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Riverbed and Near-Surface Water Quality Data, Hanford Reach, Columbia River, February 2021 - April 2022

This dataset contains longitudinal profiles of natural groundwater tracers (temperature, electrical conductivity (EC), and Radon-222 (Rn)) collected to identify locations of hydrologic exchange flows along a 75-km reach of the Columbia River near Richland, Washington.A jetboat was used to tow a 30 foot long, weighted tether that had one set of temperature and EC sensors just below the water surface and one set of temperature and EC sensors on the end (just above the river bed). The dataset contains profiles of temperature/EC data (resolution: 1 second) collected along the left and right banks during three different sampling events. During the final sampling event, we revisited some "deep holes" where the river was more than 30 feet deep with a weighted tether that was 75 feet long. The dataset has been processed to remove data collected when the sensors were not in the water, or when the water was too deep to keep the tether on the riverbed. We collected grab samples of water for analysis of dissolved Rn in places where the temperature/EC was different from the background river water. The Rn data was joined to the temperature/EC profiles based on the sample times.This dataset is comprised of one main data folder containing (1) file-level metadata; (2) data dictionary; (3) installation methods; (4) February 2021 data; (5) July 2021 data; (6) April 2022 data; and (7) April 2022 deep hole data. All files are .csv format and can be opened with a spreadsheet program (such as Microsoft Excel or OpenOffice).

54 ENVIRONMENTAL SCIENCES↗

Geothermometry Mapping of Deep Hydrothermal Reservoirs in Southeastern Idaho: Final Report

The Eastern Snake River Plain (ESRP) in southern Idaho is a region of high heat flow. Sustained volcanic activities in the wake of the passage of Yellowstone Hotspot have turned this region into an area with great potential for geothermal resources. Numerous hot springs with temperatures up to 75 ºC are scattered along the margins of the plain. Similarly, several hot-water producing wells and few hot springs are also present within the plain. The geothermal reservoirs in the area are likely to be hosted at depth in the felsic volcanic rocks underneath the thick sequences of basalts within the plain and the Paleozoic rocks underneath both basalts and felsic volcanic rocks along the margins. The heat source to these geothermal resources is thought to be the mid-crustal sill complex which sustains high heat flow in the ESRP. Several thermal anomaly areas are believed to be associated with the local thermal perturbation because of the presence of favorable structural settings. However, it is hypothesized that the pervasive presence of an overlying groundwater aquifer in the region effectively masks thermal signatures of deep-seated geothermal resources. The dilution of deeper thermal water and re-equilibration at lower temperatures are significant challenges for the evaluation of potential resource areas in the ESRP. To address this issue, this project, led by the Idaho National Laboratory (INL), aimed at applying advanced geothermometry tools including temperature-dependent mineral and isotopic equilibria with mixing models that account for processes such as boiling and dilution with shallow groundwater that could affect calculated temperatures of underlying deep thermal waters. Over the past several years, we collected approximately 100 water samples from springs/wells for chemical analysis as well as assembled existing water chemistry data from literature. We applied several geothermometric and geochemical modeling tools to the compositions of ESRP water samples. Geothermometric calculations based on principle of multicomponent equilibrium geothermometry with inverse geochemical modeling capability (e.g., Reservoir Temperature Estimator, RTEst) have been useful for evaluation of reservoir temperatures. Similarly, sulfate-water oxygen isotope geothermometry was also applied to several samples in tandem with RTEst. In summary, geothermometric calculations of ESRP thermal water samples indicated numerous potential geothermal areas with elevated reservoir temperatures. Specifically, areas around southern/southwestern side of the Mount Bennet Hills and within the Camas Prairie in the southwestern portion of the ESRP suggest temperatures 140-190 °C. In the northern portion of the ESRP, Lidy Hot Springs, Ashton, Newdale, and areas east of Idaho Falls have expected reservoir temperature ?140 °C. In the southern ERSP, areas near Buhl and Twin Falls are found to have elevated temperatures as high as 160 °C. These areas are likely to host potentially economic geothermal resources; however, further detailed study is warranted to each site to evaluate hydrothermal suitability for economic use.

58 GEOSCIENCES↗

Advanced low-cost intermetallic coatings for molten salt pump impeller. Final report

1.0 INTRODUCTION The use of solar energy to produce steam for the generation of electricity is an established technology, with multiple designs for concentrating solar power (CSP) steam generators available commercially and a number of such units currently operating. As part of the effort to improve the overall efficiency of this technology, designers have developed thermal energy storage (TES) mechanisms that employ molten salt to capture a portion of the solar energy available during the day-time hours and make that energy available for steam generation during periods when the sun’s energy is not available. In current TES systems, nitrate salts are used extensively as the medium of energy exchange. However, due to the chemical instability of these salts at temperatures above 600ºC, they are not being considered for use in the next more efficient generation of CSP plants, in which the energy exchange fluids will be required to operate in the range 600ºC to 800ºC. At these higher temperatures, molten chloride salts, such as KCl-MgCl2, are viewed as promising alternatives to the nitrate salts, but, of course, these chloride-based salts are highly corrosive to the engineering alloys commonly used in the construction of pressure vessels, heat exchangers, pumps and piping at high temperatures. Hence, a solution to the problem of material degradation is essential if the next generation of CSP plants is to be commercially viable (Ref 1). In order to effectively address the problem of molten chloride salt corrosion at high temperatures (≥ 750ºC) and thereby to reduce the cost of molten salt tanks and related apparatus, various options have been considered, with one of the most promising being an innovative coatings system developed and applied onto low alloy ferritic or austenitic stainless material. Development of such a coating system would reduce the capital costs of thermal energy storage (TES) and increase the overall efficiency of the concentrating solar power (CSP) system. Although high strength nickel base alloys are well suited for many high temperature applications, they have significant limitations when exposed to molten chloride salt conditions and they are very expensive. Monolithic materials such as the austenitic stainless steels (e.g., SS310 and SS347) and the more common nickel base alloys (e.g., IN625 and IN800H) have exhibited high corrosion rates in molten chloride salt environments (Ref1). Molten salt corrosion of IN625 in salts containing chlorides showed a catastrophic corrosion rate of 1.111”/year at 800ºC (Ref1). Hence, an innovative coatings solution to mitigate molten chloride salt corrosion is needed. Of the many coating techniques available in the market, no one technique appears to be capable of producing effective protection from molten salt corrosion at a relatively low cost. There are thermal spray coating techniques, in particular the twin wire-arc technology, that are attractive when considering the coating of large molten salt tanks because the coatings application rate is high, the overall application cost is low and repairs to the coating can readily be made either in the shop or in the field. However, the twin wire arc coating materials currently available in the market have certain inherent limitations, including porosity and mechanical bonding to the substrate, which limit their effectiveness in providing protection against molten chloride salt corrosion at high temperatures. What is required is an approach that combines the economy and versatility of thermal spray wire-arc technology with the superior resistance of advanced alloys to effectively combat molten chloride salt corrosion at elevated temperatures. Applied Thermal Coatings, Inc. (ATC) has investigated an innovative coatings technology to produce dual layer cermet/ceramic coatings that will be dense and impervious to any corrosive molten salts/fluids, thereby resisting the effects of molten salt corrosion. Development of coatings using this technology will make it possible to fabricate the molten salt systems using a relatively low-cost base material, which will reduce the overall cost of the power generating system and enhance the commercial viability of solar power. 2.0 PROJECT OVERVIEW Of the coating technologies available for the production of a cost-effective method of protection from the effects of high-temperature chloride attack, there are two that offer specific advantages for the CSP/TES application, particularly when used in a way that effectively complements the strengths of each coating method. The first of these is electroless nickel plating, through which a nickel-rich layer is produced on the surface of a component through chemical reduction. The second coating technology is aluminizing, in which aluminum is diffused into the surface of a component and, depending on the composition of the substrate, aluminides are formed. The objective of this project has been to demonstrate the feasibility of a low- cost coating for the protection of pump impellers used for the inductive transport of molten salts. Initial interest focused on an inter-metallic nickel aluminide coating and subsequently, for economic reasons, consideration also was given to a coating composed of iron aluminides. Applying such coatings to low alloy or austenitic stainless-steel substrates will provide the superior resistance to molten chloride salt corrosion and blade tip erosion that normally can be achieved only through the use of more highly alloyed – and, therefore, more expensive - nickel-base materials and it will do so at a substantially lower cost. The coating systems evaluated were designed to take advantage either of the combined strengths of the electroless nickel plating process and the aluminizing process to produce a nickel-aluminide intermetallic coating or to rely solely on the aluminizing process to produce, in conjunction with iron from the substrate, a protective layer consisting of iron aluminides. With respect to the nickel aluminide protection, this has been achieved by first depositing a layer of nickel-rich alloy onto an austenitic stainless substrate using the electroless nickel plating process and then aluminizing the nickel- coated surface to produce the nickel aluminide coating. With respect to the iron aluminide protection, this has been achieved by aluminizing bare austenitic stainless steel. Testing has supported the conclusion that the developed coatings will resist attack by molten chloride salts at high temperatures (550°C to 750°C) and have sufficient hardness to resist blade tip erosion.The steps taken to develop either a low-cost intermetallic nickel aluminide coating using electroless nickel plating and aluminizing or a low-cost iron aluminide coating using aluminizing only, and to demonstrate the quality of the coatings and their effectiveness against molten chloride salt corrosion and erosion at high temperatures (≤ 750ºC) have been as follows: 1. Design of the optimum electroless nickel plating and aluminizing mixtures using advanced thermodynamic analysis; 2. Development of processing parameters for the electroless nickel plating and aluminizing processes, to be used either together or separately; 3. Production of specimens coated using the designated electroless nickel plating process; 4. Production of samples with intermetallic nickel aluminide coating or iron aluminide coating; 5. Investigation of the performance of the intermetallic nickel aluminide coating and the iron aluminide coating when subjected to molten chloride salts corrosion at high temperatures (≥ 750ºC); and, 6. Metallurgical characterization of the microstructure of the coatings, both prior to and after exposure to molten salt environments at elevated temperatures.

14 SOLAR ENERGY↗

Successes and Lessons Learned in the Design, Construction and Use of a Tool for Sampling the WR-1 Reactor Core - 1900z

Whiteshell Laboratories, located in Manitoba, Canada, provided research facilities for the Canadian nuclear industry since the early 1960's. The facility was centered on an organically cooled, heavy water moderated nuclear reactor in the world, Whiteshell Reactor 1 (WR-1). WR-1 was safely shut down in 1985, de-fueled and drained, and has been in a safe Storage with Surveillance (Deferred Decommissioning) state until present day. In support of an Environmental Assessment for proposed In Situ Decommissioning of WR-1, detailed characterization information regarding the reactor core components (calandria, calandria tubes, pressure tubes) was necessary to verify inventory estimates and neutron activation models produced circa 1992. The Fuel Channels of WR-1 (pressure tubes) were expected to be the most activated components of the reactor core, and to account for about 60% of the reactor core inventory. The interior surface of the fuel channel is also the most readily accessible surface within the reactor core. As such the decision was made to collect a sample of one of each of 3 Fuel Channel material types (Stainless Steel, Zr-2.5Nb, Ozhenite) for comparison to modelled estimates of neutron activation of various core materials. The expected dose rates from the samples were between 5 and 100 mSv (500-10,000 mrem) on contact based on a 200 mg sample. Sampling the Fuel Channels required verification the Fuel Channel was drained of organic coolant, removal of residual organic coolant from the Fuel Channel wall, removal of material from the interior surface of the Fuel Channel at the centerline of flux (∼25' below the operator position), and extraction of that material from the core. Several options for existing tooling and technologies, including hydraulic scrapers, and horizontal drill technologies, were looked at for their applicability to WR-1. While suitable approaches to higher radiation field sample collection, they were overly complex, costly and time consuming for the configuration at WR-1. In an effort to reduce costs, and to simplify the sampling process, a specialized and simple tool was developed to mechanically scrape a small sample of the interior wall of the Fuel Channels, collect the sample into a vial, and remove the sample from the core for analysis. The tool was designed, constructed, tested and implemented in the field entirely by Canadian Nuclear Laboratories Staff. The tool required no powered components, or electronic guidance or measurements; and was designed and constructed for a fraction of the cost of other commercially available technologies for sample collection within reactor cores. The samples were successfully collected without any injuries, contamination events or equipment failures. The tool remains operational in the event additional sampling is required. This paper provides additional details of the design, construction, testing and use of the tool and the successes and lessons learned during the various stages of design and implementation. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Neutron tomographic analysis: Material characterization of silver and electrum coins from the 6th and 5th centuries BCE

Neutron tomography was applied to study a set of Greek silver coins and a single Lydian electrum coin minted in the 6 and 5th centuries BCE. The investigation was conducted at the new neutron imaging station DINGO at ANSTO in order to demonstrate capabilities and to explore limits of this non-invasive method in characterizing numismatic materials. From the reconstructed volume of each sample, the morphology, porosity, inclusions, and presence of composite structures can be revealed and evaluated. This information can be used to elucidate ancient minting technology and to prove authenticity. Moreover, the state of conservation, corrosion processes and attempts to repair the coins can be determined. - Highlights: •Pre-Roman coins •Non-destructive analysis •Neutron tomography: limit and capability.

36 MATERIALS SCIENCE↗

Documentation of operational protocol for the use of MAMA software

Image analysis of Scanning Electron Microscope (SEM) micrographs is a complex process that can vary significantly between analysts. The factors causing the variation are numerous, and the purpose of Task 2b is to develop and test a set of protocols designed to minimize variation in image analysis between different analysts and laboratories, specifically using the MAMA software package, Version 2.1. The protocols were designed to be “minimally invasive”, so that expert SEM operators will not be overly constrained in the way they analyze particle samples. The protocols will be tested using a round-robin approach where results from expert SEM users at Los Alamos National Laboratory, Lawrence Livermore National Laboratory, Pacific Northwest National Laboratory, Savannah River National Laboratory, and the National Institute of Standards and Testing will be compared. The variation of the results will be used to quantify uncertainty in the particle image analysis process. The round-robin exercise will proceed with 3 levels of rigor, each with their own set of protocols, as described below in Tasks 2b.1, 2b.2, and 2b.3. The uncertainty will be developed using NIST standard reference material SRM 1984 “Thermal Spray Powder – Particle Size Distribution, Tungsten Carbide/Cobalt (Acicular)” [Reference 1]. Full details are available in the Certificate of Analysis, posted on the NIST website (http://www.nist.gov/srm/).

97 MATHEMATICS AND COMPUTING↗

Prediction of Subsidence of Buildings as a Result of Earthquakes by Gaussian Process Regression

Subsidence of a foundation caused by liquefaction of sandy soil is a critical step in the creation of a building. Seven measured indicators of soil liquefaction in the course of an earthquake were selected as key impacting indicators that predict subsidence of a building. These indicators are the earthquake intensity EI, length–height ratio L/H, average contact pressure p, width-depth ratio, relative density D{sub R}, thickness of the non-liquefied layer, and the depth of the gravel layer. Furthermore, as a probabilistic machine learning kernel and powerful tool for solving highly nonlinear problems, Gaussian process regression (GPR model) can meet execution time requirements and assure a high level of precision of subsidence prediction due to the intrinsic defects of theoretical analysis and numerical calculation. A total of 41 groups of typical cases were first selected as a training sample and 20 groups of typical cases were defined as a test sample, where these cases possess input test value and were obtained at the yield of the prediction accuracy from the GPR model. Next, the PLS method, MLR method and LSSVM method were selected to verify the validity and reliability of the GPR model. Finally, the simulation result shows that the GPR model can improve the prediction precision for the case of the above problems and is of great value for practical applications.

58 GEOSCIENCES↗

Results of Hg speciation testing on DWPF SMECT-8, OGCT-1, AND OGCT-2 samples

The Savannah River National Laboratory (SRNL) was tasked with preparing and shipping samples for Hg speciation by Eurofins Frontier Global Sciences, Inc. in Seattle, WA on behalf of the Savannah River Remediation (SRR) Mercury Task Team. The sixteenth shipment of samples was designated to include a Defense Waste Processing Facility (DWPF) Slurry Mix Evaporator Condensate Tank (SMECT) sample from Sludge Receipt and Adjustment Tank (SRAT) Batch 738 processing and two Off-Gas Condensate Tank (OGCT) samples, one following Batch 736 and one following Batch 738. The DWPF sample designations for the three samples analyzed are provided. The Batch 738 ‘End of SME Cycle’ SMECT sample was taken at the conclusion of Slurry Mix Evaporator (SME) operations for this batch and represents the fourth SMECT sample examined from Batch 738. Batch 738 experienced a sludge slurry carryover event, which introduced sludge solids to the SMECT that were particularly evident in the SMECT-5 sample, but less evident in the ‘End of SME Cycle’ SMECT-8 sample.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Disentangling Time-series Spectra with Gaussian Processes: Applications to Radial Velocity Analysis

Measurements of radial velocity variations from the spectroscopic monitoring of stars and their companions are essential for a broad swath of astrophysics; these measurements provide access to the fundamental physical properties that dictate all phases of stellar evolution and facilitate the quantitative study of planetary systems. The conversion of those measurements into both constraints on the orbital architecture and individual component spectra can be a serious challenge, however, especially for extreme flux ratio systems and observations with relatively low sensitivity. Gaussian processes define sampling distributions of flexible, continuous functions that are well-motivated for modeling stellar spectra, enabling proficient searches for companion lines in time-series spectra. We introduce a new technique for spectral disentangling, where the posterior distributions of the orbital parameters and intrinsic, rest-frame stellar spectra are explored simultaneously without needing to invoke cross-correlation templates. To demonstrate its potential, this technique is deployed on red-optical time-series spectra of the mid-M-dwarf binary LP661-13. We report orbital parameters with improved precision compared to traditional radial velocity analysis and successfully reconstruct the primary and secondary spectra. We discuss potential applications for other stellar and exoplanet radial velocity techniques and extensions to time-variable spectra. The code used in this analysis is freely available as an open-source Python package.

79 ASTRONOMY AND ASTROPHYSICS↗

2015 Annual Report

For more than 20 years, the Atmospheric Radiation Measurement (ARM) Climate Research Facility has blazed the trail in providing the world’s atmospheric scientists with continuous observations of cloud and aerosol properties and their impacts on Earth’s energy balance. The result is an unprecedented data set that has proved invaluable for understanding the atmosphere and improving the predictive capabilities of earth system models. Over the past year, plans have been put in place to create a powerful new capability for furthering ARM’s mission and achieving the vision outlined in the Decadal Vision, the strategy for the ARM Facility for the next 5 to 10 years. The result is a significant reconfiguration for the ARM Facility that will provide more complete data sets to support process studies and model development. The combined observational and modeling elements will enable a new level of scientific inquiry. To achieve the next-generation ARM Facility, plans underway include the following: Establishing observation “megasites” at the Southern Great Plains and North Slope of Alaska sites Enhancing ARM measurement excellence to support U.S. Department of Energy climate science research Producing routine high-resolution model simulations over domains coincident with ARM sites Developing data products and software tools that facilitate analysis and enable the evaluation of models using ARM data Strengthening interactions with the broader atmospheric science and modeling communities. Changes being implemented at the megasites will provide additional spatial sampling of atmospheric and surface information to support a new ARM capability—the routine operation of high-resolution process models at ARM sites. The result will be even more comprehensive scientific data and the processes and tools for scientists to more easily use ARM data for climate model development.

Southern Great Plains, North Slope of Alaska, larg↗

Multimodal imaging and machine learning to enhance microscope images of shale

A machine learning based image processing workflow is presented to enhance shale source rock microscopic images obtained using diverse imaging platforms. Images were acquired from a 30 μm diameter cylindrical Vaca Muerta shale sample using both nondestructive Transmission X-Ray Microscopy (TXM, alternately referred to as nano computed tomography) and destructive Focused Ion Beam-Scanning Electron Microscopy (FIB-SEM). Output cross-sectional images from each modality were aligned using a combination of manual and automated registration techniques to create a registered image dataset. We then apply this dataset for two image processing tasks: prediction of image cross sections with SEM-like resolution from nondestructive TXM data and repair of charged region artifacts (localized accumulation of electrons) within SEM images. The image processing algorithms for both tasks use deep learning models, specifically image-to-image Convolutional Neural Networks (CNNs) and conditional Generative Adversarial Networks (cGANs). In the image enhancement tasks, we are able to achieve significant qualitative and quantitative improvement in TXM images. Here, the best model reaches an average Peak Signal to Noise Ratio (PSNR) of 15.8 dB. Conditioning on TXM data is also shown to reduce artifacts from SEM charging, achieving an average PSNR of 25.8 dB. Furthermore, our results suggest that properly trained and validated networks are capable of significant enhancement of images obtained using nondestructive techniques, thereby improving interpretation of two- and three-dimensional images while preserving samples for future use.

58 GEOSCIENCES↗

Rapid dissolution without elemental fractionation by laser driven hydrothermal processing

Traditional dissolution of geologic samples often requires a significant time investment. Here, we present an alternative method for the dissolution of geologic materials using laser-driven hydrothermal processing (LDHP). LDHP uses laser energy directed onto a submerged sample, which increases the temperature and pressure at the liquid–sample interface and drives the hydrothermal dissolution coupled with photomechanical spallation, an ablative process. This uses focused 527 nm laser energy at 40 W average power, 1 kHz pulse repetition rate, and 115 ns pulse duration. Importantly, when LDHP is performed on basalt geostandards (BCR-2 and BHVO-2) using the conditions outlined, we show that LDHP does not produce significant elemental fractionation and, thus, can be considered an alternative processing method to traditional mechanical crushing and acid digestion. Additionally, it is possible using LDHP to utilize the spatially confined beam to target and selectively isolate individual phases in a rock, potentially alleviating the need for mechanical separation of inclusions that are difficult to physically isolate. Furthermore, using this outlined method of LDHP, we demonstrate full dissolution of 120 mg of obsidian in 85 minu, meaning that LDHP is a potentially very useful method when sample processing is time sensitive.

58 GEOSCIENCES↗

Updraft dynamics and microphysics: on the added value of the cumulus thermal reference frame in simulations of aerosol–deep convection interactions

One fundamental question about atmospheric moist convection processes that remains debated is whether, or under which conditions, a relevant variability in background aerosol concentrations may have a significant dynamical impact on convective clouds and their associated precipitation. Furthermore, current climate models must parameterize both the microphysical and the cumulus convection processes, but this is usually implemented separately, whereas in nature there is a strong coupling between them. As a first step to improve our understanding of these two problems, we investigate how aerosol concentrations modify key properties of updrafts in eight large-eddy-permitting regional simulations of a case study of scattered convection over Houston, Texas, in which convection is explicitly simulated and microphysical processes are parameterized. Dynamical and liquid-phase microphysical responses are investigated using the following two different reference frames: static cloudy updraft grid cells versus tracked cumulus thermals. In both frameworks, we observe the expected microphysical responses to higher aerosol concentrations, such as higher cloud number concentrations and lower rain number concentrations. In terms of the dynamical responses, both frameworks indicate weak impacts of varying aerosol concentrations relative to the noise between simulations over the observationally derived range of aerosol variability for this case study. On the other hand, results suggest that thermals are more selective than cloudy updraft grid cells in terms of sampling the most active convective air masses. For instance, vertical velocity from thermals is significantly higher at upper levels than when sampled from cloudy updraft grid points, and several microphysical variables have higher average values in the cumulus thermal framework than in the cloudy updraft framework. In addition, the thermal analysis is seen to add rich quantitative information about the rates and covariability of microphysical processes spatially and throughout tracked thermal lifecycles, which can serve as a stronger foundation for improving subgrid-scale parameterizations.

54 ENVIRONMENTAL SCIENCES↗

Analysis of the MST-16 crimp process on cold 304L stainless steel

The purpose of this report is to assess the quality of the MST-16 crimp process through a series of cold experiments and metallographic analyses. Common practice in MST-16 uses a mechanical crimp on 304L stainless steel tubes to prevent radioactive contamination from escaping the tube interior for a variety of experiments. The crimped tubes must pass a leak check to demonstrate their radiological safety. Cold (i.e. non-radioactive samples) 304L tube samples were tested using two different approved crimping devices to assess the quality of the crimps. Each crimp was leak checked, followed by destructive metallographic analysis to determine the quality of both the crimp equipment and procedures used by MST-16 personnel.

42 ENGINEERING↗

Simple Analysis of Primary and Secondary Bile Salt Hydrolysis in Mouse and Human Gut Microbiome Samples by Using Fluorogenic Substrates

Animals produce bile to maximize absorption of lipophilic nutrients in the gut. The physical properties of bile are largely dictated by amphipathic bile salt molecules, which also act as signaling molecules that bind host receptors and modulate physiological processes. Upon excretion of bile salts into the gut, the gut microbiome can create metabolites that have modified signaling capabilities. The category and magnitude of bile salt metabolism can affect the host either positively or negatively. A key modification is bile salt hydrolysis, which is a prerequisite step for all additional microbial modification. Here, we have synthesized five different fluorogenic bile salts that report on hydrolysis activity using a simple, continuous assay. We profile activity in both murine and human fecal samples. Our data demonstrate that most gut microbiomes have the highest capacity for hydrolysis of host-produced bile salts, but some microbially modified bile salts also display significant turnover.

59 BASIC BIOLOGICAL SCIENCES↗

Structural Analysis of N- and O-glycans Using ZIC-HILIC/Dialysis Coupled to NMR Detection

Protein glycosylation, an important and complex post-translational modification (PTM), is involved in various biological processes including the receptor-ligand and cell-cell interaction, and plays a crucial role in many biological functions. However, little is known about the glycan structures of important biological complex samples, and the conventional glycan enrichment strategy (i.e., size-exclusion column [SEC] separation,) prior to nuclear magnetic resonance (NMR) detection is time-consuming and tedious. In this study, we employed SEC, Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC), and ZIC-HILIC coupled with dialysis strategies to enrich the glycopeptides from the pronase E digests of RNase B, followed by NMR analysis of the glycoconjugate. Our results suggest that the ZIC-HILIC enrichment coupled with dialysis is the most efficient, which was thus applied to the analysis of biological complex sample, the pronase E digest of the secreted proteins from the fungi Aspergillus niger. The NMR spectra revealed that the secreted proteins from A. niger contain both N-linked glycans with a high-mannose core and O-linked glycans bearing mannose and glucose with 1->3 and 1->6 linkages. In all, our study provides compelling evidence that ZIC-HILIC separation coupled to dialysis is superior to the commonly used SEC separation to prepare glycopeptides for the downstream NMR analysis, which could greatly facilitate the future NMR-based glycoproteomics research.

A.niger↗

Comprehensive Quantitative Analysis of Ovarian and Breast Cancer Tumor Peptidomes

Aberrant degradation of proteins is associated with many pathological states, including cancers. Mass spectrometric analysis of tumor peptidomes, the intracellular and intercellular products of protein degradation, has the potential to provide biological insights on proteolytic processing in cancer. However, attempts to use the information on these smaller protein degradation products from tumors for biomarker discovery and cancer biology studies have been fairly limited to date, largely due to the lack of effective approaches for robust peptidomics identification and quantification, and the prevalence of confounding factors and biases associated with sample handling and processing. Herein, we have developed an effective and robust analytical platform for comprehensive analyses of tissue peptidomes, which is suitable for high throughput quantitative studies. The reproducibility and coverage of the platform, as well as the suitability of clinical ovarian tumor and patient-derived breast tumor xenograft samples with post-excision delay of up to 60 min before freezing for peptidomics analysis, have been demonstrated. Moreover, our data also show that the peptidomics profiles can effectively separate breast cancer subtypes, reflecting tumor-associated protease activities. Peptidomics complements results obtainable from conventional bottom-up proteomics, and provides insights not readily obtainable from such approaches.

Environmental Molecular Sciences Laboratory↗

Plastic recovery and self-healing in longitudinally twinned SiGe nanowires

This paper reports on plastic recovery and self-healing behavior in longitudinally-twinned and [112] orientated SiGe nanowire (NW) beams when they are subjected to large bending strains. The NW alloys are comprised of lamellar nanotwin platelet(s) sandwiched between two semi-cylindrical twins. The loading curves, which are acquired from atomic force microscope (AFM) based three-point bending tests, reveal the onset of plastic deformation at a characteristic stress threshold, followed by further straining of the NWs. This ductility is attributed to dislocation activity within the semi-cylindrical crystal portions of the NW, which are hypothesized to undergo a combination of elastic and plastic straining. On the other hand, the lamellar nanoplatelets undergo purely elastic stretching. During the unloading process, the release of internal elastic stresses enables dislocation backflow and escape at the surface. As a result, the dislocations are predominantly annihilated and the NW samples evidenced self-healing via plastic recovery even at ultra-large strains, which are estimated using finite-element models at 16.3% in one of the tested devices. Finite element analysis also establishes the independence of the observed nanomechanical behavior on the relative orientation of the load with respect to the nanoplatelet. This first observation of reversible plasticity in the SiGe material system, which is aided by a concurrent evolution of segmented elastic and plastic deformation within its grains during the loading process, presents an important new pathway for mechanical stabilization of technologically important group-IV semiconductor nanomaterials.

36 MATERIALS SCIENCE↗