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At least 199 records · Page 11
All sourcesNASA NTRSDOE OSTI
DOE OSTI2025

New Mobility Technology Assessment - Benefits, Risks, and Safety

The age of vehicle automation is upon us. The fundamental perception and control technology is maturing as we speak (and write) with robo-taxis proliferating in select US cities. The news cycle is no longer filled with technological skepticism, but rather societal commentary on our collective response and use of the technology. This maturing of the technology is also accompanied by a renaissance in new types of urban movement technology to more effectively link people with goods, services, and employment, particularly in dense development areas where vehicle congestion (even automated vehicles) prevents high quality mobility. The National Renewable Energy Laboratory, through the DOE Clean Energy to Communities (C2C) program has worked with two jurisdictions to assess the benefits, risks, and safety concerns for a new product offering based on suspended cable technology which is fully automated and proposes an aerial network of self-propelled gondolas. The recognized need, independent of the technology, is to fairly assess the benefits of the proposed transportation system, assess its technical maturity, as well as risks and safety concerns. Using this new technology as an example, a technology represented by Swyft Cities, NREL developed a succession of mini-studies that characterized the technical maturity level, assessed the proposed benefits for Greenville, SC, examined the risks involved in moving forward with an emerging mobility technology, and determined the governing safety protocols and standards that are applicable to the space. These mini-case studies, though centered on Swyft Cities aerial automated gondola technology, are highly transferable to any type of new mobility technology and lays out a repeatable process for assessing new technology within an city or community context.

33 ADVANCED PROPULSION SYSTEMS↗
DOE OSTI2024

Mechanical forces orchestrate the metabolism of the developing oilseed rape embryo

The initial free expansion of the embryo within a seed is at some point inhibited by its contact with the testa, resulting in its formation of folds and borders. Although less obvious, mechanical forces appear to trigger and accelerate seed maturation. However, the mechanistic basis for this effect remains unclear. Manipulation of the mechanical constraints affecting either the in vivo or in vitro growth of oilseed rape embryos was combined with analytical approaches, including magnetic resonance imaging and computer graphic reconstruction, immunolabelling, flow cytometry, transcriptomic, proteomic, lipidomic and metabolomic profiling. Our data implied that, in vivo, the imposition of mechanical restraints impeded the expansion of testa and endosperm, resulting in the embryo's deformation. An acceleration in embryonic development was implied by the cessation of cell proliferation and the stimulation of lipid and protein storage, characteristic of embryo maturation. The underlying molecular signature included elements of cell cycle control, reactive oxygen species metabolism and transcriptional reprogramming, along with allosteric control of glycolytic flux. Constricting the space allowed for the expansion of in vitro grown embryos induced a similar response. The conclusion is that the imposition of mechanical constraints over the growth of the developing oilseed rape embryo provides an important trigger for its maturation.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Moving the needle: Employing deep reinforcement learning to push the boundaries of coarse-grained vaccine models

Highly mutable infectious disease pathogens (hm-IDPs) such as HIV and influenza evolve faster than the human immune system can contain them, allowing them to circumvent traditional vaccination approaches and causing over one million deaths annually. Agent-based models can be used to simulate the complex interactions that occur between immune cells and hm-IDP-like proteins (antigens) during affinity maturation—the process by which antibodies evolve. Compared to existing experimental approaches, agent-based models offer a safe, low-cost, and rapid route to study the immune response to vaccines spanning a wide range of design variables. However, the highly stochastic nature of affinity maturation and vast sequence space of hm-IDPs render brute force searches intractable for exploring all pertinent vaccine design variables and the subset of immunization protocols encompassed therein. To address this challenge, we employed deep reinforcement learning to drive a recently developed agent-based model of affinity maturation to focus sampling on immunization protocols with greater potential to improve the chosen metrics of protection, namely the broadly neutralizing antibody (bnAb) titers or fraction of bnAbs produced. Using this approach, we were able to coarse-grain a wide range of vaccine design variables and explore the relevant design space. Our work offers new testable insights into how vaccines should be formulated to maximize protective immune responses to hm-IDPs and how they can be minimally tailored to account for major sources of heterogeneity in human immune responses and various socioeconomic factors. Our results indicate that the first 3 to 5 immunizations, depending on the metric of protection, should be specially tailored to achieve a robust protective immune response, but that beyond this point further immunizations require only subtle changes in formulation to sustain a durable bnAb response.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2024

Algae to HEFA : Economics and potential deployment in the United States

Abstract To reach the goals set by the US Department of Energy's Sustainable Aviation Fuel (SAF) Grand Challenge, currently available feedstocks may be insufficient. Giving priority to developing, prototyping and reducing the cost of algal feedstock before investing and lining up locations is important. As the production of algal feedstocks advances, a simplified conversion approach using more mature technologies can help reduce the investment risk for algae‐based fuels. Reducing process complexity to the steps described here [namely, conversion of lipids to HEFA (hydroprocessed esters and fatty acids) fuels and relegating the remainder of the biomass to anaerobic digestion or food/feed production] enables the near‐term production of algal SAF but presents challenging economics depending on achievable cultivation costs and compositional quality. However, these economics can be improved by present‐day policy incentives. With these incentives, the modeled algae‐to‐HEFA pathway could reach a minimum fuel selling price as low as $4.7 per gasoline gallon equivalent depending on the carbon intensity reduction that can be achieved compared with petroleum. Uncertainty about algal feedstock production maturity in the current state of technology and the future will play a large role in determining the economic feasibility of building algae‐to‐HEFA facilities. For example, if immaturity increases the feedstock price by even 10%, SAF production in 2050 is about 58% of the production which could have been achieved with mature feedstock. Additionally, growth in this conversion pathway can be notably boosted through the inclusion of subsidies, and also through higher‐value coproducts or higher lipid yields beyond the scope of the process considered here.

09 BIOMASS FUELS↗
DOE OSTI2022

Beyond carbon flux partitioning: Carbon allocation and nonstructural carbon dynamics inferred from continuous fluxes

Carbon (C) allocation and nonstructural carbon (NSC) dynamics play essential roles in plant growth and survival under stress and disturbance. However, quantitative understanding of these processes remains limited. Here, in this work, we propose a framework where we connect commonly measured carbon cycle components (eddy covariance fluxes of canopy CO 2 exchange, soil CO 2 efflux, and allometry-based biomass and net primary production) by a simple mass balance model to derive ecosystem-level NSC dynamics (NSC i ), C translocation (dC i ), and the biomass production efficiency (BPE i ) in above- and belowground plant (i = agp and bgp) compartments. We applied this framework to two long-term monitored loblolly pine (Pinus taeda) plantations of different ages in North Carolina and characterized the variations of NSC and allocation in years under normal and drought conditions. The results indicated that the young stand did not have net NSC flux at the annual scale, whereas the mature stand stored a near-constant proportion of new assimilates as NSC every year under normal conditions, which was comparable in magnitude to new structural growth. Roots consumed NSC in drought and stored a significant amount of NSC post drought. The above- and belowground dC i and BPE i varied more from year to year in the young stand and approached a relatively stable pattern in the mature stand. The belowground BPE bgp differed the most between the young and mature stands and was most responsive to drought. With the internal C dynamics quantified, this framework may also improve biomass production estimation, which reveals the variations resulting from droughts. Overall, these quantified ecosystem-scale dynamics were consistent with existing evidence from tree-based manipulative experiments and measurements and demonstrated that combining the continuous fluxes as proposed here can provide additional information about plant internal C dynamics. Given that it is based on broadly available flux data, the proposed framework is promising to improve the allocation algorithms in ecosystem C cycle models and offers new insights into observed variability in soil–plant–climate interactions.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2023

Molecular structures reveal the origin of spectral variation in cryptophyte light harvesting antenna proteins

In addition to their membrane-bound chlorophyll a/c light-harvesting antenna, the cryptophyte algae have evolved a unique phycobiliprotein antenna system located in the thylakoid lumen. The basic unit of this antenna consists of two copies of an αβ protomer where the α and β subunits scaffold different combinations of a limited number of linear tetrapyrrole chromophores. While the β subunit is highly conserved, encoded by a single plastid gene, the nuclear-encoded α subunits have evolved diversified multigene families. It is still unclear how this sequence diversity results in the spectral diversity of the mature proteins. By careful examination of three newly determined crystal structures in comparison with three previously obtained, we show how the α subunit amino acid sequences control chromophore conformations and hence spectral properties even when the chromophores are identical. Previously we have shown that α subunits control the quaternary structure of the mature αβ.αβ complex (either open or closed), however, each species appeared to only harbor a single quaternary form. Here we show that species of the Hemiselmis genus contain expressed α subunit genes that encode both distinct quaternary structures. Finally, we have discovered a common single-copy gene (expressed into protein) consisting of tandem copies of a small α subunit that could potentially scaffold pairs of light harvesting units. Together, our results show how the diversity of the multigene α subunit family produces a range of mature cryptophyte antenna proteins with differing spectral properties, and the potential for minor forms that could contribute to acclimation to varying light regimes.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Litter removal impacts on soil biodiversity and eucalypt plantation development in the seasonal tropics

The little layer of tree plantations provides primary nutrients for uptake, buffers changes in soil moisture, and provides habitat and substrate to soil epigeic fauna. However, this layer in eucalypt plantations is often removed to reduce fuel load during the fire season in the Brazilian savanna (Cerrado). Therefore, it is necessary to quantify the effects of changes in litter dynamics on the function of these plantations, on key nutrient cycling processes and on epigeic fauna diversity and abundance. In two adjacent stands (one juvenile and one mature), the consequences of two years of litter removal were quantified as monthly litterfall, leaf and fine wood litter decomposition, epigeic fauna abundance and diversity, soil biogeochemical variables, and tree diameter and basal area increments. Monthly litterfall rates in juvenile and mature stands did not change with litter removal over the study period. Annual litterfall ranged from 4.1 to 4.9 Mg ha -1 a -1 in litter removal plots and from 3.9 to 4.8 Mg ha -1 a -1 in control plots. Fine wood litter decomposition was slower in litter removal plots compared to controls, while leaf decomposition rates were similar in both. Two years of litter removal in the juvenile stand did not affect topsoil biogeochemical parameters but decreased available phosphorus at 20–40 cm depth relative to controls. In the mature stand, total cation exchange capacity (0–20 cm) was higher in controls (6.4 cmol c dm -3 ) relative to litter removal plots (6.3 cmol c dm -3 ), while soil moisture (0–40 cm depth) was lower in litter removal (25.45 m 3 m -3 ) compared to control plots (26 m 3 m -3 ) in the dry season. A non-metric multidimensional scaling ordination revealed an increased homogeneity in epigeic fauna where litter was removed. Litterfall, decomposition, diameter increment, four soil physical parameters and fourteen chemical parameters at 0–20 and 20–40 cm depth explained the differences in soil epigeic fauna composition between litter removal and control plots. Diameter increment decreased with litter removal only in the juvenile stand, which had reached its growth peak. The results indicate that removing excess litter to decrease fuel volume can alter soil biodiversity and edaphic conditions that negatively affect nutrient cycling and tree growth.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2022

Unmasking the Conformational Stability and Inhibitor Binding to SARS-CoV-2 Main Protease Active Site Mutants and Miniprecursor

We recently demonstrated that inhibitor binding reorganizes the oxyanion loop of a monomeric catalytic domain of SARS CoV-2 main protease (MPro) from an unwound (E) to a wound (active, E*) conformation, independent of dimerization. Here we assess the effect of the flanking N-terminal residues, to imitate the MPro precursor prior to its autoprocessing, on conformational equilibria rendering stability and inhibitor binding. Thermal denaturation (T m ) of C145A mutant, unlike H41A, increases by 6.8 °C, relative to wild-type mature dimer. An inactivating H41A mutation to maintain a miniprecursor containing TSAVL[Q or E] of the flanking nsp4 sequence in an intact form [ (-6) MPro H41A and (-6*) MPro H41A , respectively], and its corresponding mature MPro H41A were systematically examined. While the H41A mutation exerts negligible effect on T m and dimer dissociation constant (K dimer ) of MPro H41A , relative to the wild type MPro, both miniprecursors show a 4–5 °C decrease in T m and > 85-fold increase in K dimer as compared to MPro H41A . The K d for the binding of the covalent inhibitor GC373 to (-6*) MPro H41A increases ~12-fold, relative to MPro H41A , concomitant with its dimerization. While the inhibitor-free dimer exhibits a state in transit from E to E* with a conformational asymmetry of the protomers’ oxyanion loops and helical domains, inhibitor binding restores the asymmetry to mature-like oxyanion loop conformations (E*) but not of the helical domains. Finally, disorder of the terminal residues 1–2 and 302–306 observed in both structures suggest that N-terminal autoprocessing is tightly coupled to the E-E* equilibrium and stable dimer formation.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Episodic hydrocarbon charge in tight Mississippian reservoirs of Central Oklahoma, USA: Insights from oil inclusion geochemistry

The unforeseen discovery of substantial hydrocarbons in ultra-tight Mississippian reservoirs in Oklahoma testifies to the complexity of the hydrocarbon charge across the flanks of the Nemaha Uplift. Based on crude oil and fluid-inclusion chemistry, the hydrocarbons at the western side of the Nemaha Uplift consist of highly-mature, thermally-cracked and less-mature uncracked hydrocarbons (0.50–1.4% calculated vitrinite reflectance, R c ). The oils are less mature on the eastern flank of the structure (0.5–0.7% R c ). Based on distributions of biomarkers and higher diamondoid (composed of three adamantane cages or more) distributions, the hydrocarbons entrapped within reservoir fluid-inclusions are sourced from the Woodford Shale and also from Mississippian carbonates. To our knowledge, this is the first work to describe oil correlations using diamondoids inside fluid inclusions. Petroleum compositions in the ultra-tight Mississippian reservoirs are highly variable, which highlights the need for a thorough assessment of mechanisms controlling charge. By integrating oil and fluid inclusion geochemistry and fluid-inclusion microthermometry with basin modeling, we show that the primary controls are: (1) proximity to the source-rock kitchen(s), (2) episodic charging from Permian through Cretaceous time, and (3) mixing.

58 GEOSCIENCES↗
DOE OSTI2023

Unique interface and dynamics of the complex of HSP90 with a specialized cochaperone AIPL1

Photoreceptor phosphodiesterase PDE6 is central for visual signal transduction. Maturation of PDE6 depends on a specialized chaperone complex of HSP90 with aryl hydrocarbon receptor-interacting protein-like 1 (AIPL1). Disruption of PDE6 maturation underlies a severe form of retina degeneration. Here, we report a 3.9 Å cryoelectron microscopy (cryo-EM) structure of the complex of HSP90 with AIPL1. This structure reveals a unique interaction of the FK506-binding protein (FKBP)-like domain of AIPL1 with HSP90 at its dimer interface. Unusually, the N terminus AIPL1 inserts into the HSP90 lumen in a manner that was observed previously for HSP90 clients. Deletion of the 7 N-terminal residues of AIPL1 decreased its ability to cochaperone PDE6. Multi-body refinement of the cryo-EM data indicated large swing-like movements of AIPL1-FKBP. Modeling the complex of HSP90 with AIPL1 using crosslinking constraints indicated proximity of the mobile tetratricopeptide repeat (TPR) domain with the C-terminal domain of HSP90. Our study establishes a framework for future structural studies of PDE6 maturation.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2021

Construction of extracellular matrix-based 3D hydrogel and its effects on cardiomyocytes

Highlights: • SOD and Triton X-100 could efficiently decellularize cells to get cardiac ECM. • Main proteins of cECM were fibronectin, collagen, elastin, fibrillin, laminin, etc. • 75% cECM-containing 3D hydrogel promoted cell proliferation and anti-apoptosis. • cECM3DH facilitated the differentiation of ESCs into CMs. • APcECM3DH was more conducive to NCM and ESC-derived CM maturation. Some discoveries resulted from 2-dimensional (2D) cultured cardiac cells have been disqualified in animal testing and later clinical trials. Extracellular matrix (ECM) plays a vital role in cardiac homeostasis, cardiac ECM (cECM)-based 3D cell cultures can mimics the physiological and pathological conditions in vivo closely, it is hopeful of addressing this challenge. Construction of cECM-based 3-dimensional (3D) hydrogel (cECM3DH) and its effects on cell behaviors were studied here. The results indicated that cellular compartments could be efficiently removed from heart tissue via sodium dodecyl sulfonate (SDS)- and Triton X-100-mediated decellularization, remaining the natural fibrous network structure and major proteins. 3D hydrogel consisted of 1 × 10{sup 7} cells/mL cells and 75% cECM could promote the proliferation and anti-apoptosis ability of human embryonic kidney (HEK)-293T cells. 0.25% trypsin or 0.20% collagenase was suitable to retrieve these cells from 3D hydrogel for further researches. Compared with 2D culture system, cECM3DH could significantly increase the proportion of GATA 4{sup +} cardiomyocytes (CMs) derived from heart tissue of neonatal mouse or induced differentiation of embryonic stem cells (ESCs) (P < 0.05) The expression levels of mature genes including cTnT, JCN, CaV1.2, MYL2, CASQ2, NCX1, and Cx43 of these CMs in adult pig cECM-based 3D hydrogel (APcECM3DH) were significantly higher than that in 2D culture system and in newborn piglet cECM-based 3D hydrogel (NPcECM3DH), respectively (P < 0.05). Therefore, cECM3DH supports the generation of primary CMs and ESC-derived CMs, APcECM3DH was more conducive to promoting CM maturation, which contributes to building 3D model for pathogenesis exploration, drug screening, and regenerative medicine of heart diseases.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2025

Soil moisture gradients strengthen mesoscale convective systems by increasing wind shear

Mesoscale convective systems are a class of storm linked to extensive flooding and other destructive hazards in many regions globally. In West Africa, soil moisture impacts provide a valuable source of predictability for mature storm hazards, but little is known about mature storm sensitivity to soil moisture in other climatic regions. Here we use a storm track dataset, satellite observations and reanalysis fields to investigate the response of mature storms to soil moisture in seven global storm hotspots—West Africa, India, South America, South Africa, Australia and the United States Great Plains. We demonstrate that mesoscale soil moisture gradients (~500 km) can enhance storms by driving increased vertical wind shear conditions, a crucial ingredient for storm organization, through the strengthening of atmospheric temperature gradients. This is evidenced by a 10–30% increase in precipitation feature size and rainfall for the largest storms on days with favourable soil moisture gradients compared with unfavourable gradients. Global simulations confirm that soil moisture gradients influence wind shear. The results demonstrate the importance of soil moisture feedbacks for accurate forecasting of mesoscale convective systems and future projections of extreme events under climate change.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2021

Electrochemically induced metal- vs. ligand-based redox changes in mackinawite: identification of a Fe 3+ - and polysulfide-containing intermediate

Under anaerobic conditions, ferrous iron reacts with sulfide producing FeS, which can then undergo a temperature, redox potential, and pH dependent maturation process resulting in the formation of oxidized mineral phases, such as greigite or pyrite. A greater understanding of this maturation process holds promise for the development of iron-sulfide catalysts, which are known to promote diverse chemical reactions, such as H + , CO 2 and NO 3 - reduction processes. Hampering the full realization of the catalytic potential of FeS, however, is an incomplete knowledge of the molecular and redox processess ocurring between mineral and nanoparticulate phases. Here, we investigated the chemical properties of iron-sulfide by cyclic voltammetry, Raman and X-ray absorption spectroscopic techniques. Tracing oxidative maturation pathways by varying electrode potential, nanoparticulate n(Fe 2+ S 2- ) (s) was found to oxidize to a Fe 3+ containing FeS phase at -0.5 V vs. Ag/AgCl (pH = 7). In a subsequent oxidation, polysulfides are proposed to give a material that is composed of Fe 2+ , Fe 3+ , S 2- and polysulfide (S n 2- ) species, with its composition described as Fe 2+ 1-3x Fe 3+ 2x S 2- 1-y (S n 2- ) y . Thermodynamic properties of model compounds calculated by density functional theory indicate that ligand oxidation occurs in conjunction with structural rearrangements, whereas metal oxidation may occur prior to structural rearrangement. These findings together point to the existence of a metastable FeS phase located at the junction of a metal-based oxidation path between FeS and greigite (Fe 2+ Fe 3+ 2 S 2- 4 ) and a ligand-based oxidation path between FeS and pyrite (Fe 2+ (S 2 ) 2- ).

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2025

Structural insights into VRC01-class bnAb precursors with diverse light chains elicited in the IAVI G001 human vaccine trial

The development of germline-targeting vaccines represents a potentially transformative strategy to elicit broadly neutralizing antibodies (bnAbs) against HIV and other antigenically diverse pathogens. Here, we report on structural characterization of vaccine-elicited VRC01-class bnAb precursors in the IAVI G001 Phase 1 clinical trial with the eOD-GT8 60mer nanoparticle as immunogen. High-resolution X-ray structures of eOD-GT8 monomer complexed with Fabs of five VRC01-class bnAb precursors with >90% germline identity revealed a conserved mode of binding to the HIV CD4-binding site via IGHV1-2-encoded heavy chains, mirroring mature bnAb interactions. The light-chain V-gene diversity emulated VRC01 bnAbs and stabilized antigen engagement, while their conserved five-residue LCDR3 motifs prevented steric clashes. Notably, the VRC01-class bnAb precursors accommodated the N276 glycan, a key barrier in HIV Env recognition, through structural rearrangements in HCDR3 or LCDR1, despite its absence in the immunogen. Surface plasmon resonance analysis showed that 87% of elicited antibodies retained glycan binding capacity, albeit with reduced affinity. These findings validate the ability of eOD-GT8 60mer nanoparticles to prime VRC01-class bnAb precursors with native-like paratopes but with intrinsic glycan adaptability. Structural mimicry of mature bnAbs was observed even with limited somatic hypermutation, indicating that critical features are encoded in the germline repertoire. The structures highlight how germline-encoded features drive bnAb-like recognition at early stages. This work provides molecular evidence supporting germline targeting in humans and provides guidance for designing booster immunogens to shepherd affinity maturation toward broad neutralization.

Science & Technology - Other Topics↗
DOE OSTI2023

Status and future plans for C 3 R&D

C 3 is an opportunity to realize an e + e - collider for the study of the Higgs boson at √s = 250 GeV, with a well defined upgrade path to 550 GeV while staying on the same short facility footprint. C 3 is based on a fundamentally new approach to normal conducting linear accelerators that achieves both high gradient and high efficiency at relatively low cost. Given the advanced state of linear collider designs, the key system that requires technical maturation for C 3 is the main linac. This paper presents the staged approach towards a facility to demonstrate C 3 technology with both Direct (source and main linac) and Parallel (beam delivery, damping ring, ancillary component) R&D. The primary goal of the C 3 Demonstration R&D Plan is to reduce technical and cost risk by building and operating the key components of C 3 at an adequate scale. This R&D plan starts with the engineering design, and demonstration of one cryomodule and will culminate in the construction of a 3 cryomodule linac with pre-production prototypes. This R&D program would also demonstrate the linac rf fundamentals including achievable gradient and gradient stability over a full electron bunch train and breakdown rates. It will also investigate beam dynamics including energy spread, wakefields, and emittance growth. This work will be critical to confirm the suitability of the C 3 beam parameters for the physics reach and detector performance in preparation for a Conceptual Design Report (CDR), as well as for follow-on technology development and industrialization. The C 3 Demonstration R&D Plan will open up significant new scientific and technical opportunities based on development of high-gradient and high-efficiency accelerator technology. It will push this technology to operate both at the GeV scale and mature the technology to be reliable and provide high-brightness electron beams. The timeline for progressing with C 3 technology development will be governed by practical limitations on both the technical progress and resource availability. It consists of four stages: Stage 0) Ongoing fundamental R&D on structure prototypes, damping and vibrations. Stage 1) Advancing the engineering maturity of the design and developing start-to-end simulations including space-charge and wakefield effects. This stage will include testing of strucutres operating at cryogenic temperatures. Beam tests would be performed with high beam current to test full beam loading. Stage 2) Production and testing of the first cryomodule at cryogenic temperatures. This would provide sufficient experimental data to compile a CDR and it is anticipated for Stage 2 to last 3 years and to culminate with the transport of photo-electrons through the first cryomodule. Stage 3) Updates to the engineering design of the cryomodules, production of the second and third cryomodule and their installation. Lower charge and lower emittance beams will be used to investigate emittance growth. The successful full demonstration of the 3 cryomodules to deliver up to a 3 GeV beam and achieve the C 3 five gradient will allow a comprehensive and robust evaluation of the technical design of C 3 as well as mitigate technical, schedule, and cost risks required to proceed with a Technical Design Report (TDR).

radiation hardened magnets↗
DOE OSTI2022

Optimization of sporulation and purification methods for sporicidal efficacy assessment on Bacillus spores

Abstract Validating the efficacy of sporicidal agents is a critical step in current good manufacturing practices for disinfection requirements. A limitation is that the poor quality of spores can lead to false positive sporicidal results. The aim of this study was to explore optimal sporulation and purification methods in Bacillus spores. Spores of 7 Bacillus strains were produced in 5 different sporulation media. After density centrifugation, spore yields were measured by phase-contrast microscopy and enumeration assays. Effects of purification methods including heat, sonication and lysozyme, and maturation on spore qualities were determined by sodium hypochlorite sporicidal assay. Difco sporulation media was identified as the preferred sporulation medium for 4 out of 7 tested Bacillus strains. Sporulation rates in B. cereus, B. sphaericus, and B. thuringiensis were higher at 30°C than the rates at 37°C at a difference of 5%, 65%, and 20%, respectively. Bacillus licheniformis favored Mn2+-amended 10% Columbia Broth at 37°C for sporulation with 40–72% higher sporulation rates than other media. The maximum sporulation rates of B. cereus and B. thuringiensis were observed on double-strength Schaeffer's-glucose broth. All studied purification methods improved the spore purity with strain variations. However, intense heat (80°C for 20 min) and lysozyme (100 μg/mL) treatment impaired the spore quality of specific Bacillus strains by sensitizing them against sodium hypochlorite. The length of the maturation period had an impact on the spore resistance, and the most optimal maturation periods ranged from 7 to 21 days in Bacillus strains. The results of this study will pave the way for further evaluation of the sporicidal activity of disinfectants.

Biotechnology & Applied Microbiology↗
DOE OSTI2023

The structure of the high-affinity nickel-binding site in the Ni,Zn-HypA•UreE2 complex

Abstract The maturation pathway for the nickel-dependent enzyme urease utilizes the protein UreE as a metallochaperone to supply Ni(II) ions. In Helicobacter pylori urease maturation also requires HypA and HypB, accessory proteins that are commonly associated with hydrogenase maturation. Herein we report on the characterization of a protein complex formed between HypA and the UreE2 dimer. Nuclear magnetic resonance (NMR) coupled with molecular modelling show that the protein complex apo, Zn-HypA•UreE2, forms between the rigorously conserved Met-His-Glu (MHE motif) Ni-binding N-terminal sequence of HypA and the two conserved His102A and His102B located at the dimer interface of UreE2. This complex forms in the absence of Ni(II) and is supported by extensive protein contacts that include the use of the C-terminal sequences of UreE2 to form additional strands of β-sheet with the Ni-binding domain of HypA. The Ni-binding properties of apo, Zn-HypA•UreE2 and the component proteins were investigated by isothermal titration calorimetry using a global fitting strategy that included all of the relevant equilibria, and show that the Ni,Zn-HypA•UreE2 complex contains a single Ni(II)-binding site with a sub-nanomolar KD. The structural features of this novel Ni(II) site were elucidated using proteins produced with specifically deuterated amino acids, protein point mutations, and the analyses of X-ray absorption spectroscopy, hyperfine shifted NMR features, as well as molecular modeling coupled with quantum-mechanical calculations. The results show that the complex contains a six-coordinate, high-spin Ni(II) site with ligands provided by both component proteins.

Zambelli, Barbara (ORCID:0000000238760051)↗
DOE OSTI2020

The older plant gets the sun: Age-related changes in Miscanthus × giganteus phenology

Age-related changes are usually overlooked in perennial grass research; when they are considered it is usually as a change in plant size (e.g., biomass). Whether other physiological or developmental aspects change as stands age, and how those aspects may impact long-term stand dynamics, remains unclear. Conventional experimental designs study a single stand over multiple growing seasons and thereby confound age-related changes with growing season conditions. Here we used a staggered-start experimental design with three repeated planting years over two growing seasons to isolate growing season effects. We studied changes in Miscanthus × giganteus phenology during its yield-building stage (first 3 years) and estimated age, growing season and nitrogen (N) effects on development using nonlinear regression parameters. Stand age clearly changed plant growth; faster developmental rates were usually seen in 1-year-old stands (young), but because 2- and 3-year-old stands (mature) emerged 3 months earlier than newly planted stands they produced ~30% more stems with 30%–60% more leaves. Nitrogen fertilization modulated some age-related phenological changes. Fertilized 2-year-old stands reached similar stem densities as unfertilized 3-year-old stands and had fewer number of senesced leaves like 1-year-old stands. In addition, N fertilization had no effect on young M. × giganteus , but extended mature stands’ growing season more than 2 weeks by hastening emergence and delaying senescence. It also delayed flowering regardless of stand age. Our results suggest that, along with changes in size, M. × giganteus stands showed shifts in developmental strategies: young stands emerged later and developed faster, while mature stands grew for longer but more slowly. In temperate regions, where hard frost events are likely to interrupt development in late autumn, rapid early development is critical to plant survival. Nonlinear regression parameter differences proved effective in identifying phenological shifts.

59 BASIC BIOLOGICAL SCIENCES↗