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Biochemical inhibition of acid phosphatase activity in two mountain spruce forest soils

The product inhibition of the 4-Methylumbelliferyl phosphate (MUB-P) decay rate, a measure of potential acid phosphatase activity, has not been considered in most of the published kinetic studies. The aim of this study was to determine the type and strength of the product inhibition in order to better define reaction conditions at which the acid phosphatase activity assay produces unbiased results. The MUB-P decay rate was measured in the forest floor and topsoil organic horizons of two spruce forest catchments at different initial MUB-P and P-PO4 3- concentrations. Furthermore, the type and strength of the inhibition was analyzed by non-linear regression. We found that MUB-P decay was competitively inhibited by the P-PO4 3- but also by dissolved organic P. By using estimated kinetic parameters, we calculated an underestimation of potential acid phosphatase activity by up to ~20% at concentrations of added MUB-P as high as 300 µmol g-1 dry weight and an incubation time one hour due to background inorganic and organic P concentrations in range of units of µmol per gram of dry soil. We discuss approaches that can be used to minimize the underestimation and show that the previously defined recommendations might not be always sufficient to avoid the bias. Therefore, we recommend to analyze progress curves at a wide range of initial MUB-P concentrations using non-linear methods when the enzyme assay is optimized for a given soil. When changes in inorganic and/or organic P concentrations are expected, we further recommend measuring background inorganic and organic P concentrations.

59 BASIC BIOLOGICAL SCIENCES↗

Critical and Californium Source-Driven Noise Analysis Subcritical Measurements with an Unreflected Cylindrical Tank of Mixed Uranium-Plutonium Nitrate Solution

The reported experiments were performed by Oak Ridge National Laboratory (ORNL) at the Battelle Northwest Laboratory’s (now Pacific Northwest National Laboratory) critical experiments facility at Hanford, Washington in 1981 and used 16 days of critical facility time, not including 10 days for setup and removal of ORNL equipment. These measurements were to assess the capability of the Cf source-driven noise analysis (CSDNA) method to measure the subcriticality (keff) of mixed U-Pu nitrate solutions. In addition to the CSNDA measurements, measurements were also performed near delayed criticality where CSDNA measurements cannot be performed. This report documents the experiments that were not reported at that time by presenting the ORNL experimental results and any online analysis performed during and shortly after the measurements. The mixed nitrate solution had a U concentration at 188 grams per liter (g/L), a Pu concentration of 280 g/L, free acid normality of 2.80, H ion molarity of 5.9 and a specific gravity of 1.754 g/cm 3 , a 240 Pu isotopic content of 7.981 wt. %, and a 235 U isotopic content of 0.724 wt. %. The stainless-steel tank for the solution had an inside diameter of 35.38 cm, an outside diameter of 35.53 cm, a height of 56.72 cm, and bottom thickness of 0.9525 cm. A Zircaloy pipe with a 3.1496 cm outside diameter, a 2.7788 cm inside diameter, and bottom thickness of 0.635 cm was available for insertion of the Cf source in the center of the fissile solution. The Cf source was also located at the outside surface of the tank (solution height varied from 10 to 53 cm) and in the center of the solution (solution height varied from 10 to 60.7 cm). The CSDNA measurements were not analyzed online to determine the subcritical neutron multiplication factors. At all subcritical states, the break frequency noise analysis data was fitted to obtain the prompt neutron decay constant. The neutron multiplication factors were determined for the two configurations of the measurements near delayed criticality. The subcritical neutron multiplication factors from the CNSDA measurements can be obtained with further analysis. However, the near delayed critical configuration, the prompt neutron decay constants, the count rates, and the measured cross and auto power spectral densities can be calculated directly for benchmarking. Much of data presented in this report are from ORNL notes—not in the ORNL logbooks. For the final benchmark analysis, the data from the Battelle Northwest Laboratory (which operated the critical facility in 1981) critical facility logbook should be consulted and be incorporated where appropriate. The purpose of this report is to document the experimental information for the measurements performed so that at a later date researchers could perform the required uncertainty and calculational analyses and documentation to use these data for an International Criticality Safety Benchmark Program (ICSBEP) or Nuclear Energy Agency benchmark. The data from these measurements are available from the ORNL Records Management Services Department, and the logbook is available from ICSBEP at Idaho National Laboratory. Preparation of the present report is part of a larger cooperative effort between Idaho National Laboratory (INL) and Oak Ridge National Laboratory (ORNL) to document more than 15 undocumented critical and subcritical experiments enumerated in ORNL/TM-2019/18 and performed by ORNL at ORCEF and other USDOE critical experiments facilities using more than 500 operational days of critical facility time.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Total Measurement Uncertainty in Neutron Coincidence Multiplicity Analysis

Neutron multiplicity counting is the most commonly used nondestructive assay technique for determining the plutonium mass within containers of scrap PuO 2 or mixed oxide (MOX). In multiplicity analysis, the 240 Pu eff mass, leakage multiplication, and alpha ratio (the ratio of [α, n]-to-spontaneous fission neutron production) are the three primary unknown sample properties. They must be determined simultaneously. To solve for these three unknowns in a multiplicity assay, three measured values are needed: the singles, doubles, and triples neutron count rates. While the analysis is limited to solving for three unknowns, there are many additional factors that impact the observed count rates and contribute to the measurement uncertainty. In this study we investigate the various uncertainty contributors for the multiplicity analysis through a combination of traditional uncertainty propagation techniques supplemented by Monte Carlo simulations to address the dependences not explicitly expressed by the point source model. Uncertainties arising from counting statistics, calibration parameters, calibration method, nuclear data, and various material characteristics (isotopic abundances, chemical form, density, and impurities) are considered. A Total Measurement Uncertainty (TMU) estimate is then developed from these uncertainty contributors. This study is confined to multiplicity analysis of items commonly encountered in international safeguards applications. That is, the study focused on Pu oxides and MOX materials for the masses ranging up to 4000 grams total Pu. Multiplicity measurements were simulated using MCNP V6 based on the Plutonium Scrap Multiplicity Counter (PSMC), Epithermal Multiplicity Counter (ENMC), Pyrochemical Multiplicity Counter, and Large Epithermal Multiplicity Counter (LEMC) for this study; however, this report focuses on the parameterization of the uncertainties for the PSMC. The performance differences between the PSMC and the other multiplicity counting systems are relatively small, primarily manifesting in the impact on measurement precision so that the evaluation developed for the PSMC can be applied to the other multiplicity counting systems. Finally an analysis tool, the Multiplicity TMU Estimator, was developed from this study to serve as an aid for evaluation of the total measurement uncertainty of multiplicity assay results obtained from the commonly used INCC acquisition and analysis software.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Carbon Dioxide Utilization for Plasma Nanosynthesis of Carbon

This DOE Phase I SBIR project has developed a glow discharge-based plasma process to convert carbon dioxide to sold carbon and water. By adding natural gas, the additional enthalpy can be used to reform the output carbon dioxide to value added carbon and EPA-compliance-grade water, thus creating a process that has no gaseous products at all, such as: CO 2 + CH 4 → 2C + 2H 2 O, which is simply the exothermic reverse of the syngas reaction. During the Phase I work, several milestones were accomplished successfully. a. First, a synthesis reactor was designed, assembled, and successfully operated using the combined simultaneous operation of glow discharge and microwaves. DC discharge and microwaves can also be operated individually, allowing multiple configurations to be used This is one of the first reactors of its kind, and perhaps the first ever. b. Second, the research team successfully produced carbon nanomaterials using carbon dioxide as the source of carbon via CO 2 + 2H 2 → C + 2H 2 O. This is certainly one of the first processes to use plasma to convert carbon dioxide to nanocarbon, and perhaps the first ever demonstration. c. Third, it was also shown that carbon monoxide and carbon dioxide can be used with methane to reduce the synthesis temperature of ASI graphene decorated carbon nanotubes (GDCNT). Hitherto GDCNT were only fabricated above 1200 °C. GDCNT was synthesized at 780 °C. CO 2 + C → 2CO; 2 CO + CH 4 → 3 C + 2H 2 O, and also, CH 4 + CO 2 → 2 C + 2H 2 O. d. Fourth, closed cycle synthesis of nanomaterial was demonstrated with over 80% conversion of carbon dioxide to carbon and water was achieved, on the basis of pressure drop during glow discharge. e. Fifth, plasma operation was achieved via glow discharge and microwaves at 500 Torr, demonstrating a path forward to atmospheric pressure operation f. Sixth, using carbon monoxide and carbon dioxide in an electrothermal furnace, we produced 350 grams per hour of high-quality nanotubes in a single furnace, compared to 200 grams per hour for the commercial process. Controlling the enthalpy prevents self-extinguishing hot gas pyrolysis. Thus, carbon dioxide enhances the properties and reduces the production cost. This is a near term advance that can go into trial production in the near term. g. Seventh, economic analysis suggests an economic plan forward, producing carbon nanotubes for less than $50 per pound. The use of hydrogen and carbon dioxide as the feedstocks is acceptable from a cost basis, and in fact hydrogen is less expensive than carbon monoxide. Niche applications are available in the near term, though quantities relevant to sequestration remain a far-term goal. h. Eighth, though it was not the research team’s intention to create a new process for producing carbon monoxide, it was realized that almost by accident we demonstrated plasma reforming of carbon dioxide via a plasma version of the reverse Boudouard reaction. Accordingly, we have contacted a major gas manufacturer (Matheson Gas) to determine whether there is interest in creating an industrial version of the process. i. Ninth, a long-range strategy is identified in which a progressively more carbon-conscious world will seek to create solid carbon as a means of sequestering carbon dioxide, impeding its return to the atmosphere. In the long term the price must drop by orders of magnitude with larger production, and assuming success, solid carbon could be the preferred form for sequestration rather than underground storage. If carbon can be made as cheaply as dirt, or nearly so, then it can serve a useful purpose as structural material for housing, soil amendments in agriculture and fillers to modify low areas.

03 NATURAL GAS↗

Sphingomonas clade and functional distribution with simulated climate change

Microbes are essential for the functioning of all ecosystems, and as global warming and anthropogenic pollution threaten ecosystems, it is critical to understand how microbes respond to these changes. We investigated the climate response of Sphingomonas, a widespread gram-negative bacterial genus, during an 18-month microbial community reciprocal transplant experiment across a Southern California climate gradient. We hypothesized that after 18 months, the transplanted Sphingomonas clade and functional composition would correspond with site conditions and reflect the Sphingomonas composition of native communities. We extracted Sphingomonas sequences from metagenomic data across the gradient and assessed their clade and functional composition. Representatives of at least 12 major Sphingomonas clades were found at varying relative abundances along the climate gradient, and transplanted Sphingomonas clade composition shifted after 18 months. Site had a significant effect (PERMANOVA; P < 0.001) on the distribution of both Sphingomonas functional (R 2 = 0.465) and clade composition (R 2 = 0.400), suggesting that Sphingomonas composition depends on climate parameters. Additionally, for both Sphingomonas clade and functional composition, ordinations revealed that the transplanted communities shifted closer to the native Sphingomonas composition of the grassland site compared with the site they were transplanted into. Overall, our results indicate that climate and substrate collectively determine Sphingomonas clade and functional composition.

59 BASIC BIOLOGICAL SCIENCES↗

The calculation of light element impurity (α, n) yield curves in a PuO 2 matrix and associated specific yield coefficients: Influence of the reaction cross sections

Most of the Pu separated from irradiated commercial nuclear fuel is stored as PuO 2 . The primary quantitative nondestructive measurement technique used to verify the amount of Pu in storage containers is passive neutron correlation counting. An important physical property of the oxide material is the ratio, α, of the rate of (α, n) neutrons produced inside the item to the rate of neutrons produced by spontaneous fission. This ratio influences the precision of the correlated counting method and affects the interpretation of the data because of how it changes both the primary total neutron production rate and the rate of induced fission events taking place inside the item. In addition to the main O(α, n) contribution, additional contributions come from α-particle interactions with light element impurities that are inevitably present. In this work, we calculate specific (α, n) yield coefficients, expressed in units of neutrons per second per gram of α-emitting nuclide per part per million by mass of the specified impurity element distributed in a pure PuO 2 matrix, for some key α-emitting actinides commonly present in reprocessed Pu ( 238–242 Pu+ 241 Am). These coefficients are directly applicable to nuclear safeguards verification work in which the α ratio is often calculated from the Pu-isotopic composition and chemical information obtained by other means. Further, they also provide a convenient up-to-date reference set against which values generated by other methods can be compared. Results are presented for impurities with atomic number from 3 to 17 inclusive, plus K and Fe. In most cases, these coefficients are not expected to change by more than 5%–10% at any time in the future. However, as new data become available, changes as large as 20% may be needed for some targets (e.g., F). The present yield calculations are limited by the general shortage of quality experimental total (α, n) reaction cross section data, which, together with unexplained variation between determinations, means that an objective and coherent evaluation is not possible. The situation is even less satisfactory for the partial differential cross section needed to calculate neutron spectra.

(𝛼, n) reactions↗

Enhanced phagocytosis and complement-mediated killing of Mannheimia haemolytica serotype 1 following in-frame CMP-sialic acid synthetase ( neuA ) gene deletion

ABSTRACT Mannheimia haemolyticais the most significant bacterial pathogen associated with the bovine respiratory disease complex. Although sialic acid is a known virulence factor in other members of Pasteurellaceae, such asHistophilus somniandPasteurella multocida, the significance of sialic acid to the virulence ofM. haemolyticais currently unknown. Therefore, the role of sialic acid as a virulence determinant ofM. haemolyticawas investigated by constructing an in-frameneuA[CMP-N-acetylneuraminic acid (Neu5Ac/sialic) synthetase] mutant, which was shown by high-performance anion exchange chromatographic analysis (HPAEC) to be devoid of sialic acid on the lipopolysaccharide (LPS). Both theneuAmutant and wild-type parent strains exhibited similar growth rates in the growth curve assay. Real-time qPCR and ELISA evaluation showed no differences in proinflammatory cytokine expressions (IL-1β, IL-6, and IL-8) between theneuAmutant and parent strain when peripheral blood mononuclear cells were incubated with LPS. Interestingly, theneuAmutant was three to four logs more sensitive to a whole-blood bacterial killing assay than the parent strain. Similar results were also observed in plasma and serum bacterial killing assays. Flow cytometry analyses showed higher uptake ofneuAmutant by phagocytes, compared to the parent strain, in the whole-blood phagocytosis assay; however, no difference in reactive oxygen species production in neutrophils or monocytes was detected for either strain. Taken together, these results indicate that sialylation ofM. haemolyticaLPS plays a vital role in reducing complement-mediated and phagocytic killing. IMPORTANCE The Gram-negative coccobacillusMannheimia haemolyticais a natural inhabitant of the upper respiratory tract in ruminants and the most common bacterial agent involved in bovine respiratory disease complex development. Key virulence factors harbored byM. haemolyticaare leukotoxin, lipopolysaccharide, capsule, adhesins, and neuraminidase which are involved in evading innate and adaptive immune responses. In this study, we have shown that CMP-sialic acid synthetase (neuA) is necessary for the incorporation of sialic acid onto the membrane, and inactivation ofneuAresults in increased phagocytosis and complement-mediated killing ofM. haemolytica,thus demonstrating that sialylation contributes to the virulence ofM. haemolytica.

Microbiology↗

Assessing Gas-Phase Ion Reactivity of 50 Elements with NO and the Direct Application for 239 Pu in Complex Matrices Using ICP-MS/MS

Understanding the reactivity of metal cations with various reaction gases in ICP-MS/MS is important to determine the best gas to use for a given analyte/interference pair. In this study, nitric oxide (NO) was investigated as the reaction gas following previous experimental designs. The reactions with 50 elements were investigated to examine periodic trends in reactivity, validate theoretical modeling of reaction enthalpies as a method to screen reactant gases, and provide a baseline for potential in-line gas separation methods. ICP-MS/MS studies involving actinides are typically limited to Th, U and Pu, with analyses of Np and Am rarely reported in the literature. To date, only two previous methods have investigated the use of NO in ICP-MS/MS analyses. To showcase the utility of NO, a method was developed to measure 239 Pu in the presence of environmental matrix constituent and other actinides, like what could be expected from post-detonation debris, with no chemical separation prior to analysis. 239 Pu + was reacted to form 239 Pu 16 O + , eliminating interferences derived from the sample matrix by measuring 239 Pu + intensity at m/z = 255 ( 239 Pu 16 O + ). To validate NO for 238 U 1 H + interference removal in environmental matrices, standard reference materials were diluted to 1 mg per gram of solution and spiked to 0.05 pg/g of 239 Pu and 1 µg/g 238 U (Pu/U = 5 x 10 -8 ). Measured 239 Pu concentrations were within 6% of the spiked value. In conclusion, these results demonstrate that reliable 239 Pu measurements can be made at levels relevant to nuclear forensics without the need for extensive chemical matrix separation prior to analysis.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Parametric Study of 239 Pu and 235 U Mass Equivalency in Spherical Geometries

In this technical document, parametric studies were performed to determine if a bounding mass equivalency can be used to simplify multi-material models in MCNP6 for criticality evaluations. The studies explored the simplification of MCNP6 models by substituting materials, particularly 239 Pu for 235 U. The initial hypothesis for the material simplification is based on the subcritical mass limits from the ANSI/ANS- 8.1 standard which states that for water reflected metal spheres the limits are 20.1 kg 235 U and 5.0 kg 239 Pu. Using those masses, it was hypothesized that a 20.1 kg sphere of 235 U and a 5.0 kg sphere of 239 Pu could be considered to have the same multiplication. Given that equivalency, 4kg 235 U could be modeled by 1kg 239 Pu (i.e., a mass ratio of 4:1). The parametric studies compare the materials 239 Pu and 235 U independently and in multi-material spherical models. Critical mass, critical thickness, critical mass ratio and critical thickness ratio values are presented as functions of inner radius. keff values are presented as a function of inner radius at different mass ratios and total masses. Results indicate that the conservative bounding mass equivalency is one. That is, if substituting 239 Pu for 235 U it should be done on a gram for gram basis.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

South Africa’s Appliance Energy Efficiency Standards and Labeling Program

Electricity consumption in South Africa comes with a hefty environmental cost to the society. For every kilowatt-hour (kWh) produced, 1 kilogram (kg) of carbon dioxide (CO 2 ) is emitted, 1.4 liters of water are used, and 0.37 grams of particulate emissions are released in the atmosphere. These environmental implications result from the large share of electricity produced from coal (91 percent in 2015). While planned new capacity will ramp up renewable energy, the integrated resource plan for the country still projects the share of col to be 64 percent in 2030. Energy savings provide environmental benefits as well as economic benefits. Energy efficiency standards and labeling (S&L) programs are a policy measure proven to save energy. Such programs have been implemented in many countries to remove inefficient technologies and transform markets to more efficient technologies. In this study, we describe the methodology, assumptions and results of a stock turnover modeling tool that estimates the energy savings achievable by South Africa’s S&L program in the residential sector. We show that if regulations are passed in 2020 and effective in 2021 for 10 major end-use adopting international standards best practices, 6 terawatt-hours (TWh) will be saved in 2030 and 9.5 TWh will be saved in 2040, representing a CO 2 mitigation of 3.7 megatonnes (Mt) in 2030 and 5.8 Mt in 2040, which will contribute to the South African government’s international engagement in fighting against climate change through its National Determined Contribution (NDC). Additional environmental benefits include saving of 6.5 billion liters of water in 2030, representing approximately 100 liters per capita in 2030. Air quality will also improve, as 4 kilotons (kt) of particulate emissions will be avoided, as well as 4.3 Mt of sulfur oxides (SOx) and 23 kt of nitrogen oxides (NOx) in 2030.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Sulfonamidoboronic Acids as “Cross-Class” Inhibitors of an Expanded-Spectrum Class C Cephalosporinase, ADC-33, and a Class D Carbapenemase, OXA-24/40: Strategic Compound Design to Combat Resistance in Acinetobacter baumannii

Acinetobacter baumannii is a Gram-negative organism listed as an urgent threat pathogen by the World Health Organization (WHO). Carbapenem-resistant A. baumannii (CRAB), especially, present therapeutic challenges due to complex mechanisms of resistance to β-lactams. One of the most important mechanisms is the production of β-lactamase enzymes capable of hydrolyzing β-lactam antibiotics. Co-expression of multiple classes of β-lactamases is present in CRAB; therefore, the design and synthesis of “cross-class” inhibitors is an important strategy to preserve the efficacy of currently available antibiotics. To identify new, nonclassical β-lactamase inhibitors, we previously identified a sulfonamidomethaneboronic acid CR167 active against Acinetobacter-derived class C β-lactamases (ADC-7). The compound demonstrated affinity for ADC-7 with a K i = 160 nM and proved to be able to decrease MIC values of ceftazidime and cefotaxime in different bacterial strains. Herein, we describe the activity of CR167 against other β-lactamases in A. baumannii: the cefepime-hydrolysing class C extended-spectrum β-lactamase (ESAC) ADC-33 and the carbapenem-hydrolyzing OXA-24/40 (class D). These investigations demonstrate CR167 as a valuable cross-class (C and D) inhibitor, and the paper describes our attempts to further improve its activity. Five chiral analogues of CR167 were rationally designed and synthesized. The structures of OXA-24/40 and ADC-33 in complex with CR167 and select chiral analogues were obtained. The structure activity relationships (SARs) are highlighted, offering insights into the main determinants for cross-class C/D inhibitors and impetus for novel drug design.

60 APPLIED LIFE SCIENCES↗

Nuclear Criticality Safety Assessment of Criticality Control Containers without Moderation Control at the Waste Isolation Pilot Plant

The Waste Isolation Pilot Plant (WIPP) provides for safe, permanent disposal of government-owned transuranic (TRU) and TRU mixed wastes. Receipt and disposal of waste at the WIPP site began in March 1999. The Sandia report, Consideration of Nuclear Criticality When Disposing of Transuranic Waste at the Waste Isolation Pilot Plant, addressed potential nuclear criticality safety issues based on the projected inventory characteristics known at the time [1]. As designs for inventory, waste forms, and disposal packages have changed, new analyses have been performed, and updates have been made to address any potential effects to the WIPP safety basis. New analyses performed include Saylor 2017 [2] and Brickner 2019 [3], which address certain waste containers with specified loadings under post-closure conditions. Both examined several hypothetical scenarios and included analyses to bound (from a criticality potential standpoint) credible configurations that could occur at WIPP during the repository regulatory post-closure disposal time period for feature, event, and process (FEP) considerations—10,000 years. During this post-closure period at WIPP, the screening of FEPs is governed by the risk-based standards and implementing regulations of the US Environmental Protection Agency (EPA) (i.e., 40 CFR 191 and 40 CFR 194, respectively) [4,5]. An FEP screening can be based on either a low-consequence or low-probability rationale. A low-probability rationale includes either (a) a qualitative rationale that the FEP is not credible or (b) a quantitative demonstration that the probability is less than 10-4 in 104 years. In this evaluation, a qualitative lowprobability rationale of not credible is used by demonstrating that bounding configurations of the waste are not critical. The demonstration of subcriticality is through quantitative calculations, but a probability of criticality is not evaluated. Rather, the rationale for this evaluation is that bounding configurations with an effective neutron multiplication factor (keff) well below the upper subcriticality limit (USL) make criticality incredible. Reference [2] documented a nuclear criticality assessment of the WIPP repository for disposal of dilute surplus plutonium materials using the Dilute and Dispose Approach and packaging in criticality control overpacks (CCOs). The CCO is the waste disposal container recently designed to allow for up to 380 fissile gram equivalent (FGE) 239 Pu per drum, which is a higher fissile loading than typical waste containers. The CCO consists of a criticality control container (CCC) positioned by upper and lower plywood spacers within a standard 55 gal drum. The CCC is used to establish a geometry control for fissile materials during transportation and WIPP emplacement operations. The current WIPP waste acceptance criteria for CCO payloads limit beryllium to less than or equal to 1% by weight of the waste contents and require the waste form to be non-machine compacted. Reference [2] considered two scenario progressions—room closure from salt creep, hereafter referred to as the reconfigured dry scenario, and flooding with brine, hereafter referred to as the reconfigured wet scenario. The subsequent drying out of the reconfigured wet scenarios was also considered. For all scenarios, subcriticality was maintained when 50 g of B 4 C (acting as a neutron absorber) per CCC was intermixed within the plutonium disposition waste form. The analysis used a waste form description that limits the amount of moderation that could be present within the waste form (i.e., it limits the amount of water and polyethylene that could be present based on planned processing conditions). This analysis to evaluate increased limits on the amount of moderation that could be present was performed as a companion to Reference [2] to address concerns associated with verifying moisture and/or plastic contents of waste materials following packaging of dilute surplus plutonium in the CCO. To that end, this analysis used the models and methods from Reference [2] to evaluate a more generic base waste form consisting of water and polyethylene that is more similar (and nearly identical) to the generic waste forms utilized in other models/analyses supporting the TRU Package Transporter Model II (TRUPACTII) safety analysis [6] (all are without moderation controls). The waste form in this analysis uses a base mixture of 75% water and 25% polyethylene, the total amount of which is varied to determine the optimum moderation to fissile material (H/Pu) ratio. The fissile loading is maintained at up to 380 FGE 239 Pu (modeled as PuO 2 ) per CCO with an additional 545 g of beryllium (to bound the 1% by weight contents restriction) and 50 g of B 4 C intermixed per CCO. The beryllium content (1% by weight) is based on the total allowed waste weight (this does not include packaging and container weights). Figures ES-1 and ES-2 display summary results, showing that with this model including 50 g of B 4 C per CCO, the system keff remains under 0.85 for all moderator amounts and provides a significant margin against post-closure criticality under postulated bounding conditions for compaction. Figure ES-1 compares an infinite model with a room model at the initial emplacement spacing and under full radial compaction. Full radial compaction places each CCC in direct contact and does not credit any anticipated spacing associated with current post-closure geomechanical modeling of the repository [7]. The effects of variations in the H/Pu ratio were evaluated by varying the amount of the water/polyethylene component of the waste model, with fissile loading maintained at 380 239 Pu FGE. Similarly, Figure ES-2 illustrates how various amounts of B 4 C per CCO influence k eff at different radial compactions, all at the H/Pu ratio of 200 (in the room array model). Therefore, while the results from Saylor 2017 [2] modeled more realistic process limits associated with packaging of dilute surplus plutonium, this analysis demonstrates that limits on moderation (plastic and water content) are not necessary to ensure subcriticality in the WIPP repository, provided the requisite B 4 C absorber is present.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Identification and characterization of mono- and bifunctional galactan synthases in the pediatric pathogen Kingella kingae

The emerging pediatric pathogen Kingella kingae elaborates a lipopolysaccharide (LPS) that is extended with a galactofuranose homopolymer called galactan, which is a key virulence determinant that contributes to resistance to complement-mediated and neutrophil-mediated killing. Previous work has demonstrated that the pamABCDE locus is required for galactan synthesis. In this study, mutational studies suggested that the pamC gene product is a UDP-galactofuranose (Galf) transferase and is the galactan synthase. Analysis of genome sequence data revealed two distinct pamC alleles designated pamC1 and pamC2, which correlate with the two galactan structures in K. kingae. Examination of isogenic mutants expressing either pamC1 or pamC2 demonstrated that the pamC alleles are the determinants of galactan structure. Experiments with recombinant PamC1 and PamC2 in vitro established that these proteins are galactan synthases capable of extending synthetic Galf disaccharide acceptors in the presence of UDP-Galf. Homology analysis identified critical amino acids that are essential for PamC1 and PamC2 enzymatic activity both in vitro and in K. kingae. Structural analysis of the in vitro-modified synthetic acceptors implicated PamC1 as a monofunctional enzyme capable of generating a β-(1 → 5) Galf linkage and PamC2 as a bifunctional enzyme capable of generating β-(1 → 3) and β-(1 → 6) Galf linkages. This study advances our understanding of the GT2 family of UDP-galactofuranosyltransferases.

60 APPLIED LIFE SCIENCES↗

The Snow Albedo Evolution (SALVO) Campaign at the North Slope of Alaska Field Campaign Report

The springtime surface-albedo transition in the Alaskan Arctic, along with the forces that determine its duration and nature, was the focus of the Snow ALbedo eVOlution (SALVO) campaign. The SALVO team investigated the reasons and durations of the stages of spring melt, during which albedo values decrease from 0.8 to 0.1, signifying the year’s largest and most significant radiative energy change. SALVO II (2022, 2024) built on findings from the successful SALVO I (2019) melt season campaign and previous research on the melt transition period conducted by Grenfell and Perovich (2004). SALVO used the ARM NSA observatory, thus aligning with a core principle of the ARM Decadal Vision to “provide comprehensive and impactful field measurements to support scientific advancement of atmospheric process understanding.” SALVO fieldwork was conducted in the spring near the ARM NSA Central Facility in Utqiagvik, Alaska (Figure 1). Comprehensive data sets were collected in 2019, 2022, and 2024, which included spectral and broadband albedos, snow and meltwater depths, snow stratigraphy, snow grain size data, and aerial imagery. In April of each project year, before the snow began to melt, we established survey lines at three or four locations: inland tundra at NSA E12, coastal tundra at the NSA C1 site, Elson Lagoon, and offshore on the Chukchi Sea sea ice (2022 only). We set up a 200-meter-long line, marked every five meters, to enable repeated measurements of surface albedo and snow depth at each location. During most site visits, the SALVO II team dug at least one snow pit to assess the characteristics of the vertical snow layers, including the types and sizes of snow grains, snow density, and the distribution of liquid water. We evaluated the characteristics of flowing or pooled water at the bottom of the snowpack. Watching snow melt in the Arctic has been compared by some to being as exciting as watching paint dry, but it is far more exciting and dynamic than that. Initially, changes happen slowly while the snow cover remains above 80%, but then reductions in the albedo of the landscape, from about 0.8 to 0.4, combined with rising spring temperatures, begin to accelerate the melt. It happens so quickly that no matter how hard the field team works, they cannot keep up with documenting the changes. At first, liquid water is scarce—only a few wet layers of snow in the snowpack. But then, water becomes ubiquitous, transporting melt energy and creating an extremely heterogeneous albedo and melt landscape. At the end of each field season, the SALVO team is exhausted, relieved to see the snow gone, and joyful to play in the melt ponds on the sea ice under the midnight sun. Overall, the 2024 SALVO field campaign was a highlight, reflecting numerous lessons learned from 2019 and 2022. Daily measurements were taken more than 10 times at each site, resulting in a comprehensive time series of snow conditions and albedo. The team deployed using snowmachines and then on foot when snowmachine travel was no longer permitted on the melting tundra. Instrument mounts and packaging were optimized for quick deployment, regardless of the mode of transportation.

54 ENVIRONMENTAL SCIENCES↗