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DOE OSTI2019

Persistent Viral Reservoirs in Lymphoid Tissues in SIV-Infected Rhesus Macaques of Chinese-Origin on Suppressive Antiretroviral Therapy

Understanding HIV latent reservoirs in tissues is essential for the development of new strategies targeting these sites for eradication. Here, we assessed the size of latent reservoirs and the source of residual viruses in multiple lymphoid tissues of SIV-infected and fully suppressed rhesus macaques of Chinese-origin (cRMs). Eight cRMs were infected with SIVmac251 and treated with tenofovir and emtricitabine daily for 24 weeks initiated 4 weeks post-infection. Four of the eight animals reached sustained full viral suppression with undetectable viremia. The levels of cell-associated SIV DNA varied in peripheral blood mononuclear cells (PBMCs) and multiple lymphoid tissues, but with higher levels in the mesenteric lymph nodes (MesLNs). The levels of cell-associated SIV RNA also varied in different tissues. The higher frequency of viral RNA detection in the MesLNs was also observed by in situ hybridization. Consistently, the infection unit per million cells (IUPM) in the MesLNs was higher than in PBMCs and other tested lymphoid tissues by quantitative viral outgrowth assay (QVOA). Furthermore, env gp120 from tissue SIV RNA was amplified by single genome amplification. Phylogenetic analysis revealed diverse variants from tissues parallel to the viral inoculum in all viral suppressed animals. These results demonstrate that the latency and viral reservoirs in the lymphoid tissues still exist in aviremic macaques under full suppressive therapy. Moreover, the size of viral latent reservoirs differs in various lymphoid tissues with a relatively larger size in the MesLNs.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2018

Benchtop-compatible sample processing workflow for proteome profiling of < 100 mammalian cells

Extending proteomics to smaller samples can enable the mapping of protein expression across tissues with high spatial resolution and can reveal sub-group heterogeneity. However, despite the continually improving sensitivity of LC-MS instrumentation, in-depth profiling of samples containing low-nanogram amounts of protein has remained challenging due to analyte losses incurred during preparation and analysis. To address this, we recently developed nanodroplet processing in one pot for trace samples (nanoPOTS), a robotic/microfluidic platform that generates ready-to-analyze peptides from cellular material in ~200 nL droplets with greatly reduced sample losses. In combination with ultrasensitive LC-MS, nanoPOTS has enabled >3000 proteins to be confidently identified from as few as 10 cultured human cells and ~700 proteins from single cells. However, the nanoPOTS platform requires a highly skilled operator and a costly in-house-built robotic nanopipetting instrument. In this work, we sought to evaluate the extent to which the benefits of nanodroplet processing could be preserved when upscaling reagent dispensing volumes by a factor of 10 to those addressable by commercial micropipette. We characterized the resulting platform, termed microdroplet processing in one pot for trace samples (µPOTS), for the analysis of as few as ~25 cultured HeLa cells (4 ng total protein) or 50 µm square mouse liver tissue thin sections and found that ~1800 and ~1200 unique proteins were respectively identified with high reproducibility. As such, the reduced equipment requirements should facilitate broad dissemination of nanoproteomics workflows by obviating the need for a capital-intensive custom liquid handling system.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2017

Study of the specific features of single-crystal boron microstructure

A complex study of the structure of β-boron single crystal grown by the floating-zone method, with sizes significantly exceeding the analogs known in the literature, has been performed. The study includes X-ray diffraction analysis and X-ray diffractometry (measurement of pole figures and rocking curves), performed on both laboratory and synchrotron sources; atomic-resolution scanning transmission electron microscopy with spherical aberration correction; and energy-dispersive microanalysis. X-ray diffraction analysis using synchrotron radiation has been used to refine the β-boron structure and find impurity Si atoms. The relative variations in the unit-cell parameters a and c for the crystal bulk are found to be δa/a ≈ 0.4 and δc/c ≈ 0.1%. X-ray diffractometry has revealed that the single-crystal growth axis coincides with the [22̄013] crystallographic axis and makes an angle of 21.12° with the [0001] threefold axis. Electron microscopy data have confirmed that the sample under study is a β-boron crystal, which may contain 0.3–0.4 at % Si as an impurity. Planar defects (stacking faults and dislocations) are found. The results of additional measurements of the temperature dependence of the thermal conductivity of the crystal in the range of 50–300 K are indicative of its high structural quality.

36 MATERIALS SCIENCE↗

NASA NTRS2014

High Efficiency Quantum Well Waveguide Solar Cells and Methods for Constructing the Same

Photon absorption, and thus current generation, is hindered in conventional thin-film solar cell designs, including quantum well structures, by the limited path length of incident light passing vertically through the device. Optical scattering into lateral waveguide structures provides a physical mechanism to increase photocurrent generation through in-plane light trapping. However, the insertion of wells of high refractive index material with lower energy gap into the device structure often results in lower voltage operation, and hence lower photovoltaic power conversion efficiency. The voltage output of an InGaAs quantum well waveguide photovoltaic device can be increased by employing a III-V material structure with an extended wide band gap emitter heterojunction. Analysis of the light IV characteristics reveals that non-radiative recombination components of the underlying dark diode current have been reduced, exposing the limiting radiative recombination component and providing a pathway for realizing solar-electric conversion efficiency of 30% or more in single junction cells.

Welser, Roger E.↗

NASA NTRS2018

High Efficiency Quantum Well Waveguide Solar Cells and Methods for Constructing the Same

Welser, Roger E.↗

DOE OSTI2019

High spatial resolution imaging of biological tissues using nanospray desorption electrospray ionization mass spectrometry

Mass spectrometry imaging (MSI) enables label-free spatial mapping of hundreds of biomolecules in tissue sections. This capability provides valuable information on tissue heterogeneity that is difficult to obtain using population-averaged assays. Despite substantial developments in both instrumentation and methodology, MSI of tissue samples at single-cell resolution remains challenging. In our research, we describe a protocol for robust imaging of tissue sections with a high (better than 10-μm) spatial resolution using nanospray desorption electrospray ionization (nano-DESI) mass spectrometry, an ambient ionization technique that does not require sample pretreatment before analysis. In this protocol, mouse uterine tissue is used as a model system to illustrate both the workflow and data obtained in these experiments. We provide a detailed description of the nano-DESI MSI platform, fabrication of the nano-DESI and shear force probes, shear force microscopy experiments, spectral acquisition, and data processing. A properly trained researcher (e.g., technician, graduate student, or postdoc) can complete all the steps from probe fabrication to data acquisition and processing within a single day. We further describe a new strategy for acquiring both positive- and negative-mode imaging data in the same experiment. This is achieved by alternating between positive and negative data acquisition modes during consecutive line scans. Using our imaging approach, hundreds of high-quality ion images were obtained from a single uterine section. This protocol enables sensitive and quantitative imaging of lipids and metabolites in heterogeneous tissue sections with high spatial resolution, which is critical to understanding biochemical processes occurring in biological tissues.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2013

Evaluation of the use of color-set geometry during lattice physics constants generation for boiling water reactor simulation

Current methods for BWR nuclear design and analysis consist of using lattice physics neutron transport methods to generate the two-group homogenized cross-sections that are then used in a nodal diffusion theory code. The lattice transport solutions are performed for a single assembly with reflective boundary conditions, which is a practical approximation. A method is developed to account for assembly exposure distributions (environment) in the core within the lattice transport calculations with the use of color-sets (2x2) geometry. The loading pattern is examined and an appropriate number of characteristic color-set cells are selected for analysis. Treatment of the co-resident exposed fuel within this method is also presented. The calculation process was followed for a recent BWR cycle design with comparisons being performed on both a lattice and core-wide basis to evaluate the proposed method. The lattice based comparisons show noticeable differences in the pin power distribution predictions, which require further investigation to see how this translates into core performance calculations. The core-wide comparisons show minor differences and are generally in a good agreement, which is expected with this small perturbation. A slight improvement was noticed in the reduction of the power distribution uncertainty. However, given the additional amount of work and computer run time increase, further evaluation, especially of core pin power predictions, is needed to consider this method for production level design and safety analysis calculations. (authors)

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

DOE OSTI2023

Mechanically induced thermal runaway severity analysis for Li-ion batteries

Thermal runaway is one of the most important safety concerns in the deployment of Li-ion batteries. For this study, we developed a standardized single-side indentation test protocol to induce an internal short-circuit. Cell voltage, temperature, and applied compressive force are monitored as a function of time. Each cell is given an observed hazard severity (OHS in five categories) modified from the EUCAR table. Meanwhile a calculated hazard severity (CHS) is obtained solely based on temperature and voltage curves. The calculation formula considers the cell temperature, rate of temperature increase, cell capacity, state of charge (SOC), voltage drop, and voltage drop rate. Each term is assigned with an appropriate weight to scale the calculated score from 0 to100. This method was applied to >100 Li-ion batteries with various SOCs and chemistries, and their OHS and CHS were displayed against SOC. The purpose is to provide battery designers, manufacturers, and end-users a clear comparison of thermal runaway severity of different batteries.

25 ENERGY STORAGE↗

DOE OSTI2018

First Measurements of the Parity-Violating and Beam-Normal Single-Spin Asymmetries in Elastic Electron-Aluminum Scattering

The Q weak collaboration has made the first measurements of the elastic parity-violating and beam-normal single-spin asymmetries from the 27 Al nucleus. Both are the result of ancillary measurements conducted during the Q weak experiment at Jefferson Lab. The goal of the experimental was to determine the proton's weak charge, Q p W , via a measurement of the elastic parity-violating electron-proton scattering asymmetry. During the experiment, ancillary measurements were made with different beam configurations on a separate aluminum alloy target, in an effort to directly measure the aluminum background coming from the experiment's liquid hydrogen target cell. This dissertation discusses three primary results: the parity-violating 27 Al asymmetry analysis used to correct for the aluminum target background in the final Q weak analysis, its extended analysis leading to the extraction of the pure elastic parity-violating 27 Al asymmetry, and the determination of the elastic beam-normal single-spin 27 Al asymmetry. The parity-violating result was also used to make a semi model-independent determination of 27 Al neutron distribution radius, an important test for models used to describe neutron-rich matter. The beam-normal single-spin asymmetry stands to possibly shed light on an observed disagreement between theory and a previous measurement performed on 208 Pb, as 27 Al is the next highest atomic mass nucleus to have this observable measured. The elastic parity-violating 27 Al asymmetry was found to be 1.927 +/- 0.173 ppm at (Q 2 ) = 0.0236 +/- 0.0001 GeV 2 . This measured parity-violating asymmetry implies a 27 Al neutron distribution radius of 3.024 +/- 0.104 fm. Calculating the difference between this radius and the 27 Al proton distribution radius yields the neutron skin, which was found to be 0.092 +/- 0.104 fm. This skin value is consistent with zero, within its uncertainty, and it confirms the naive expectation for a light nucleus like 27 Al. The beam-normal single-spin 27 Al asymmetry was found to be -16.322 +/- 2.679 ppm at &lang;Q&rang; = 0.154 GeV. This value agrees with the previous observed trend of beam-normal single-spin asymmetries measured from light nuclei, which motivates the need for future measurements of higher atomic mass nuclei.

Source record↗

DOE OSTI2021

Scaling-up up-flow microbial electrolysis cells with a compact electrode configuration for continuous hydrogen production

Maintaining high current densities is a key challenge in scaling-up microbial electrolysis cell (MEC) reactors. In this study, a novel 10 L MEC reactor with a total electrode surface area > 1 m 2 was designed and evaluated to maximize the current density and H 2 recovery. Performances of the reactor suggest that the longitudinal structure with parallel vertical orientation of the electrodes encouraged high fluid mixing and the sheet metal electrode frames provided distributed electrical connection. Results also demonstrated that the electrode pairs located next to reactor walls decreased current density, as did separating the electrodes with separators. High volumetric H 2 production rate of 5.9 L/L/d was achieved at a volumetric current density of 970 A/m 3 (34 A/m 2 ). Furthermore, the observed current densities of the large reactor were accurately predicted based on the internal resistance analysis of small scale MECs (0.15 L), demonstrating the scalability of the single chamber MEC design.

08 HYDROGEN↗

DOE OSTI2016

Synthesis and characterization of nanostructured titanium carbide for fuel cell applications

Titanium carbide (TiC) nanoparticles have been successfully synthesized by carbo-thermic reaction of titanium and acetone at 800 °C. This method is relatively low temperature synthesis route. It can be used for large scale production of TiC. The synthesized nanoparticles have been characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM) and differential thermal analyzer (DTA) techniques. XRD analysis confirmed the formation of single phase TiC. XRD analysis confirmed that the particles are spherical in shape with an average particle size of 13 nm. DTA analysis shows that the phase is stable upto 900 °C and the material can be used for high temperature applications.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

DOE OSTI2017

Design of the First-in-Class, Highly Potent Irreversible Inhibitor Targeting the Menin-MLL Protein–Protein Interaction

The structure-based design of M-525 as the first-in-class, highly potent, irreversible small-molecule inhibitor of the menin-MLL interaction is presented. M-525 targets cellular menin protein at sub-nanomolar concentrations and achieves low nanomolar potencies in cell growth inhibition and in the suppression of MLL-regulated gene expression in MLL leukemia cells. M-525 demonstrates high cellular specificity over non-MLL leukemia cells and is more than 30 times more potent than its corresponding reversible inhibitors. Mass spectrometric analysis and co-crystal structure of M-525 in complex with menin firmly establish its mode of action. A single administration of M-525 effectively suppresses MLL-regulated gene expression in tumor tissue. An efficient procedure was developed to synthesize M-525. This study demonstrates that irreversible inhibition of menin may be a promising therapeutic strategy for MLL leukemia.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2020

Complete genome of Polaromonas vacuolata KCTC 22033 T isolated from beneath Antarctic Sea ice

Polaromonas vacuolata KCTC 22033 T is an obligate aerobic, Gram-negative, psychrophilic and rod-shaped bacterium isolated from beneath the sea ice off the coast of the Palmer Peninsula, Anvers Islands, Antarctica. P. vacuolata is the type species of Polaromonas genus and the first example of gas vacuolate Betaproteobacteria isolated from marine habitats. In this study, we report a complete genome of P. vacuolata KCTC 22033 T , which consists of 3,837,686 bp (G + C content of 52.07%) with a single chromosome, 3461 protein-coding genes, 56 tRNAs and 6 rRNA operons. Genomic analysis revealed the presence of genes involved in bacterial adaptation under saline conditions, cold adaptation via the production of gas vesicles and cell adhesion proteins, and a photoheterotrophic lifestyle when challenged by starvation. Intriguingly, several of these genes were likely acquired from species outside the Polaromonas genus. The genomic information therefore describes the unique evolution and adaptation of P. vacuolata to its extraordinary habitat, i.e., beneath the Antarctic sea ice.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

A multimodal and integrated approach to interrogate human kidney biopsies with rigor and reproducibility: guidelines from the Kidney Precision Medicine Project

Comprehensive and spatially mapped molecular atlases of organs at a cellular level are a critical resource to gain insights into pathogenic mechanisms and personalized therapies for diseases. The Kidney Precision Medicine Project (KPMP) is an endeavor to generate three-dimensional (3-D) molecular atlases of healthy and diseased kidney biopsies by using multiple state-of-the-art omics and imaging technologies across several institutions. Obtaining rigorous and reproducible results from disparate methods and at different sites to interrogate biomolecules at a single-cell level or in 3-D space is a significant challenge that can be a futile exercise if not well controlled. Here we describe a “follow the tissue” pipeline for generating a reliable and authentic single-cell/region 3-D molecular atlas of human adult kidney. Our approach emphasizes quality assurance, quality control, validation, and harmonization across different omics and imaging technologies from sample procurement, processing, storage, shipping to data generation, analysis, and sharing. We established benchmarks for quality control, rigor, reproducibility, and feasibility across multiple technologies through a pilot experiment using common source tissue that was processed and analyzed at different institutions and different technologies. A peer review system was established to critically review quality control measures and the reproducibility of data generated by each technology before their being approved to interrogate clinical biopsy specimens. The process established economizes the use of valuable biopsy tissue for multiomics and imaging analysis with stringent quality control to ensure rigor and reproducibility of results and serves as a model for precision medicine projects across laboratories, institutions and consortia.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2012

Spatial and Temporal Measurements of Temperature and Cell Viability in Response to Nanoparticle Mediated Photothermal Therapy

Aim: Nanoparticle enhanced photothermal therapy is a promising alternative to tumor resection. However, quantitative measurements of cellular response to these treatments are limited. This paper introduces a Bimodal Enhanced Analysis of Spatiotemporal Temperature (BEAST) algorithm to rapidly determine the viability of cancer cells in vitro following photothermal therapy alone or in combination with nanoparticles. Materials & Methods: To illustrate the capability of the BEAST viability algorithm, single wall carbon nanohorns were added to renal cancer (RENCA) cells in vitro and time-dependent spatial temperature maps measured with an infrared camera during laser therapy were correlated with post-treatment cell viability distribution maps obtained by cell-staining fluorescent microscopy. Conclusion: The BEAST viability algorithm accurately and rapidly determined the cell viability as function of time, space, and temperature.

laser↗

NASA NTRS2000

Development of Electrostatically Clean Solar Array Panels

Certain missions require Electrostatically Clean Solar Array (ECSA) panels to establish a favorable environment for the operation of sensitive scientific instruments. The objective of this program was to demonstrate the feasibility of an ECSA panel that minimizes panel surface potential below 100mV in LEO and GEO charged particle environments, prevents exposure of solar cell voltage and panel insulating surfaces to the ambient environment, and provides an equipotential, grounded structure surrounding the entire panel. An ECSA panel design was developed that uses a Front Side Aperture-Shield (FSA) that covers all inter-cell areas with a single graphite composite laminate, composite edge clips for connecting the FSA to the panel substrate, and built-in tabs that interconnect the FSA to conductive coated coverglasses using a conductive adhesive. Analysis indicated the ability of the design to meet the ECSA requirements. Qualification coupons and a 0.5m x 0.5m prototype panel were fabricated and tested for photovoltaic performance and electrical grounding before and after exposure to acoustic and thermal cycling environments. The results show the feasibility of achieving electrostatic cleanliness with a small penalty in mass, photovoltaic performance and cost, with a design is structurally robust and compatible with a wide range of current solar panel technologies.

Stern, Theodore G.↗

DOE OSTI2024

Modeling dynamics of acute HIV infection incorporating density-dependent cell death and multiplicity of infection

Understanding the dynamics of acute HIV infection can offer valuable insights into the early stages of viral behavior, potentially helping uncover various aspects of HIV pathogenesis. The standard viral dynamics model explains HIV viral dynamics during acute infection reasonably well. However, the model makes simplifying assumptions, neglecting some aspects of HIV infection. For instance, in the standard model, target cells are infected by a single HIV virion. Yet, cellular multiplicity of infection (MOI) may have considerable effects in pathogenesis and viral evolution. Further, when using the standard model, we take constant infected cell death rates, simplifying the dynamic immune responses. Here, we use four models—1) the standard viral dynamics model, 2) an alternate model incorporating cellular MOI, 3) a model assuming density-dependent death rate of infected cells and 4) a model combining (2) and (3)—to investigate acute infection dynamics in 43 people living with HIV very early after HIV exposure. We find that all models qualitatively describe the data, but none of the tested models is by itself the best to capture different kinds of heterogeneity. Instead, different models describe differing features of the dynamics more accurately. For example, while the standard viral dynamics model may be the most parsimonious across study participants by the corrected Akaike Information Criterion (AICc), we find that viral peaks are better explained by a model allowing for cellular MOI, using a linear regression analysis as analyzed by R 2 . These results suggest that heterogeneity in within-host viral dynamics cannot be captured by a single model. Depending on the specific aspect of interest, a corresponding model should be employed.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2018

Effect of Infiltration of Barium Carbonate Nanoparticles on the Electrochemical Performance of La 0.6 Sr 0.4 Co 0.2 Fe 0.8 O 3–δ Cathodes for Protonic Ceramic Fuel Cells

In this work BaCO 3 nanoparticles were infiltrated into a La 0.6 Sr 0.4 Co 0.2 Fe 0.8 O 3– δ (LSCF) electrode as a synergistic catalyst to enhance the performance of proton conducting solid oxide fuel cells (H-SOFCs). Electrochemical impedance analysis showed that the polarization resistance was dramatically reduced by nearly 75% from 1.123 Ω cm 2 to 0.293 Ω cm 2 at 700°C after infiltration of BaCO 3 nanoparticles. The chemical stability between the BaCO 3 and LSCF electrode was investigated by running a long-term 300-h test, during which the polarization resistance exhibited only minor degradation (2.22–2.20 Ω cm 2 ). In addition, single cells with infiltrated LSCF electrode and BaCe 0.7 Zr 0.1 Y 0.1 Yb 0.1 O 3– δ (BCZYYb) electrolyte yielded a maximum power density of 404 mW cm –2 at 700°C, much higher than cells with a bare LSCF electrode (268 mW cm –2 at 700°C). BaCO 3 demonstrated promising performance enhancements of LSCF electrodes for H-SOFCs and warrants further development.

25 ENERGY STORAGE↗