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At least 721 records · Page 40

Enhancing Critical Infrastructure and Key Resources (CIKR) Level-0 Physical Process Security Using Field Device Distinct Native Attribute Features

Here, the need for improved Critical Infrastructure and Key Resource (CIKR) security is unquestioned and there has been minimal emphasis on Level-0 (PHY Process) improvements. Wired Signal Distinct Native Attribute (WS-DNA) Fingerprinting is investigated here as a non-intrusive PHY-based security augmentation to support an envisioned layered security strategy. Results are based on experimental response collections from Highway Addressable Remote Transducer (HART) Differential Pressure Transmitter (DPT) devices from three manufacturers (Yokogawa, Honeywell, Endress+Hauer) installed in an automated process control system. Device discrimination is assessed using Time Domain (TD) and Slope-Based FSK (SB-FSK) fingerprints input to Multiple Discriminant Analysis, Maximum Likelihood (MDA/ML) and Random Forest (RndF) classifiers. For 12 different classes (two devices per manufacturer at two distinct set points), both classifiers performed reliably and achieved an arbitrary performance benchmark of average cross-class percent correct of %C > 90%. The least challenging cross-manufacturer results included near-perfect %C ≈ 100%, while the more challenging like-model (serial number) discrimination results included 90%< %C < 100%, with TD Fingerprinting marginally outperforming SB-FSK Fingerprinting; SB-FSK benefits from having less stringent response alignment and registration requirements. The RndF classifier was most beneficial and enabled reliable selection of dimensionally reduced fingerprint subsets that minimize data storage and computational requirements. The RndF selected feature sets contained 15% of the full-dimensional feature sets and only suffered a worst case %CΔ = 3% to 4% performance degradation.

60 APPLIED LIFE SCIENCES↗

Three Confinement Systems—Spherical Tokamak, Standard Tokamak, and Stellarator: A Comparison of Key Component Cost Elements

Since the 1950s “next-step” fusion devices and power plant studies have been developed for a number of magnetic confinement systems but an open question remains: can a magnetic fusion device be simplified to the point where it will be cost competitive and operate with high availability? Concept designs based on the ARIES advanced tokamak, spherical tokamak (ST), and the quasi-axisymmetric (QAS) stellarator option have progressed in recent years through a series of Princeton Plasma Physics Laboratory (PPPL) studies with an underlying intent to improve the engineering feasibility of each, giving special attention to concepts that simplify the device configuration and improve maintenance features. For the ST option, design details centered on a 3-m Fusion Nuclear Science Facility that evolved to incorporate vertical maintenance, high temperature superconductor magnets, a small inboard duel coolant lead lithium blanket, and a liquid metal divertor. In collaboration with the Korean fusion demonstration reactor (K-DEMO) and CFETR concept study teams the tokamak design has evolved to increase plasma component access within a vertical maintenance approach using enlarged toroidal field coils incorporating a low- and high-field Nb 3 Sn winding pack that provide a peak field of 16 T. A recent PPPL stellarator study focused on simplifying the stellarator winding topology to improve access to in-vessel components; combining coil optimization with winding surfaces that incorporated geometry constraints specified by engineering. This paper centered on a 1000-MW power plant design with a tokamak like vertical maintenance scheme that allows access to remove large internal blanket sectors. Results of three confinement studies (PPPL developed ST, K-DEMO, and QAS stellarator) will be presented to highlight concepts that simplify each device configuration and improved their maintenance features. Furthermore, scaling each option to a common 1000-MW net electric power plant mission allows comparisons to be made of key cost elements such as the size of major core components, sizing of the reactor hall, or external facilities needed to handle and store activated in-vessel components.

70 PLASMA PHYSICS AND FUSION TECHNOLOGY↗

Scavenging organic nitrogen and remodelling lipid metabolism are key survival strategies adopted by the endophytic fungi, Serendipita vermifera and Serendipita bescii to alleviate nitrogen and phosphorous starvation in vitro

Summary Serendipitaceae represents a diverse fungal group in the Basidiomycota that includes endophytes and lineages that repeatedly evolved ericoid, orchid and ectomycorrhizal lifestyle. Plants rely upon both nitrogen and phosphorous, for essential growth processes, and are often provided by mycorrhizal fungi. In this study, we investigated the cellular proteome of Serendipita vermifera MAFF305830 and closely related Serendipita vermifera subsp. bescii NFPB0129 grown in vitro under (N) ammonium and (P) phosphate starvation conditions. Mycelial growth pattern was documented under these conditions to correlate growth‐specific responses to nutrient starvation. We found that N‐starvation accelerated hyphal radial growth, whereas P‐starvation accelerated hyphal branching. Additionally, P‐starvation triggers an integrated starvation response leading to remodelling of lipid metabolism. Higher abundance of an ammonium transporter known to serve as both an ammonium sensor and stimulator of hyphal growth was detected under N‐starvation. Additionally, N‐starvation led to strong up‐regulation of nitrate, amino acid, peptide, and urea transporters, along with several proteins predicted to have peptidase activity. Taken together, our finding suggests S. bescii and S. vermifera have the metabolic capacity for nitrogen assimilation from organic forms of N compounds. We hypothesize that the nitrogen metabolite repression is a key regulator of such organic N assimilation.

54 ENVIRONMENTAL SCIENCES↗

Identification of key bacterial populations affecting early embryonic development in cattle uterus

Abstract Superovulation is an important animal breeding biotechnology, while the quality of embryos obtained from superovulation is unstable in cattle. The relationship between the microorganisms in the cattle uterus and embryo qualities was determined to identify the key bacterial populations affecting early embryonic development. A total of 10 Xia Nan cows underwent superovulation, we collected cervical mucus and flush samples to investigated by 16S rDNA sequencing. Results showed that there were abundant microorganisms in cervical mucus, but no obvious relationship with the quality of embryos. The clustering results of flush samples were consistent with the grouping of embryo quality. Proteobacteria accounted for more than 95% of the total bacterial community in group A with the best embryo quality (qualified embryo ratio above 0.8), and as embryo quality decreased, the Proteobacteria proportion also decreased. In contrast to the proportion of Proteobacteria, the proportions of Firmicutes and Bacteroidetes significantly increased as embryo quality decreased. For group C with the worst embryo quality, the proportions of Firmicutes and Bacteroidetes increased to 4.7 times and 12.3 times of group A, respectively. These results showed that the quantities and proportions of Firmicutes and Bacteroidetes may be related to early embryonic development in cattle.

Lyu, Chenchen↗

When evolution is the solution to pollution: Key principles, and lessons from rapid repeated adaptation of killifish ( Fundulus heteroclitus ) populations

Abstract For most species, evolutionary adaptation is not expected to be sufficiently rapid to buffer the effects of human‐mediated environmental changes, including environmental pollution. Here we review how key features of populations, the characteristics of environmental pollution, and the genetic architecture underlying adaptive traits, may interact to shape the likelihood of evolutionary rescue from pollution. Large populations of Atlantic killifish (Fundulus heteroclitus) persist in some of the most contaminated estuaries of the United States, and killifish studies have provided some of the first insights into the types of genomic changes that enable rapid evolutionary rescue from complexly degraded environments. We describe how selection by industrial pollutants and other stressors has acted on multiple populations of killifish and posit that extreme nucleotide diversity uniquely positions this species for successful evolutionary adaptation. Mechanistic studies have identified some of the genetic underpinnings of adaptation to a well‐studied class of toxic pollutants; however, multiple genetic regions under selection in wild populations seem to reflect more complex responses to diverse native stressors and/or compensatory responses to primary adaptation. The discovery of these pollution‐adapted killifish populations suggests that the evolutionary influence of anthropogenic stressors as selective agents occurs widely. Yet adaptation to chemical pollution in terrestrial and aquatic vertebrate wildlife may rarely be a successful “solution to pollution” because potentially adaptive phenotypes may be complex and incur fitness costs, and therefore be unlikely to evolve quickly enough, especially in species with small population sizes.

Evolutionary Biology↗

Bioenergy sorghum’s deep roots: A key to sustainable biomass production on annual cropland

Abstract Bioenergy sorghum has high biomass yield potential, drought resilience, good nitrogen use efficiency, and a root system that contributes to the accumulation of soil organic carbon. In this study, field grown bioenergy sorghum root systems were analyzed during the growing season to characterize their depth, biomass, morphology, anatomy, and gene expression profiles. Bioenergy sorghum roots grew continuously during a 155‐day growing season producing ~175 nodal roots, accumulating ~7 Mg of dry biomass per hectare, and reaching >2 m deep in the soil profile. Nodal roots within 20 cm of the stem were 1–5 mm in diameter, whereas roots deeper in soil profiles were enriched in lateral roots with small diameters (~30–500 µm) enabling growth through soil macropores. In older field‐grown plants, roots with intact endodermal, vascular and inner root tissues were surrounded by degraded or aerenchyma‐filled epidermal and cortical cell layers. Transcriptome analysis of nodal, surface, and deep roots identified >2,500 differentially expressed genes involved in root growth, transport, adaptation, defense, and AMF–root interaction. Deep roots (180–240 cm) differentially expressed genes that regulate lateral root growth. Surface roots (0–20 cm) located mid‐row differentially expressed genes involved in nitrate transport, whereas ammonium transport genes were expressed in surface and deep roots and genes involved in phosphate transport were expressed in nodal, surface, and deep roots. Overall, bioenergy sorghum's long growing season enables root systems to grow deeper and accumulate more biomass than annual grain crops such as maize, attributes that could help restore annual cropland soil organic carbon levels and improve soil productivity. Deep roots active in nutrient transport are positioned to take‐up fertilizer leached deep into soil profiles mitigating potential nutrient run‐off. Bioenergy sorghum's large and deep root system is a key to sustainable production of biofuels, biopower, and bioproducts on annual cropland.

59 BASIC BIOLOGICAL SCIENCES↗

SWEET 11 and 15 as key players in seed filling in rice

Summary Despite the relevance of seed‐filling mechanisms for crop yield, we still have only a rudimentary understanding of the transport processes that supply the caryopsis with sugars. We hypothesized that SWEET sucrose transporters may play important roles in nutrient import pathways in the rice caryopsis. We used a combination of mRNA quantification, histochemical analyses, translational promoter–reporter fusions and analysis of knockout mutants created by genomic editing to evaluate the contribution of SWEET transporters to seed filling. In rice caryopses, SWEET 11 and 15 had the highest mRNA levels and proteins localized to four key sites: all regions of the nucellus at early stages; the nucellar projection close to the dorsal vein; the nucellar epidermis that surrounds the endosperm; and the aleurone. ossweet11 ; 15 double knockout lines accumulated starch in the pericarp, whereas caryopses did not contain a functional endosperm. Jointly, SWEET 11 and 15 show all the hallmarks of being necessary for seed filling with sucrose efflux functions at the nucellar projection and a role in transfer across the nucellar epidermis/aleurone interface, delineating two major steps for apoplasmic seed filling, observations that are discussed in relation to observations made in rice and barley regarding the relative prevalence of these two potential import routes.

59 BASIC BIOLOGICAL SCIENCES↗

The ectomycorrhizal basidiomycete Laccaria bicolor releases a secreted β‐1,4 endoglucanase that plays a key role in symbiosis development

Summary In ectomycorrhiza, root ingress and colonization of the apoplast by colonizing hyphae is thought to rely mainly on the mechanical force that results from hyphal tip growth, but this could be enhanced by secretion of cell‐wall‐degrading enzymes, which have not yet been identified. The sole cellulose‐binding module ( CBM 1) encoded in the genome of the ectomycorrhizal Laccaria bicolor is linked to a glycoside hydrolase family 5 ( GH 5) endoglucanase, Lb GH 5‐ CBM 1. Here, we characterize Lb GH 5‐ CBM 1 gene expression and the biochemical properties of its protein product. We also immunolocalized Lb GH 5‐ CBM 1 by immunofluorescence confocal microscopy in poplar ectomycorrhiza. We show that Lb GH 5‐ CBM 1 expression is substantially induced in ectomycorrhiza, and RNA i mutants with a decreased Lb GH 5‐ CBM 1 expression have a lower ability to form ectomycorrhiza, suggesting a key role in symbiosis. Recombinant Lb GH 5‐ CBM 1 displays its highest activity towards cellulose and galactomannans, but no activity toward L. bicolor cell walls. In situ localization of Lb GH 5‐ CBM 1 in ectomycorrhiza reveals that the endoglucanase accumulates at the periphery of hyphae forming the Hartig net and the mantle. Our data suggest that the symbiosis‐induced endoglucanase Lb GH 5‐ CBM 1 is an enzymatic effector involved in cell wall remodeling during formation of the Hartig net and is an important determinant for successful symbiotic colonization.

Zhang, Feng↗

Multiple mechanisms explain loss of anthocyanins from betalain‐pigmented Caryophyllales, including repeated wholesale loss of a key anthocyanidin synthesis enzyme

Summary In this study, we investigate the genetic mechanisms responsible for the loss of anthocyanins in betalain‐pigmented Caryophyllales, considering our hypothesis of multiple transitions to betalain pigmentation. Utilizing transcriptomic and genomic datasets across 357 species and 31 families, we scrutinize 18 flavonoid pathway genes and six regulatory genes spanning four transitions to betalain pigmentation. We examined evidence for hypotheses of wholesale gene loss, modified gene function, altered gene expression, and degeneration of the MBW (MYB‐bHLH‐WD40) trasnscription factor complex, within betalain‐pigmented lineages. Our analyses reveal that most flavonoid synthesis genes remain conserved in betalain‐pigmented lineages, with the notable exception of TT19 orthologs, essential for the final step in anthocyanidin synthesis, which appear to have been repeatedly and entirely lost. Additional late‐stage flavonoid pathway genes upstream of TT19 also manifest strikingly reduced expression in betalain‐pigmented species. Additionally, we find repeated loss and alteration in the MBW transcription complex essential for canonical anthocyanin synthesis. Consequently, the loss and exclusion of anthocyanins in betalain‐pigmented species appear to be orchestrated through several mechanisms: loss of a key enzyme, downregulation of synthesis genes, and degeneration of regulatory complexes. These changes have occurred iteratively in Caryophyllales, often coinciding with evolutionary transitions to betalain pigmentation.

59 BASIC BIOLOGICAL SCIENCES↗

Understanding the dynamic nature of plant lipid anabolic and catabolic metabolism is key to sustainable oilseed engineering

Plant-derived oils are essential sources of reduced carbon and various fatty acid (FA) structures for food, biofuels, and the oleochemical industry. Despite extensive efforts, engineering mainstream oilseed crops to produce high levels of industrially valuable unusual FAs (UFAs) remains challenging. This review synthesizes recent advances in the understanding of lipid metabolic networks, emphasizing how species-specific regulation of FA synthesis, activation, and delivery influences triacylglycerol (TAG) assembly to govern the efficiency of UFA accumulation. Key insights reveal that acyl flux through anabolic and catabolic branches of lipid metabolism is tightly controlled by enzyme substrate selectivities, diacylglycerol (DAG) pool compartmentalization, and metabolic context, including lipid remodeling and degradation pathways. Engineering success is often constrained by incompatibilities between UFA biosynthetic enzymes and endogenous host metabolism, leading to flux imbalances, futile cycles, and undesired phenotypes. We highlight emerging strategies to overcome these barriers, such as the use of UFA-selective acyltransferases, coordinated manipulation of DAG source pools, suppression of competing endogenous enzymes, and exploitation of TAG remodeling mechanisms. This integrated synthesis provides a conceptual framework for logic-based engineering of oilseeds with enhanced UFA content by offering new avenues for sustainable biomanufacturing of valuable lipids.

acyltransferase specificity↗

Response Surfaces for Key Controlled Variables in a Hybrid Solid Oxide Fuel Cell/Gas Turbine System

Hybrid generation systems have been extensively modeled as a first step toward the development of automatic controls for the system. In most cases, it is impossible to validate mathematical models against real hardware because only a handful of hardware systems exist in the world. Data taken from the existing hardware has demonstrated significant nonlinearity, complex coupling between controlled variables, and sometimes non-intuitive behavior. This work exploits the capability of the HyPer hardware test bed at the National Energy Technology Laboratory (NETL) to generate data from a real recuperated gas turbine coupled with hardware simulations of a fuel cell cathode and appropriate ancillary equipment. Prior work has characterized the system only over a limited range of its operating envelope, due to the inability to manipulate multiple control inputs simultaneously. The work presented here fills the gaps using data from a 34 factorial experiment to generate quasi-continuous response surfaces describing the operating state space of the HyPer system. Polynomial correlation functions have been fitted to the data with excellent agreement. Relationships between the control inputs and critical state variables such as cathode mass flow, cathode temperature, turbine inlet and exhaust temperatures and other key system parameters are presented.

30 DIRECT ENERGY CONVERSION↗

WE-F-201-01: Identify Key Clinical Applications Needing Advanced Dose Calculation in Brachytherapy

With the recent introduction of heterogeneity correction algorithms for brachytherapy, the AAPM community is still unclear on how to commission and implement these into clinical practice. The recently-published AAPM TG-186 report discusses important issues for clinical implementation of these algorithms. A charge of the AAPM-ESTRO-ABG Working Group on MBDCA in Brachytherapy (WGMBDCA) is the development of a set of well-defined test case plans, available as references in the software commissioning process to be performed by clinical end-users. In this practical medical physics course, specific examples on how to perform the commissioning process are presented, as well as descriptions of the clinical impact from recent literature reporting comparisons of TG-43 and heterogeneity-based dosimetry. Learning Objectives: Identify key clinical applications needing advanced dose calculation in brachytherapy. Review TG-186 and WGMBDCA guidelines, commission process, and dosimetry benchmarks. Evaluate clinical cases using commercially available systems and compare to TG-43 dosimetry.

60 APPLIED LIFE SCIENCES↗

Isolation of bis(copper) key intermediates in Cu-catalyzed azide-alkyne “click reaction”

The copper-catalyzed 1,3-dipolar cycloaddition of an azide to a terminal alkyne (CuAAC) is one of the most popular chemical transformations, with applications ranging from material to life sciences. However, despite many mechanistic studies, direct observation of key components of the catalytic cycle is still missing. Initially, mononuclear species were thought to be the active catalysts, but later on, dinuclear complexes came to the front. We report the isolation of both a previously postulated π,σ-bis(copper) acetylide and a hitherto never-mentioned bis(metallated) triazole complex. We also demonstrate that although mono- and bis-copper complexes promote the CuAAC reaction, the dinuclear species are involved in the kinetically favored pathway.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Biosensor-driven strain engineering reveals key cellular processes for maximizing isoprenol production in Pseudomonas putida

Synthetic biology generates vast combinatorial designs, yet high-throughput analytical methods to screen them are poorly matched to interrogate this search space. We address this challenge by developing a biosensor-driven, growth-coupled selection strategy in Pseudomonas putida for isoprenol, a potential aviation fuel precursor. We found and characterized a noncanonical signaling pathway, revealing a functional and physical complex between a hybrid histidine kinase and an alcohol dehydrogenase, whose activity is tuned by heterodimerization. Leveraging this biosensor in a pooled CRISPRi library selection, we identified key host limitations. Iterative combinatorial strain engineering derived from these hits yielded a 36-fold titer increase to ~900 milligrams per liter. Integrated omics analysis revealed that metabolic rewiring toward amino acid catabolism was crucial for this improvement. This observation was found to be beneficial by technoeconomic analysis. Our modular workflow provides a powerful strategy for optimizing complex heterologous pathways and uncovering emergent host biology.

CRISPRi↗

The Transcription Factor Roc1 Is a Key Regulator of Cellulose Degradation in the Wood-Decaying Mushroom Schizophyllum commune

Wood-decaying fungi of the class Agaricomycetes (phylum Basidiomycota) are saprotrophs that break down lignocellulose and play an important role in nutrient recycling. They secrete a wide range of extracellular plant cell wall degrading enzymes that break down cellulose, hemicellulose, and lignin, the main building blocks of plant biomass. Although the production of these enzymes is regulated mainly at the transcriptional level, no activating regulators have been identified in any wood-decaying fungus in the class Agaricomycetes. We studied the regulation of cellulase expression in the wood-decaying fungus Schizophyllum commune. Comparative genomics and transcriptomics on two wild isolates revealed a Zn 2 Cys 6 -type transcription factor gene (roc1) that was highly upregulated during growth on cellulose, compared to glucose. It is only conserved in the class Agaricomycetes. A roc1 knockout strain showed an inability to grow on medium with cellulose as sole carbon source, and growth on cellobiose and xylan (other components of wood) was inhibited. Growth on non-wood-related carbon sources was not inhibited. Cellulase gene expression and enzyme activity were reduced in the Δroc1 strain. ChIP-Seq identified 1474 binding sites of the Roc1 transcription factor. Promoters of genes involved in lignocellulose degradation were enriched with these binding sites, especially those of LPMO (lytic polysaccharide monooxygenase) CAZymes, indicating that Roc1 directly regulates these genes. A conserved motif was identified as the binding site of Roc1, which was confirmed by a functional promoter analysis. Together, Roc1 is a key regulator of cellulose degradation and the first identified in wood-decaying fungi in the phylum Basidiomycota.

59 BASIC BIOLOGICAL SCIENCES↗

Quantum Key Distribution with Temporal Coding of Time-Bin Qubits

In this study, we demonstrate the possibility of realizing the quantum key distribution system by temporal coding of time-bin qubits that are prepared and measured using unbalanced Mach-Zehnder interferometers. We also show that, to exclude a clear a priori identification of time-bin qubits with equal bases, their encoding with the symbols 0 and 1 can be accomplished by delaying one of them by half the clock interval. The uniqueness of measurements of qutrits in this algorithm is shown to be based on the ability of quantum particles in β-states to interfere with the probability amplitudes at the ports of the system interferometers, as well as the absence of collisions of signaling states of the qutrits in adjacent clock intervals. We describe a method for controlling such collisions, including those that are based on qubits prepared from quasi-one-photon coherent laser states.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗