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At least 793 records · Page 44

Hybrid nanocapsules for in situ TEM imaging of gas evolution reactions in confined liquids

Liquid cell transmission electron microscopy (TEM) enables the direct observation of dynamic physical and chemical processes in liquids at the nanoscale. Quantitative investigations into reactions with fast kinetics and/or multiple reagents will benefit from further advances in liquid cell design that facilitate rapid in situ mixing and precise control over reagent volumes and concentrations. This work reports the development of inorganic–organic nanocapsules for high-resolution TEM imaging of nanoscale reactions in liquids with well-defined zeptoliter volumes. These hybrid nanocapsules, with 48 nm average diameter, consist of a thin layer of gold coating a lipid vesicle. As a model reaction, the nucleation, growth, and diffusion of nanobubbles generated by the radiolysis of water is investigated inside the nanocapsules. When the nanobubbles are sufficiently small (10–25 nm diameter), they are mobile in the nanocapsules, but their movement deviates from Brownian motion, which may result from geometric confinement by the nanocapsules. Gases and fluids can be transported between two nanocapsules when they fuse, demonstrating in situ mixing without using complex microfluidic schemes. The ability to synthesize nanocapsules with controlled sizes and to monitor dynamics simultaneously inside multiple nanocapsules provides opportunities to investigate nanoscale processes such as single nanoparticle synthesis in confined volumes and biological processes such as biomineralization and membrane dynamics.

36 MATERIALS SCIENCE↗

Isotopic, petrologic and biogeochemical investigations of banded iron-formations

It is recognized that the first occurrence of banded iron-formations (BIFs) clearly predates biological oxygenation of the atmosphere-hydrosphere system and that their last occurrences extend beyond plausible dates of pervasive biological oxygenation. For this reason, and because enormous quantities of oxidizing power have been sequestered in them, it is widely thought that these massive, but enigmatic, sediments must encode information about the mechanism and timing of the rise of atmospheric O2. By coupling isotopic analyses of iron-formation carbonates with biogeochemical and petrologic investigations, we are studying (1) the mechanism of initial sedimentation of iron; (2) the role of iron in microbially mediated diagenetic processes in fresh iron-formation sediments; and (3) the logical integration of mechanisms of deposition with observed levels of banding. Thus far, it has been shown that (1) carbonates in BIFs of the Hamersley Group of Western Australia are isotopically inhomogenous; (2) the nature and pattern of isotopic ordering is not consistent with a metamorphic origin for the overall depletion of C-13 observed in the carbonates; (3) if biological, the origin of the C-13 depleted carbonate could be either respiratory or fermentative; (4) iron may have been precipitate d as Fe(3+), then reduced to Fe(2+) within the sediment; and (5) sedimentary biogeochemical systems may have been at least partially closed to mass transport of carbonate species.

Hayes, J. M.↗

An Algorithm for the Transport of Anisotropic Neutrons

One major obstacle to human space exploration is the possible limitations imposed by the adverse effect of long-term exposure to the space environment. Even before human spaceflight began, the potentially brief exposure of astronauts to the very intense random solar particle events (SPE) were of great concern. A new challenge appears in deep space exploration from exposure to the low-intensity heavy-ion flux of the galactic cosmic rays (GCR) since the missions are of long duration and the accumulated GCR exposures can be high. Because cancer induction rates increase behind low to rather large thicknesses of aluminum shielding, according to available biological data on mammalian exposures to GCR like ions, the shield requirements for a Mars mission are prohibitively expensive in terms of mission launch costs. Therefore, a critical issue in the Human Exploration and Development of Space enterprise is cost effective mitigation of risk associated with ionizing radiation exposure. In order to estimate astronaut risk to GCR exposure and associated cancer risks and health hazards, it is necessary to do shield material studies. To determine an optimum radiation shield material it is necessary to understand nuclear interaction processes such as fragmentation and secondary particle production which is a function of energy dependent cross sections. This requires knowledge of material transmission characteristics either through laboratory testing or improved theoretical modeling. Here ion beam transport theory is of importance in that testing of materials in the laboratory environment generated by particle accelerators is a necessary step in materials development and evaluation for space use. The approximations used in solving the Boltzmann transport equation for the space setting are often not sufficient for laboratory work and those issues are a major emphasis of the present work.

Tweed, J.↗

A mycobacterial ABC transporter mediates the uptake of hydrophilic compounds

Mycobacterium tuberculosis (Mtb) is an obligate human pathogen and the causative agent of tuberculosis. Although Mtb can synthesize vitamin B 12 (cobalamin) de novo, uptake of cobalamin has been linked to pathogenesis of tuberculosis. Mtb does not encode any characterized cobalamin transporter; however, the gene rv1819c was found to be essential for uptake of cobalamin. This result is difficult to reconcile with the original annotation of Rv1819c as a protein implicated in the transport of antimicrobial peptides such as bleomycin. In addition, uptake of cobalamin seems inconsistent with the amino acid sequence, which suggests that Rv1819c has a bacterial ATP-binding cassette (ABC)-exporter fold. In this paper, we present structures of Rv1819c, which reveal that the protein indeed contains the ABC-exporter fold, as well as a large water-filled cavity of about 7,700 Å, which enables the protein to transport the unrelated hydrophilic compounds bleomycin and cobalamin. On the basis of these structures, we propose that Rv1819c is a multi-solute transporter for hydrophilic molecules, analogous to the multidrug exporters of the ABC transporter family, which pump out structurally diverse hydrophobic compounds from cells.

59 BASIC BIOLOGICAL SCIENCES↗

Universal energy-speed-accuracy trade-offs in driven nonequilibrium systems

The connection between measure theoretic optimal transport and dissipative nonequilibrium dynamics provides a language for quantifying nonequilibrium control costs, leading to a collection of thermodynamic speed limits, which rely on the assumption that the target probability distribution is perfectly realized. This is almost never the case in experiments or numerical simulations, so here we address the situation in which the external controller is imperfect. We obtain a lower bound for the dissipated work in generic nonequilibrium control problems that (1) is asymptotically tight and (2) matches the thermodynamic speed limit in the case of optimal driving. Along with analytically solvable examples, we refine this imperfect driving notion to systems in which the controlled degrees of freedom are slow relative to the nonequilibrium relaxation rate, and identify independent energy contributions from fast and slow degrees of freedom. Furthermore, we develop a strategy for optimizing minimally dissipative protocols based on optimal transport flow matching, a generative machine learning technique. Furthermore, this latter approach ensures the scalability of both the theoretical and computational framework we put forth. Crucially, we demonstrate that we can compute the terms in our bound numerically using efficient algorithms from the computational optimal transport literature and that the protocols we learn saturate the bound.

59 BASIC BIOLOGICAL SCIENCES↗

Methane Consuming Biocatalysts in Gas-Solid Reactor System

Carbon dioxide and methane are the two most abundant greenhouse gases with methane having a higher global warming potential (GWP) than carbon dioxide. Use of methanotrophs as a biocatalyst is a green alternative to chemical conversion of methane. Methanotrophs were immobilized in a 4-arm poly (-ethylene glycol)-tetra-acrylate, PEGTA, hydrogel that is supported by a 3D printed lattice containing micropores. The consumption rate of methane was 2.5 and 42 g CH 4 /L-hr in samples that were at 1 and 3 atm, respectively. A small increase in pressure of the system improves the consumption rate by almost 20x with minimal changes to the reactor system. This system relies on the mass transport of gas through a solid-like material rather than dissolving the gas in a liquid. Immobilization of cells in hydrogel allows for higher cell density and overall increased conversion rates compared to traditional stirred tank reactors. There are many other applications of the use of immobilized biological materials in polymers that require gas-phase reactions.

36 MATERIALS SCIENCE↗

Structural insights into regulation of CNNM-TRPM7 divalent cation uptake by the small GTPase ARL15

Cystathionine-β-synthase (CBS)-pair domain divalent metal cation transport mediators (CNNMs) are an evolutionarily conserved family of magnesium transporters. They promote efflux of Mg 2+ ions on their own and influx of divalent cations when expressed with the transient receptor potential ion channel subfamily M member 7 (TRPM7). Recently, ADP-ribosylation factor-like GTPase 15 (ARL15) has been identified as CNNM-binding partner and an inhibitor of divalent cation influx by TRPM7. Here, we characterize ARL15 as a GTP and CNNM-binding protein and demonstrate that ARL15 also inhibits CNNM2 Mg 2+ efflux. The crystal structure of a complex between ARL15 and CNNM2 CBS-pair domain reveals the molecular basis for binding and allowed the identification of mutations that specifically block binding. A binding deficient ARL15 mutant, R95A, failed to inhibit CNNM and TRPM7 transport of Mg 2+ and Zn 2+ ions. Structural analysis and binding experiments with phosphatase of regenerating liver 2 (PRL2 or PTP4A2) showed that ARL15 and PRLs compete for binding CNNM to coordinate regulation of ion transport by CNNM and TRPM7.

59 BASIC BIOLOGICAL SCIENCES↗

P-MASS and P-GBA: Two new hardware developments for growing plants in space

Plant growth, and especially plant performance experiments in microgravity are limited by the currently available plant growth facilities (low light levels, inadequate nutrient delivery and atmosphere conditioning systems, insufficient science instrumentation, infrequent flight opportunities). In addition, mission durations of 10 to 14 days aboard the NSTS Space Shuttle allow for only brief periods of microgravity exposure with respect to the life cycle of a plant. Based on seed germination experiments, using the Generic BioProcessing Apparatus hardware (GBA), two new payloads have been designed specifically for plant growth. These payloads provide new opportunities for plant gravitational and space biology research and emphasize the investigation of plant performance (photosynthesis, biomass accumulations) in microgravity. The Plant-Module for Autonomous Space Support (P-MASS) was designed to utilize microgravity exposure times in excess of 30 days on the first flight of the recoverable COMET satellite (Commercial Experiment Transporter). The Plant-Generic Bioprocessing Apparatus (P-GBA), is designed for the National Space Transportation System (NSTS) Space Shuttle middeck and the SPACEHAB environment. The P-GBA is an evolution from the GBA hardware and P-MASS (plant chamber and instrumentation). The available light levels of both payloads more than double currently available capabilities.

Hoehn, Alexander↗

Phosphorylation of insulin receptor substrate-1 serine 307 correlates with JNK activity in atrophic skeletal muscle

c-Jun NH(2)-terminal kinase (JNK) has been shown to negatively regulate insulin signaling through serine phosphorylation of residue 307 within the insulin receptor substrate-1 (IRS-1) in adipose and liver tissue. Using a rat hindlimb suspension model for muscle disuse atrophy, we found that JNK activity was significantly elevated in atrophic soleus muscle and that IRS-1 was phosphorylated on Ser(307) prior to the degradation of the IRS-1 protein. Moreover, we observed a corresponding reduction in Akt activity, providing biochemical evidence for the development of insulin resistance in atrophic skeletal muscle.

Non-NASA Center↗

Oxidative stress inhibits caveolin-1 palmitoylation and trafficking in endothelial cells

During normal and pathological conditions, endothelial cells (ECs) are subjected to locally generated reactive oxygen species, produced by themselves or by other vessel wall cells. In excess these molecules cause oxidative injury to the cell but at moderate levels they might modulate intracellular signalling pathways. We have investigated the effect of oxidative stress on the palmitoylation and trafficking of caveolin-1 in bovine aortic ECs. Exogenous H2O2 did not alter the intracellular localization of caveolin-1 in ECs. However, metabolic labelling experiments showed that H2O2 inhibited the trafficking of newly synthesized caveolin-1 to membrane raft domains. Several mechanisms potentially responsible for this inhibition were examined. Impairment of caveolin-1 synthesis by H2O2 was not responsible for diminished trafficking. Similarly, the inhibition was independent of H2O2-induced caveolin-1 phosphorylation as shown by the markedly different concentration dependences. We tested the effect of H2O2 on palmitoylation of caveolin-1 by the incorporation of [3H]palmitic acid. Exposure of ECs to H2O2 markedly inhibited the palmitoylation of caveolin-1. Comparable inhibition was observed after treatment of cells with H2O2 delivered either as a bolus or by continuous delivery with glucose and glucose oxidase. Kinetic studies showed that H2O2 did not alter the rate of caveolin-1 depalmitoylation but instead decreased the 'on-rate' of palmitoylation. Together these results show for the first time the modulation of protein palmitoylation by oxidative stress, and suggest a cellular mechanism by which stress might influence caveolin-1-dependent cell activities such as the concentration of signalling proteins and cholesterol trafficking.

NASA Discipline Cell Biology↗

Root gravitropism: a complex response to a simple stimulus?

Roots avoid depleting their immediate environment of essential nutrients by continuous growth. Root growth is directed by environmental cues, including gravity. Gravity sensing occurs mainly in the columella cells of the root cap. Upon reorientation within the gravity field, the root-cap amyloplasts sediment, generating a physiological signal that promotes the development of a curvature at the root elongation zones. Recent molecular genetic studies in Arabidopsis have allowed the identification of genes that play important roles in root gravitropism. Among them, the ARG1 gene encodes a DnaJ-like protein involved in gravity signal transduction, whereas the AUX1 and AGR1 genes encode proteins involved in polar auxin transport. These studies have important implications for understanding the intra- and inter-cellular signaling processes that underlie root gravitropism.

Review, Tutorial↗

Decoding Ca2+ signals in plants

Different input signals create their own characteristic Ca2+ fingerprints. These fingerprints are distinguished by frequency, amplitude, duration, and number of Ca2+ oscillations. Ca(2+)-binding proteins and protein kinases decode these complex Ca2+ fingerprints through conformational coupling and covalent modifications of proteins. This decoding of signals can lead to a physiological response with or without changes in gene expression. In plants, Ca(2+)-dependent protein kinases and Ca2+/calmodulin-dependent protein kinases are involved in decoding Ca2+ signals into phosphorylation signals. This review summarizes the elements of conformational coupling and molecular mechanisms of regulation of the two groups of protein kinases by Ca2+ and Ca2+/calmodulin in plants.

Non-NASA Center↗

Radiation Beamline Testbeds for the Simulation of Planetary and Spacecraft Environments for Human and Robotic Mission Risk Assessment

The Center for Radiation Engineering and Science for Space Exploration (CRESSE) at Prairie View A&M University, Prairie View, Texas, USA, is establishing an integrated, multi-disciplinary research program on the scientific and engineering challenges faced by NASA and the international space community caused by space radiation. CRESSE focuses on space radiation research directly applicable to astronaut health and safety during future long term, deep space missions, including Martian, lunar, and other planetary body missions beyond low earth orbit. The research approach will consist of experimental and theoretical radiation modeling studies utilizing particle accelerator facilities including: 1. NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory; 2. Proton Synchrotron at Loma Linda University Medical Center; and 3. Los Alamos Neutron Science Center (LANSCE) at Los Alamos National Laboratory. Specifically, CRESSE investigators are designing, developing, and building experimental test beds that simulate the lunar and Martian radiation environments for experiments focused on risk assessment for astronauts and instrumentation. The testbeds have been designated the Bioastronautics Experimental Research Testbeds for Environmental Radiation Nostrum Investigations and Education (BERT and ERNIE). The designs of BERT and ERNIE will allow for a high degree of flexibility and adaptability to modify experimental configurations to simulate planetary surface environments, planetary habitats, and spacecraft interiors. In the nominal configuration, BERT and ERIE will consist of a set of experimental zones that will simulate the planetary atmosphere (Solid CO2 in the case of the Martian surface.), the planetary surface, and sub-surface regions. These experimental zones can be used for dosimetry, shielding, biological, and electronic effects radiation studies in support of space exploration missions. BERT and ERNIE are designed to be compatible with the experimental areas associated with the above facilities. CRESSE has broad expertise in space radiation in the areas of space radiation environment modeling, Monte-Carlo radiation transport modeling, space radiation instrumentation and dosimetry, radiation effects on electronics, and multi-functional composite shielding materials. The BERT and ERNIE testbeds will be utilized in individual and collaborative research incorporating this expertise. The research goal is to maximize the technical readiness level (TRL) of radiation instrumentation for human and robotic missions, optimizing the return value of CRESSE for NASA exploration and international co-operative missions. Outcomes and knowledge from research utilizing BERT and ERNIE will be applied to a variety of scientific and engineering disciplines vital for safe and reliable execution of future space exploration missions, which can be negatively impacted by the space radiation environment. The testbeds will be central to a variety of university educational activities and educational goals of NASA. Specifically, BERT and ERNIE will enhance educational opportunities in science, technology, engineering and mathematics (STEM) disciplines for engineering and science students at PVAMU, a historically black college/university. Preliminary data on prototype testbed configurations, including simulated lunar regolith (JSC-1A stimulant based on Apollo 11 samples), regolith/polyethylene composites, and dry ice, will be presented to demonstrate the usefulness of BERT and ERNIE in radiation beam line experiments.

Wilkins, Richard↗

Installation and commissioning of the ion source systems for the new spallation neutron source 2.5 MeV injector

The U.S. Spallation Neutron Source (SNS) is a state-of-the-art neutron scattering facility delivering the world’s most intense pulsed neutron beams to a wide array of instruments, which are used to conduct investigations in many fields of engineering, physics, chemistry, material science, and biology. Neutrons are produced by spallation of liquid Hg by the bombardment of short (∼1 µs), intense (∼35 A) pulses of protons delivered at 60 Hz by an accumulator ring which is fed by a high-intensity, 1 GeV, H− LINAC (linear accelerator). This facility has operated nearly continuously since 2006 but has recently undergone a 4-month maintenance period, which featured a complete replacement of the 2.5 MeV injector feeding the LINAC. The new injector was developed at ORNL in an off-line beam test facility and consists of an ion source, low energy beam transport, and a Radio Frequency Quadrupole (RFQ). This report first describes the installed configuration of the new injector detailing the ion source system. The first beam current, RFQ transmission, emittance, and energy measurements from the injector installed on the SNS are reported. These data not only show a significant performance improvement for our existing facility but will also make accessible the higher beam current requirements for future SNS upgrade projects: the proton power upgrade and second target station.

Welton, R. F.↗

Beyond Component Optimization: Systems Level Biodesign for Lanthanide Recovery

Global demand for lanthanides (Ln) is projected to rise sharply over the next decade, while geographically concentrated supply chains and the low concentrations and matrix complexity of secondary feedstocks limit the reach of conventional hydro- and pyrometallurgical separation. Engineered biological systems offer a selective, low-energy alternative, and component-level advances in Ln-binding proteins, AI-designed selective scaffolds, and cell-surface display platforms now rival synthetic chelators in affinity and selectivity. These components, however, remain functionally isolated. Currently, there are no engineered chassis coupling recognition, intracellular trafficking, accumulation, and controlled release into an end-to-end pipeline. Here, we outline how new biodesign strategies and chassis selection must move beyond bioleaching to encompass the full recovery pathway. Achieving this requires integrating AI/ML-guided design, genome-scale build tools, high-throughput phenotyping, and biophysical transport modeling within a Design–Build–Test–Learn cycle tuned to recognition, trafficking, accumulation, and release.

Biodesign↗

Metabolic Model of the Nitrogen-Fixing Obligate Aerobe Azotobacter vinelandii Predicts Its Adaptation to Oxygen Concentration and Metal Availability

There is considerable interest in promoting biological nitrogen fixation (BNF) as a mechanism to reduce the inputs of nitrogenous fertilizers in agriculture, but considerable fundamental knowledge gaps still need to be addressed. BNF is catalyzed by nitrogenase, which requires a large input of energy in the form of ATP and low potential electrons. Diazotrophs that respire aerobically have an advantage in meeting the ATP demands of BNF but face challenges in protecting nitrogenase from inactivation by oxygen. Here, we constructed a genome-scale metabolic model of the nitrogen-fixing bacterium Azotobacter vinelandii, which uses a complex respiratory protection mechanism to consume oxygen at a high rate to keep intracellular conditions microaerobic. Our model accurately predicts growth rate under high oxygen and substrate concentrations, consistent with a large electron flux directed to the respiratory protection mechanism. While a partially decoupled electron transport chain compensates for some of the energy imbalance under high-oxygen conditions, it does not account for all substrate intake, leading to increased maintenance rates. Interestingly, the respiratory protection mechanism is required for accurate predictions even when ammonia is supplemented during growth, suggesting that the respiratory protection mechanism might be a core principle of metabolism and not just used for nitrogenase protection. We have also shown that rearrangement of flux through the electron transport system allows A. vinelandii to adapt to different oxygen concentrations, metal availability, and genetic disruption, which cause an ammonia excretion phenotype. Accurately determining the energy balance in an aerobic nitrogen-fixing metabolic model is required for future engineering approaches.

59 BASIC BIOLOGICAL SCIENCES↗

Bottom‐Up Synthesized Nanoporous Graphene Transistors

Abstract Nanoporous graphene (NPG) can exhibit a uniform electronic band gap and rationally‐engineered emergent electronic properties, promising for electronic devices such as field‐effect transistors (FETs), when synthesized with atomic precision. Bottom‐up, on‐surface synthetic approaches developed for graphene nanoribbons (GNRs) now provide the necessary atomic precision in NPG formation to access these desirable properties. However, the potential of bottom‐up synthesized NPG for electronic devices has remained largely unexplored to date. Here, FETs based on bottom‐up synthesized chevron‐type NPG (C‐NPG), consisting of ordered arrays of nanopores defined by laterally connected chevron GNRs, are demonstrated. C‐NPG FETs show excellent switching performance with on–off ratios exceeding 10 4 , which are tightly linked to the structural quality of C‐NPG. The devices operate as p‐type transistors in the air, while n‐type transport is observed when measured under vacuum, which is associated with reversible adsorption of gases or moisture. Theoretical analysis of charge transport in C‐NPG is also performed through electronic structure and transport calculations, which reveal strong conductance anisotropy effects in C‐NPG. The present study provides important insights into the design of high‐performance graphene‐based electronic devices where ballistic conductance and conduction anisotropy are achieved, which could be used in logic applications, and ultra‐sensitive sensors for chemical or biological detection.

Mutlu, Zafer↗