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At least 703 records · Page 39

Standard Analysis Report INV-SAR-81, Revision 0 Chemical and Cement Components 2023 Inventory Estimates for the Interim State Performance Assessment of the Waste Isolation Pilot Plant

This standard analysis report provides the estimates for the chemical (oxyanions and complexing agents) and cement components with a data collection cut-off date of December 31, 2023. These estimates will be included in a Performance Assessment Inventory Report developed for the U.S. Department of Energy (DOE) interim state repository configuration performance assessment (PA) for the 2026 Compliance Recertification Application (CRA).

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Isolation and Conversion of Electrolyte Components into a Value-Added Product

The overall objective of this work was to develop a process for effectively (i) recycling the electrolyte components from the end-of-life batteries and reduce the environmental hazard, (ii) understand the fundamental differences between pristine and recycled electrolyte components, (iii) electrochemically and analytically evaluate the recycled electrolyte components and (iv) establish deviations from the pristine electrolyte, and effectively separation of value-added product. We focused on determining scaling up the electrolyte recovery process by distillation of end-of-life batteries to get yield enough to use it for electrolyte formulation of reuse in the battery cell. The used cells from the ORNL battery manufacture facility (BMF) as well as received from the industry partner, Austin Elements Inc., was scaled to 10 used multilayered pouch cells in each batch of distillation and systematically separated and identified the electrolyte components of the used cells with FTIR and NMR studies. We were able to recovery 10g of solvent mass from the used pouch cells. NMR spectroscopy showed that the solvent was pure EMC, DMC and the sample was used to make a new battery cell. The end-of-life battery, it was noted that some cells were visually drier than the other cells and were more advanced in aged. In that case, even though the same distillation conditions were utilized, no usable mass of electrolyte solvent was recovered. It is expected that most of the solvent was decomposed during the battery charge-discharge cycling. The solid salt residue was separated by chemical and water treatment processes. Components of resulting solid product was analyzed by XRD and XPS measurements. The solid products are very much related to the salts used in the electrolyte solution.

25 ENERGY STORAGE↗

KBase Narrative - BIT 295 MRA Isolate 106- Final

Arthrobactor is a bacteria that commonly resides in soil, plants, and wastewater (Gobbetti 2014). It is commonly used in the agricultural industry for degrading pesticides which will detox the soil and allow wildlife not to be harmed by said pesticides. The sample that is being tested came from an acid drain from a mine. The test sample was able to live and thrive in a metal ridden environment which could lead it to having some genes that could aid in e-waste recycling. A species of arthrobacter known as Arthrobacter citreus can be used for polyamide waste (Baxi 2019). Polyamide is used in fishing nets and industrial applications and the waste generated by this impacts ocean environments and ecosystems (Rietzler et all... 2021). Based on the resilience of arthrobacter as a genus, the hopes for this sample to aid in e-waste recycling are high.

Chadha, Aditya↗

Targeted mutagenesis and high-throughput screening of diversified gene and promoter libraries for isolating gain-of-function mutations

Targeted mutagenesis of a promoter or gene is essential for attaining new functions in microbial and protein engineering efforts. In the burgeoning field of synthetic biology, heterologous genes are expressed in new host organisms. Similarly, natural or designed proteins are mutagenized at targeted positions and screened for gain-of-function mutations. Here, we describe methods to attain complete randomization or controlled mutations in promoters or genes. Combinatorial libraries of one hundred thousands to tens of millions of variants can be created using commercially synthesized oligonucleotides, simply by performing two rounds of polymerase chain reactions. With a suitably engineered reporter in a whole cell, these libraries can be screened rapidly by performing fluorescence-activated cell sorting (FACS). Within a few rounds of positive and negative sorting based on the response from the reporter, the library can rapidly converge to a few optimal or extremely rare variants with desired phenotypes. Library construction, transformation and sequence verification takes 6–9 days and requires only basic molecular biology lab experience. Screening the library by FACS takes 3–5 days and requires training for the specific cytometer used. Further steps after sorting, including colony picking, sequencing, verification, and characterization of individual clones may take longer, depending on number of clones and required experiments.

59 BASIC BIOLOGICAL SCIENCES↗

Pulse Protein Isolates as Competitive Food Ingredients: Origin, Composition, Functionalities, and the State-of-the-Art Manufacturing

The ever-increasing world population and environmental stress are leading to surging demand for nutrient-rich food products with cleaner labeling and improved sustainability. Plant proteins, accordingly, are gaining enormous popularity compared with counterpart animal proteins in the food industry. While conventional plant protein sources, such as wheat and soy, cause concerns about their allergenicity, peas, beans, chickpeas, lentils, and other pulses are becoming important staples owing to their agronomic and nutritional benefits. However, the utilization of pulse proteins is still limited due to unclear pulse protein characteristics and the challenges of characterizing them from extensively diverse varieties within pulse crops. To address these challenges, the origins and compositions of pulse crops were first introduced, while an overarching description of pulse protein physiochemical properties, e.g., interfacial properties, aggregation behavior, solubility, etc., are presented. For further enhanced functionalities, appropriate modifications (including chemical, physical, and enzymatic treatment) are necessary. Among them, non-covalent complexation and enzymatic strategies are especially preferable during the value-added processing of clean-label pulse proteins for specific focus. This comprehensive review aims to provide an in-depth understanding of the interrelationships between the composition, structure, functional characteristics, and advanced modification strategies of pulse proteins, which is a pillar of high-performance pulse protein in future food manufacturing.

59 BASIC BIOLOGICAL SCIENCES↗

An Immunomodulatory Polysaccharide–Protein Complex Isolated from the Polypore Fungus Royoporus badius

Many wild edible polypore mushrooms have medicinal value. In this study, we investigate the potential medicinal properties of the wild polypore mushroom Royoporus badius collected from north-central British Columbia, Canada. Water extract from R. badius was found to exhibit potent immunomodulatory activity. The extract was purified using DEAE-Sephadex anion-exchange chromatography as well as Sephacryl S-500 and HPLC BioSEC5 size-exclusion chromatography, to yield a novel polysaccharide-protein complex (IMPP-Rb).IMPP-Rb has a peak maxima molecular weight (M p ) of 950 kDa. GC-MS analyses showed that IMPP-Rb is composed predominantly of glucose (49.2%), galactose (11.3%), mannose (10.8%), rhamnose (9.6%), and galacturonic acid (8.2%), with smaller amounts of xylose (5.2%), fucose (2.8%), N-acetyl glucosamine (1.8%), and arabinose (1.2%). IMPP-Rb has multiple linkages, with 4-Glcp, 4-Manp, 6-Manp, 3,4-Manp, 4-Xylp, and 2-Rhap being the most prominent. IMPP-Rb is capable of inducing many cytokines in vitro and the protein component is indispensable for its immunomodulatory activity. IMPP-Rb has potential application as an immuno-stimulatory agent with pharmaceutical value.

59 BASIC BIOLOGICAL SCIENCES↗

Wild Isolates of Neurospora crassa Reveal Three Conidiophore Architectural Phenotypes

The vegetative life cycle in the model filamentous fungus, Neurospora crassa, relies on the development of conidiophores to produce new spores. Environmental, temporal, and genetic components of conidiophore development have been well characterized; however, little is known about their morphological variation. We explored conidiophore architectural variation in a natural population using a wild population collection of 21 strains from Louisiana, United States of America (USA). Our work reveals three novel architectural phenotypes, Wild Type, Bulky, and Wrap, and shows their maintenance throughout the duration of conidiophore development. Furthermore, we present a novel image-classifier using a convolutional neural network specifically developed to assign conidiophore architectural phenotypes in a high-throughput manner. To estimate an inheritance model for this discrete complex trait, crosses between strains of each phenotype were conducted, and conidiophores of subsequent progeny were characterized using the trained classifier. Our model suggests that conidiophore architecture is controlled by at least two genes and has a heritability of 0.23. Additionally, we quantified the number of conidia produced by each conidiophore type and their dispersion distance, suggesting that conidiophore architectural phenotype may impact N. crassa colonization capacity.

59 BASIC BIOLOGICAL SCIENCES↗

Isolating the Source Region of Infrasound Travel Time Variability Using Acoustic Sensors on High-Altitude Balloons

High-altitude balloons carrying infrasound sensor payloads can be leveraged toward monitoring efforts to provide some advantages over other sensing modalities. On 10 July 2020, three sets of controlled surface explosions generated infrasound waves detected by a high-altitude floating sensor. One of the signal arrivals, detected when the balloon was in the acoustic shadow zone, could not be predicted via propagation modeling using a model atmosphere. Considering that the balloon’s horizontal motion showed direct evidence of gravity waves, we examined their role in infrasound propagation. Implementation of gravity wave perturbations to the wind field explained the signal detection and aided in correctly predicting infrasound travel times. Our results show that the impact of gravity waves is negligible below 20 km altitude; however, their effect is important above that height. The results presented here demonstrate the utility of balloon-borne acoustic sensing toward constraining the source region of variability, as well as the relevance of complexities surrounding infrasound wave propagation at short ranges for elevated sensing platforms.

47 OTHER INSTRUMENTATION↗

Deletion of African Swine Fever Virus Histone-like Protein, A104R from the Georgia Isolate Drastically Reduces Virus Virulence in Domestic Pigs

African swine fever virus (ASFV) is the etiological agent of a frequently lethal disease, ASF, affecting domestic and wild swine. Currently, ASF is causing a pandemic affecting pig production in Eurasia. There are no vaccines available, and therefore control of the disease is based on culling infected animals. We report here that deletion of the ASFV gene A104R, a virus histone-like protein, from the genome of the highly virulent ASFV-Georgia2010 (ASFV-G) strain induces a clear decrease in virus virulence when experimentally inoculated in domestic swine. A recombinant virus lacking the A104R gene, ASFV-G-ΔA104R, was developed to assess the role of the A104R gene in disease production in swine. Domestic pigs were intramuscularly inoculated with 10 2 HAD 50 of ASFV-G-ΔA104R, and compared with animals that received a similar dose of virulent ASFV-G. While all ASFV-G inoculated animals developed a fatal form of the disease, animals receiving ASFV-G-ΔA104R survived the challenge, remaining healthy during the 28-day observational period, with the exception of only one showing a protracted but fatal form of the disease. ASFV-G-ΔA104R surviving animals presented protracted viremias with reduced virus titers when compared with those found in animals inoculated with ASFV-G, and all of them developed a strong virus-specific antibody response. This is the first report demonstrating that the A104R gene is involved in ASFV virulence in domestic swine, suggesting that A104R deletion may be used to increase the safety profile of currently experimental vaccines.

59 BASIC BIOLOGICAL SCIENCES↗

Evaluation of the Deletion of the African Swine Fever Virus Gene O174L from the Genome of the Georgia Isolate

African swine fever virus (ASFV) is a structurally complex, double-stranded DNA virus, which causes African swine fever (ASF), a contagious disease affecting swine. ASF is currently affecting pork production in a large geographical region, including Eurasia and the Caribbean. ASFV has a large genome, which harbors more than 160 genes, but most of these genes’ functions have not been experimentally characterized. One of these genes is the O174L gene which has been experimentally shown to function as a small DNA polymerase. Here, we demonstrate that the deletion of the O174L gene from the genome of the virulent strain ASFV Georgia2010 (ASFV-G) does not significantly affect virus replication in vitro or in vivo. A recombinant virus, having deleted the O174L gene, ASFV-G-ΔO174L, was developed to study the effect of the O174L protein in replication in swine macrophages cultures in vitro and disease production when inoculated in pigs. The results demonstrated that ASFV-G-ΔO174L has similar replication kinetics to parental ASFV-G in swine macrophage cultures. In addition, animals intramuscularly inoculated with 10 2 HAD 50 of ASFV-G-ΔO174L presented a clinical form of the disease that is indistinguishable from that induced by the parental virulent strain ASFV-G. All animals developed a lethal disease, being euthanized around day 7 post-infection. Therefore, although O174L is a well-characterized DNA polymerase, its function is apparently not critical for the process of virus replication, both in vitro and in vivo, or for disease production in domestic pigs.

59 BASIC BIOLOGICAL SCIENCES↗