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567 records · Page 32

Single-Cell Genomics of Novel Actinobacteria With the Wood–Ljungdahl Pathway Discovered in a Serpentinizing System

Serpentinite-hosted systems represent modern-day analogs of early Earth environments. In these systems, water-rock interactions generate highly alkaline and reducing fluids that can contain hydrogen, methane, and low-molecular-weight hydrocarbons-potent reductants capable of fueling microbial metabolism. In this study, we investigated the microbiota of Hakuba Happo hot springs (~50°C; pH~10.5–11), located in Nagano (Japan), which are impacted by the serpentinization process. Analysis of the 16S rRNA gene amplicon sequences revealed that the bacterial community comprises Nitrospirae (47%), “Parcubacteria” (19%), Deinococcus-Thermus (16%), and Actinobacteria (9%), among others. Notably, only 57 amplicon sequence variants (ASV) were detected, and fifteen of these accounted for 90% of the amplicons. Among the abundant ASVs, an early-branching, uncultivated actinobacterial clade identified as RBG-16-55-12 in the SILVA database was detected. Ten single-cell genomes (average pairwise nucleotide identity: 0.98–1.00; estimated completeness: 33–93%; estimated genome size: ~2.3 Mb) that affiliated with this clade were obtained. Taxonomic classification using single copy genes indicates that the genomes belong to the actinobacterial class-level clade UBA1414 in the Genome Taxonomy Database. Based on metabolic pathway predictions, these actinobacteria are anaerobes, capable of glycolysis, dissimilatory nitrate reduction and CO 2 fixation via the Wood–Ljungdahl (WL) pathway. Several other genomes within UBA1414 and two related class-level clades also encode the WL pathway, which has not yet been reported for the Actinobacteria phylum. For the Hakuba actinobacterium, the energy metabolism related to the WL pathway is likely supported by a combination of the Rnf complex, group 3b and 3d [NiFe]-hydrogenases, [FeFe]-hydrogenases, and V-type (H + /Na + pump) ATPase. The genomes also harbor a form IV ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) complex, also known as a RubisCO-like protein, and contain signatures of interactions with viruses, including clustered regularly interspaced short palindromic repeat (CRISPR) regions and several phage integrases. This is the first report and detailed genome analysis of a bacterium within the Actinobacteria phylum capable of utilizing the WL pathway. The Hakuba actinobacterium is a member of the clade UBA1414/RBG-16-55-12, formerly within the group “OPB41.” We propose to name this bacterium ‘Candidatus Hakubanella thermoalkaliphilus.'

59 BASIC BIOLOGICAL SCIENCES↗

Rugged, Portable, Real-Time Optical Gaseous Analyzer for Hydrogen Fluoride

Hydrogen fluoride (HF) is a primary evolved combustion product of fluorinated and perfluorinated hydrocarbons. HF is produced during combustion by the presence of impurities and hydrogen- containing polymers including polyimides. This effect is especially dangerous in closed occupied volumes like spacecraft and submarines. In these systems, combinations of perfluorinated hydrocarbons and polyimides are used for insulating wiring. HF is both highly toxic and short-lived in closed environments due to its reactivity. The high reactivity also makes HF sampling problematic. An infrared optical sensor can detect promptly evolving HF with minimal sampling requirements, while providing both high sensitivity and high specificity. A rugged optical path length enhancement architecture enables both high HF sensitivity and rapid environmental sampling with minimal gaseous contact with the low-reactivity sensor surfaces. The inert optical sample cell, combined with infrared semiconductor lasers, is joined with an analog and digital electronic control architecture that allows for ruggedness and compactness. The combination provides both portability and battery operation on a simple camcorder battery for up to eight hours. Optical detection of gaseous HF is confounded by the need for rapid sampling with minimal contact between the sensor and the environmental sample. A sensor is required that must simultaneously provide the required sub-parts-permillion detection limits, but with the high specificity and selectivity expected of optical absorption techniques. It should also be rugged and compact for compatibility with operation onboard spacecraft and submarines. A new optical cell has been developed for which environmental sampling is accomplished by simply traversing the few mm-thick cell walls into an open volume where the measurement is made. A small, low-power fan or vacuum pump may be used to push or pull the gaseous sample into the sample volume for a response time of a few seconds. The optical cell simultaneously provides for an enhanced optical interaction path length between the environmental sample and the infrared laser. Further, the optical cell itself is comprised of inert materials that render it immune to attack by HF. In some cases, the sensor may be configured so that the optoelectronic devices themselves are protected and isolated from HF by the optical cell. The optical sample cell is combined with custom-developed analog and digital control electronics that provide rugged, compact operation on a platform that can run on a camcorder battery. The sensor is inert with respect to acidic gases like HF, while providing the required sensitivity, selectivity, and response time. Certain types of combustion events evolve copious amounts of HF, very little of other gases typically associated with combustion (e.g., carbon monoxide), and very low levels of aerosols and particulates (which confound traditional smoke detectors). The new sensor platform could warn occupants early enough to take the necessary countermeasures.

Pilgrim, Jeffrey↗

1025:1 Area Ratio Nozzle Evaluated at High Combustion Chamber Pressure

A recently completed experimental test program obtained performance data on an optimally contoured nozzle with an exit-to-throat area ratio of 1025:1 and on a truncated version of this nozzle with an area ratio of 440:1. The nozzles were tested with gaseous hydrogen and liquid oxygen propellants at combustion chamber pressures of 12.4 to 16.5 mPa (1800 to 2400 psia). Testing was conducted in the altitude test capsule at the NASA Lewis Research Center's Rocket Engine Test Facility (RETF), and results were compared with analytical performance predictions. This testing builds on previous work with this nozzle at Lewis, where testing was completed at nominal chamber pressure of 350 psia. High-area-ratio nozzles have long been sought as a means to increase the performance of spacebased rocket engines. However, as the area ratio increases, the physical size and weight of the nozzle also increase. As a result, engine and vehicle designers must make tradeoffs between nozzle size and performance enhancement. Until this test program, very little experimental data existed on the performance of the high-area-ratio nozzles used in rocket engine designs. The computer codes being used by rocket engine designers rely on data extrapolated from tests of low-area-ratio nozzles, and these extrapolations do not always provide the accuracy needed for a reliable design assessment. Therefore, we conducted this high-area-ratio nozzle testing program to provide performance data for use in rocket engine design and analysis computer codes. The nozzle had a nominal 2.54-cm- (1-in.-) diameter throat, an exit diameter of 81.3-cm (32.0-in.) at an exit-to-throat area ratio of 1025, and a length of 128.6 cm (50.6 in.). Testing was conducted in an altitude test capsule to simulate the static pressure at altitude by vacuum pumping. Data such as propellant mass flow, oxidizer-to-fuel mixture, and thrust were measured. These measurements were then used to calculate performance factors such as the thrust coefficient, the characteristic exhaust velocity efficiency, and the vacuum specific impulse. In addition, the nozzle temperature was measured to calculate the amount of heat transferred from the combustion gases to the nozzle.

Source record↗

Engine Cycle Comparison for Alternative Propellant Nuclear Thermal Propulsion Engines

A modular Nuclear Thermal Propulsion (NTP) engine modeling suite was coded in Simulink to analyze various engine cycle configurations and propellants. This model was validated and used for previous studies involving alternative propellants for NTP engines. The current study compared hydrogen-based NTP (H-NTP) with ammonia-based Alternative propellant NTP (A-NTP) engines with expander and bleed cycle configurations. Based on prior work, the bleed cycle configuration was modified to yield a feasible result. The assumption that was made was that the Testing Reference Design (TRD) reactor will be used for all cases and that the difference in fuel loading fractions will be enough to offset any differences in the neutronics caused by switching the propellant. This work provided and compared propellant state points throughout the cycles as well as the key performance parameters. The conclusion was that bleed cycles for H-NTP engines are advantageous over expander cycles in terms of maximum system pressure, cycle simplicity, and potentially engine mass. Conversely, expander cycles are more advantageous for A-NTP engines according to the same scope of parameters. Future work will analyze reactor designs that are different from the TRD and use neutronics software integrated with the X-NTP model to provide higher fidelity results.

Nuclear↗

Passive Cooling in Aerogel-Based Insulation Systems for Liquid Hydrogen Upper Stage Launch Vehicle Tanks

Spray-On Foam Insulation (SOFI) is typically used to protect the windward-facing side of flight tanks for space launch vehicle liquid hydrogen (LH2) stages. SOFI is an excellent insulator in ambient pressure environments, however, its performance pales in comparison to reflective-type systems such as Multi-Layer Insulation in the thermal radiation dominated vacuum environment of space. If a windward insulation system employing radiation shields could be devised to replace or supplement SOFI, the on-orbit heat load could be drastically reduced, and the residual propellants could then be used to facilitate secondary missions. Such insulation systems, utilizing aerogel blanket insulation, have been explored by the Cryogenics Test Laboratory at NASA Kennedy Space Center. Owing to its nano-porous structure, aerogel is an excellent adsorbent as well as an insulator at cryogenic temperatures and will readily uptake condensable background gasses such as air when protecting a surface near LH2 temperatures. When an adsorbed blanket is rapidly exposed to a vacuum, such as during the ascent of a rocket, it will release the background gas, producing a passive cooling effect that could potentially reduce or eliminate the heat load on the propellant tank for a time. This work discusses the setup and results of a test program carried out to determine the effectiveness and impact of this cooling effect as it pertains to LH2 upper stage tanks. A vertical-cylindrical liquid nitrogen (LN2) calorimeter test instrument acted as the upper stage analog. The cold-mass was wrapped with two layers of 10 mm thick aerogel blanket material, with and without aluminum heat shields depending on the test, and interspersed with numerous thermocouple temperature sensors. Gaseous argon was used as a stand-in for air as the condensable background gas. The test procedure mimicked vehicle cryo-loading/stabilization, and ascent phases, with the ascent phase accomplished using parallel vacuum pumps. Results show that the desorption cooling effect was enough to not only eliminate the heat load on the LN2 cold-mass entirely, but further refrigerated the liquid below its boiling point.

A.M. Swanger↗

Atmospheric H2O2 measurement: comparison of cold trap method with impinger bubbling method

Collection of atmospheric H2O2 was performed by a cold trap method using dry ice-acetone as the refrigerant. The air was drawn by a pump into a glass gas trap immersed in the dry ice-acetone slush in a dewar flask at a flow rate of 2.5 l min-1 for approximately 2 h. Collection efficiency was > 99% and negligible interferences by O3, SO2 or organic matter with the collected H2O2 in the trap were observed. This method was compared with the air impinger bubbling method which has been previously described (Kok et al., 1978a, b, Envir. Sci. Technol. 12, 1072-1080). The measured total peroxide (H2O2 + organic peroxide) values in a series of aim samples collected by the impinger bubbling method (0.06-3.7 ppb) were always higher than those obtained by the cold trap method (0.02-1.2 ppb). Laboratory experiments suggest that the difference in values between the two methods probably results from the aqueous phase generation of H2O2 and organic peroxide in the impinger solution by a reaction of atmospheric O3 with olefinic and aromatic compounds. If these O3-organic compound reactions which occur in the impinger also occur in aqueous droplets in the atmosphere, the process could be very important for aqueous phase generation of H2O2 in clouds and rainwater.

NASA Discipline Exobiology↗

Energetic and structural control of polyspecificity in a multidrug transporter

Multidrug efflux pumps are dynamic molecular machines that drive antibiotic resistance by harnessing ion gradients to export chemically diverse substrates. Despite their clinical importance, the molecular principles underlying multidrug promiscuity and energy efficiency remain poorly understood. Using multiparametric deep mutational scanning across eight substrates and two energy conditions, we deconvolute the contributions of substrate recognition, energetic coupling, and protein stability, providing an integrated, high-resolution view of multidrug transport. We find that substrate specificity arises from a distributed network of residues extending beyond the binding site, with mutations that reshape binding, coupling, conformational flexibility, and membrane interactions. Further, we apply a pH-based selection scheme to measure the effect of mutation on pH-dependent transport efficiency. By integrating these data, we reveal a fundamental relationship between efficiency and promiscuity: Highly efficient variants exhibit broad substrate profiles, while inefficient variants are narrower. In conclusion, these findings establish a direct link between energy coupling and polyspecificity, uncovering the biochemical logic underlying multidrug transport.

Biological Sciences↗

Results of Speciation Determination Measurements of Insulating Sulfur Hexafluoride Gas

Under the direction of the NNSA NA-231 Mo-99 Program, SRNL has provided technical assistance to U.S. companies under cooperative agreements. One such area of technical assistance has been for the recommendation for contamination management methods in the eventuality that tritium contaminates the insulating gas of a high-voltage ion source used in the accelerator-based process developed by SHINE Medical Technologies to produce Mo-99. The SHINE accelerator process uses tritium as the target of a deuterium ion beam, where the ion beam is produced within a high-voltage ion source insulated with sulfur hexafluoride (SF 6 ), and is separated from the tritium target by an extensive pump train. Normal operating conditions preclude the conditions necessary for contamination of the insulating gas, however certain atypical conditions could result in a tritium contamination event. Accelerator based processes for medical isotope production represent an anomalous tritium contamination challenge due to the presence of SF 6 . SF 6 decomposes through normal use as an insulator gas, and the exact speciation of the decomposition byproducts depends on a variety of factors. This creates a scenario where the anticipation of species present in the insulating gas mixture is difficult. In turn, the fate of tritium within such a chemical system is unclear. This document discusses the results of measurements made in-situ of the SF 6 insulating gas in use within a SHINE Medical Technology’s high-voltage ion source. This was done in an effort to determine the chemical speciation of the gas, allowing for the recommendation of management strategies in the event that tritium contaminates the insulating gas. The results of the measurements reported in this work suggest that tritium contamination in the insulating gas may result in two chemical forms of tritium in the system, tritiated water and molecular tritium. These two forms of tritium would be captured by two methods. Tritiated water may be captured and removed from the SF 6 by an adsorbent canister already present within the pressure vessel that houses the insulating gas while in use. It may also be separated from the SF 6 during routine purification of the SF 6 , which is performed by SHINE personnel routinely. The purification cart will perform all non-embedded systems cleanup necessary to remediate tritiated species in the system. The purification cart passes the insulating gas through several filters, followed by the liquefication of the SF 6 . The filters will separate tritiated water from the SF 6 at this stage. It is expected that molecular tritium will accumulate in the gas-phase during the liquification step, which is separated and stored. Tritium may be recovered from this mixture through more conventional hydrogen purification methods such as a palladium diffuser.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Ion-Atom Cold Collisions and Atomic Clocks

Collisions between ultracold neutral atoms have for some time been the subject of investigation, initially with hydrogen and more recently with laser cooled alkali atoms. Advances in laser cooling and trapping of neutral atoms in a Magneto-Optic Trap (MOT) have made cold atoms available as the starting point for many laser cooled atomic physics investigations. The most spectacularly successful of these, the observation of Bose-Einstein Condensation (BEC) in a dilute ultra-cold spin polarized atomic vapor, has accelerated the study of cold collisions. Experimental and theoretical studies of BEC and the long range interaction between cold alkali atoms is at the boundary of atomic and low temperature physics. Such studies have been difficult and would not have been possible without the development and advancement of laser cooling and trapping of neutral atoms. By contrast, ion-atom interactions at low temperature, also very difficult to study prior to modern day laser cooling, have remained largely unexplored. But now, many laboratories worldwide have almost routine access to cold neutral atoms. The combined technologies of ion trapping, together with laser cooling of neutrals has made these studies experimentally feasible and several very important, novel applications might come out of such investigations . This paper is an investigation of ion-atom interactions in the cold and ultra-cold temperature regime. Some of the collisional ion-atom interactions present at room temperature are very much reduced in the low temperature regime. Reaction rates for charge transfer between unlike atoms, A + B(+) approaches A(+) + B, are expected to fall rapidly with temperature, approximately as T(sup 5/2). Thus, cold mixtures of atoms and ions are expected to coexist for very long times, unlike room temperature mixtures of the same ion-atom combination. Thus, it seems feasible to cool ions via collisions with laser cooled atoms. Many of the conventional collisional interactions, exploited as a useful tool at room temperature and higher, are greatly enhanced at low energy. For example, collisional spin transfer from one species of polarized atoms to another has long been a useful method for polarizing a sample of atoms where no other means was available. Because optical pumping cannot be used to polarize the nuclear spin of Xe-129 or He-3 (for use in nmr imaging of the lungs), the nuclear spins are polarized via collisions with an optically pumped Rb vapor in a cell containing both gases. In another case, a spin polarized thermal Cs beam was used to polarize the hyperfine states of trapped He(+)-3 ions in order to measure their hyperfine clock transition frequency. The absence of an x-ray light source to optically pump the ground state of the He(+)-3 ion necessitated this alternative state preparation. Similarly, Cd(+) and Sr(+) ions were spin-oriented via collisions in a cell with optically pumped Rb vapor. Resonant RF spin changing transitions in the ground state of the ions were detected by changes in the Rb resonance light absorption. Because cold collision spin exchange rates scale with temperature as T(sup -1/2) this technique is expected to be a far more powerful tool than the room temperature counterpart. This factor of 100 or more enhancement in spin exchange reaction rates at low temperatures is the basis for a novel trapped ion clock where laser cooled neutrals will cool, state select and monitor the ion clock transition. The advantage over conventional direct laser cooling of trapped ions is that the very expensive and cumbersome UV laser light sources, required to excite the ionic cooling transition, are effectively replaced by simple diode lasers.

Prestage, John D.↗