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DOE OSTI2017

Spin Unrestricted Nonradiative Relaxation Dynamics of Cobalt-Doped Anatase Nanowire

Spin-resolved charge transfer dynamics at the interface of a Co(NH3)2-doped (001) anatase TiO2 nanowire and liquid water calculations based on density functional theory and density matrix formalism are considered. Three models with the same stoichiometry but different electronic structure are explored. While one model had no change to electron count and spin count (neutral model), the other two models were assigned a charge of 2+, one in a doublet and the other a quartet spin configuration. Co2+ is the most probable state for dopant in all models and Co acts as an electron acceptor. The optical absorption spectra show a rather unique pattern post 400 nm where the α and β absorptions happen independently at different frequency ranges. Essentially, the relaxation dynamics can be controlled as when an α electron is excited, there is a near zero probability of a β electron also undergoing an excitation and vice versa. The isolated exemption is between 400 and 650 nm in the neutral model. All the models absorb light in the visible range, while the electrons and holes have drastic differences in relaxation rates. The spatial charge separation occurs upon excitation and subsequent trapping, designating the considered system a prospect for electrochemical cell applications. (2+) models trap states more effectively slow down electron relaxation, making the charged models slightly better options for application compared to the neutral model.

Jensen, Stephanie J.↗

DOE OSTI2015

Synergy between pair coupled cluster doubles and pair density functional theory

Pair coupled cluster doubles (pCCD) has been recently studied as a method capable of accounting for static correlation with low polynomial cost. We present three combinations of pCCD with Kohn–Sham functionals of the density and on-top pair density (the probability of finding two electrons on top of each other) to add dynamic correlation to pCCD without double counting. With a negligible increase in computational cost, these pCCD+DFT blends greatly improve upon pCCD in the description of typical problems where static and dynamic correlations are both important. We argue that—as a black-box method with low scaling, size-extensivity, size-consistency, and a simple quasidiagonal two-particle density matrix—pCCD is an excellent match for pair density functionals in this type of fusion of multireference wavefunctions with DFT.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

DOE OSTI2019

Experimental Evaluation of the Fast-Neutron Passive Collar (FNPC) Performance (Interim Report)

This report represents a deliverable in the Safeguards Technology WBS # Project Title, 24.1.3.1, Task 3 (Report on laboratory-based performance testing of the FNPC). A new, simplified method for the verification of the 235 U mass in fresh LEU fuel assemblies that is relatively independent of burnable poison content is being developed. LWR fresh fuel is currently verified using the uranium neutron coincidence collar (UNCL), which relies on an AmLi source to induce fission in the 235 U. AmLi neutrons are slowed down in polyethylene, so the induced fission is primarily due to thermal neutrons. Burnable poisons, which are added to nuclear fuel assemblies to extend the lifetime of the fuel, absorb neutrons that would otherwise have induced fission in the fuel, thereby reducing the neutron count rate and thus the measured 235 U mass. To remove this bias factor a correction is applied based on the operator’s declared burnable poison content. The new technique uses 238 U spontaneous fission in LWR fresh fuel rods to self-interrogate the 235 U mass. The 238 U spontaneous fission neutrons have a hard neutron energy spectrum, which means that the technique is less sensitive to burnable poisons than the AmLi based systems. This project has continued the work done over the past two years to study this new technique using the optimized 3 He based fastneutron passive collar (FNPC). Fabrication of the FNPC has been completed and measurements are in progress using the LANL mockup PWR fuel assembly. Also, preliminary evaluations of advanced analysis techniques and unattended mode operation have been performed. In FY18, the 3 He tube based detector was compared with a 10 B sealed cell based detector for the same set of PWR fuel assemblies in the Rodeo-II program. The results were that the optimized 3 He detector had higher efficiency and less weight than the 10 B based system where both systems had about the same cost. Thus, the FY19 work continued exclusively with the 3 He based detector; although, either system had the potential for future applications. However, the high efficiency of the FPNC (~24%) is critical for precise measurements of the triples rate necessary in the multiplicity advanced analysis technique which can solve for the burnable poison content.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

DOE OSTI2017

Enhancing Mo:BiVO 4 Solar Water Splitting with Patterned Au Nanospheres by Plasmon-Induced Energy Transfer [Rational Nanopositioning for BiVO 4 Solar Water Splitting by Plasmon-induced Energy Transfer]

Here, plasmonic metal nanostructures have been extensively investigated to improve the performance of metal oxide photoanodes for photoelectrochemical (PEC) solar water splitting cells. Most of these studies have focused on the effects of those metal nanostructures on enhancing light absorption and enabling direct energy transfer via hot electrons. However, several recent studies have shown that plasmonic metal nanostructures can improve the PEC performance of metal oxide photoanodes via another mechanism known as plasmon–induced resonant energy transfer (PIRET). However, this PIRET effect has not yet been tested for the molybdenum–doped bismuth vanadium oxide (Mo:BiVO 4 ), regarded as one of the best metal oxide photoanode candidates. Here, this study constructs a hybrid Au nanosphere/Mo:BiVO 4 photoanode interwoven in a hexagonal pattern to investigate the PIRET effect on the PEC performance of Mo:BiVO 4 . This study finds that the Au nanosphere array not only increases light absorption of the photoanode as expected, but also improves both its charge transport and charge transfer efficiencies via PIRET, as confirmed by time–correlated single photon counting and transient absorption studies. As a result, incorporating the Au nanosphere array increases the photocurrent density of Mo:BiVO 4 at 1.23 V versus RHE by ≈2.2–fold (2.83 mA cm –2 ).

36 MATERIALS SCIENCE↗

DOE OSTI2022

Upgrade of Gamma Spectrometry Systems for ORNL TRISO Fuel PIE

Gamma spectrometry is a key element in much of the post-irradiation examination (PIE) work performed under the Advanced Gas Reactor Fuel Development and Qualification (AGR) Program (Demkowicz et al. 2015; Stempien et al. 2021). Gamma spectrometers are integrated into three major capabilities used at the Oak Ridge National Laboratory (ORNL) Irradiated Fuels Examination Laboratory (IFEL) for PIE of tristructural-isotropic (TRISO) coated particles and fuel compacts: the Core Conduction Cooldown Test Facility (CCCTF), the Vertical Counting System (VCS), and the Irradiated Microsphere Gamma Analyzer (IMGA). The CCCTF includes liquid-nitrogen-cooled traps to extract 85 Kr out of the He sweep gas that passes through the furnace in which the fuel compacts are heated during safety testing. Analysis of the 85 Kr activity in the traps is the primary indicator for TRISO failure during safety testing. The VCS is a system used to accurately measure gamma emission from components placed in a lead-shielded chamber. It is used to count the CCCTF deposition cups after removal from furnace. Each cup resides in the CCCTF furnace for typically 12–24 h and is periodically replaced with a fresh cup throughout the safety test. Metallic fission products collect on the water-cooled cups and several gamma-emitting isotopes ( 110 mAg, 134 Cs, 137 Cs, 154 Eu, and 155 Eu) are often measured and provide indication of the retention performance of the TRISO coatings. The VCS is also used to measure the presence of these isotopes on the CCCTF tantalum liner and sweep gas inlet tube for the determination of cup collection efficiency, as well as support other gamma spectrometry needs related to calibration of the 85 Kr fission gas traps and various other special PIE tasks. The IMGA uses gamma spectrometry to measure the inventory of gamma-emitting isotopes in individual TRISO particles. An automated particle handling system within the IMGA hot cell removes each particle from a source vial and positions it in front of a gamma detector, and output from the gamma spectrometer is used by the IMGA software to determine a destination vial such that particles are sorted according to their inventory and retention characteristics. At the conclusion of the AGR-1 and AGR-2 PIE campaigns, the gamma spectrometer systems used at ORNL to support that PIE had reached the end of its life cycle due to gradual obsolescence of the hardware and software. Upgrade of the Canberra Genie 2000 software used by these systems to a Windows 10 version was not a viable option, because the newest Windows 10 version offered by Mirion (the new owner of the Canberra technology) did not include the dynamic-link libraries (DLLs) needed for integration with the custom PIE software used with the CCCTF and IMGA, and Mirion had no current plans for development and release of Windows 10 versions of these DLLs with the Model S560 Genie 2000 Programming Library. Ultimately a switch was made to ORTEC gamma spectrometry systems, which appeared to be a more sustainable solution due to more proactive vendor support. The ORTEC conversion involved replacing the aging detector preamplifier and multichannel analyzer (MCA) hardware, upgrading the obsolete Windows 7 computers to Windows 10 compatible models, adopting ORTEC GammaVision software, and extensive modification of the ORNL-developed Visual Basic .NET (VB.NET) programs that provide the CCCTF and IMGA user interfaces.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

DOE OSTI2016

Prostate extracellular vesicles in patient plasma as a liquid biopsy platform for prostate cancer using nanoscale flow cytometry

Background: Extracellular vesicles released by prostate cancer present in seminal fluid, urine, and blood may represent a non-invasive means to identify and prioritize patients with intermediate risk and high risk of prostate cancer. We hypothesize that enumeration of circulating prostate microparticles (PMPs), a type of extracellular vesicle (EV), can identify patients with Gleason Score≥4+4 prostate cancer (PCa) in a manner independent of PSA. Patients and Methods: Plasmas from healthy volunteers, benign prostatic hyperplasia patients, and PCa patients with various Gleason score patterns were analyzed for PMPs. We used nanoscale flow cytometry to enumerate PMPs which were defined as submicron events (100-1000nm) immunoreactive to anti-PSMA mAb when compared to isotype control labeled samples. Levels of PMPs (counts/µL of plasma) were also compared to CellSearch CTC Subclasses in various PCa metastatic disease subtypes (treatment naïve, castration resistant prostate cancer) and in serially collected plasma sets from patients undergoing radical prostatectomy. Results: PMP levels in plasma as enumerated by nanoscale flow cytometry are effective in distinguishing PCa patients with Gleason Score≥8 disease, a highrisk prognostic factor, from patients with Gleason Score≤7 PCa, which carries an intermediate risk of PCa recurrence. PMP levels were independent of PSA and significantly decreased after surgical resection of the prostate, demonstrating its prognostic potential for clinical follow-up. CTC subclasses did not decrease after prostatectomy and were not effective in distinguishing localized PCa patients from metastatic PCa patients. Conclusions: PMP enumeration was able to identify patients with Gleason Score ≥8 PCa but not patients with Gleason Score 4+3 PCa, but offers greater confidence than CTC counts in identifying patients with metastatic prostate cancer. CTC Subclass analysis was also not effective for post-prostatectomy follow up and for distinguishing metastatic PCa and localized PCa patients. Nanoscale flow cytometry of PMPs presents an emerging biomarker platform for various stages of prostate cancer.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2021

Blinking statistics and molecular counting in direct stochastic reconstruction microscopy (dSTORM)

Abstract Motivation Many recent advancements in single-molecule localization microscopy exploit the stochastic photoswitching of fluorophores to reveal complex cellular structures beyond the classical diffraction limit. However, this same stochasticity makes counting the number of molecules to high precision extremely challenging, preventing key insight into the cellular structures and processes under observation. Results Modelling the photoswitching behaviour of a fluorophore as an unobserved continuous time Markov process transitioning between a single fluorescent and multiple dark states, and fully mitigating for missed blinks and false positives, we present a method for computing the exact probability distribution for the number of observed localizations from a single photoswitching fluorophore. This is then extended to provide the probability distribution for the number of localizations in a direct stochastic optical reconstruction microscopy experiment involving an arbitrary number of molecules. We demonstrate that when training data are available to estimate photoswitching rates, the unknown number of molecules can be accurately recovered from the posterior mode of the number of molecules given the number of localizations. Finally, we demonstrate the method on experimental data by quantifying the number of adapter protein linker for activation of T cells on the cell surface of the T-cell immunological synapse. Availability and implementation Software and data available at https://github.com/lp1611/mol_count_dstorm. Supplementary information Supplementary data are available at Bioinformatics online.

Patel, Lekha↗

DOE OSTI2018

Molecular Properties That Define the Activities of Antibiotics in Escherichia coli and Pseudomonas aeruginosa

The permeability barrier of Gram-negative cell envelopes is the major obstacle in the discovery and development of new antibiotics. In Gram-negative bacteria, these difficulties are exacerbated by the synergistic interaction between two biochemically distinct phenomena, the low permeability of the outer membrane (OM) and active multidrug efflux. In this study, we used Pseudomonas aeruginosa and Escherichia coli strains with controllable permeability barriers, achieved through hyperporination of the OMs and varied efflux capacities, to evaluate the contributions of each of the barriers to protection from antibacterials. We analyzed antibacterial activities of β-lactams and fluoroquinolones, antibiotics that are optimized for targets in the periplasm and the cytoplasm, respectively, and performed a machine learning-based analysis to identify physicochemical descriptors that best classify their relative potencies. Our results show that the molecular properties selected by active efflux and the OM barriers are different for the two species. Antibiotic activity in P. aeruginosa was better classified by electrostatic and surface area properties, whereas topology, physical properties, and atom or bond counts best capture the behavior in E. coli . In several cases, descriptor values that correspond to active antibiotics also correspond to significant barrier effects, highlighting the synergy between the two barriers where optimizing for one barrier promotes strengthening of the other barrier. Furthermore, both barriers should be considered when optimizing antibiotics for favorable OM permeability, efflux evasion, or both.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS2007

Laser Sounder for Global Measurement of CO2 Concentrations in the Troposphere from Space

Measurements of tropospheric CO2 abundance with global-coverage, a few hundred km spatial and monthly temporal resolution are needed to quantify processes that regulate CO2 storage by the land and oceans. The Orbiting Carbon Observatory (OCO) is the first space mission focused on atmospheric CO2 for measuring total column CO, and O2 by detecting the spectral absorption in reflected sunlight. The OCO mission is an essential step, and will yield important new information about atmospheric CO2 distributions. However there are unavoidable limitations imposed by its measurement approach. These include best accuracy only during daytime at moderate to high sun angles, interference by cloud and aerosol scattering, and limited signal from CO2 variability in the lower tropospheric CO2 column. We have been developing a new laser-based technique for the remote measurement of the tropospheric CO2 concentrations from orbit. Our initial goal is to demonstrate a lidar technique and instrument technology that will permit measurements of the CO2 column abundance in the lower troposphere from aircraft. Our final goal is to develop a space instrument and mission approach for active measurements of the CO2 mixing ratio at the 1-2 ppmv level. Our technique is much less sensitive to cloud and atmospheric scattering conditions and would allow continuous measurements of CO2 mixing ratio in the lower troposphere from orbit over land and ocean surfaces during day and night. Our approach is to use the 1570nm CO2 band and a 3-channel laser absorption spectrometer (i.e. lidar used an altimeter mode), which continuously measures at nadir from a near polar circular orbit. The approach directs the narrow co-aligned laser beams from the instrument's lasers toward nadir, and measures the energy of the laser echoes reflected from land and water surfaces. It uses several tunable fiber laser transmitters which allowing measurement of the extinction from a single selected CO2 absorption line in the 1570 nm band. This band is free from interference from other gases and has temperature insensitive absorption lines. During the measurement the lasers are tuned on- and off- a selected CO2 line near 1572 nm and a selected O2 line near 768 nm in the Oxygen A band at kHz rates. The lasers use tunable diode seed lasers followed by fiber amplifiers, and have spectral widths much narrower than the gas absorption lines. The receiver uses a 1-m diameter telescope and photon counting detectors and measures the background light and energies of the laser echoes from the surface. The extinction and column densities for the CO2 and O2 gases are estimated from the ratio of the on and offline surface echo via the differential optical absorption technique. Our technique rapidly alternates between several on-line wavelengths set to the sides of the selected gas absorption lines. It exploits the atmospheric pressure broadening of the lines to weight the measurement sensitivity to the atmospheric column below 5 km. This maximizes sensitivity to CO2 in the boundary layer, where variations caused by surface sources and sinks are largest. Simultaneous measurements of O2 column will use an identical approach with an O2 line. Thee laser frequencies are tunable and have narrow (MHz) line widths. In combination with sensitive photon counting detectors these enables much higher spectral resolution and precision than is possible with passive spectrometer. 1aser backscatter profiles are also measured, which permits identifying measurements made to cloud tops and through aerosol layers. The measurement approach using lasers in common-nadir-zenith path allows retrieving CO2 column mixing ratios in the lower troposphere irrespective of sun angle. Pulsed laser signals, time gated receiver and a narrow receiver field-of-view are used to isolate the surface laser echo signals and to exclude photons scattered from clouds and aerosols. Nonetheless, the optical absorption change due to a change of a few ppO2 is small, <1 % which makes achieving the needed measurement sensitivities and stabilities quite challenging. Measurement SNRs and stabilities of >600:1 are needed to estimate CO2 mixing ratio at the 1-2 ppm level. We have calculated characteristics of the technique and have demonstrated aspects of the laser, detector and receiver approaches in th e laboratory We have also measured O2 in an absorption cell, and made C02 measurements over a 400 m long (one way) horizontal path using a sensor breadboard. We will describe these and more details of our approach in the paper.

Abshire, James B.↗

DOE OSTI2020

Maximising the investment returns of a grid‐connected battery considering degradation cost

Energy storage systems (ESSs) are being deployed widely due to numerous benefits including operational flexibility, high ramping capability, and decreasing costs. This study investigates the economic benefits provided by battery ESSs when they are deployed for market‐related applications, considering the battery degradation cost. A comprehensive investment planning framework is presented, which estimates the maximum revenue that the ESS can generate over its lifetime and provides the necessary tools to investors for aiding the decision making process regarding an ESS project. The applications chosen for this study are energy arbitrage and frequency regulation. Lithium‐ion batteries are considered due to their wide popularity arising from high efficiency, high energy density, and declining costs. A new degradation cost model based on energy throughput and cycle count is developed for Lithium‐ion batteries participating in electricity markets. The lifetime revenue of ESS is calculated considering battery degradation and a cost–benefit analysis is performed to provide investors with an estimate of the net present value, return on investment and payback period. The effect of considering the degradation cost on the estimated revenue is also studied. The proposed approach is demonstrated on the IEEE Reliability Test System and historical data from PJM Interconnection.

25 ENERGY STORAGE↗

DOE OSTI2016

SU-G-IeP4-08: Initial Investigations of Up-Converting Nanoparticles (UCNP) for 3D Tissue Imaging in Optical-ECT

Purpose: Near-IR absorptive up-converting nanoparticles (UCNPs) is a novel contrast for optical-ECT that allows auto-fluorescence-free 3D imaging of labeled cells in a matrix of large (∼1cm{sup 3}) unsectioned normal tissue. This has the potential to image small metastases or dormant cells that is difficult with down-converting fluorescing dyes due to auto-fluorescence. The feasibility of imaging UCNP in agarose phantoms and a mouse lung is demonstrated, aided by a 3D-printed optical-ECT stage designed to excite UCNP in a mouse lung. Methods: The UCNP, NaYF{sub 4}:Yb/Er (20/2%), studied in this work up-converts 980nm light to visible light peaking sharply at ∼540nm. To characterize the UCNP emission as a function of UCNP concentration, cylindrical 2.5%wt agarose phantoms infused with UCNP at concentrations of 25µg/mL and 50µg/mL were exposed to 1.5W 980nm laser coupled to an optical fiber. The fiber was held stably at 1cm above the stage via a custom 3D-printed stage. An optically cleared lung harvested from a BALBc mice was then injected with 100µL of 1mg/mL UCNP solution ex vivo. Tomographic imaging of the UCNP emission in lung was performed. Results: The laser beam tract is visualized within the agarose phantom. A line profile of UCNP emission at 25µg/mL versus 50µg/mL shows that increasing the UCNP concentration increases emission count. UCNPs injected into a cleared mouse lung disperse throughout the respiratory tract, allowing for visualization and 3D reconstruction. Excitation before and after UCNP injection shows the tissue exhibits no auto-fluorescence at 980nm, allowing clear view of the UCNP without any obscuring features such as conventional down-converting fluorescent tags. Conclusion: We confirm that up-conversion in tissue circumvents completely tissue auto-fluorescence, which allowed background-free 3D reconstruction of the UCNP distribution. We also confirm that raising the UCNP concentration increases emission and that UCNPs are retained in agarose samples during the optical clearing process.

60 APPLIED LIFE SCIENCES↗

NASA NTRSDate not supplied

Center for Isotope Cosmochemistry and Geochronology at NASA Johnson Space Center

Ten years in the making the Center for Isotope Cosmochemistry and Geochronology (CICG) at NASA Johnson Space Center is designed to be a cooperative analytical facility where novel techniques are developed and performed. Our team works collaboratively to operate this integrated lab suite comprising sample preparation, metal-free clean chemistry, and mass spectrometers to measure a wide variety of elements and isotopes in astromaterials. Contamination control is enacted throughout by a positive pressure cascade of HEPA filtered air, weekly surface cleaning and particle counting. Astromaterials processing is performed with application-specific mortar and pestles, heavy liquid separation, a Frantz magnetic separator, as well as hand-picking using optical microscopes in laminar flow boxes. Microsampling is also carried out using a New Wave MicroMill. Elements for isotopic analysis are isolated from acid-digested samples using ion-exchange chromatography. Cross-contamination is minimized with dedicated laminar flow boxes and labware for different purification protocols. Instrumentation includes a ThermoFisher Element XR inductively coupled plasma mass spectrometer (ICP-MS) for quantification of elemental concentrations in dissolved sample solutions. For in situ analysis, a Teledyne Photon Machines Analyte Excite+ laser ablation system is coupled to the Element XR. The ThermoFisher Triton thermal ionization mass spectrometer (TIMS) is used to measure the isotopic composition of elements such as CA, K, Cr, Rb, Sr, Sm, Nd at high precision in processed samples. For a more diverse range of isotopic analyses, CICG houses a Nu Sapphire 1700 multi-collector inductively coupled plasma mass spectrometer (MC-ICP-MS), which is a large geometry instrument with a multi-pole collision cell for interference removal. An Applied Spectra Instruments J200 femto-second laser is planned for install this Fall and will be coupled to the Nu Sapphire 1700. NASA’s Internal Scientist Funding Model (ISFM)supports this facility. Advanced analytical geochemistry techniques are applied to targeted astromaterials to address important problems in planetary sciences, with a longer-term goal of preparing for samples collected and returned by space missions. CICG is open to collaborations on projects that utilize the facility’s unique capabilities and are within its research scope. We are particularly eager to support and share expertise with the next generation of planetary scientists.

Justin I Simon↗

DOE OSTI2018

Virologic outcome among patients receiving antiretroviral therapy at five hospitals in Haiti

Introduction Viral load (VL) assessment is the preferred method for diagnosing and confirming virologic failure for patients on antiretroviral therapy (ART). We conducted a retrospective cross-sectional study to evaluate the virologic suppression rate among patients on ART for 6 months in five hospitals around Port-au-Prince, Haiti. Methods Plasma VL was measured and patients with VL <1,000 copies/mL were defined as virologically suppressed. A second VL test was performed within at least six months of the first test. Factors associated with virologic suppression were analyzed using logistic regression models accounting for site-level clustering using complex survey procedures. Results Data were analyzed for 2,313 patients on ART for six months or longer between July 2013 and February 2015. Among them, 1,563 (67.6%) achieved virologic suppression at the first VL test. A second VL test was performed within at least six months for 718 (31.0%) of the patients. Of the 459 patients with an initial HIV-1 RNA <1,000 copies/mL who had a second VL performed, 394 (85.8%) maintained virologic suppression. Virologic suppression was negatively associated with male gender (adjusted odds ratio [aOR]: 0.80, 95% CI: 0.74– 0.0.86), 23 to 35 months on ART (aOR:0.72[0.54–0.96]), baseline CD4 counts of 201–500 cells/mm 3 and 200 cells/mm 3 or lower (aORs: 0.77 [0.62–0.95] and 0.80 [0.66–0.98], respectively), poor adherence (aOR: 0.69 [0.59–0.81]), and TB co-infection (aOR: 0.73 [0.55–0.97]). Conclusions This study showed that over two-thirds of the patients in this evaluation achieved virologic suppression after six months on ART and the majority of them remained suppressed. These results reinforce the importance of expanding access to HIV-1 viral load testing in Haiti for monitoring ART outcomes.

59 BASIC BIOLOGICAL SCIENCES↗

NASA NTRS1991

Acridine orange staining reaction as an index of physiological activity in Escherichia coli

The assumption that the acridine orange (AO) color reaction may be used as an index of physiological activity was investigated in laboratory grown Escherichia coli. Spectrofluorometric observations of purified nucleic acids, ribosomes and the microscopic color of bacteriophage-infected cells stained with AO confirmed the theory that single-stranded nucleic acids emit orange to red fluorescence while those that are double-stranded fluoresce green in vivo. Bacteria growing actively in a rich medium could be distinguished from cells in stationary phase by the AO reaction. Cells from log phase appeared red, whereas those in stationary phase were green. However, this differentiation was not seen when the bacteria were grown in a minimal medium or when a variation of the staining method was used. Also, shifting bacteria in stationary phase to starvation conditions rapidly changed their AO staining reaction. Boiling and exposure to lethal concentrations of azide and formalin resulted in stationary-phase cells that appeared red after staining but bacteria killed with chlorine remained green. These findings indicate that the AO staining reaction may be suggestive of physiological activity under defined conditions. However, variables in staining and fixation procedures as well as uncertainties associated with mixed bacterial populations in environmental samples may produce results that are not consistent with the classical interpretation of this reaction. The importance of validating the putative physiological implications of this staining reaction is stressed.

NASA Discipline Number 04-10↗

NASA NTRS2022

Terrestrial Exospheric Dayside H-density Profile at 3 - 15 RE from UVIS/HDAC and TWINS Lyman-alpha Data Combined

Terrestrial ecliptic dayside observations of the exospheric Lyman-α column intensity between 3–15 Earth radii (RE) by UVIS/HDAC (UVIS – ultraviolet imaging spectrograph; HDAC – hydrogen-deuterium absorption cell) Lyman-α photometer at CASSINI have been analyzed to derive the neutral exospheric H-density profile at the Earth’s ecliptic dayside in this radial range. The data were measured during CASSINI’s swing-by maneuver at the Earth on 18 August 1999 and are published by Werner et al. (2004). In this study the dayside HDAC Lyman-α observations published by Werner et al. (2004) are compared to calculated Lyman-α intensities based on the 3D H-density model derived from TWINS (Two Wide-angle Imaging Neutral-atom Spectrometers) Lyman-α observations between 2008–2010 (Zoennchen et al., 2015). It was found that both Lyman-α profiles show a very similar radial dependence in particular between 3–8RE. Between 3.0–5.5RE impact distance Lyman-α observations of both TWINS and UVIS/HDAC exist at the ecliptic dayside. In this overlapping region the cross-calibration of the HDAC profile against the calculated TWINS profile was done, assuming that the exosphere there was similar for both due to comparable space weather conditions. As a result of the cross-calibration the conversion factor between counts per second and rayleigh, fc = 3.285 counts s−1R−1, is determined for these HDAC observations. Using this factor, the radial H-density profile for the Earth’s ecliptic dayside was derived from the UVIS/HDAC observations, which constrained the neutral H density there at 10RE to a value of 35 cm-3. Furthermore, a faster radial H-density decrease was found at distances above 8RE (≈r−2.37) compared to the lower distances of 3–7RE (≈r−2.37). This increased loss of neutral H above 8RE might indicate a higher rate of H ionization in the vicinity of the magnetopause at 9–11RE (near subsolar point) and beyond, because of increasing charge exchange interactions of exospheric H atoms with solar wind ions outside the magnetosphere.

Jochen H Zoennchen↗

NASA NTRS1998

Effects of starvation on physiological activity and chlorine disinfection resistance in Escherichia coli O157:H7

Escherichia coli O157:H7 can persist for days to weeks in microcosms simulating natural conditions. In this study, we used a suite of fluorescent, in situ stains and probes to assess the influence of starvation on physiological activity based on membrane potential (rhodamine 123 assay), membrane integrity (LIVE/DEAD BacLight kit), respiratory activity (5-cyano-2,3-di-4-tolyl-tetrazolium chloride assay), intracellular esterase activity (ScanRDI assay), and 16S rRNA content. Growth-dependent assays were also used to assess substrate responsiveness (direct viable count [DVC] assay), ATP activity (MicroStar assay), and culturability (R2A agar assay). In addition, resistance to chlorine disinfection was assessed. After 14 days of starvation, the DVC values decreased, while the values in all other assays remained relatively constant and equivalent to each other. Chlorine resistance progressively increased through the starvation period. After 29 days of starvation, there was no significant difference in chlorine resistance between control cultures that had not been exposed to the disinfectant and cultures that had been exposed. This study demonstrates that E. coli O157:H7 adapts to starvation conditions by developing a chlorine resistance phenotype.

NASA Discipline Environmental Health↗

NASA NTRS2011

Standardization of Spore Inactivation Method for PMA-PhyloChip Analysis

In compliance with the Committee on Space Research (COSPAR) planetary protection policy, National Aeronautics and Space Administration (NASA) monitors the total microbial burden of spacecraft as a means for minimizing the inadvertent transfer of viable contaminant microorganisms to extraterrestrial environments (forward contamination). NASA standard assay-based counts are used both as a proxy for relative surface cleanliness and to estimate overall microbial burden as well as to assess whether forward planetary protection risk criteria are met for a given mission, which vary by the planetary body to be explored and whether or not life detection missions are present. Despite efforts to reduce presence of microorganisms from spacecraft prior to launch, microbes have been isolated from spacecraft and associated surfaces within the extreme conditions of clean room facilities using state of the art molecular technologies. Development of a more sensitive method that will better enumerate all viable microorganisms from spacecraft and associated surfaces could support future life detection missions. Current culture-based (NASA standard spore assay) and nucleic-acid-based polymerase chain reaction (PCR) methods have significant shortcomings in this type of analysis. The overall goal of this project is to evaluate and validate a new molecular method based on the use of a deoxyribonucleic acid (DNA) intercalating agent propidium monoazide (PMA). This is used in combination with DNA microarray (PhyloChip) which has been shown to identify very low levels of organisms on spacecraft associated surfaces. PMA can only penetrate the membrane of dead cells. Once penetrated, it intercalates the DNA and, upon photolysis using visible light it produces stable DNA monoadducts. This allows DNA to be unavailable for further PCR analysis. The specific aim of this study is to standardize the spore inactivation method for PMA-PhyloChip analysis. We have used the bacterial spores Bacillus subtilis 168 (standard laboratory isolate) as a test organism.

spore inactivation↗

DOE OSTI2026

Online Alpha Monitoring of High Cs-137 Hanford and SRS High Level Waste with Tensioned Metastable Fluid Detectors

The Department of Energy’s Hanford and Savannah River Sites maintain millions of gallons of caustic supernate and salt high activity waste in their high-level waste (HLW) tank farm inventories. The Savannah River Site is currently treating this waste with a calixarene-based solvent extraction of Cs-137 to reduce these inventories. Hanford is employing an at-tank crystalline silico titanate (CST) based solid phase extraction methodology to reduce their liquid HLW inventories. Due to the high solubility of Cs-137 and the relative insolubility of the actinides in these caustic waste forms, the beta to alpha radioactivity ratio can often exceed six orders of magnitude in the feed solutions to these treatment processes. This unique characteristic leads to significant technical challenges in making rapid gross alpha measurements in the presence of the overwhelming beta, gamma, as well as dissolved sodium salt in these HLW matrices. Conventional radioanalytical techniques, such as liquid scintillation analysis or gas flow proportional counting require significant radiochemistry preparation prior to the radiometric measurements for gross alpha activity. These required pretreatments render these technologies untenable for rapid quantification of gross alpha activity that could be required to support on or at-line measurements ensuring a waste stream will meet regulatory requirements. The radiation measurement properties of Tensioned Metastable Fluid Detectors (TMFDs) have been studied by Purdue University’s Taleyarkhan research group for well over a decade. Fluids tensioned to the appropriate degree will rupture when struck by radiation, resulting in a measurable cavitation event. The negative pressure generating this tension can be adjusted by centrifugal rotation or by acoustic means in such a way that these cavitation events can be generated from alpha radiation but will not be generated by beta or gamma radiation. Purdue University and the Savannah River National Laboratory are currently collaborating to develop a gamma/beta blind, spectroscopic alpha measurement system based on the Tensioned Metastable Fluid Detector technology to provide a potential solution for performing rapid gross alpha measurements on these high gamma/beta sample matrices. Measurements using the Indirect Drive Acoustically Tensioned Metastable Fluid Detectors developed as part of this collaboration were performed with an alpha emitting radionuclide. Successful determination of gross alpha activity was observed, indicating a potential pathway for rapid gross alpha measurements in remote-handled shielded cells or in process situations requiring online alpha monitoring. Measurements using this system have been conducted on high beta activity solutions, demonstrating the beta blind capability of this system. Measurements are currently underway to test the system’s capability to measure gross alpha activity on Savannah River Site high level waste high Cs-137 samples that have been measured by the SRNL radiochemistry team. This work was supported by the DOE EM Technology Development program.

DiPrete, David [Savannah River National Laboratory↗