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At least 415 records · Page 23

Improved quality metrics for association and reproducibility in chromatin accessibility data using mutual information

Correlation metrics are widely utilized in genomics analysis and often implemented with little regard to assumptions of normality, homoscedasticity, and independence of values. This is especially true when comparing values between replicated sequencing experiments that probe chromatin accessibility, such as assays for transposase-accessible chromatin via sequencing (ATAC-seq). Such data can possess several regions across the human genome with little to no sequencing depth and are thus non-normal with a large portion of zero values. Despite distributed use in the epigenomics field, few studies have evaluated and benchmarked how correlation and association statistics behave across ATAC-seq experiments with known differences or the effects of removing specific outliers from the data. Here, we developed a computational simulation of ATAC-seq data to elucidate the behavior of correlation statistics and to compare their accuracy under set conditions of reproducibility. Using these simulations, we monitored the behavior of several correlation statistics, including the Pearson’s R and Spearman’s ρ coefficients as well as Kendall’s τ and Top–Down correlation. We also test the behavior of association measures, including the coefficient of determination R 2 , Kendall’s W, and normalized mutual information. Our experiments reveal an insensitivity of most statistics, including Spear man’s ρ, Kendall’s τ, and Kendall’s W, to increasing differences between simulated ATAC-seq replicates. The removal of co-zeros (regions lacking mapped sequenced reads) between simulated experiments greatly improves the estimates of correlation and association. After removing co-zeros, the R 2 coefficient and normalized mutual information display the best performance, having a closer one-to-one relationship with the known portion of shared, enhanced loci between simulated replicates. When comparing values between experimental ATAC-seq data using a random forest model, mutual information best predicts ATAC-seq replicate relationships. Collectively, this study demonstrates how measures of correlation and association can behave in epigenomics experiments. We provide improved strategies for quantifying relationships in these increasingly prevalent and important chromatin accessibility assays.

59 BASIC BIOLOGICAL SCIENCES↗

Direct tests of cytochrome c and c 1 functions in the electron transport chain of malaria parasites

The mitochondrial electron transport chain (ETC) of Plasmodium malaria parasites is a major antimalarial drug target, but critical cytochrome (cyt) functions remain unstudied and enigmatic. Parasites express two distinct cyt c homologs (c and c-2) with unusually sparse sequence identity and uncertain fitness contributions. P. falciparum cyt c-2 is the most divergent eukaryotic cyt c homolog currently known and has sequence features predicted to be incompatible with canonical ETC function. We tagged both cyt c homologs and the related cyt c1 for inducible knockdown. Translational repression of cyt c and cyt c1 was lethal to parasites, which died from ETC dysfunction and impaired ubiquinone recycling. In contrast, cyt c-2 knockdown or knockout had little impact on blood-stage growth, indicating that parasites rely fully on the more conserved cyt c for ETC function. Biochemical and structural studies revealed that both cyt c and c-2 are hemylated by holocytochrome c synthase, but UV-vis absorbance and EPR spectra strongly suggest that cyt c-2 has an unusually open active site in which heme is stably coordinated by only a single axial amino acid ligand and can bind exogenous small molecules. These studies provide a direct dissection of cytochrome functions in the ETC of malaria parasites and identify a highly divergent Plasmodium cytochrome c with molecular adaptations that defy a conserved role in eukaryotic evolution.

60 APPLIED LIFE SCIENCES↗

Comparative physiological and genomic characterization of a novel Nitrobacter vulgaris strain from a nitrate-contaminated subsurface

Nitrite-oxidizing bacteria (NOB) represent a crucial node in the global nitrogen cycle. By catalyzing the second step of nitrification—the oxidation of nitrite to nitrate to generate energy for growth—NOB activity controls the fate of nitrite (NO 2 - ) in aerobic environments. Despite thriving in diverse environments, including soils, freshwater, marine ecosystems, subsurface habitats, and water treatment systems, organisms capable of nitrite oxidation are confined to Nitrobacter, Nitrospira, Nitrospina, Nitrotoga, and a few other specific lineages. The genus Nitrobacter, recognized for its facultative heterotrophic metabolism, is often associated with high-nitrogen environments. Here, we report the physiological characterization of a novel strain, Nitrobacter vulgaris strain MLSD-S22, isolated from a nitrate- and heavy-metal-contaminated subsurface. Growth inhibition experiments revealed that strain MLSD-S22 and the N. vulgaris type strain Z exhibited similar sensitivities to nitrite and nitrate, with nitrite being the most inhibitory. Microrespirometry demonstrated that the two N. vulgaris strains and Nitrobacter winogradskyi Nb-255 possessed higher affinities for nitrite and oxygen than previously reported for Nitrobacter, suggesting potential to compete in low-substrate environments. Long-read DNA sequencing provided a complete genome for strain MLSD-S22, revealing two plasmids and an intact nitrous oxide (N 2 O) reduction operon—an unexpected feature for Nitrobacter. While N 2 O reduction activity was not observed under the tested conditions, this discovery raises questions about the contribution of Nitrobacter NOB to the N 2 O sink. These findings broaden the physiological and genomic diversity of Nitrobacter, offering new insights into their adaptation strategies and providing a framework for future evaluation of their potential roles in nitrogen loss.

Nitrobacter↗

Assessment of Sodium Thermal Stratification Models Utilizing the TSTF Benchmark

As a result of certain transient scenarios, a thermally stratified layer of liquid sodium can develop in the bulk coolant volumes of a sodium-cooled fast reactor (SFR). In addition to the effects a stratification layer has on the temperature of the heat transport system, a stratification layer can also influence the transition to and establishment of natural circulation flow, which plays an important role in passive cooling and the inherent safety of a pool-type SFR. Therefore, the ability to accurately capture thermal stratification phenomena is important when demonstrating the safety basis of a pool-type SFR during transient sequences. The present work assesses various computational models with different fidelities in their ability to predict thermal stratification in the upper plenum of an SFR. Each computational model will be assessed using the data generated at the Thermal Stratification Test Facility (TSTF) located at the University of Wisconsin-Madison. Using measured flow rate and inlet temperature data, the measured temperature distributions of the tests are compared to the predictions of the lumped volume-based models in SAS4A/SASSYS-1, a 1D-based model in SAM, and a 3-D computational fluid dynamics (CFD) model using STAR-CCM+. The relative performance of the various computational methods is assessed with respect to key metrics such as bulk coolant temperature distribution and plenum exit temperature. A total of eight tests are analyzed, covering different combinations of flow rates (3 and 10 GPM) and upper internal structure (UIS) configurations (none, solid, porous, and open) The perfect mixing model of SAS4A/SASSYS-1 provides the highest accuracy when the flow rate is high and there is no UIS in the test vessel, as high flow rate injection promotes thermal mixing of the sodium in the test vessel. For most of the analyzed tests, the stratified volume model of SAS4A/SASSYS-1 is able to predict the delay in the outlet temperature drop and temperature distribution in the test vessel by a small number of layers to represent thermal stratification. However, the stratified volume model can only simulate a maximum of three temperature layers within a volume and when a layer approaches the elevation of the outlet, the predicted outlet temperature can demonstrate rapid, non-physical changes. The 1-D axial mixing model of SAM provides results that agree reasonably well with the measured data in the prediction of the temporal evolution of the outlet temperature with the exception of the case with a high flow rate and no UIS. The SAM 1-D model has a similar level of accuracy to CFD results when it comes to predicting the outlet temperature. CFD shows overall good agreement in predicting the temperature distribution in the test vessel and outlet temperature. As CFD can model the test vessel geometry in detail, it performs well in the cases of complex geometries such as tests that included a UIS and internal flow through the UIS resulting in active mixing of the coolant in the test vessel. Each of the models discussed in the present work has the potential to be useful during the various stages of reactor design, analysis, and licensing. The lumped-volume approach can be applied for fast turnaround safety calculations to obtain overall reactor behavior during transients. The 1-D models provide improved accuracy when stratification is expected for a relatively low increase in the computational cost. The CFD model can be utilized for confirmatory analysis of the 1-D model, when experimental measurements are not available.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

Evaluating a Laboratory Flume Microbiome as a Window Into Natural Riverbed Biogeochemistry

Riverbeds are hotspots for microbially-mediated reactions that exhibit pronounced variability in space and time. It is challenging to resolve biogeochemical mechanisms in natural riverbeds, as uncontrolled settings complicate data collection and interpretation. To overcome these challenges, laboratory flumes are often used as proxies for natural riverbed systems. Flumes capture spatiotemporal variability and thus allow for controlled investigations of riverbed biogeochemistry. These investigations implicitly rely on the assumption that the flume microbiome is similar to the microbiome of natural riverbeds. However, this assumption has not been tested and it is unknown how the microbiome of a flume compares to natural aquatic settings, including riverbeds. To evaluate the fundamental assumption that a flume hosts a microbiome similar to natural riverbed systems, we used 16s rRNA gene sequencing and publicly available data to compare the sediment microbiome of a single large laboratory flume to a wide variety of natural ecosystems including lake and marine sediments, river, lake, hyporheic, soil, and marine water, and bank and wetland soils. Richness and Shannon diversity metrics, analyses of variance, Bray-Curtis dissimilarity, and analysis of the common microbiomes between flume and river sediment all indicated that the flume microbiome more closely resembled natural riverbed sediments than other ecosystems, supporting the use of flume experiments for investigating natural microbially-mediated biogeochemical processes in riverbeds.

54 ENVIRONMENTAL SCIENCES↗

Hierarchical semi-Markov models with duration-aware dynamics for activity sequences

Residential electricity demand at granular scales is driven by what people do and for how long. Accurately forecasting this demand for applications like microgrid management and demand response therefore requires generative models for activities that can produce realistic daily activity sequences, capturing both the timing and duration of human behavior. This paper develops a generative model of human activity sequences using nationally representative time-use diaries at a 10-min resolution. We use this model to quantify which demographic factors are most critical for improving predictive performance. We propose a hierarchical semi-Markov framework that addresses two key modeling challenges. First, a time-inhomogeneous Markov router learns the patterns of “which activity comes next.” Second, a semi-Markov hazard component explicitly models activity durations, capturing “how long” activities realistically last. To ensure statistical stability when data are sparse, the model pools information across related demographic groups and time blocks. The entire framework is trained and evaluated using survey design weights to ensure our findings are representative of the U.S. population. On a held-out test set, we demonstrate that explicitly modeling durations with the hazard component provides a substantial and statistically significant improvement over purely Markovian models. Furthermore, our analysis reveals a clear hierarchy of demographic factors: Sex, Day-Type, and Household Size provide the largest predictive gains, while Region and Season, though important for energy calculations, contribute little to predicting the activity sequence itself. The result is an interpretable and robust generator of synthetic activity traces, providing a high-fidelity foundation for downstream energy systems modeling.

24 POWER TRANSMISSION AND DISTRIBUTION↗

Polymerase chain reaction for the in vitro detection of the pESI plasmid associated with the globally circulating Salmonella Infantis outbreak strain

Abstract A globally circulating strain of Salmonella enterica serotype Infantis containing the pESI plasmid has increased in prevalence in poultry meat samples and cases of human infections. In this study, a polymerase chain reaction (PCR) protocol was designed to detect the pESI plasmid and confirm the Infantis serotype of Salmonella isolates. Primers were tested bioinformatically to predict specificity, sensitivity, and precision. A total of 54 isolates of Salmonella serotypes Infantis, Senftenberg, and Alachua were tested, with and without the pESI plasmid carriage. Isolates of 31 additional serotypes were also screened to confirm specificity to Infantis. Specificity, sensitivity, and precision of each primer were >0.95. All isolates tested produced the expected band sizes. This PCR protocol provides a rapid and clear result for the detection of the pESI plasmid and serotype Infantis and will allow for the in vitro detection for epidemiological studies where whole-genome sequencing is not available.

McMillan, Elizabeth A. (ORCID:000000027503514X)↗

Scalability of high-performance PDE solvers

Performance tests and analyses are critical to effective high-performance computing software development and are central components in the design and implementation of computational algorithms for achieving faster simulations on existing and future computing architectures for large-scale application problems. In this article, we explore performance and space-time trade-offs for important compute-intensive kernels of large-scale numerical solvers for partial differential equations (PDEs) that govern a wide range of physical applications. We consider a sequence of PDE-motivated bake-off problems designed to establish best practices for efficient high-order simulations across a variety of codes and platforms. We measure peak performance (degrees of freedom per second) on a fixed number of nodes and identify effective code optimization strategies for each architecture. In addition to peak performance, we identify the minimum time to solution at 80% parallel efficiency. The performance analysis is based on spectral and p-type finite elements but is equally applicable to a broad spectrum of numerical PDE discretizations, including finite difference, finite volume, and h-type finite elements.

97 MATHEMATICS AND COMPUTING↗

Multichannel Deconvolution of Vibrational Shock Signals: An Inverse Filtering Approach

When transporting critical systems of national security interest, out-of-the ordinary, impulsive events that can potentially be undetected and affect overall system performance are of great concern. Impulsive events that can occur are essentially pulse-like, transient signals of short duration that evolve from various phenomena. Here the event can be created by either the dropping of a test object, the system, subjecting it to a compact high-energy blow or being struck unintentionally during transit resulting in potential damage. The intensity and location of the strike can cause an inoperability condition that is unacceptable in a national security environment. Therefore, it is essential to detect, classify and localize damage of any test object subjected to an impulsive-event. This effort was targeted to evaluate the vibrational response of test objects that are subjected to “transport” shocks and roadway vibrations during shipping and handling. Any potential damage that could be inflicted during transportation must not only be detected, but also be evaluated to determine the operational readiness of a test object before and after transport. This event is a critical task that must be addressed as part of the Lawrence Livermore National Laboratory (LLNL) national security mission. The estimation of excitation signals from noisy data is termed the deconvolution problem in the signal processing literature. The deconvolution problem is based on recovering the input excitation signal from a system characterized by its impulse response sequence. Using this model of the system, an “inverse” representation or filter is developed to remove the system from the measured data and recover the input. Deconvolution techniques have existed for a long-time; however, transient deconvolution presents a few uncommon problems, since the signal has a finite-length time duration resulting in a limited amount of data containing information about the excitation process. The transient is wideband in the frequency domain relative to any measurement sensor implying that the smaller bandwidth sensor system “filters” the excitation eliminating some of its essential information for recovery. This fact, coupled with the filtering effect of the test object itself makes this excitation recovery (deconvolution) problem a challenge for signal processing.

42 ENGINEERING↗

Oxidation of Glycolate in the Defense Waste Processing Facility (DWPF) Recycle Collection Tank - 20305

The Savannah River Site's Defense Waste Processing Facility (DWPF) operations are being upgraded with the introduction of the Nitric-Glycolic Flowsheet. Glycolic acid has been shown superior to formic acid as the reducing acid used during chemical processing. The new flowsheet improves or maintains necessary parameters such as 1) reduction of mercury, 2) adjustment of feed rheology and 3) adjustment of melter oxidation/reduction potential. Further, the potential for catalytic hydrogen generation in DWPF processing is virtually eliminated. DWPF process condensates are collected and returned to the SRS Concentration, Storage and Transfer Facilities (CSTF). The Recycle Collection Tank (RCT) collects off-gas condensate during chemical processing, vitrification, and other unit operations performed in DWPF and is the singular return vessel delivering recycle effluent back to CSTF. Each batch of recycle may contain a small amount of glycolate from chemical processing and melter off-gas condensates. To avoid potential flammability issues due to thermolysis of glycolate in the CSTF, chemical oxidation within the RCT has been investigated as an option for mitigating the transfer of glycolate. Sodium permanganate has been down-selected as the best option for oxidation of glycolate. Testing was performed using both 2-L and 22-L reactors (16,800:1 and 1,530:1 scale by volume) with non-radioactive waste simulants to approximate the expected RCT compositions. RCT simulants were evaluated at various process pH and temperature conditions. Also, RCT operations, namely the sequence of addition of corrosion inhibitors (NaOH and NaNO{sub 2}) versus a permanganate strike, were evaluated. Glycolate was introduced via a sludge simulant to mimic both expected entrainment and abnormal process foam-over conditions - the range being between 68 and 5100 mg/kg glycolate. Glycolate destruction was monitored by ion chromatography (IC). The corresponding manganese behavior was monitored in real-time using in situ ultraviolet-visible (UV-Vis) spectroscopy. RCT glycolate content can be reduced to below the IC detection limit within 90 minutes for all concentrations investigated. Ion Chromatography analysis revealed that under alkaline conditions, glycolate is primarily oxidized to oxalate with no significant formation of CO{sub 2} or carbonate, and nitrite is not oxidized to nitrate. Initially, complete oxidation of organics species and nitrite was assumed. Determination of the mechanistic chemical reaction has allowed the required amount of permanganate to be more accurately predicted and the total addition to be significantly reduced. UV-Vis measurements reveal that permanganate (Mn{sup 7+}) is reduced to manganate (Mn{sup 6+}) in the RCT. The oxidant stoichiometry is defined by using the initial permanganate to glycolate (P/G) molar ratio. At low initial glycolate concentration (68 and 140 mg/kg), the minimum required initial permanganate to glycolate (P/G) molar ratio was found to be 5-6. With high initial glycolate concentrations (5100 mg/kg) a lower (P/G) molar ratio of ∼2.5 was needed. The final portion of this effort supporting the nitric/glycolic flowsheet will be to test actual (fully radioactive) RCT samples as per the above simulant tests. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

TICC Clustering Library v.1.0

SAND2024-01234O TICC is a clustering algorithm that labels a sequence of data points according to numerical properties. This library is a Python implementation of the algorithm described in "Toeplitz Inverse Covariance-Based Clustering of Multivariate Time Series Data" (Hallac et al. 2017). It includes documentation, performance improvements, examples, and test coverage. This library allows users to automatically segment a series of multivariate data points according to their covariance—that is, the way the values at each data point are changing in relation to one another. This is useful for identifying periods in which a system is behaving. For example, if a sensor is measuring a car's velocity, steering wheel angle, braking and acceleration, TICC can determine when the car was stopped, beginning/exiting a turn, slowing or accelerating at an intersection, or driving on straight or curved roads. TICC can be applied to measure multiple quantities at known times. Sandia National Laboratories is a multimission laboratory managed and operated by National Technology & Engineering Solutions of Sandia, LLC, a wholly owned subsidiary of Honeywell International Inc., for the U.S. Department of Energy’s National Nuclear Security Administration under contract DE-NA0003525

Dalbey, Keith↗

Multimodal Approaches for Leveraging Domain Knowledge with State-of-the-Art Machine Learning to Engineer Biocatalysts

This grant aimed to accelerate the development of specialized enzymes—biological catalysts essential for sustainable manufacturing and medicine—by integrating traditional laboratory evolution with cutting-edge artificial intelligence. To achieve this, we developed a suite of high-throughput sequencing tools and a centralized database to bridge the gap between a protein’s genetic "code" and its physical function. By training machine learning models on large datasets, we also demonstrated the ability to move beyond slow, trial-and-error testing to a "generative" approach, where AI can independently design new, versatile enzymes like tryptophan synthases. Ultimately, these findings demonstrate that combining laboratory data with computer-guided design enables the engineering of highly efficient biological tools with unprecedented speed and precision.

59 BASIC BIOLOGICAL SCIENCES↗

Geology of the One Earth Energy Site

The One Earth Energy site is one of two sites in the Illinois Storage Corridor (ISC) project. The objectives of the ISC project is to accelerate commercial deployment of carbon capture utilization and storage at two individual sites and receive approvals for Underground Injection Control (UIC) Class VI permits for construction at each site. At the One Earth Energy site, an extensive data collection program was undertaken, which included the drilling of a test well (One Earth Energy #1 [OEE #1]), four 2D seismic lines, and a small 3D seismic survey. The OEE #1 well was drilled in 2022 and acquired extensive core, log, and testing data to characterize the subsurface geology of the site. Coring was focused on the storage interval, the Mt. Simon Sandstone, and the confining interval, the Eau Claire Formation. The core and log data were used to evaluate the sedimentology and sequence stratigraphy, as well as to develop the conceptual geologic model. This report includes the geological summaries of the Mt. Simon Sandstone and the Eau Claire Formation. The extensive analysis of the log data is included in the petrophysical section, showing ranges of porosity, estimated pore size, and the mineral content of selected zones in the well. The separate petrographic technical report entitled “Petrographic and Advanced Geologic Characterization Report on One Earth Energy #1 (API# 1211325373)”, report number DOE-UIUC-0031892-04, details thin section point-counting analysis that includes mineralogical and pore space analysis, including grain size analysis, annotated thin section photomicrographs, scanning electron microscopy (SEM) with energy dispersive X-ray spectroscopy (EDS), and statistics of grain size analysis on Mt. Simon thin sections from OEE #1. The final OEE #1 well data to be included in this geology report is the routine core analysis of both whole core plugs and rotary sidewall core plugs. In addition to the OEE #1 well, four 2D seismic lines and a small 3D survey were acquired as part of the overall subsurface geological characterization. This geology report references the seismic interpretation report, entitled “One Earth Energy Site Seismic Interpretation Task 5.0”, report number DOE-UIUC-0031892-07. This report details the stratigraphic and structural interpretation of the 2D and 3D seismic data acquired at the One Earth Energy site. The 2D seismic data was acquired in 2019 and 2021, and the 3D survey was acquired in 2022. The objectives of the seismic programs were to contribute to the subsurface characterization of the Mt. Simon-Eau Claire Storage Complex by evaluating the continuity of potential storage reservoirs and containment intervals across the project area, and to determine if any geologic features are present that would increase containment risk to the proposed carbon storage project.

09 BIOMASS FUELS↗

DL-TODA: A Deep Learning Tool for Omics Data Analysis

Metagenomics is a technique for genome-wide profiling of microbiomes; this technique generates billions of DNA sequences called reads. Given the multiplication of metagenomic projects, computational tools are necessary to enable the efficient and accurate classification of metagenomic reads without needing to construct a reference database. The program DL-TODA presented here aims to classify metagenomic reads using a deep learning model trained on over 3000 bacterial species. A convolutional neural network architecture originally designed for computer vision was applied for the modeling of species-specific features. Using synthetic testing data simulated with 2454 genomes from 639 species, DL-TODA was shown to classify nearly 75% of the reads with high confidence. The classification accuracy of DL-TODA was over 0.98 at taxonomic ranks above the genus level, making it comparable with Kraken2 and Centrifuge, two state-of-the-art taxonomic classification tools. DL-TODA also achieved an accuracy of 0.97 at the species level, which is higher than 0.93 by Kraken2 and 0.85 by Centrifuge on the same test set. Application of DL-TODA to the human oral and cropland soil metagenomes further demonstrated its use in analyzing microbiomes from diverse environments. Compared to Centrifuge and Kraken2, DL-TODA predicted distinct relative abundance rankings and is less biased toward a single taxon.

59 BASIC BIOLOGICAL SCIENCES↗

Nitrogen Metabolism in Pseudomonas putida : Functional Analysis Using Random Barcode Transposon Sequencing

Pseudomonas putida KT2440 has long been studied for its diverse and robust metabolisms, yet many genes and proteins imparting these growth capacities remain uncharacterized. Using pooled mutant fitness assays, we identified genes and proteins involved in the assimilation of 52 different nitrogen containing compounds. To assay amino acid biosynthesis, 19 amino acid drop-out conditions were also tested. From these 71 conditions, significant fitness phenotypes were elicited in 672 different genes including 100 transcriptional regulators and 112 transport-related proteins. We divide these conditions into 6 classes, and propose assimilatory pathways for the compounds based on this wealth of genetic data. To complement these data, we characterize the substrate range of three promiscuous aminotransferases relevant to metabolic engineering efforts in vitro. So we examine the specificity of five transcriptional regulators, explaining some fitness data results and exploring their potential to be developed into useful synthetic biology tools. In addition, we use manifold learning to create an interactive visualization tool for interpreting our BarSeq data, which will improve the accessibility and utility of this work to other researchers.

59 BASIC BIOLOGICAL SCIENCES↗

Exploring drought-responsive crucial genes in Sorghum

Drought severely affects global food production. Sorghum is a typical drought-resistant model crop. Based on RNA-seq data for Sorghum with multiple time points and the gray correlation coefficient, this paper firstly selects candidate genes via mean variance test and constructs weighted gene differential co-expression networks (WGDCNs); then, based on guilt-by-rewiring principle, the WGDCNs and the hidden Markov random field model, drought-responsive crucial genes are identified for five developmental stages respectively. Enrichment and sequence alignment analysis reveal that the screened genes may play critical functional roles in drought responsiveness. A multilayer differential co-expression network for the screened genes reveals that Sorghum is very sensitive to pre-flowering drought. Furthermore, a crucial gene regulatory module is established, which regulates drought responsiveness via plant hormone signal transduction, MAPK cascades, and transcriptional regulations. The proposed method can well excavate crucial genes through RNA-seq data, which have implications in breeding of new varieties with improved drought tolerance.

60 APPLIED LIFE SCIENCES↗

Activity-based cell sorting reveals responses of uncultured archaea and bacteria to substrate amendment

Metagenomic studies have revolutionized our understanding of the metabolic potential of uncultured microorganisms in various ecosystems. However, many of these genomic predictions have yet to be experimentally tested, and the functional expression of genomic potential often remains unaddressed. In order to obtain a more thorough understanding of cell physiology, novel techniques capable of testing microbial metabolism under close to in situ conditions must be developed. Here, we provide a benchmark study to demonstrate that bioorthogonal non-canonical amino acid tagging (BONCAT) in combination with fluorescence-activated cell sorting (FACS) and 16S rRNA gene sequencing can be used to identify anabolically active members of a microbial community incubated in the presence of various growth substrates or under changing physicochemical conditions. We applied this approach to a hot spring sediment microbiome from Yellowstone National Park (Wyoming, USA) and identified several microbes that changed their activity levels in response to substrate addition, including uncultured members of the phyla Thaumarchaeota, Acidobacteria, and Fervidibacteria. Because shifts in activity in response to substrate amendment or headspace changes are indicative of microbial preferences for particular growth conditions, results from this and future BONCAT-FACS studies could inform the development of cultivation media to specifically enrich uncultured microbes. Most importantly, BONCAT-FACS is capable of providing information on the physiology of uncultured organisms at as close to in situ conditions as experimentally possible.

59 BASIC BIOLOGICAL SCIENCES↗

Testing of a Line Driver With Configurable Pre-Emphasis on Lossy Transmission Lines

Rare-event physics experiments such as the Deep Underground Neutrino Experiment (DUNE) or the next Enriched Xenon Observatory (nEXO) experiment search for rare, low-energy events, detected by sensitive detectors immersed in a cryogenic noble liquid (e.g., liquid argon or xenon). Readout electronics used within such detectors must consume minimal power while operating reliably in cryogenic environments. Furthermore, in the case of nEXO, maximizing the radiopurity of the environment is vital to minimize background noise, thus placing strict limits on the volume of dielectric materials, leading to high-loss data cables spanning distances up to 12 m. Such cables cause high attenuation and intersymbol interference (ISI), resulting in a high bit-error rate (BER). These issues were addressed by developing an integrated line driver with configurable pre-emphasis in a 65-nm CMOS process. The pre-emphasis parameters can be programmed to minimize BER for specific cables and data rates under power constraints. Here, the driver was tested at both room and cryogenic temperatures. In both cases, the output BER was found to be strongly correlated with the pre-emphasis settings. Furthermore, analysis and simulation showed that adapting the pre-emphasis settings based on the incoming bit sequence can further improve performance with minimal changes to the current solution.

47 OTHER INSTRUMENTATION↗