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At least 163 records · Page 9

A distinct subpopulation of membrane vesicles in Pseudomonas putida is enriched in enzymes for lignin catabolism

Bacterial membrane vesicles (MVs) mediate diverse microbial processes and are emerging as powerful biomedical tools, but MV population heterogeneity remains an open question. Here, we separate, enumerate, and characterize two MV populations from the soil bacterium Pseudomonas putida during growth with or without lignin-derived carbon, a major carbon source from plant cells in the rhizosphere. Small MVs (MV-S, diameter ~100 nm) were produced from all cultures, whereas large MVs (MV-L, diameter ~300 nm) were observed during the late stationary phase of lignin cultivations. MV-S contained selectively packaged proteins with diverse physiological functions, whereas the MV-L proteome was smaller and largely enriched in outer membrane proteins. Interestingly, enzymes known to mediate the catabolism of lignin-derived aromatic compounds were enriched in MV-S. Overall, this study highlights the need for careful consideration of MV populations in microbial systems.

59 BASIC BIOLOGICAL SCIENCES↗

Enrichable consortia of microbial symbionts degrade macroalgal polysaccharides in Kyphosus fish

ABSTRACT Coastal herbivorous fishes consume macroalgae, which is then degraded by microbes along their digestive tract. However, there is scarce genomic information about the microbiota that perform this degradation. This study explores the potential of Kyphosus gastrointestinal microbial symbionts to collaboratively degrade and ferment polysaccharides from red, green, and brown macroalgae through in silico study of carbohydrate-active enzyme and sulfatase sequences. Recovery of metagenome-assembled genomes (MAGs) from previously described Kyphosus gut metagenomes and newly sequenced bioreactor enrichments reveals differences in enzymatic capabilities between the major microbial taxa in Kyphosus guts. The most versatile of the recovered MAGs were from the Bacteroidota phylum, whose MAGs house enzyme collections able to decompose a variety of algal polysaccharides. Unique enzymes and predicted degradative capacities of genomes from the Bacillota (genus Vallitalea ) and Verrucomicrobiota (order Kiritimatiellales ) highlight the importance of metabolic contributions from multiple phyla to broaden polysaccharide degradation capabilities. Few genomes contain the required enzymes to fully degrade any complex sulfated algal polysaccharide alone. The distribution of suitable enzymes between MAGs originating from different taxa, along with the widespread detection of signal peptides in candidate enzymes, is consistent with cooperative extracellular degradation of these carbohydrates. This study leverages genomic evidence to reveal an untapped diversity at the enzyme and strain level among Kyphosus symbionts and their contributions to macroalgae decomposition. Bioreactor enrichments provide a genomic foundation for degradative and fermentative processes central to translating the knowledge gained from this system to the aquaculture and bioenergy sectors. IMPORTANCE Seaweed has long been considered a promising source of sustainable biomass for bioenergy and aquaculture feed, but scalable industrial methods for decomposing terrestrial compounds can struggle to break down seaweed polysaccharides efficiently due to their unique sulfated structures. Fish of the genus Kyphosus feed on seaweed by leveraging gastrointestinal bacteria to degrade algal polysaccharides into simple sugars. This study reconstructs metagenome-assembled genomes for these gastrointestinal bacteria to enhance our understanding of herbivorous fish digestion and fermentation of algal sugars. Investigations at the gene level identify Kyphosus guts as an untapped source of seaweed-degrading enzymes ripe for further characterization. These discoveries set the stage for future work incorporating marine enzymes and microbial communities in the industrial degradation of algal polysaccharides.

59 BASIC BIOLOGICAL SCIENCES↗

Eev (enrich Enforce Validate) With Cpefinder

This code is designed to take an existing STIX bundle with vulnerability data and enrich it with additional potential vulnerabilities to provide further insight during threat analysis. It also acts as a launch platform for other enrichments tools. The additional tools include, WAVgraph and STIXEnforcer.

Beckman, BryanR [Idaho National Laboratory (INL), ↗

Recycling Isotopically Enriched Mo from the RadioGenix Waste Stream

NorthStar’s RadioGenix generator utilizes an aqueous biphasic extraction chromatography column to separate Tc from low-specific-activity 99 Mo. During this separation process, a fraction of the source Mo material can be found in the waste. This is not of concern when natural Mo targets are used. However, the introduction of isotopically enriched 98 Mo for neutron capture or 100 Mo for accelerator-based production requires recovery of this valuable enriched Mo material. The generator waste is absorbed and solidified in a superabsorbent hydrogel polymer. In this contribution, we report results from our small-scale experimental study on development of a separation process for recovery of molybdenum from the polymeric gel waste using common chemicals and simple chemical techniques. Our investigation confirms that the acidic deswelling of the alkaline hydrogel with concentrated hydrochloric acid converts the occluded molybdate to a neutral molybdenyl dichloride species that is freely released from the gel. Mo can then be concentrated on a commercial tri-n-butyl phosphate resin, eluted with ammonium hydroxide, and crystallized as ammonium heptamolybdate for conversion back to Mo metal. Typical Mo recoveries are 90-95%.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Development and Optimization of a Purification Process to Recover 99 Mo from Low-enriched Uranium

Supported by the DOE-NNSA Office of Material Management & Minimization (M3), a number of domestic entities are pursuing non-highly enriched uranium (non-HEU) production of 99 Mo. As the production technologies of 99Mo pivot toward low-enriched uranium (LEU) or molybdenum targets, new reaction channels and accelerators are being evaluated. Superconducting electron linear accelerators (LINACs) with high-Z converter targets can generate bremsstrahlung photons and neutron fluxes that are capable of inducing photonuclear reactions and LEU fission. A particular advantage of a LINAC is that it does not rely on HEU-fueled reactor cores (which are currently slated for LEU conversion) and can operate on an almost continuous basis. Regarding the chemical purification of 99 Mo from irradiated uranium targets under acidic digestion, there exists a procedure known as Cintichem or modifications thereof with respect to LEU (LEU Modified Cintichem process, LMC). The process relies on a number of selective precipitation steps and column chromatography to purify Mo. It is important to note that LMC prescribes the addition of stable Mo to carry 99 Mo on alpha benzoin oxime, which reduces the specific activity of 99 Mo. This is especially important for processing 99 Mo batches with lower activities (~33 Ci of 99 Mo per batch).

07 ISOTOPE AND RADIATION SOURCES↗

High Flux Isotope Reactor Low-Enriched Uranium Low Density Silicide Fuel Design Parameters

High Flux Isotope Reactor (HFIR) highly enriched uranium (HEU) to low-enriched uranium (LEU) conversion activities are ongoing as part of the Department of Energy (DOE) National Nuclear Security Administration (NNSA)’s nuclear nonproliferation mission. Design activities studying the conversion of HFIR from HEU to LEU fuel explored different fuel design features and shapes with a low density uranium-silicide dispersion (U 3 Si 2 -Al) fuel, which has a uranium density of 4.8 gU/cm 3 . The goal of these studies is to generate several HFIR LEU fuel designs of varying fuel fabrication complexity that meet the current HEU performance metrics and safety requirements. The documented designs will serve as references for fuel fabrication and qualification activities. Recent advancements in modeling and simulation tools enable quick prototyping of fuel designs. Shift, a Monte Carlo neutron transport and depletion tool optimized for high-performance computing (HPC) architectures, is used for efficient fuel cycle and performance metrics calculations. The HFIR Steady State Heat Transfer Code (HSSHTC) is used to vet the thermal safety margin. Also, a new automation tool that connects all fuel design analysis steps, named Python HFIR Analysis and Measurement Engine (PHAME), has been developed to expedite the design study in an efficient and reproducible manner. Leveraging these tools, several candidate fuel designs were selected for varying fabrication complexity. This report provides design feature details for four selected HFIR LEU low density U 3 Si 2 -Al fuel designs and their corresponding performance and safety metrics. Nominal, best-estimate design parameters and irradiation conditions, including fission rate densities, power densities, heat fluxes, and cumulative fission densities are provided for candidate fuel designs relevant to framing irradiation experiments to support fuel qualification efforts. Simulations show that the low density U 3 Si 2 -Al, with design features to enhance safety, can meet HEU core performance metrics and safety requirements if the reactor power is increased from 85 MW (HEU) to 95 MW (LEU) and if the active fuel length is increased from 50.80 cm (HEU) to 55.88 cm (LEU).

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

MITR Low-Enriched Uranium Conversion Fluid-Structure Interaction Preliminary Design Verification

The U.S. National Nuclear Security Administration (NNSA) Office of Material Management and Minimization (M 3 ) has developed and is pursuing an integrated approach to address the persistent threat posed by unintentional proliferation of nuclear materials. The NNSA M 3 approach reduces the risk of highly enriched uranium (HEU) and plutonium falling into the hands of non-state actors by minimizing the use of and, when possible, eliminating weapons-usable nuclear material around the world. In this geopolitical context, most research and test reactors, both domestic and international, have completed or started a program of conversion from the use of HEU to low-enriched uranium (LEU) as fuel. Conversion of civilian research reactors from HEU to LEU, and the return of the HEU to the country of origin, are important components of the NNSA non-proliferation program. Worldwide, 71 reactors have been converted to the use of LEU fuel, and an additional 32 have been confirmed to be permanently shut down. The U.S., with 20 reactor conversions, is among the 39 countries on six continents where conversions have occurred. With recent conversions in Ghana and Nigeria, an important milestone was reached in completing conversion of all reactors on the continent of Africa to LEU fuel. Africa thus becomes the third continent to have completed conversion of all HEU reactors to LEU, following Australia and South America

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

The Large Enriched Germanium Experiment for Neutrinoless ββ Decay (LEGEND-1000 Preconceptual Design Report)

We propose the construction of LEGEND-1000, the ton-scale Large Enriched Germanium Experiment for Neutrinoless ββ Decay. This experiment is an international project designed to answer one of the highest priority questions in fundamental physics. It consists of 1000 kg of Ge detectors enriched to more than 90% in the 76 Ge isotope of interest operated in a liquid argon active shield at a deep underground lab. The experiment is designed to achieve a discovery potential that covers the inverted-ordering neutrino mass scale region. The baseline design assumes installation in the SNOLAB cryopit. A similar experimental setup could also be realized at the LNGS alternative site.

72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS↗

Delayed Critical Enriched Uranium Metal, 7-in.-diam. Cylinder with Thin Stainless Steel Top and Bottom Reflectors

Three 7 in. diameter highly enriched uranium (HEU; 93.17 wt % 235 U) metal cylinders were assembled on the vertical assembly machine of the Oak Ridge Critical Experiments Facility (ORCEF) with thin stainless steel reflectors (0.5, 1, and 2 in. thick) on the top and bottom. These experiments were performed during January 13–25, 1965, and used seven operational days at ORCEF. Before these experiments, unreflected and unmoderated 7 in. diameter HEU metal cylinders had been assembled to delayed criticality at ORCEF, and the results were benchmarked in HEU-MET-FAST-051. In addition to the critical experiments, the prompt neutron decay constant was measured via the Rossi-α technique, and those results are also presented in this report. To achieve near-delayed-critical systems, 5 × 5 × 0.03125 in. rectangular uranium metal parts were used on top of 7 in. diameter cylinders inside the top reflector. The delayed critical configuration from the three experiments described herein are acceptable for use as criticality safety benchmark experiments for the International Criticality Safety Benchmark Evaluation Program (ICSBEP), coordinated by the Nuclear Energy Agency, once the uncertainty analysis is completed. Based on previous ICSBEP benchmarks with this enriched uranium metal at ORCEF, the uncertainty in $k_{eff}$ is expected to be as low as ±0.0002. The prompt neutron time decay data could be the basis of an International Reactor Physics Evaluation Program benchmark.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗

Transition Core Modeling for Extended Enrichment & Accident-Tolerant Fuels Using Polaris/PARCS

Commercial light water reactor (LWR) operators and fuel vendors are currently interested in increasing the low-enriched uranium (LEU) fuel enrichments from the current limit of 5.0 $^w/_o$ $^{235}U$ up to 10 $^ w/_o$ $^{235}U$ (referred to as "LEU+") in their current fleets; they are also interested in using accident-tolerant fuel (ATF) with both LEU and LEU+ fuel. This report aims to identify modeling challenges and accuracy concerns in transition core analysis using the SCALE Polaris lattice physics code and U.S. Nuclear Regulatory Commission core simulator PARCS. At the time this study was started, no publicly available LEU+ core designs existed for boiling water reactor (BWR) or pressurized water reactor (PWR) systems. Therefore, fuel lattices were shuffled within a multi-assembly model to mimic neutronically challenging lattice combinations seen in transition cores, such as a fresh LEU+ lattice next to depleted LEU lattices. In addition to multi-assembly models, whole-core BWR transition core calculations were performed for ATF and LEU+ fuel using an existing Hatch-1 Cycle 3 core model. A whole-core BWR model was chosen due to the more heterogeneous core designs compared to those for a PWR core. Since the original core is an old checkerboard core design and no core or fuel design optimization was performed for the modeled fuel types, these core calculations were intended only to provide: (1) Comparisons of core characteristics of interest, such as the pin power distributions and peaking factors, Doppler temperature coefficients (DTCs), and control blade worths (CBWs) under challenging core designs, (2) Identification of reactor physics challenges in modeling LEU+ and ATF cores, and (3) A stress test for the Polaris/PARCS two-step modeling approach, including characterization of the relative accuracy for predicting characteristics of interest such as pin power distributions.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Scalable Enrichment of 48 Ca at the Solid/liquid Interface by Chemical and Electrochemical Methods

This award targets to develop methods to enrich 48 Ca, which is a critical isotope for synthesizing superheavy elements and testing the standard model through neutrinoless double beta decay. The team first tested chemical exchange-based separation between solids and liquids, which is based on the free energy change due to the different vibrational frequencies caused by Ca isotopes in a material. However, the separation factor (alpha), which is defined as the ratio of 40 Ca/ 48 Ca ratios in the two phases, only reach ~1.01. The team then developed liquid centrifugation-based isotope separation, where a Ca salt aqueous solution is centrifuged at a speed of ~60 kRPM, and 48 Ca is enriched at the bottom of a centrifuge tube due to its larger mass. A high α of ~1.2-1.4 is achieved for 40 Ca/ 48 Ca at 40 °C. This method is further approved to be generic for any isotope that can be dissolved in a liquid solution or form liquid chemicals near room temperature. The experimental results also align well with modeling prediction. The team further develop a model to evaluate isotope separation in countercurrent liquid centrifugation. The team found that the countercurrent configuration can also enhance isotope separation in liquids, similar with gas centrifugation, which boost separation for isotopes which are difficult to be gasified near room temperature.

07 ISOTOPE AND RADIATION SOURCES↗

High Flux Isotope Reactor Low-Enriched Uranium High Density Silicide Fuel Preliminary Design Update: System Transient Analysis

As a part of conversion efforts from highly enriched uranium (HEU) to low-enriched uranium (LEU) fuel under direction of the National Nuclear Security Administration of the U.S. Department of Energy, multiple proposed designs of the High Flux Isotope Reactor (HFIR) have been created and assessed regarding reactor physics performance metrics, including designs utilizing uranium silicide dispersion fuel (U3Si2-Al). This report updates the previous analyses that evaluated the nuclear safety performance of LEU fuel designs with respect to selected accident events from the HFIR Safety Analysis Report (SAR). Both the Low Density (LD) and High Density (HD) Optimized designs’ reactivity initiated accident fuel performance improved relative to the HEU fuel, attributed to greater 238 U negative Doppler feedback. However, the thermal margins for primary coolant system accidents were reduced with some acceptance criteria unable to be met. The need to resolve reduced thermal margin, open modeling items, and unresolved assumptions was identified.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Gamma Spectrometry Code Rodeo for Uranium Enrichment—FY 2021 Report

In FY22, the participating Department of Energy (DOE) laboratories continued the work on the project “Gamma Spectrometry Code Rodeo for Uranium Enrichment”. Oak Ridge National Laboratory (ORNL) and Lawrence Livermore National Laboratory (LLNL) continued their efforts to acquire spectra using uranium sources. A collimator (with side and back shields) was custom designed and constructed at ORNL. Spectra were acquired at ORNL using the uranium enrichment standards supplied by New Brunswick Laboratory (NBL) and the collimated M400 detector. Spectra were also acquired using steel absorbers of different thickness placed between the source and the collimated M400 detector. The plan was to ship the collimator to LLNL so that spectra could be acquired using the LLNL detector in collimated geometries. This activity was suspended since problems were noticed with the performance of the LLNL M400 detector. Also, it was brought to light to the project team that some of the aspects of the M400 detector had been updated by the vendor H3D in response to requests by the IAEA. In February 2021, the ORNL and LLNL M400 detectors were returned to the H3D factory in order to resolve the problems with the LLNL detector and to update ORNL and LLNL detectors to the same configuration as the M400 supplied to the IAEA. H3D repaired the LLNL M400 and performed the IAEA updates on both the ORNL and LLNL detectors. ORNL and LLNL received the detectors in April 2021. Data acquisition was continued using the upgraded detectors. ORNL re-collected the spectra using point sources and the NBL standards in uncollimated and collimated geometries. The collimator was shipped from ORNL to LLNL. Spectra were acquired at LLNL through Q4 of FY22. When the ORNL spectra were examined closely, it became apparent that there were spurious artifacts present in some of the spectra. This rendered suspect the ORNL spectra collected in June/July 2021 time frame. ORNL contacted H3D and per H3D’s advice, the M400 detector was returned to H3D for repairs in mid-August 2021. H3D diagnosed the problem and established that the root cause was the extra tight packaging that created a strain on the CZT crystals, causing one of them to break down. The repaired M400 was returned to ORNL in early September 2021. ORNL re-started data acquisition for the third time. Data was continued to be acquired at ORNL and LLNL through September 2021. The ORNL and LLNL data will be shared with the principal investigators from the analyzing laboratories. The GADRAS code and FRAM have been developed to have the capability to analyze the M400 spectra. Good quality spectra from FY21 have been analyzed using modified versions of GADRAS and FRAM. The spectra have also been analyzed using the GEM code. The analysis results from GADRAS, FRAM and GEM codes are presented and discussed in this report. Some preliminary results from FRAM and GEM analysis were presented in February 2021 at the U-Pu Isotopics Workshop sponsored by the IAEA. CZTU code is being modified to analyze M400 spectra. Data collection, code development and analysis will be continued in FY22 as per the FY22 Project Work Plan (PWP).

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Cyclotron production of 43Sc and 44gSc from enriched 42CaO, 43CaO, and 44CaO targets

Introduction: 43 Sc and 44g Sc are both positron-emitting radioisotopes of scandium with suitable half-lives and favorable positron energies for clinical positron emission tomography (PET) imaging. Irradiation of isotopically enriched calcium targets has higher cross sections compared to titanium targets and higher radionuclidic purity and cross sections than natural calcium targets for reaction routes possible on small cyclotrons capable of accelerating protons and deuterons. Methods: In this work, we investigate the following production routes via proton and deuteron bombardment on CaCO 3 and CaO target materials: 42 Ca(d,n) 43 Sc, 43 Ca(p,n) 43 Sc, 43 Ca(d,n) 44g Sc, 44 Ca(p,n) 44g Sc, and 44 Ca(p,2n) 43 Sc. Radiochemical isolation of the produced radioscandium was performed with extraction chromatography using branched DGA resin and apparent molar activity was measured with the chelator DOTA. The imaging performance of 43 Sc and 44g Sc was compared with 18 F, 68 Ga, and 64 Cu on two clinical PET/CT scanners. Discussion: The results of this work demonstrate that proton and deuteron bombardment of isotopically enriched CaO targets produce high yield and high radionuclidic purity 43 Sc and 44g Sc. Laboratory capabilities, circumstances, and budgets are likely to dictate which reaction route and radioisotope of scandium is chosen.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Cascade Dielectrophoretic Separation for Selective Enrichment of Polyhydroxybutyrate (PHB)-Producing Cyanobacterium Synechocystis sp. PCC 6803

Maintaining favorable biological productivities in photosynthetic biomanufacturing systems, especially when the risk of contamination with competing microbes is high, remains a challenge to achieve while maintaining economic feasibility. This study presents a dielectrophoresis (DEP)-based microfluidic approach for isolating a desired strain within a co-culture. The cyanobacterium Synechocystis sp. PCC 6803 (a strain capable of producing the bioplastic precursor polyhydroxybutyrate, or PHB) was enriched from mixed cultures containing the competing cyanobacterium Synechococcus elongatus PCC 7942 (which does not naturally produce PHB). A DEP cascade electrode system was established to increase purification efficiency through sequential enrichment, which leveraged inherent differences in cell morphology and dielectric properties, to achieve the selective separation of these strains under physiological conditions. A substantial increase in the relative abundance of PHB-producing cells was assessed by optical microscopy and flow cytometry characterization, confirming more than five-fold reduction of the Synechococcus fraction in the refined cell mix. The presented electrokinetic platform offers a scalable and effective approach for selectively enhancing desired microbial components within microbial biomanufacturing systems, leading towards improved product yields.

60 APPLIED LIFE SCIENCES↗

Bioengineered algal lipids enriched in structured medium- and long-chain triacylglycerols, linoleate, and sn -2 palmitate for human milk fat substitutes

Human milk fat (HMF) contains triacylglycerol (TAG) as its primary component, providing over 50% of the calories for infant nutrition, along with structural and bioactive lipids that are important for immune and nervous system development. Palmitic acid, comprising 20-25% of the fatty acid complement of HMF, is predominantly esterified to the sn -2 position on the glycerol backbone. This regiospecific positioning facilitates absorption as 2-palmitoyl-monoacylglycerol after hydrolysis of the fatty acids at sn -1 and sn -2 by gut lipases. Other features of HMF include enrichment in structured medium- and long-chain triglycerides (MLCTs), and variation in the ratio of oleic acid to linoleic acid with maternal diet and geography. We have engineered Auxenochlorella, an oleaginous green alga, for biosynthesis of an MLCT- and sn -2 palmitate-enriched HMF substitute for infant formula, matching the regioisomeric composition and proportions of the most abundant fatty acids in HMF.

Lin, Jon Y-T [University of California, Berkeley;]↗

Spent enrichment media for growth of gene-matched microorganisms

A method for amplifying and detecting microorganisms, such as species of Listeria, is described. The method utilizes gene-matched enrichment media and PCR-based detection. The enrichment media is spent media produced using a modified microorganism containing a plurality of mutations in a selected gene such that the modified microorganism does not contain the PCR signature. Thus, PCR detects only the amplified microorganism of interest, not the modified microorganism. Exemplary methods and kits for amplification and detection of Listeria species are described.

Ozanich, Richard M.↗

Emulating Variable Uranium Enrichments With Radiation Signature Training Devices (RSTD)

Radiological signature training devices (RSTDs) are sealed sources (or assemblies of sources) that emulate the radiation signature of larger masses of special nuclear material for passive detection while maintaining a minimal logistical footprint in security, radiological safety, criticality safety, and transportation requirements. Previous work focused on emulation of large quantities of highly enriched uranium for search and field identification with radioisotope identification detectors and mobile systems. New work has expanded into investigating use of the RSTDs with laboratory safeguards tools, such as MGAU and FRAM, and emulating uranium spectra of multiple enrichments over more energy regions (particularly the x-ray region).

Blessinger, Christopher S.↗