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At least 1,423 records · Page 79

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ~1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ~4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io, where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

Trubl, Gareth↗

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ~1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ~4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io, where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

Trubl, Gareth↗

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ∼1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ∼4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io , where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

dsDNA viruses↗

Distinction of saffron cod ( Eleginus gracilis ) from several other gadid species by using microsatellite markers

Nine microsatellite loci isolated in saffron cod ( Eleginus gracilis ) have potential applications for population genetics. Polymerase chain reaction products of samples of E. gracilis from northwestern Alaska amplified reliably, produced only one or two microsatellite bands, and had no apparent homozygote excess. A collection of E. gracilis sampled in the Gulf of Alaska (GOA) near Kodiak Island did not amplify reliably at one locus, and allele frequency profiles clustered distinctly (with principal component analysis [PCA]) from the northwestern Alaska collection. Northwestern Alaska and GOA E. gracilis collections were genetically different (on the basis of a standardized genetic differentiation measure [ G ' ST ]=0.313, chord distance [ D chord ]=0.078, P<0.0001) and differed in expected aver-age heterozygosities at shared loci (0.859 and 0.689, respectively). We tested the microsatellite primers on other gadid species endemic to the northern Pacific Ocean, Bering Sea, and Arctic Ocean for cross-species amplification. Not all loci amplified reliably in navaga ( E. nawaga ), Pacific tomcod ( Microgadus proximus ), Arctic cod ( Boreogadus saida ), Pacific cod ( Gadus macrocephalus ), or walleye pollock ( G. chalcogrammus ). Reliable loci varied in microsatellite size profiles and produced distinct PCA clusters and accurate genotype assignments that allowed accurate species identification. Furthermore the identifications support previous morphological and genetically determined systematic classifications and distinguished the geographically separated collections of E. gracilis .

59 BASIC BIOLOGICAL SCIENCES↗

Burning plasma achieved in inertial fusion

The achievement of obtaining a burning plasma is a critical step toward self-sustaining fusion energy. A burning plasma is a fusion plasma where the alpha-particles created by the deuterium-tritium (DT) fusion reactions are the primary source of heating in the plasma, which is necessary to sustain and propagate the fusion reaction to enable high energy gain. After decades of fusion research, a burning plasma state has finally been achieved. Herein, we report upon the first burning-plasma experiments; this state was achieved using a strategy to increase the capsule spatial scale via two different implosion concepts, on the US National Ignition Facility. These experiments show energies from self-heating in excess of the mechanical work injected into the implosions satisfying several burning plasma metrics, the last experiment additionally shows that the fusion self-heating is greater than losses from radiation and heat conduction. These experiments triple the fusion yield performance and show significantly higher yield amplification from self-heating than prior results; remaining degradations can be reduced for even higher fusion performance.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Modeling of the interaction of a volumetric metallic metamaterial structure with a relativistic electron beam

We present the design of a volumetric metamaterial (MTM) structure and its interaction with a relativistic electron beam. This novel structure has promising applications in particle beam diagnostics, acceleration and microwave generation. The volumetric MTM has a cubic unit cell allowing structures of arbitrary size to be configured as an array of identical cells. This structure allows the exploration of the properties of a metamaterial structure without having to consider substrates or other supporting elements. The dispersion characteristics of the unit cell are obtained using eigenmode simulations in the HFSS code and also using an effective medium theory with spatial dispersion. Good agreement is obtained between these two approaches. The lowest order mode of the MTM structure is found to have a negative group velocity in all directions of propagation. The frequency spectrum of the radiation from a relativistic electron beam passing through the MTM structure is calculated analytically and also calculated with the CST code, with very good agreement. The radiation pattern from the relativistic electron beam is found to be backward Cherenkov radiation, which is a promising tool for particle diagnostics. Calculations are also presented for the application of a MTM-based wakefield accelerator as a possible all-metal replacement for the conventional dielectric wakefield structure. The proposed structure may also be useful for MTM-based vacuum electron devices for microwave generation and amplification.

43 PARTICLE ACCELERATORS↗

Weakly Magnetized, Hall Dominated Plasma Couette Flow

A novel plasma equilibrium in the high-β, Hall regime that produces centrally-peaked, high Mach number Couette flow is described. Flow is driven using a weak, uniform magnetic field and large, cross field currents. Large magnetic field amplification (factor 20) due to the Hall effect is observed when electrons are flowing radially inward, and near perfect field expulsion is observed when the flow is reversed. A dynamic equilibrium is reached between the amplified (removed) field and extended density gradients.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Time-resolved turbulent dynamo in a laser plasma

Understanding magnetic-field generation and amplification in turbulent plasma is essential to account for observations of magnetic fields in the universe. A theoretical framework attributing the origin and sustainment of these fields to the so-called fluctuation dynamo was recently validated by experiments on laser facilities in low-magnetic-Prandtl-number plasmas (Pm<1). However, the same framework proposes that the fluctuation dynamo should operate differently when Pm≳1, the regime relevant to many astrophysical environments such as the intracluster medium of galaxy clusters. This paper reports an experiment that creates a laboratory Pm≳1 plasma dynamo. We provide a time-resolved characterization of the plasma’s evolution, measuring temperatures, densities, flow velocities, and magnetic fields, which allows us to explore various stages of the fluctuation dynamo’s operation on seed magnetic fields generated by the action of the Biermann-battery mechanism during the initial drive-laser target interaction. The magnetic energy in structures with characteristic scales close to the driving scale of the stochastic motions is found to increase by almost three orders of magnitude and saturate dynamically. It is shown that the initial growth of these fields occurs at a much greater rate than the turnover rate of the driving-scale stochastic motions. Our results point to the possibility that plasma turbulence produced by strong shear can generate fields more efficiently at the driving scale than anticipated by idealized magnetohydrodynamics (MHD) simulations of the nonhelical fluctuation dynamo; this finding could help explain the large-scale fields inferred from observations of astrophysical systems.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Aneuploidy induces premature aging in yeast due to defects in Ribosome Quality Control [RNASeqData]

Premature aging is a hallmark of Down syndrome, caused by trisomy of human chromosome 21; but the reason is unclear and difficult to study in humans. We used an aneuploid model in wild yeast to show that chromosome amplification disrupts nutrient-induced cell-cycle arrest, quiescence entry, and healthy aging, across genetic backgrounds and amplified chromosomes. We discovered that these defects are due in part to aneuploidy-induced dysfunction in Ribosome Quality Control (RQC). Aneuploids entering quiescence display aberrant ribosome profiles, accumulate RQC intermediates, and harbor an increased load of protein aggregates. Although they have normal proteasome capacity, aneuploids show signs of ubiquitin dysregulation, which impacts cyclin abundance to disrupt arrest. Remarkably, inducing ribosome stalling in euploids produces similar aberrations, while up-regulating limiting RQC subunits or proteins in ubiquitin metabolism alleviates many of the aneuploid defects. Our results raise major implications for other aneuploidy disorders including Down syndrome.

aneuploidy↗

Aneuploidy induces premature aging in yeast due to defects in Ribosome Quality Control [MoBYSeqData]

Premature aging is a hallmark of Down syndrome, caused by trisomy of human chromosome 21; but the reason is unclear and difficult to study in humans. We used an aneuploid model in wild yeast to show that chromosome amplification disrupts nutrient-induced cell-cycle arrest, quiescence entry, and healthy aging, across genetic backgrounds and amplified chromosomes. We discovered that these defects are due in part to aneuploidy-induced dysfunction in Ribosome Quality Control (RQC). Aneuploids entering quiescence display aberrant ribosome profiles, accumulate RQC intermediates, and harbor an increased load of protein aggregates. Although they have normal proteasome capacity, aneuploids show signs of ubiquitin dysregulation, which impacts cyclin abundance to disrupt arrest. Remarkably, inducing ribosome stalling in euploids produces similar aberrations, while up-regulating limiting RQC subunits or proteins in ubiquitin metabolism alleviates many of the aneuploid defects. Our results raise major implications for other aneuploidy disorders including Down syndrome.

aneuploidy↗

Premature aging in aneuploid yeast is influenced by aneuploidy-induced defects in Ribosome Quality Control [RiboSeq]

Premature aging is a hallmark of Down syndrome, caused by trisomy of human chromosome 21; but the reason is unclear and difficult to study in humans. We used an aneuploid model in wild yeast to show that chromosome amplification disrupts nutrient-induced cell-cycle arrest, quiescence entry, and healthy aging, across genetic backgrounds and amplified chromosomes. We discovered that these defects are due in part to aneuploidy-induced dysfunction in Ribosome Quality Control (RQC). Aneuploids entering quiescence display aberrant ribosome profiles, accumulate RQC intermediates, and harbor an increased load of protein aggregates. Although they have normal proteasome capacity, aneuploids show signs of ubiquitin dysregulation, which impacts cyclin abundance to disrupt arrest. Remarkably, inducing ribosome stalling in euploids produces similar aberrations, while up-regulating limiting RQC subunits or proteins in ubiquitin metabolism alleviates many of the aneuploid defects. Our results raise major implications for other aneuploidy disorders including Down syndrome.

aneuploidy↗

Temperature transducer has high output, is time stable

Compact, lightweight temperature transducer requires no amplification of its output signal and is time stable. It uses the temperature-dependent characteristics of a silicon transistor to provide a zero-to-five-volt signal proportional to temperature.

Follett, W. H.↗

High-performance RC bandpass filter is adapted to miniaturized construction

Miniaturized bandpass filter with RC networks is suitable for use in integrated circuits. The circuit consists of three stages of amplification with additional resistive and capacitive components to obtain the desired characteristics. The advantages of the active RC filter network are the reduction in size and weight and elimination of magnetic materials.

Source record↗

Digital system detects binary code patterns containing errors

System of square loop magnetic cores associated with code input registers to react to input code patterns by reference to a group of control cores in such a manner that errors are canceled and patterns containing errors are accepted for amplification and processing. This technique improves reception capabilities in PCM telemetry systems.

Muller, R. M.↗