will die within the first 10 days do not differ in these two strains. This indicates that genetic regulation of the size of the proliferating population is independent of the genetic regulation of cell death among those newly produced cells. The dynamic changes in the number of labeled cells as revealed by the SSM protocol also indicate that neither single nor repeated daily injections of BUdR accurately measure 'proliferation.'.

Non-NASA Center↗

Excited quantum anomalous and spin Hall effect: dissociation of flat-bands-enabled excitonic insulator state

Quantum anomalous Hall effect (QAHE) and quantum spin Hall effect (QSHE) are two interesting physical manifestations of 2D materials that have an intrinsic nontrivial band topology. In principle, they are ground-state equilibrium properties characterized by Fermi level lying in a topological gap, below which all the occupied bands are summed to a non-zero topological invariant. In this work, we propose theoretical concepts and models of ‘excited’ QAHE (EQAHE) and EQSHE generated by dissociation of an excitonic insulator (EI) state with complete population inversion (CPI), a unique many-body ground state enabled by two yin-yang flat bands (FBs) of opposite chirality hosted in a diatomic Kagome lattice. The two FBs have a trivial gap in between, i.e. the system is a trivial insulator in the single-particle ground-state, but nontrivial gaps above and below, so that upon photoexcitation the quasi-Fermi levels of both electrons and holes will lie in a nontrivial gap achieved by the CPI-EI state, as demonstrated by exact diagonalization calculations. Then dissociation of singlet and triplet EI state will lead to EQAHE and EQSHE, respectively. Realizations of yin-yang FBs in real materials are also discussed.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

Quantitation of amiodarone and N-desethylamiodarone in single HepG2 cells by single-cell printing-liquid vortex capture-mass spectrometry

Quantitative measure of a drug and its associated metabolite(s) with single-cell resolution is often limited by sampling throughput or other compromises that limit broad use. Here, we demonstrate the use of single-cell printing-liquid vortex capture-mass spectrometry (SCP-LVC-MS) to quantitatively measure the intracellular concentrations of amiodarone (AMIO) and its metabolite, N-desethylamiodarone (NDEA), from thousands of single cells across several AMIO incubation concentrations ranging from 0 to 10 μM. Concentrations obtained by SCP-LVC-MS were validated through comparison with average assays and traditional measurement of cells in bulk. Average of SCP-LVC-MS measurements and aggregate vial collection assay the concentrations differed by < 5%. Both AMIO and NDEA had clear log-normal distributions with similar standard deviation of concentrations in the cell population. The mean of both AMIO and NDEA intracellular concentrations were positively correlated with AMIO incubation concentration, increasing from 0.026 to 0.520 and 0.0055 to 0.048 mM for AMIO and NDEA, respectively. The standard deviation of AMIO and NDEA log-normal distribution fits were relatively similar in value across incubation concentrations, 0.15–0.19 log 10 (mM), and exhibited a linear trend with respect to each other. The single cell-resolved conversion ratio of AMIO to NDEA increased with decreasing incubation concentration, 7 ± 2%, 18 ± 3%, and 20 ± 7% for 10.0, 1.0, and 0.1 μM AMIO incubation concentrations, respectively. Finally, association with simultaneously measured lipids had several ions with statistically significant difference in intensity but no clear correlations with AMIO intracellular content was observed.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Structure-Based Design and Antigenic Validation of Respiratory Syncytial Virus G Immunogens

Respiratory syncytial virus (RSV) is a leading cause of severe lower respiratory tract disease of children, the elderly, and immunocompromised individuals. Currently, there are no FDA-approved RSV vaccines. The RSV G glycoprotein is used for viral attachment to host cells and impairment of host immunity by interacting with the human chemokine receptor CX3CR1. Antibodies that disrupt this interaction are protective against infection and disease. Nevertheless, development of an RSV G vaccine antigen has been hindered by its low immunogenicity and safety concerns. A previous study described three engineered RSV G proteins containing single-point mutations that induce higher levels of IgG antibodies and have improved safety profiles compared to wild-type RSV G. However, it is unclear if the mutations affect RSV G protein folding and display of its conformational epitopes. In this study, we show that the RSV G S177Q protein retains high-affinity binding to protective human and mouse monoclonal antibodies and has equal reactivity as wild-type RSV G protein to human reference immunoglobulin to RSV. Additionally, we determined the high-resolution crystal structure of RSV G S177Q protein in complex with the anti-RSV G antibody 3G12, further validating its antigenic structure. These studies show for the first time that an engineered RSV G protein with increased immunogenicity and safety retains conformational epitopes to high-affinity protective antibodies, supporting its further development as an RSV vaccine immunogen. Respiratory syncytial virus (RSV) causes severe lower respiratory diseases of children, the elderly, and immunocompromised populations. There currently are no FDA-approved RSV vaccines. Most vaccine development efforts have focused on the RSV F protein, and the field has generally overlooked the receptor-binding antigen RSV G due to its poor immunogenicity and safety concerns. However, single-point mutant RSV G proteins have been previously identified that have increased immunogenicity and safety. In this study, we investigate the antibody reactivities of three known RSV G mutant proteins. We show that one mutant RSV G protein retains high-affinity binding to protective monoclonal antibodies, is equally recognized by anti-RSV antibodies in human sera, and forms the same three-dimensional structure as the wild-type RSV G protein. Our study validates the structure-guided design of the RSV G protein as an RSV vaccine antigen.

59 BASIC BIOLOGICAL SCIENCES↗

Influenza A viral burst size from thousands of infected single cells using droplet quantitative PCR (dqPCR)

An important aspect of how viruses spread and infect is the viral burst size, or the number of new viruses produced by each infected cell. Surprisingly, this value remains poorly characterized for influenza A virus (IAV), commonly known as the flu. In this study, we screened tens of thousands of cells using a microfluidic method called droplet quantitative PCR (dqPCR). The high-throughput capability of dqPCR enabled the measurement of a large population of infected cells producing progeny virus. By measuring the fully assembled and successfully released viruses from these infected cells, we discover that the viral burst sizes for both the seasonal H3N2 and the 2009 pandemic H1N1 strains vary significantly, with H3N2 ranging from 10 1 to 10 4 viruses per cell, and H1N1 ranging from 10 1 to 10 3 viruses per cell. Some infected cells produce average numbers of new viruses, while others generate extensive number of viruses. In fact, we find that only 10% of the single-cell infections are responsible for creating a significant portion of all the viruses. This small fraction produced approximately 60% of new viruses for H3N2 and 40% for H1N1. On average, each infected cell of the H3N2 flu strain produced 709 new viruses, whereas for H1N1, each infected cell produced 358 viruses. This novel method reveals insights into the flu virus and can lead to improved strategies for managing and preventing the spread of viruses.

60 APPLIED LIFE SCIENCES↗

Application of prophage sequence analysis to investigate a disease outbreak involving Salmonella Adjame, a rare serovar and implications for the population structure

Introduction Outbreak investigation of foodborne salmonellosis is hindered when the food source is contaminated by multiple strains of Salmonella , creating difficulties matching an incriminated organism recovered from patients with the specific strain in the suspect food. An outbreak of the rare Salmonella Adjame was caused by multiple strains of the organism as revealed by single-nucleotide polymorphism (SNP) variation. The use of highly discriminatory prophage analysis to characterize strains of Salmonella should enable a more precise strain characterization and aid the investigation of foodborne salmonellosis. Methods We have carried out genomic analysis of S. Adjame strains recovered during the course of a recent outbreak and compared them with other strains of the organism ( n = 38 strains), using SNPs to evaluate strain differences present in the core genome, and prophage sequence typing (PST) to evaluate the accessory genome. Phylogenetic analyses were performed using both total prophage content and conserved prophages. Results The PST analysis of the S. Adjame isolates showed a high degree of strain heterogeneity. We observed small clusters made up of 2-6 isolates ( n = 27) and singletons ( n = 11) in stark contrast with the three clusters observed by SNP analysis. In total, we detected 24 prophages of which only four were highly prevalent, namely: Entero_p88 (36/38 strains), Salmon_SEN34 (35/38 strains), Burkho_phiE255 (33/38 strains) and Edward_GF (28/38 strains). Despite the marked strain diversity seen with prophage analysis, the distribution of the four most common prophages matched the clustering observed using core genome. Discussion Mutations in the core and accessory genomes of S. Adjame have shed light on the evolutionary relationships among the Adjame strains and demonstrated a convergence of the variations observed in both fractions of the genome. We conclude that core and accessory genomes analyses should be adopted in foodborne bacteria outbreak investigations to provide a more accurate strain description and facilitate reliable matching of isolates from patients and incriminated food sources. The outcomes should translate to a better understanding of the microbial population structure and an 46 improved source attribution in foodborne illnesses.

Gao, Ruimin↗

Experimental evidence for recovery of mercury-contaminated fish populations

Anthropogenic releases of mercury (Hg) are a human health issue because the potent toxicant methylmercury (MeHg), formed primarily by microbial methylation of inorganic Hg in aquatic ecosystems, bioaccumulates to high concentrations in fish consumed by humans. Predicting the efficacy of Hg pollution controls on fish MeHg concentrations is complex because many factors influence the production and bioaccumulation of MeHg. Here we conducted a 15-year whole-ecosystem, single-factor experiment to determine the magnitude and timing of reductions in fish MeHg concentrations following reductions in Hg additions to a boreal lake and its watershed. During the seven-year addition phase, we applied enriched Hg isotopes to increase local Hg wet deposition rates fivefold. The Hg isotopes became increasingly incorporated into the food web as MeHg, predominantly from additions to the lake because most of those in the watershed remained there. Thereafter, isotopic additions were stopped, resulting in an approximately 100% reduction in Hg loading to the lake. The concentration of labelled MeHg quickly decreased by up to 91% in lower trophic level organisms, initiating rapid decreases of 38–76% of MeHg concentration in large-bodied fish populations in eight years. Although Hg loading from watersheds may not decline in step with lowering deposition rates, this experiment clearly demonstrates that any reduction in Hg loadings to lakes, whether from direct deposition or runoff, will have immediate benefits to fish consumers.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Observation of Long-Lived UV-Induced Fluorescence from Environmental Materials Using the HVeV Detector as Developed for SuperCDMS

Here, we describe recent experiments using a SuperCDMS HVeV single-charge sensitive detector illuminated with an ultraviolet LED (275 nm) and a monochromatic laser(650 nm) using a dual fiber optic system installed in a small dilution refrigerator at Stanford University. We observed a population of fluorescence background events after UV exposure but not after exposure to the laser source. The fluorescence was likely due to scattered UV photons absorbed outside the detector. We discuss the possibility of fluorescence being a contributor to the low energy excess background observed in above-ground Dark Matter experiments.

36 MATERIALS SCIENCE↗

Single-cell measurement of plasmid copy number and promoter activity

Abstract Accurate measurements of promoter activities are crucial for predictably building genetic systems. Here we report a method to simultaneously count plasmid DNA, RNA transcripts, and protein expression in single living bacteria. From these data, the activity of a promoter in units of RNAP/s can be inferred. This work facilitates the reporting of promoters in absolute units, the variability in their activity across a population, and their quantitative toll on cellular resources, all of which provide critical insights for cellular engineering.

59 BASIC BIOLOGICAL SCIENCES↗

Jet quenching in the glasma phase: Medium-induced radiation

Inspired by the recent considerations of parton momentum broadening in the glasma phase, we study the medium-induced soft gluon radiation of jet partons at early times in heavy-ion collisions. The glasma state is assumed to be comprised of independent color domains with homogenous longitudinal fields that vary event by event, and we further complete this model with an event-averaging procedure accounting for the finite correlation length. Using this description, we evaluate the rate of medium-induced radiation from an energetic parton at midrapidity in the glasma phase. We mainly focus on SU(2) color fields for simplicity, also referring to the U(1) case and comparing with the Baier, Dokshitzer, Mueller, Peigné, Schiff, and Zakharov rate to gain further insight. Our results show that there is an intricate interplay of the synchrotron-like radiation in a single color domain with the destructive interference between different color domains, after the medium averaging is performed. Thus, we find that the emission rate is sensitive to the matter structure, decreasing for a glasma state populated by smaller color domains—i.e., for a glasma with a larger characteristic saturation scale. Our approach can be applied to more realistic backgrounds, and it sets the stage for the modeling of jet evolution in the early stages of heavy-ion collisions.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

Cosmological Simulations of Stellar Halos with Gaia Sausage–Enceladus Analogs: Two Sausages, One Bun?

Observations of the Milky Way’s stellar halo find that it is predominantly comprised of a radially biased population of stars, dubbed the Gaia Sausage–Enceladus, or GSE. These stars are thought to be debris from dwarf galaxy accretion early in the Milky Way’s history. Though typically considered to be from a single merger, it is possible that the GSE debris has multiple sources. To investigate this possibility, we use the IllustrisTNG50 simulation to identify stellar accretion histories in 98 Milky Way analogs—the largest sample for which such an identification has been performed—and find GSE-like debris in 32, with two-merger GSEs accounting for a third of these cases. Distinguishing single-merger GSEs from two-merger GSEs is difficult in common kinematic spaces, but differences are more evident through chemical abundances and star formation histories. This is because single-merger GSEs are typically accreted more recently than the galaxies in two-merger GSEs: the median infall times (with 16th and 84th percentiles) are $5.{9}_{-2.0}^{+3.3}$ and $10.{7}_{-3.7}^{+1.2}$ Gyr ago for single- and two-merger scenarios, respectively. The systematic shifts in abundances and ages that occur as a result suggest that efforts in modeling these aspects of the stellar halo prove ever-important in understanding its assembly.

galactic archaeology↗

Suprathermal electrons produced by beam-plasma-discharge

Experiments conducted with a low energy plasma lens, HARP, in the electron beam of the large vacuum chamber at Johnson Space Center indicate that an enhanced population of 50 to 300 volt electrons appear when the beam goes into the Beam-Plasma Discharge (BPD) mode. Below the BPD instability the electron distribution appears to be characterized as non-energized single particle scattering and energy loss. At 100 cm from the beam core in the BPD mode the fluxes parallel to the beam are reduced by a factor of 20 with respect to the fluxes at 25 cm. Some evidence for isotropy near the beam core is presented.

Sharp, W. E.↗

Role of zircon in tracing crustal growth and recycling

Single crystal ion probe ages of zircons is discussed, which allow much better time resolution compared to other geochronological methods, although the technique is not without problems. Rocks from two areas that contain composite zircon populations, including true magmatic zircons as well as a variety of xenocrystic types are described. It is often difficult to distinguish these; xenocrystic zircons, for example, cannot always be identified on the basis of morphology alone. Additional evidence is needed before making age interpretations. Evidence is also presented of zircon growth long after the original time of crystallization, in some cases apparently at temperatures less than 300 C. The spectacular discovery of 4.1 to 4.2 Ga detrital zircons in metaquartzites from the Mount Narryer area of Western Australia is described. Similar zircons with ages as old as 4276 Ma have been found in the nearby Jack Hills area. The source areas or parent lithologies of these zircons have not yet been determined, but the author expects that they may be unrecognized or buried antecedents of the K rich Narryer gneisses. U or Th concentrations of zircon cannot be used to discriminate between felsic and mafic source rocks.

Compston, W.↗

Influence of multiple ion species on low-frequency electromagnetic wave instabilities

The effect of multiple (singly ionized) coexisting newborn ion species on the stability of low-frequency electromagnetic waves was investigated using a plasma model in which solar wind magnetoplasma is made up of isotropic Maxwellian electron and proton populations with a common number density of 4.95/cu cm and temperatures equal to 17.2 eV and 6.9 eV, respectively. It is shown that the effect of multiple ions on wave growth, for given background magnetoplasma conditions and relative densities, depends not only on their mass but also on the physical nature of the wave modes. If the ion masses are disparate, each one of the coexisting ion beams tends to stimulate instabilities without undue influence from the other species. If the masses of newborn ions are similar, they can strongly catalyze wave growth of fluidlike nonresonant modes, but bring about weak growth enhancements in cyclotron resonant instabilities.

Brinca, Armando L.↗

Improving the Representation of Snow Crystal Properties within a Single-Moment Microphysics Scheme

The assumptions of a single-moment microphysics scheme (NASA Goddard) were evaluated using a variety of surface, aircraft and radar data sets. Fixed distribution intercepts and snow bulk densities fail to represent the vertical variability and diversity of crystal populations for this event. Temperature-based equations have merit, but they can be adversely affected by complex temperature profiles that are inverted or isothermal. Column-based approaches can mitigate complex profiles of temperature but are restricted by the ability of the model to represent cloud depth. Spheres are insufficient for use in CloudSat reflectivity comparisons due to Mie resonance, but reasonable for Rayleigh scattering applications. Microphysics schemes will benefit from a greater range of snow crystal characteristics to accommodate naturally occurring diversity.

Molthan, Andrew L.↗

Lessons from Earth Bioaerosol Sampling for Venus Habitability & Biosignature Science

Venus’s clouds are often discussed as a potential habitat. Several high-priority science goals – cloud aerosol composition, internal radiative flux, and aerosol residence time and circulation models – will improve our understanding of Venus in an astrobiology context. Lessons from Earth’s aerobiosphere can inform these requirements and interpretation of results. A stable Venus aerobiosphere would need microorganisms to reproduce faster than they settle out due to gravity. This is a joint constraint of the potential nutrients and energy influx, bioenergetic costs such as desiccation and radiation damage, and aerosol dynamics. Though life cycles involving S- and Fe-based redox metabolism have been proposed, at an optimistic estimate of 75% H2SO4, Venus aerosol water activity (aw) is still ~0.02, far below the observed microbial growth limit of ~0.6. Long-term desiccation with brief spurts of repair, growth and reproduction in response to transient water influx (perhaps volcanism) is the most likely model for Earth-like life on Venus – a ‘desert bloom’ scenario. In Earth’s troposphere, clouds can carry 103 – 105 cells/mL, some metabolically active. A better analogue may be Earth’s stratospheric sulfate aerosol layer: supercooled sulfuric acid aerosols (acid weight fraction 0.6 – 0.85, 0.1 – 1 µm diameter, 0.1 – 1 cm-3) with little water activity, long residence times, high UV radiation, and only sporadic influx from surface particle sources. Viable cells isolated from this region are rare (~102 cells/m3), though ‘hot spots’ at tropospheric mixing occur. Cells are primarily in inactive forms such as spores, and it is unclear if any are associated with sulfate aerosols or simply co-located. Given the heterogeneities in Venus clouds and the model of a sparse, largely dormant ecosystem, a single transect on descent would most likely pass through a low-water, inactive region, missing potential signs of habitability or biosignatures. Even a passive aerial platform (balloon) may end up following a single air mass over its lifetime. A better strategy for Venus would target an aerial region with some upwelling from surface sources, and take multiple transects separated in time and space. Earth’s stratospheric bioaerosol population could provide a starting point for estimating the required lower volume and detection limits.

Earth↗

Application of wideband pulsed high-frequency EPR to molecular quantum spin science

This presentation provides an overview of a range of new applications in the area of molecular quantum spin science that are now possible at the US National High Magnetic Field Laboratory as a result of the development of a wideband, high-power 94 GHz pulsed EPR spectrometer. Here, this instrument allows true Fourier-transform detected experiments spanning a 1 GHz instantaneous bandwidth, akin to what has been possible in NMR for several decades. The presentation will begin by describing experiments on a nitroxide radical using chirped pulses, enabling acquisition of the full spectrum in a single sequence, thus providing a very efficient and convenient means to measure the field orientation dependence of the spin-lattice relaxation time, T 1 . This will be followed by a discussion of a coherent population transfer protocol involving the 2S + 1 = 8 Zeeman levels associated with the spin S = 7/2 Gd 3+ ion, potentially paving the way towards implementation of well-known quantum search algorithms. It will also be shown how the use of chirped pulses provides a route to achieving highly non-thermal spin populations, suggesting novel initialization schemes.

47 OTHER INSTRUMENTATION↗