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At least 127 records · Page 7

Dimension statistics of rain cell cores and associated rain rate isopleths derived from radar measurements in the mid-Atlantic coast of the United States

Employing a multi-year radar database of precipitation in a mid-Atlantic coast region, the authors have characterized rain cell size statistics. The measurements were made with a high resolution, high power radar in which equicircle diameter contours of resolutions of 1 km and greater were identified out to ranges of 100 km from the radar. The rain cell was constructed by its core intensity isopleth and a family of enveloping contours having defined rain rate intervals in the regions between them. The isopleth statistics were extracted from a culled database whose cores had intensities of RLN = 7 (18-24 mm/h) and greater. This implies a bias of the statistics toward various types of convective rains generally associated with thunderstorms in the mid-Atlantic coast. Since the radar measurements were made in the mid-Atlantic coast region, the isopleth statistics may not be directly applicable to other geographic regions (such as the tropics).

Goldhirsh, Julius↗

Slab reformer

Slab-shaped high efficiency catalytic reformer configurations particularly useful for generation of fuels to be used in fuel cell based generation systems. A plurality of structures forming a generally rectangular peripheral envelope are spaced about one another to form annular regions, an interior annular region containing a catalytic bed and being regeneratively heated on one side by a hot comubstion gas and on the other side by the gaseous products of the reformation. An integrally mounted combustor is cooled by impingement of incoming oxidant.

Spurrier, Francis R.↗

Slab reformer

Slab-shaped high efficiency catalytic reformer configurations particularly useful for generation of fuels to be used in fuel cell based generation systems. A plurality of structures forming a generally rectangular peripheral envelope are spaced about one another to form annular regions, an interior annular region containing a catalytic bed and being regeneratively heated on one side by a hot combustion gas and on the other side by the gaseous products of the reformation. An integrally mounted combustor is cooled by impingement of incoming oxidant.

Spurrier, Francis R.↗

A Structural Perspective on the Alphavirus Life Cycle

Alphaviruses are mosquito-borne, enveloped viruses with a positive-sense, single-stranded RNA genome. Alphaviruses enter host cells via receptor-mediated endocytosis, using various cellular surface receptors such as matrix remodeling-associated protein 8 (MXRA8), low-density lipoprotein receptor class A domain-containing 3 (LDLRAD3), and very low-density lipoprotein receptor (VLDLR), which facilitate binding to the viral glycoproteins. Following entry, viral proteins are expressed and nonstructural proteins assemble into replication complexes in host cells, driving RNA synthesis and genome replication. Viral assembly occurs at the plasma membrane, where nascent virions bud from the host cell in a process driven by capsid and spike proteins. Recent combinatorial structural studies have provided detailed molecular insights into various steps of the alphavirus life cycle. These structural insights into the alphavirus life cycle enhance our understanding of viral replication and assembly, with significant implications for antiviral strategies and the development of alphavirus-based vaccine vectors.

RNA virus↗

Identification of a calmodulin-regulated Ca2+-ATPase in the endoplasmic reticulum

A unique subfamily of calmodulin-dependent Ca2+-ATPases was recently identified in plants. In contrast to the most closely related pumps in animals, plasma membrane-type Ca2+-ATPases, members of this new subfamily are distinguished by a calmodulin-regulated autoinhibitor located at the N-terminal instead of a C-terminal end. In addition, at least some isoforms appear to reside in non-plasma membrane locations. To begin delineating their functions, we investigated the subcellular localization of isoform ACA2p (Arabidopsis Ca2+-ATPase, isoform 2 protein) in Arabidopsis. Here we provide evidence that ACA2p resides in the endoplasmic reticulum (ER). In buoyant density sucrose gradients performed with and without Mg2+, ACA2p cofractionated with an ER membrane marker and a typical "ER-type" Ca2+-ATPase, ACA3p/ECA1p. To visualize its subcellular localization, ACA2p was tagged with a green fluorescence protein at its C terminus (ACA2-GFPp) and expressed in transgenic Arabidopsis. We collected fluorescence images from live root cells using confocal and computational optical-sectioning microscopy. ACA2-GFPp appeared as a fluorescent reticulum, consistent with an ER location. In addition, we observed strong fluorescence around the nuclei of mature epidermal cells, which is consistent with the hypothesis that ACA2p may also function in the nuclear envelope. An ER location makes ACA2p distinct from all other calmodulin-regulated pumps identified in plants or animals.

NASA Discipline Plant Biology↗

Antibody Recognition of CD4-Induced Open HIV-1 Env Trimers

Human immunodeficiency virus type 1 (HIV-1) envelope (Env), a heterotrimer of gp120-gp41 subunits, mediates fusion of the viral and host cell membranes after interactions with the host receptor CD4 and a coreceptor. CD4 binding induces rearrangements in Env trimer, resulting in a CD4-induced (CD4i) open Env conformation. Structural studies of antibodies isolated from infected donors have defined antibody-Env interactions, with one class of antibodies specifically recognizing the CD4i open Env conformation. In this study, we characterized a group of monoclonal antibodies isolated from HIV-1 infected donors (V2i MAbs) that displayed characteristics of CD4i antibodies. Binding experiments demonstrated that the V2i MAbs preferentially recognize CD4-bound open Env trimers. Structural characterizations of V2i MAb-Env-CD4 trimer complexes using single-particle cryo-electron microscopy showed recognition by V2i MAbs using different angles of approach to the gp120 V1V2 domain and the β2/β3 strands on a CD4i open conformation Env with no direct interactions of the MAbs with CD4. We also characterized CG10, a CD4i antibody that was raised in mice immunized with a gp120-CD4 complex, bound to an Env trimer plus CD4. CG10 exhibited characteristics similar to those of the V2i antibodies, i.e., recognition of the open Env conformation, but showed direct contacts to both CD4 and gp120. Structural comparisons of these and previously characterized CD4i antibody interactions with Env provide a suggested mechanism for how these antibodies are elicited during HIV-1 infection.

60 APPLIED LIFE SCIENCES↗

Spatial and temporal control of lysis by the lambda holin

The infection cycle of phage λ terminates in lysis mediated by three types of lysis proteins, each disrupting a layer in the bacterial envelope: the S105 holin, the R endolysin, and the Rz/Rz1 spanin complex targeting the inner membrane, cell wall or peptidoglycan, and the outer membrane, respectively. Video microscopy has shown that in most infections, lysis occurs as a sudden, explosive event at a cell pole, such that the initial product is a less refractile ghost that retains rod-shaped morphology. Here, we investigate the molecular basis of polar lysis using time-lapse fluorescence microscopy. The results indicate that the holin determines the morphology of lysis by suddenly forming two-dimensional rafts at the poles about 100 s prior to lysis. Given the physiological and biochemical similarities between the lambda holin and other class I holins, dynamic redistribution and sudden concentration may be common features of holins, probably reflecting the fitness advantage of all-or-nothing lysis regulation

Microbiology↗

Vaccination induces broadly neutralizing antibody precursors to HIV gp41

A key barrier to the development of vaccines that induce broadly neutralizing antibodies (bnAbs) against human immunodeficiency virus (HIV) and other viruses of high antigenic diversity is the design of priming immunogens that induce rare bnAb-precursor B cells. The high neutralization breadth of the HIV bnAb 10E8 makes elicitation of 10E8-class bnAbs desirable; however, the recessed epitope within gp41 makes envelope trimers poor priming immunogens and requires that 10E8-class bnAbs possess a long heavy chain complementarity determining region 3 (HCDR3) with a specific binding motif. We developed germline-targeting epitope scaffolds with affinity for 10E8-class precursors and engineered nanoparticles for multivalent display. Scaffolds exhibited epitope structural mimicry and bound bnAb-precursor human naive B cells in ex vivo screens, protein nanoparticles induced bnAb-precursor responses in stringent mouse models and rhesus macaques, and mRNA-encoded nanoparticles triggered similar responses in mice. Thus, germline-targeting epitope scaffold nanoparticles can elicit rare bnAb-precursor B cells with predefined binding specificities and HCDR3 features.

60 APPLIED LIFE SCIENCES↗

Immunological and biochemical evidence for nuclear localization of annexin in peas

Immunofluorescent localization of annexins using an anti-pea annexin polyclonal antibody (anti-p35) in pea (Pisum sativum) leaf and stem epidermal peels showed staining of the nuclei and the cell periphery. Nuclear staining was also seen in cell teases prepared from pea plumules. The amount of nuclear stain was reduced both by fixation time and by dehydration and organic solvent treatment. Observation with confocal microscopy demonstrated that the anti-p35 stain was diffusely distributed throughout the nuclear structure. Immunoblots of purified nuclei, nuclear envelope matrix, nucleolar, and chromatin fractions showed a cross-reactive protein band of 35 kDa. These data are the first to show annexins localized in plant cell nuclei where they may play a role in nuclear function.

Non-NASA Center↗

Comparative assessment of the polypeptide profiles from lateral and primary roots of Phaseolus vulgaris L

In Phaseolus vulgaris, primary roots show gravitational sensitivity soon after emerging from the seed. In contrast, lateral roots are agravitropic during early development, and become gravitropic after several cm growth. Primary and lateral root tissues were examined by polyacrylamide gel electrophoresis, coupled with western blotting techniques, to compare proteins which may contribute to the acquisition of gravitational sensitivity. Root tips and zones of cell elongation were compared for each root type, using immunological probes for calmodulin, alpha-actin, alpha-tubulin, and proteins of the plastid envelope. Lateral roots contained qualitatively less calmodulin, and showed a slightly different pattern of actin-related epitope proteins, than did primary root tissues, suggesting that polypeptide differences may contribute to the gravitational sensitivity which these root types express.

Non-NASA Center↗

Installation and Demonstration of CNC Machining for Mechanical Test Specimens in the IFEL Hot Cell

This report documents the installation and demonstration of computer numerical control (CNC) machining capabilities in the Irradiated Fuels Examination Laboratory hot cell facility at Oak Ridge National Laboratory (ORNL). A modified Tormach PCNC 440 mill was integrated into the hot cell with custom fixturing, fines management, and manipulator-compatible interfaces to enable the fabrication of axial tension test (ATT) and ring tension test (RTT) specimens from irradiated cladding. The first irradiated specimens machined included ATT and RTT geometries harvested from the high-burnup 6XV fuel rod. Dimensional inspections confirmed that machined specimens met the ±0.025 mm tolerance envelope established in prior development; deviations were consistent with expected measurement scatter and inherent specimen variability, such as wall thickness gradients and eccentricity. Comparisons with out-of-cell metrology confirmed that in-cell machining performance aligns with baseline scatter observed under ideal conditions. This work establishes reproducible, end-to-end specimen preparation at ORNL, directly supporting the US Department of Energy’s Accident-Tolerant Fuel program by enabling reliable, traceable mechanical testing of irradiated cladding.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Characteristics of statoliths from rootcaps and coleoptiles

Amyloplasts with intact envelopes were isolated from corn coleoptiles, incubated with either cationized ferritin (CF) or fluorescein isothiocyanate (FITC)-CF, and examined with epifluorescence optics. Before incubation, the mean zeta potential was found to be -19.4 mv. The net negative surface charge was confirmed ultrastructurally by the binding of CF to both amyloplasts and starch grains. Binding to the latter was reduced compared to the former. FITC-CF showed binding to some but not all amyloplasts, and was absent from individual starch granules. Membrane-bound calcium, indicated by chlorotetracycline, fluorescence, was detected throughout the cytoplasm of root cap cells. These results may reflect an affinity of CF for membrane-based charge, and suggest that a calcium shell exists on the amyloplast envelope.

Sack, F. D.↗

Orbiter Capability for Providing Water to the International Space Station according to the Most Probable Flight Attitudes

Water to be generated by, delivered to, and processed by the International Space Station (ISS) is a critical Environmental Control and Life Support (ECLS) element, especially for the early ISS missions. A significant portion of the water required by the ISS shall be provided by the Shuttle Transportation System (STS) Orbiter. The balance of water generated by the Orbiter Fuel Cells (FC), minus that water consumed by the Orbiter Flash Evaporator System (FES) and crew, is available for transfer to the ISS. During later missions, crew respired and perspired water, as well as effluent water from the Orbiter LiOH canisters, will be collected as condensate and available for transfer to the ISS. Orbiter radiator performance provides the most variance in determining the amount of net Orbiter water available for transfer to the ISS. As radiator performance decreases, the dependence upon the FES (and FC water) increases for rejecting Orbiter waste heat. Generally, radiator performance decreases as the ISS assembly size increases (especially as solar arrays are added), and also as beta angle increases. ISS solar array deployment necessitates the use of models with articulating solar arrays (for Earth local-vertical attitudes), as array position dramatically affects Orbiter radiator performance. Recent developments in the relaxation of beta angle limitations have also increased the complexity and difficulty of providing water to the ISS. Other factors that may hinder the ability to transfer water are the number of empty Contingency Water Containers (CWCs) available, duration of open-hatch time, crew activity timeline, and full CWC storage capability. A parametric study has been accomplished that provides a quick-reference table for determining expected water generation rates for ISS missions 2A.2 through 7A.1. An hourly Orbiter water generation rate is reported according to a matrix that consists of: (1) (six) significant changes in ISS assembly configuration; (2) (four) beta angles (0 deg. , +37 deg., +53 deg. , and +75 deg.); (3) the (three) most representative ISS attitudes (XPOP-O, XPOP-180 and +XVV); (4) (four) Orbiter radiator configurations (both stowed, starboard deployed, port deployed, and both deployed) and (5) the (two) conditions (radiator inlet temperatures and fuel cell power) most consistent with sleep and wake periods. Those permutations of higher probability of occurrence than others have been identified. Another parametric study has been accomplished that provides a quick-reference table for determining expected water generation rates for ISS assembly complete missions. An hourly Orbiter water generation rate is reported according to a matrix that consists of: (1) (seven) beta angles (-75 deg., -60 deg., -30 deg., 0 deg., +30 deg., +60 deg., and +75 deg.); (2) the (nine) PYR angles that define the corners of the envelope; (3) (four) Orbiter radiator configurations (both stowed, starboard deployed, port deployed, and both deployed) and (4) the (two) conditions (radiator inlet temperatures and fuel cell power) most consistent with sleep and wake periods.

Dunaway, Brian↗

Zika-specific neutralizing antibodies targeting inter-dimer envelope epitopes

Zika virus (ZIKV) is an emerging pathogen that causes devastating congenital defects. The overlapping epidemiology and immunologic cross-reactivity between ZIKV and dengue virus (DENV) pose complex challenges to vaccine design, given the potential for antibody-dependent enhancement of disease. Therefore, classification of ZIKV-specific antibody targets is of notable value. From a ZIKV-infected rhesus macaque, we identify ZIKV-reactive B cells and isolate potent neutralizing monoclonal antibodies (mAbs) with no cross-reactivity to DENV. We group these mAbs into four distinct antigenic groups targeting ZIKV-specific cross-protomer epitopes on the envelope glycoprotein. Co-crystal structures of representative mAbs in complex with ZIKV envelope glycoprotein reveal envelope-dimer epitope and unique dimer-dimer epitope targeting. All four specificities are serologically identified in convalescent humans following ZIKV infection, and representative mAbs from all four groups protect against ZIKV replication in mice. These results provide key insights into ZIKV-specific antigenicity and have implications for ZIKV vaccine, diagnostic, and therapeutic development.

59 BASIC BIOLOGICAL SCIENCES↗

Development status of solid polymer electrolyte water electrolysis for manned spacecraft life support systems

Details of the design and system verification test results are presented for a six-man-rated oxygen generation system. The system configuration incorporates components and instrumentation for computer-controlled operation with automatic start-up/shutdown sequencing, fault detection and isolation, and with self-contained sensors and controls for automatic safe emergency shutdown. All fluid and electrical components, sensors, and electronic controls are designed to be easily maintainable under zero-gravity conditions. On-board component spares are utilized in the system concept to sustain long-term operation (six months minimum) in a manned spacecraft application. The system is centered on a 27-cell solid polymer electrolyte water electrolysis module which, combined with the associated system components and controls, forms a total system envelope 40 in. high, 40 in. wide, and 30 in. deep.

Nuttall, L. J.↗

Probing the in situ volumes of Arabidopsis leaf plastids using three‐dimensional confocal and scanning electron microscopy

SUMMARY Leaf plastids harbor a plethora of biochemical reactions including photosynthesis, one of the most important metabolic pathways on Earth. Scientists are eager to unveil the physiological processes within the organelle but also their interconnection with the rest of the plant cell. An increasingly important feature of this venture is to use experimental data in the design of metabolic models. A remaining obstacle has been the limited in situ volume information of plastids and other cell organelles. To fill this gap for chloroplasts, we established three microscopy protocols delivering in situ volumes based on: (i) chlorophyll fluorescence emerging from the thylakoid membrane, (ii) a CFP marker embedded in the envelope, and (iii) calculations from serial block‐face scanning electron microscopy (SBFSEM). The obtained data were corroborated by comparing wild‐type data with two mutant lines affected in the plastid division machinery known to produce small and large mesophyll chloroplasts, respectively. Furthermore, we also determined the volume of the much smaller guard cell plastids. Interestingly, their volume is not governed by the same components of the division machinery which defines mesophyll plastid size. Based on our three approaches, the average volume of a mature Col‐0 wild‐type mesophyll chloroplasts is 93 μm 3 . Wild‐type guard cell plastids are approximately 18 μm 3 . Lastly, our comparative analysis shows that the chlorophyll fluorescence analysis can accurately determine chloroplast volumes, providing an important tool to research groups without access to transgenic marker lines expressing genetically encoded fluorescence proteins or costly SBFSEM equipment.

Knoblauch, Jan↗

An Open-Cell Environmental Transmission Electron Microscopy Technique for In Situ Characterization of Samples in Aqueous Liquid Solutions

The desire to image specimens in liquids has led to the development of open-cell and closed-cell techniques in transmission electron microscopy (TEM). The closed-cell approach is currently more common in TEM and has yielded new insights into a number of biological and materials processes in liquid environments. The open-cell approach, which requires an environmental TEM (ETEM), is technically challenging but may be advantageous in certain circumstances due to fewer restrictions on specimen and detector geometry. Here, we demonstrate a novel approach to open-cell liquid TEM, in which we use salt particles to facilitate the in situ formation of droplets of aqueous solution that envelope specimen particles coloaded with the salt. This is achieved by controlling sample temperature between 1 and 10°C and introducing water vapor to the ETEM chamber above the critical pressure for the formation of liquid water on the salt particles. Our use of in situ hydration enables specimens to be loaded into a microscope in a dry state using standard 3 mm TEM grids, allowing specimens to be prepared using trivial sample preparation techniques. Furthermore, our future aim will be to combine this technique with an in situ light source to study photocorrosion in aqueous environments.

47 OTHER INSTRUMENTATION↗

Long-Duration Low-to Medium-Altitude Solar Electric Airship Concept

This report presents the conceptual design for a solar electric lighter-than-air, unmanned aerial vehicle, based on existing technology already reduced to practice, that could carry a 600-kg (1322-lbm) payload to altitudes up to 30 kft (9000 m), continuously maintain an airspeed up to 40 kt (21 m/sec), and remain in flight for up to 100 days. The design is based on modern nonrigid airship technology, high-strength polymer fabrics and barrier films, and previously demonstrated aerospace electrical power technology, including lightweight photovoltaics and hydrogen-air polymer electrolyte membrane (PEM) fuel cells. The vehicle concept exploits the inherent synergy between the use of hydrogen as a lifting gas and the use of hydrogen-air PEM fuel-cell technology for onboard solar energy storage. In this report, the air vehicle concept is physically characterized and its estimated performance envelope is defined

Bents, David J.↗