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At least 289 records · Page 16

Evolutionary diversification of methanotrophic ANME-1 archaea and their expansive virome

' Candidatus Methanophagales' (ANME-1) is an order-level clade of archaea responsible for anaerobic methane oxidation in deep-sea sediments. The diversity, ecology and evolution of ANME-1 remain poorly understood. In this study, we use metagenomics on deep-sea hydrothermal samples to expand ANME-1 diversity and uncover the effect of virus–host dynamics. Phylogenetic analyses reveal a deep-branching, thermophilic family, ' Candidatus Methanospirareceae', closely related to short-chain alkane oxidizers. Global phylogeny and near-complete genomes show that hydrogen metabolism within ANME-1 is an ancient trait that was vertically inherited but differentially lost during lineage diversification. Metagenomics also uncovered 16 undescribed virus families so far exclusively targeting ANME-1 archaea, showing unique structural and replicative signatures. The expansive ANME-1 virome contains a metabolic gene repertoire that can influence host ecology and evolution through virus-mediated gene displacement. Our results suggest an evolutionary continuum between anaerobic methane and short-chain alkane oxidizers and underscore the effects of viruses on the dynamics and evolution of methane-driven ecosystems.

59 BASIC BIOLOGICAL SCIENCES↗

Single-molecule epitranscriptomic analysis of full-length HIV-1 RNAs reveals functional roles of site-specific m6As

Abstract Although the significance of chemical modifications on RNA is acknowledged, the evolutionary benefits and specific roles in human immunodeficiency virus (HIV-1) replication remain elusive. Most studies have provided only population-averaged values of modifications for fragmented RNAs at low resolution and have relied on indirect analyses of phenotypic effects by perturbing host effectors. Here we analysed chemical modifications on HIV-1 RNAs at the full-length, single RNA level and nucleotide resolution using direct RNA sequencing methods. Our data reveal an unexpectedly simple HIV-1 modification landscape, highlighting three predominant N 6 -methyladenosine (m 6 A) modifications near the 3′ end. More densely installed in spliced viral messenger RNAs than in genomic RNAs, these m 6 As play a crucial role in maintaining normal levels of HIV-1 RNA splicing and translation. HIV-1 generates diverse RNA subspecies with distinct m 6 A ensembles, and maintaining multiple of these m 6 As on its RNAs provides additional stability and resilience to HIV-1 replication, suggesting an unexplored viral RNA-level evolutionary strategy.

60 APPLIED LIFE SCIENCES↗

Structures of the TMC-1 complex illuminate mechanosensory transduction

The initial step in the sensory transduction pathway underpinning hearing and balance in mammals involves the conversion of force into the gating of a mechanosensory transduction channel1. Despite the profound socioeconomic impacts of hearing disorders and the fundamental biological significance of understanding mechanosensory transduction, the composition, structure and mechanism of the mechanosensory transduction complex have remained poorly characterized. Here we report the single-particle cryo-electron microscopy structure of the native transmembrane channel-like protein 1 (TMC-1) mechanosensory transduction complex isolated from Caenorhabditis elegans. The two-fold symmetric complex is composed of two copies each of the pore-forming TMC-1 subunit, the calcium-binding protein CALM-1 and the transmembrane inner ear protein TMIE. CALM-1 makes extensive contacts with the cytoplasmic face of the TMC-1 subunits, whereas the single-pass TMIE subunits reside on the periphery of the complex, poised like the handles of an accordion. A subset of complexes additionally includes a single arrestin-like protein, arrestin domain protein (ARRD-6), bound to a CALM-1 subunit. Single-particle reconstructions and molecular dynamics simulations show how the mechanosensory transduction complex deforms the membrane bilayer and suggest crucial roles for lipid–protein interactions in the mechanism by which mechanical force is transduced to ion channel gating.

59 BASIC BIOLOGICAL SCIENCES↗

Safety and antiviral activity of triple combination broadly neutralizing monoclonal antibody therapy against HIV-1: a phase 1 clinical trial

HIV-1 therapy with single or dual broadly neutralizing antibodies (bNAbs) has shown viral escape, indicating that at least a triple bNAb therapy may be needed for robust suppression of viremia. We performed a two-part study consisting of a single-center, randomized, double-blind, dose-escalation, placebo-controlled first-in-human trial of the HIV-1 V2-glycan-specific antibody PGDM1400 alone or in combination with the V3-glycan-specific antibody PGT121 in 24 adults without HIV in part 1, as well as a multi-center, open-label trial of the combination of PGDM1400, PGT121 and the CD4-binding-site antibody VRC07-523LS in five viremic adults living with HIV not on antiretroviral therapy (ART) in part 2. The primary endpoints were safety, tolerability and pharmacokinetics for both parts and antiviral activity among viremic adults living with HIV and not on ART for part 2 of the study. The secondary endpoints were changes in CD4 + T cell counts and development of HIV-1 sequence variations associated with PGDM1400, PGT121 and VRC07-523LS resistance in part 2. Intravenously administered PGDM1400 was safe and well-tolerated at doses up to 30 mg kg -1 and when given in combination with PGT121 and VRC07-523LS. A single intravenous infusion of 20 mg kg -1 of each of the three antibodies reduced plasma HIV RNA levels in viremic individuals by a maximum mean of 2.04 log10 copies per ml; however, viral rebound occurred in all participants within a median of 20 days after nadir. Rebound viruses demonstrated partial to complete resistance to PGDM1400 and PGT121 in vitro, whereas susceptibility to VRC07-523LS was preserved. Viral rebound occurred despite mean VRC07-523LS serum concentrations of 93 µg ml -1 . The trial met the pre-specified endpoints. Our data suggest that future bNAb combinations likely need to achieve broad antiviral activity, while also maintaining high serum concentrations, to mediate viral control.

60 APPLIED LIFE SCIENCES↗

MIP-4 is Induced by Bleomycin and Stimulates Cell Migration Partially via Nir-1 Receptor

Background. CC-chemokine ligand 18 also known as MIP-4 is a chemokine with roles in inflammation and immune responses. It has been shown that MIP-4 is involved in the development of several diseases including lung fibrosis and cancer. How exactly MIP-4 is regulated and exerts its role in lung fibrosis remains unclear. Therefore, in the present study, we examined how MIP-4 is regulated and whether it acts via its potential receptor Nir-1. Materials and Methods. A549 cells were grown and maintained in DMEM : F12 (1 : 1) and supplemented with 10% FBS and 1000 U of penicillin/streptomycin and maintained as recommended by the manufacturer (ATCC). Cell migration and invasion, immunohistochemistry (IHC), Western blot, qPCR, and siRNA Nir-1 were used to determine MIP-4 regulation and its role in cell migration. Results. Cell migration was increased following stimulation of cells with recombinant (r) MIP-4 and bleomycin (BLM), whereas quenching rMIP-4 with its antibody (Ab) or addition of the Ab to BLM or H 2 O 2 diminished rMIP-4-induced cell migration. Along with cell migration, rMIP-4, BLM, and H 2 O 2 induced the formation of actin filaments dynamic structures whereas costimulation with MIP-4 Ab limited BLM- and H 2 O 2 -induced effects. MIP-4 mRNA and protein were increased by BLM and H 2 O 2 , and the addition of its Ab significantly reduced treatments effect. Experiments with siRNA investigating whether Nir-1 is a potential MIR-4 receptor indicated that the inhibition of Nir-1 decreased cell migration/invasion but did not totally inhibit rMIP-4-induced cell migration. Conclusion. Therefore, our data indicate that MIP-4 is regulated by BLM and H 2 O 2 and costimulation with its Ab limits the effects on MIP-4 and that the Nir-1 receptor partially mediates MIP-4’s effects on increased cell migration. These data also evidenced that MIP-4 is regulated by fibrotic and oxidative stimuli and that quenching MIP-4 with its Ab or therapeutically targeting the Nir-1 receptor may partially limit MIP-4 effects under fibrotic or oxidative stimulation.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Cooperation between somatic mutation and germline-encoded residues enables antibody recognition of HIV-1 envelope glycans

Viral glycoproteins are a primary target for host antibody responses. However, glycans on viral glycoproteins can hinder antibody recognition since they are self glycans derived from the host biosynthesis pathway. During natural HIV-1 infection, neutralizing antibodies are made against glycans on HIV-1 envelope glycoprotein (Env). However, such antibodies are rarely elicited with vaccination. Previously, the vaccine-induced, macaque antibody DH501 was isolated and shown to bind to high mannose glycans on HIV-1 Env. Understanding how DH501 underwent affinity maturation to recognize glycans could inform vaccine induction of HIV-1 glycan antibodies. Here, we show that DH501 Env glycan reactivity is mediated by both germline-encoded residues that contact glycans, and somatic mutations that increase antibody paratope flexibility. Only somatic mutations in the heavy chain were required for glycan reactivity. The paratope conformation was fragile as single mutations within the immunoglobulin fold or complementarity determining regions were sufficient for eliminating antibody function. Taken together, the initial germline V H DJ H rearrangement generated contact residues capable of binding glycans, and somatic mutations were required to form a flexible paratope with a cavity conducive to HIV-1 envelope glycan binding. The requirement for the presence of most somatic mutations across the heavy chain variable region provides one explanation for the difficulty in inducing anti-Env glycan antibodies with HIV-1 Env vaccination.

59 BASIC BIOLOGICAL SCIENCES↗

200-IA-1 Operable Unit Human Health Risk and Kd Screen

The purpose of this environmental calculation file (ECF) is to provide the following: Document the data processing and data reduction steps taken to prepare the 200-IA-1 Operable Unit (OU) data set that will be used to calculate the sample-specific screening level human health risk evaluation; Document the data processing and data reduction steps taken to prepare the 200-IA-1 OU data set that will be used to identify analytes that could potentially impact groundwater in the future beneath the 200-IA-1 OU representative waste sites; Document the assumptions, equations, and methodologies used to calculate the screening levels for human health cancer risks and noncancer hazards for each representative waste site assigned to the 200-IA-1 OU. Individual measured soil concentrations from 0 to 4.6 m (15 ft) below ground surface (bgs) (shallow vadose zone) are used to calculate the total excess lifetime cancer risk (ELCR) and hazard index (HI) for the outdoor worker scenario to determine if there is a basis for remedial action. Individual measured soil concentrations from the ground surface to the groundwater table are used for the distribution coefficient (Kd) screen to identify analytes that could potentially impact groundwater in the future beneath the 200-IA-1 OU representative waste sites. This ECF supports DOE/RL-2020-51, 200-IA-1 OU Focused Feasibility Study, under the Comprehensive Environmental Response, Compensation, and Liability Act of 1980 (CERCLA). A risk characterization based upon the evaluation of the health risk estimates developed in this ECF will be presented in the focused feasibility study report.

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTAN↗

Caenorhabditis elegans Dicer acts with the RIG-I-like helicase DRH-1 and RDE-4 to cleave dsRNA

Invertebrates use the endoribonuclease Dicer to cleave viral dsRNA during antiviral defense, while vertebrates use RIG-I-like Receptors (RLRs), which bind viral dsRNA to trigger an interferon response. While some invertebrate Dicers act alone during antiviral defense, Caenorhabditis elegans Dicer acts in a complex with a dsRNA binding protein called RDE-4, and an RLR ortholog called DRH-1. We used biochemical and structural techniques to provide mechanistic insight into how these proteins function together. We found RDE-4 is important for ATP-independent and ATP-dependent cleavage reactions, while helicase domains of both DCR-1 and DRH-1 contribute to ATP-dependent cleavage. DRH-1 plays the dominant role in ATP hydrolysis, and like mammalian RLRs, has an N-terminal domain that functions in autoinhibition. A cryo-EM structure indicates DRH-1 interacts with DCR-1’s helicase domain, suggesting this interaction relieves autoinhibition. Our study unravels the mechanistic basis of the collaboration between two helicases from typically distinct innate immune defense pathways.

59 BASIC BIOLOGICAL SCIENCES↗

Geologic and mineral and water resources investigations in western Colorado using ERTS-1 data

The author has identified the following significant results. Geologic interpretation of ERTS-1 imagery is dependent on recognition of the distribution, continuity, trend, and geometry of key surface features. In the examination of ERTS-1 imagery, lithology must be interpreted largely from the geomorphic expression of the terrain. ERTS-1 imagery is extremely useful in detecting local structures. Most mapped structures are topographically-expressed. Consequently, ERTS-1 imagery acquired during mid-winter, when the solar illumination angle is low, provides the largest amount of structural information. Stereoscopic analyses of ERTS-1 images significantly aid geologic interpretation. Positive transparencies of ERTS-1 images (1:1,000,000) commonly contain more geologic information than can be adequately annotated during geologic interpretation.

Knepper, D. H., Jr.↗

Application of ecological, geological and oceanographic ERTS-1 imagery to Delaware's coastal resources planning

The author has identified the following significant results. Communities containing five different coastal vegetation species, developed marshlands, and fresh water impoundments have been identified in ERTS-1 images. Suspended sediment and circulation patterns in imagery from five ERTS-1 passes over Delaware Bay have been enhanced and correlated with predicted current patterns. Conclusions reached are: (1) ERTS-1 is suitable platform for observing suspended sediment patterns and water masses synoptically over large areas. (2) Suspended sediment acts as a natural tracer allowing photointerpreters to deduce gross current circulation patterns from ERTS-1 imagery. (3) Under atmospheric conditions encountered along the East Coast of the United States MSS band 5 seems to give the best representation of sediment load in upper one meter of water column. (4) In the ERTS-1 imagery the sediment patterns are delineated by three to four neighboring shades of grey. (5) Negative transparencies of the ERTS-1 images give better contrast whenever the suspended sediment tones fall within the first few steps of the grey scale. (6) Color density slicing helps delineate the suspended sediment patterns more clearly and differentiate turbidity levels.

Klemas, V.↗

The uses of ERTS-1 imagery in the analysis of landscape change

The author has identified the following significant results. The analysis of strip mining from ERTS-1 data has resulted in the mapping of landscape changes for the Cumberland Plateau Test Site. Several mapping experiments utilizing ERTS-1 data have been established for the mapping of state-wide land use regions. The first incorporates 12 frames of ERTS-1 imagery for the generalized thematic mapping of forest cover for the state of Tennessee. In another mapping effort, 14 ERTS-1 images have been analyzed for plowed ground signatures to produce a map of agricultural regions for Tennessee, Kentucky, and the northern portions of Mississippi and Alabama. Generalized urban land use categories and transportation networks have been determined from ERTS-1 imagery for the Knoxville Test Site. Finally, through the analysis of ERTS-1 imagery, short-lived phenomena such as the 1973 spring floods on the Mississippi River in western Tennessee, have been detected, monitored, and mapped.

Rehder, J. B.↗

Skylab S192 data evaluation: Comparisons with ERTS-1 results

The author has identified the following significant results. The Skylab S192 data was evaluated by: (1) comparing the classification results using S192 and ERTS-1 data over the Holt County, Nebraska agricultural study area, and (2) investigating the impact of signal-to-noise ratio on classification accuracies using registered S192 and ERTS-1 data. Results indicate: (1) The classification accuracy obtained on S192 data using its best subset of four bands can be expected to be as high as that on ERTS-1 data. (2) When a subset of four S192 bands that are spectrally similar to the ERTS-1 bands was used for classification, an obvious deterioration in the classification accuracy was observed with respect to the ERTS-1 results. (3) The thermal bands 13 and 14 as well as the near IR bands were found to be relatively important in the classification of agricultural data. Although bands 11 and 12 were highly correlated, both were invariably included in the best subsets of the band sizes, four and beyond, according to the divergence criterion. (4) The differentiation of corn from popcorn was difficult on both S192 and ERTS-1 data acquired at an early summer date. (5) The results on both sets of data indicate that it was relatively easy to differentiate grass from any other class.

Chang, C. Y.↗

Snow-extent mapping and lake ice studies using ERTS-1 MSS together with NOAA-2 VHRR

Five snow extent maps of the 5,601 sq km American River Basin were prepared using a Zoom Transfer Scope from ERTS-1 MSS band 4 imagery. The maps were generally completed within one hour. A snowmelt curve based on ERTS-1 imagery was used as a calibration standard or comparison for maps prepared from NOAA-2 VHRR imagery in the same manner. Cost comparisons with U-2 derived imagery indicate that ERTS-1 snow mapping of the basins is six times faster. Conservative estimates of comparable aircraft snow survey flights yields a cost figure 200 times that of the ERTS-1 snow map. Snow mapping attempts in the Lake Ontario Basin demonstrated that ERTS-1 is not well suited to large basins. Optimum size of basins for ERTS studies is believed to range from about 250 sq km to 30,000 sq km. The value of the ERTS-1 MSS for Great Lake ice evaluation was proved during the past winter on Lake Erie. Not only were ice features and types of ice identified, but melting ice was detected through the combined use of band 5 and band 7. Ice movement (direction and speed) was mapped by examining imagery from two successive days.

Wiesnet, D. R.↗

Monitoring coastal water properties and current circulation with ERTS-1

Imagery and digital tapes from nine successful ERTS-1 passes over Delaware Bay during different portions of the tidal cycle have been analyzed with special emphasis on turbidity, current circulation, waste disposal plumes and convergent boundaries between different water masses. ERTS-1 image radiance correlated well with Secchi depth and suspended sediment concentration. Circulation patterns observed by ERTS-1 during different parts of the tidal cycle, agreed well with predicted and measured currents throughout Delaware Bay. Convergent shear boundaries between different water masses were observed from ERTS-1. In several ERTS-1 frames, waste disposal plumes have been detected 36 miles off Delaware's Atlantic coast. The ERTS-1 results are being used to extend and verify hydrodynamic models of the bay, developed for predicting oil slick movement and estimating sediment transport.

Klemas, V.↗

Sco X-1 - A galactic radio source with an extragalactic radio morphology

VLA observations of radio emissions at 1465 and 4885 MHz, of Sco X-1 confirm the existence of a colinear triple structure. Evidence that the three components of Sco X-1 are physically associated is presented, including the morphology, spectrum, variability, volume emissivity and magnetic field strength. The possibility of a physical phenomenon occurring in Sco X-1 similar to that occurring in extragalactic radio sources is discussed, and two galactic sources are found having extended emission similar to that in extragalactic objects. The extended structure of Sco X-1 is also observed to be similar to that of the hot spots in luminous extragalactic sources, and a radio source 20 arcmin from Sco X-1 is found to lie nearly along the radio axis formed by the components of Sco X-1.

Geldzahler, B. J.↗

Space Shuttle Flight Support Motor no. 1 (FSM-1)

Space Shuttle Flight Support Motor No. 1 (FSM-1) was static test fired on 15 Aug. 1990 at the Thiokol Corporation Static Test Bay T-24. FSM-1 was a full-scale, full-duration static test fire of a redesigned solid rocket motor. FSM-1 was the first of seven flight support motors which will be static test fired. The Flight Support Motor program validates components, materials, and manufacturing processes. In addition, FSM-1 was the full-scale motor for qualification of Western Electrochemical Corporation ammonium perchlorate. This motor was subjected to all controls and documentation requirements CTP-0171, Revision A. Inspection and instrumentation data indicate that the FSM-1 static test firing was successful. The ambient temperature during the test was 87 F and the propellant mean bulk temperature was 82 F. Ballistics performance values were within the specified requirements. The overall performance of the FSM-1 components and test equipment was nominal.

Hughes, Phil D.↗

Recommended fine positioning test for the Development Test Flight (DTF-1) of the NASA Flight Telerobotic Servicer (FTS)

Test procedures are proposed for the NASA DTF (Development Test Flight)-1 positioning tests of the FTS (Flight Telerobotic Servicer). The unique problems associated with the DTF-1 mission are discussed, standard robot performance tests and terminology are reviewed and a very detailed description of flight-like testing and analysis is presented. The major technical problem associated with DTF-1 is that only one position sensor can be used, which will be fixed at one location, with a working volume which is probably smaller than some of the robot errors to be measured. Radiation heating of the arm and the sensor could also cause distortions that would interfere with the test. Two robot performance testing committees have established standard testing procedures relevant to the DTF-1. Due to the technical problems associated with DTF-1, these procedures cannot be applied directly. These standard tests call for the use of several test positions at specific locations. Only one position, that of the position sensor, can be used by DTF-1. Off-line programming accuracy might be impossible to measure and in that case it will have to be replaced by forward kinetics accuracy.

Dagalakis, N.↗

Lubrication of an 85-mm ball bearing with RP-1

A parametric experimental investigation of an 85 millimeter bore angular contact ball bearing running in RP-1 fuel was performed at speeds of 10000 to 24000 RPM. Thrust loads were varied from 4450 to 17800 Newtons (1000 to 4000 lbs.). Radial loads were varied from 1335 to 13350 Newtons (300 to 3000 lbs.). RP-1 lubrication for the bearing was provided through a stationary jet ring located adjacent to the test bearing outer ring. Increases in both the thrust and radial loads resulted in increased bearing temperature, while increases in shaft speed resulted in much more dramatic increases in bearing temperature. These trends are typical for ball bearings operating under these types of conditions. Results are given for outer ring temperatures of the test bearing at the various test conditions employed. In addition, the heat energy removed from the bearing by the RP-1 was determined by measuring the increase in temperature as the RP-1 passed through the bearing. Results showed that the amount of heat energy removed by the RP-1 increased with both shaft speed and RP-1 flow rate to the bearing.

Addy, Harold E., Jr.↗