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DOE OSTI2018

T7 Expression Systems for Inducible Production of Proteins from Cloned Genes in E. coli

Inducible T7 expression systems are capable of producing a wide range of proteins in E. coli . Improvements over common practice include: preventing unintended induction by establishing and maintaining expression strains in non-inducing growth media comprised entirely of purified components instead of complex growth media that may variably induce target proteins on approach to saturation; and expressing many target proteins in parallel by convenient and productive auto-induction in BL21(DE3) and other suitable hosts, instead of IPTG induction. From earliest days, basal expression prevented establishment of inducible strains for producing proteins that are stressful to the host. Newly developed pAL vectors now reduce basal expression to levels where coding sequences for even the most stressful proteins can be maintained and induced. In conclusion, asymmetric ligation allows simple and efficient cloning of individual coding sequences or simultaneous cloning of two or three coding sequences for co expression from a single pAL vector.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2015

A position-aware linear solid constitutive model for peridynamics

A position-aware linear solid (PALS) peridynamic constitutive model is proposed for isotropic elastic solids. The PALS model addresses problems that arise, in ordinary peridynamic material models such as the linear peridynamic solid (LPS), due to incomplete neighborhoods near the surface of a body. We improved model behavior in the vicinity of free surfaces through the application of two influence functions that correspond, respectively, to the volumetric and deviatoric parts of the deformation. Furthermore, the model is position-aware in that the influence functions vary over the body and reflect the proximity of each material point to free surfaces. Demonstration calculations on simple benchmark problems show a sharp reduction in error relative to the LPS model.

36 MATERIALS SCIENCE↗

DOE OSTI2020

Positron annihilation lifetime spectroscopy study of polyvinylpyrrolidone‐added polyvinylidene fluoride membranes: Investigation of free volume and permeation relationships

ABSTRACT Polyvinylidene fluoride (PVDF) membranes were prepared via the phase inversion method from casting solutions containing PVDF, dimethylformamide (DMF), and polyvinylpyrrolidone (PVP) as pore former. PVP was used in the casting solution in a range of 0–5 wt % and extracted. The effect on membranes of using PVP in the casting process was analyzed by X‐ray diffraction, differential scanning calorimetry, scanning electron microscopy, viscosity, and water permeability techniques. With an increase of PVP from 0 to 5 wt %, the PVDF casting solution viscosities increased from 858 to 1148 cP; the resulting PVDF membrane thickness increased; and the crystallinity of PVDF membranes decreased from 40.0 to 33.3%, which indicates that the addition of PVP inhibits the degree of crystallization in the PVDF membranes. SEM results revealed the shape and size of macropores in the membranes; these macropores changed after PVP addition to the casting solutions. The impact of structural changes on free‐volume properties was evaluated using positron annihilation lifetime spectroscopy (PALS) studies. PALS analysis indicated no effect on the average radius (~3.4 Å) of membrane free‐volume holes from the addition of PVP to the casting solution. However, the percentage of o ‐Ps pick‐off annihilation intensity, I 3 , increased from 1.7 to 5.1% with increased PVP content. Further, increasing the PVP content from 0.5 to 5% resulted in an increased final pure water permeability flux. For instance, the 210 min flux for a 14% PVDF + 0.5% PVP membrane was found to be 3.3 times greater than a control membrane having the same PVDF concentration. © 2020 Wiley Periodicals, Inc. J. Polym. Sci. 2020 , 58 , 589–598

Eren, Bilge↗

DOE OSTI2018

Degradation of Congo Red dye by a Fe 2 O 3 @CeO 2 -ZrO 2 /Palygorskite composite catalyst: Synergetic effects of Fe 2 O 3

A newly designed Fe 2 O 3 @CeO 2 -ZrO 2 /Palygorskite heterogeneous Fenton catalyst with high degradation efficiency toward Congo Red (CR) is reported in this paper. The iron oxide (Fe 2 O 3 ) was used as the originator of hydroxyl radicals ( OH), CeO 2 -ZrO 2 was used as an efficient matrix of Fe 2 O 3 to enhance the oxidation performance, and the Palygorskite (Pal) played the role of substrate and adsorbent. A synergetic effect of the catalyst was realized by enhancing the adsorption capacity and dispersibility of CeO 2 -ZrO 2 and Fe 2 O 3 nanoparticles, and also due to charge transfer between these two species. The products were characterized by XRD, FTIR, SEM, TEM, XPS, VSM and nitrogen adsorption-desorption isotherms. The catalytic activities were evaluated in terms of many reaction parameters. The removal efficiency of CR was up to 95% on Fe 2 O 3 @CeO 2 -ZrO 2 /Pal under optimized conditions (Pal accounted for 30% of the catalyst mass fraction and Ce/Zr atoms molar ratio = 8:2, T = 30 °C, pH = 3.4, 4 g L -1 catalyst, 40 mM H 2 O 2 and 500 mg/L CR), the product also has shown good reusability. Both α-Fe 2 O 3 and CeO 2 -ZrO 2 played important roles in the generation of hydroxyl radicals, which determined the catalytic efficiency of the composite catalyst. Fe 2 O 3 @CeO 2 -ZrO 2 /Pal was proven to be a promising candidate in the field of heterogeneous Fenton catalysis.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2015

Organic solution-processible electroluminescent molecular glasses for non-doped standard red OLEDs with electrically stable chromaticity

Highlights: • The D–A–D electroluminescent molecular glasses are synthesized. • Non-doped red electroluminescent film is fabricated by spin-coating. • Red OLED shows stable wavelength, luminous efficiency and chromaticity. • CIE1931 coordinate is in accord with standard red light in PAL system. - Abstract: Organic light-emitting molecular glasses (OEMGs) are synthesized through the introduction of nonplanar donor and branched aliphatic chain into electroluminescent emitters. The target OEMGs are characterized by {sup 1}H NMR, {sup 13}C NMR, IR, UV–vis and fluorescent spectra as well as elemental analysis, TG and DSC. The results indicated that the optical, electrochemical and electroluminescent properties of OEMGs are adjusted successfully by the replacement of electron-donating group. The non-doped OLED device with a standard red electroluminescent emission is achieved by spin-coating the THF solution of OEMG with a triphenylamine moiety. This non-doped red OLED device takes on an electrically stable electroluminescent performance, including the stable maximum electroluminescent wavelength of 640 nm, the stable luminous efficiency of 2.4 cd/A and the stable CIE1931 coordinate of (x, y) = (0.64, 0.35), which is basically in accord with the CIE1931 coordinate (x, y) = (0.64, 0.33) of standard red light in PAL system.

36 MATERIALS SCIENCE↗

DOE OSTI2016

Analysis of governing factors controlling gas transport through fresh and aged triptycene-based polyimide films

In this report the fundamental gas transport properties and physical aging behavior of a series of triptycene-based polyimides with various substitution groups were investigated. Wide-angle x-ray diffraction revealed that the 6FDA-1,4-triptycene polyimides exhibit two chain packing domains with d-spacing values of ~6.7 Å and ~5.3 Å, corresponding to triptycene-induced chain packing disruption and chain segments lacking the triptycene moiety, respectively. Positron annihilation lifetime spectroscopy (PALS) showed a bimodal distribution of microcavity size made up of triptycene cleft cavities that are ~3 Å diameter and larger interchain cavities with an average diameter of ~7 Å. Additionally, PALS confirmed 6FDA-1,4-trip_CH 3 has the lowest fractional free volume, primarily due to smaller contributions from the interchain cavities, and 6FDA-1,4-trip_CF 3 has the largest fractional free volume. It was also found that the addition of substituent groups had a unique and positive effect on the aging properties over nine months, in which the addition of CH 3 and CF 3 groups led to permeability increases of ~20% and ~40%, respectively. This is caused by the combination of the lowered driving force for physical aging, the substituent groups sterically hindering tighter packing with time, and the interconnectivity and/or unblocking of free volume over time through small-scale, local chain motion.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2017

Intrinsically Hierarchical Nanoporous Polymers via Polymerization-Induced Microphase Separation

The synthesis of microporous polymers generally requires postpolymerization modification via hyper-cross-linking to trap the polymeric network in a state with high void volume. An alternative approach utilizes rigid, sterically demanding monomers to inhibit efficient packing, thus leading to a high degree of free volume between polymer side groups and main chains. In this work we combine polymers of intrinsic microporosity with polymerization-induced microphase separation (PIMS), a versatile methodology for the synthesis of nanostructured materials that can be rendered mesoporous. Copolymerization of various styrenic monomers with divinylbenzene in the presence of a poly(lactide) terminated with a chain-transfer agent (PLA-CTA) results in kinetic trapping of a microphase-separated state. Subsequent etching of PLA provides a bicontinuous mesoporous network. Using equilibrium and kinetic nitrogen sorption experiments as well as positron annihilation lifetime spectroscopy (PALS), we demonstrate that variations in the steric characteristics of the styrenic monomer impart the network with microporosity, resulting in hierarchically (meso and micro) porous materials. Additionally, structure–property relationships of the styrenic monomer with total surface area and pore volume indicate that the glass transition temperature (T g ) of the corresponding styrenic homopolymers provides a reasonable measure of the steric interactions and resultant microporosity in these systems. Finally, PALS provides insight into micro- and mesoscopic void volume differences between porous monoliths containing either tert-butyl or TMS-modified styrenic monomers compared to the parent, unmodified styrene.

36 MATERIALS SCIENCE↗

DOE OSTI2022

Interface effect of Fe and Fe 2 O 3 on the distributions of ion induced defects

The stability of structural materials in extreme nuclear reactor environments—with high temperature, high radiation, and corrosive media—directly affects the lifespan of the reactor. In such extreme environments, an oxide layer on the metal surface acts as a passive layer protecting the metal underneath from corrosion. To predict the irradiation effect on the metal layer in these metal/oxide bilayers, nondestructive depth-resolved positron annihilation lifetime spectroscopy (PALS) and complementary transmission electron microscopy (TEM) were used to investigate small-scale defects created by ion irradiation in an epitaxially grown (100) Fe film capped with a 50 nm Fe 2 O 3 oxide layer. In this study, the evolution of induced vacancies was monitored, from individual vacancy formation at low doses—10 –5 dpa—to larger vacancy cluster formation at increasing doses, showing the sensitivity of positron annihilation spectroscopy technique. Furthermore, PALS measurements reveal how the presence of a metal–oxide interface modifies the distribution of point defects induced by irradiation. TEM measurements show that irradiation induced dislocations at the interface is the mechanism behind the redistribution of point defects causing their accumulation close to the interface. Finally, this work demonstrates that the passive oxide layers formed during corrosion impact the distribution and accumulation of radiation induced defects in the metal underneath and emphasizes that the synergistic impact of radiation and corrosion will differ from their individual impacts.

36 MATERIALS SCIENCE↗

DOE OSTI2012

THE CENTRAL BLUE STRAGGLER POPULATION IN FOUR OUTER-HALO GLOBULAR CLUSTERS

Using Hubble Space Telescope/Wide Field Planetary Camera 2 data, we have performed a comparative study of the Blue Straggler Star (BSS) populations in the central regions of the globular clusters (GCs) AM 1, Eridanus, Palomar 3, and Palomar 4. Located at distances R{sub GC} > 50 kpc from the Galactic center, these are (together with Palomar 14 and NGC 2419) the most distant clusters in the halo. We determine their color-magnitude diagrams and centers of gravity. The four clusters turn out to have similar ages (10.5-11 Gyr), significantly smaller than those of the inner-halo globulars, and similar metallicities. By exploiting wide-field ground-based data, we build the most extended radial density profiles from resolved star counts ever published for these systems. These are well reproduced by isotropic King models of relatively low concentration. BSSs appear to be significantly more centrally segregated than red giants in all GCs, in agreement with the estimated core and half-mass relaxation times which are smaller than the cluster ages. Assuming that this is a signature of mass segregation, we conclude that AM 1 and Eridanus are slightly dynamically more evolved than Pal 3 and Pal 4.

79 ASTRONOMY AND ASTROPHYSICS↗

DOE OSTI2022

The essential Rhodobacter sphaeroides CenKR two-component system regulates cell division and envelope biosynthesis

Bacterial two-component systems (TCSs) often function through the detection of an extracytoplasmic stimulus and the transduction of a signal by a transmembrane sensory histidine kinase. This kinase then initiates a series of reversible phosphorylation modifications to regulate the activity of a cognate, cytoplasmic response regulator as a transcription factor. Several TCSs have been implicated in the regulation of cell cycle dynamics, cell envelope integrity, or cell wall development in Escherichia coli and other well-studied Gram-negative model organisms. However, many α-proteobacteria lack homologs to these regulators, so an understanding of how α-proteobacteria orchestrate extracytoplasmic events is lacking. In this work we identify an essential TCS, CenKR ( C ell en velope K inase and R egulator), in the α-proteobacterium Rhodobacter sphaeroides and show that modulation of its activity results in major morphological changes. Using genetic and biochemical approaches, we dissect the requirements for the phosphotransfer event between CenK and CenR, use this information to manipulate the activity of this TCS in vivo , and identify genes that are directly and indirectly controlled by CenKR in Rb . sphaeroides . Combining ChIP-seq and RNA-seq, we show that the CenKR TCS plays a direct role in maintenance of the cell envelope, regulates the expression of subunits of the Tol-Pal outer membrane division complex, and indirectly modulates the expression of peptidoglycan biosynthetic genes. CenKR represents the first TCS reported to directly control the expression of Tol-Pal machinery genes in Gram-negative bacteria, and we predict that homologs of this TCS serve a similar function in other closely related organisms. We propose that Rb . sphaeroides genes of unknown function that are directly regulated by CenKR play unknown roles in cell envelope biosynthesis, assembly, and/or remodeling in this and other α-proteobacteria.

Lakey, Bryan D. (ORCID:0000000332795242)↗

DOE OSTI2023

LANL Institutional Computing Close-out Report for Project t22_ocean_time_step

This year, my team made a very productive use of LANL IC time. Four papers were published that used IC resources, and two more are under review. These publications fall into three categories: 1.) Improving tide modeling in the global ocean by adding self attraction and loading (Barton et. al. 2022), ice shelf cavities (Pal et al. 2023), and local time stepping (Lilly et al. 2023). 2.) A new sea ice numerical formulation (Capodaglio 2023). 3.) Performance comparisons, methods, and test cases for ocean model development, This includes a verification suite for ocean models (Bishnu, submitted) and a comparison between Julia and Fortran (Strauss, submitted). A new time-stepping method was introduced in Calandrini et al. (2022). These publications, and the use of LANL IC resources, go hand-in-hand with our mentoring efforts to train young scientists. Two of the lead authors are graduate students who are conducting their PhD research: Kristin Barton at the University of Michigan and Jeremy Lilly at Oregon State. These are both their first publications, and they both have DOE funding and DOE mentors. In addition, Bishnu is a post-doctoral researcher at LANL; Strauss conducted his research as a senior in high school; and Capodaglio, Calandrini and Pal are all early-career scientists who were converted to staff in 2021 or 2022. Here we highlight two publications on improvements in tidal modeling: Barton et al. 2022 and Lilly et al. 2023.

58 GEOSCIENCES↗

DOE OSTI2025

Summary of Pilot Project State Technical Assistance on Multi-Sector Analysis for Electric and Petroleum Fuels

The Oregon Energy Security Plan, (ODOE 2024) published in September 2024, builds a strong case for the state to give acute attention to the fuel supply chain. In December 2024, Pacific Northwest National Laboratory (PNNL) in partnership with Oregon Department of Energy (ODOE), announced a pilot project to conduct an analysis that synthesizes current and projected transportation fuel dynamics, supply chain risks, and risk comparators with relevant sectors, such as transportation electrification, sponsored by the Department of Energy’s (DOE) Office of Cybersecurity, Energy Security, and Emergency Response (CESER). The study, is intended to leverage existing modeling and frameworks from a recent 2024 sector coupling analysis supported by the DOEs Office of Electricity (OE) (B. Mitra, S. Pal, et al., Coupling of the Electricity and Transportation Sectors - Part I: Sector Overviews 2024) (B. Mitra, S. Pal and J. Reeve, et al. 2024). While the PNNL team set out to conduct a quantitative risk analysis driven by detailed data that synthesizes current and projected transportation fuel dynamics, supply chain risks, and risk comparators with relevant sectors. The intention was to provide an approach that could be extendable to other parts of the country. They encountered data limitations and adjusted their approach accordingly. This report summarizes PNNL's original plan for executing the study, including limitations for obtaining data requirements for fuel flows and interim products, as well as a risk matrix that can be used to identify supply chain risks.

02 PETROLEUM↗

DOE OSTI2016

Multi-sightline observation of narrow absorption lines in lensed quasar SDSS J1029+2623

We exploit the widely separated images of the lensed quasar SDSS J1029+2623 (z{sub em} = 2.197, θ = 22.″5) to observe its outflowing wind through two different sightlines. We present an analysis of three observations, including two with the Subaru telescope in 2010 February and 2014 April, separated by four years, and one with the Very Large Telescope, separated from the second Subaru observation by ∼2 months. We detect 66 narrow absorption lines (NALs), of which 24 are classified as intrinsic NALs that are physically associated with the quasar based on partial coverage analysis. The velocities of intrinsic NALs appear to cluster around values of v{sub ej} ∼ 59,000, 43,000, and 29,000 km s{sup −1}, which is reminiscent of filamentary structures obtained by numerical simulations. There are no common intrinsic NALs at the same redshift along the two sightlines, implying that the transverse size of the NAL absorbers should be smaller than the sightline distance between two lensed images. In addition to the NALs with large ejection velocities of v{sub ej} > 1000 km s{sup −1}, we also detect broader proximity absorption lines (PALs) at z{sub abs} ∼ z{sub em}. The PALs are likely to arise in outflowing gas at a distance of r ≤ 620 pc from the central black hole with an electron density of n {sub e} ≥8.7 × 10{sup 3} cm{sup −3}. These limits are based on the assumption that the variability of the lines is due to recombination. We discuss the implications of these results on the three-dimensional structure of the outflow.

79 ASTRONOMY AND ASTROPHYSICS↗

NASA NTRS1985

Oxidant abundances in rainwater and the evolution of atmospheric oxygen

H2O2 is probably the dominant oxidant in rainwater at O2 levels lower than 0.01-0.001 present atmospheric levels (PAL), and the magnitude of its dissolved flux is sensitive to the concentration of the trace gases NO, CO, and CH4 in the atmosphere, as well as to the partial pressure of atmospheric CO2. The earlier interpretation of the paleosol data in terms of the ratio of p-O2/p-CO2 in the atmosphere must be modified by taking into account the effects of photochemically produced oxidants and reactants at p-O2 of les than 0.01 PAL.

Kasting, J. F.↗

NASA NTRS1988

The origin and evolution of atmospheric oxygen

This paper discusses the chemical processes involved in the evolution of the earth's atmospheric oxygen and ozone, as well as the sources, sinks, and transfer rates of oxygen in the present atmosphere. Special attention is given the evolution of atmospheric O3 as a function of the buildup of O2, with the results of calculations presented as the vertical profiles of O3, in terms of the present atmospheric level (PAL) oxygen values. Calculations show that the total O3 column density that is approximately half of the present level was reached when atmospheric oxygen level reached 0.1 PAL. At this level of ozone, the biological shielding of the earth's surface from the UV radiation is believed to have been achieved.

Levine, Joel S.↗

NASA NTRS1992

Young globular clusters in the Milky Way Galaxy

Ruprecht 106 and Pal 12 are two known globular clusters in the Milky Way that are unequivocally younger than other clusters of similar metallicity. The Galactic coordinates of Ruprecht 106 place it near to the Magellanic Stream in projection, suggesting a tidal capture from the Magellanic Clouds. It is demonstrated that a family of orbits for both clusters can be constructed that are consistent with this capture hypothesis and that these then lead to a prediction of 3 milliarcseconds per year for the proper motions of both Ruprecht 106 and Pal 12.

Lin, D. N. C.↗

NASA NTRS1996

Linearization of an annular image by using a diffractive optic

The goal for this project is to develop the algorithms for fracturing the zones defined by the mapping transformation, and to actually produce the binary optic in an appropriate setup. In 1984 a side-viewing panoramic viewing system was patented, consisting of a single piece of glass with spherical surfaces which produces a 360 degree view of the region surrounding the lens which extends about 25 degrees in front of and 20 degrees behind the lens. The system not only produces images of good quality, it is also afocal, i.e., images stay in focus for objects located right next to the lens as well as those located far from the lens. The lens produced a panoramic view in an annular shaped image, and so the lens was called a PAL (panoramic annular lens). When applying traditional measurements to PAL images, it is found advantageous to linearize the annular image. This can easily be done with a computer and such a linearized image can be produced within about 40 seconds on current microcomputers. However, this process requires a frame-grabber and a computer, and is not real-time. Therefore, it was decided to try to perform this linearization optically by using a diffractive optic.

Matthys, Donald R.↗

NASA NTRS1996

Investigation of Polymer Liquid Crystals

The positron annihilation lifetime spectroscopy (PALS) using a low energy flux generator may provide a reasonably accurate technique for measuring molecular weights of linear polymers and characterization of thin polyimide films in terms of their dielectric constants and hydrophobity etc. Among the tested samples are glassy poly arylene Ether Ketone films, epoxy and other polyimide films. One of the proposed techniques relates the free volume cell size (V(sub f)) with sample molecular weight (M) in a manner remarkably similar to that obtained by Mark Houwink (M-H) between the inherent viscosity (eta) and molecular wieght of polymer solution. The PALS has also demonstrated that free-volume cell size in thermoset is a versatile, useful parameter that relates directly to the polymer segmental molecular weight, the cross-link density, and the coefficient of thermal expansion. Thus, a determination of free volume cell size provides a viable basis for complete microstructural characterization of thermoset polyimides and also gives direct information about the cross-link density and coefficient of expansion of the test samples. Seven areas of the research conducted are reported here.

Han, Kwang S.↗