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At least 199 records · Page 11
All sourcesNASA NTRSDOE OSTI
DOE OSTI2011

Water Velocity Measurements on a Vertical Barrier Screen at the Bonneville Dam Second Powerhouse

Fish screens at hydroelectric dams help to protect rearing and migrating fish by preventing them from passing through the turbines and directing them towards the bypass channels by providing a sweeping flow parallel to the screen. However, fish screens may actually be harmful to fish if they become impinged on the surface of the screen or become disoriented due to poor flow conditions near the screen. Recent modifications to the vertical barrier screens (VBS) at the Bonneville Dam second powerhouse (B2) intended to increase the guidance of juvenile salmonids into the juvenile bypass system (JBS) have resulted in high mortality and descaling rates of hatchery subyearling Chinook salmon during the 2008 juvenile salmonid passage season. To investigate the potential cause of the high mortality and descaling rates, an in situ water velocity measurement study was conducted using acoustic Doppler velocimeters (ADV) in the gatewell slot at Units 12A and 14A of B2. From the measurements collected the average approach velocity, sweep velocity, and the root mean square (RMS) value of the velocity fluctuations were calculated. The approach velocities measured across the face of the VBS varied but were mostly less than 0.3 m/s. The sweep velocities also showed large variances across the face of the VBS with most measurements being less than 1.5 m/s. This study revealed that the approach velocities exceeded criteria recommended by NOAA Fisheries and Washington State Department of Fish and Wildlife intended to improve fish passage conditions.

13 HYDRO ENERGY↗
DOE OSTI2016

A Water-Based Silver-Nanowire Screen-Print Ink for the Fabrication of Stretchable Conductors and Wearable Thin-Film Transistors

Silver nanowire is a very promising material for fabricating compliant conductors which are essential for stretchable/wearable electronic devices. Screen printing is a cost-effective and scalable technology to fabricate large-area thin film coatings with modest pattern resolution. The biggest challenge to prepare a screen printable silver nanowire ink stems from the low viscosity of silver nanowire dispersions and that the addition of a thickening agent could dramatically increase the inter-nanowire contact resistance in the resulting coating. Herein, we report the synthesis of a water-based silver nanowire ink, which was formulated with low solid contents, high viscosity at 0.1 s -1 shear rate, and appropriate rheological behavior suitable for screen printing. Silver nanowire coating patterns were screen printed with uniform sharp edges, ~50 μm resolution, and electrical conductivity as high as 4.67 × 10 4 S cm -1 . The screen printed silver nanowires were then used to fabricate a composite conductor that retained a conductivity greater than 10,000 S cm -1 under 70% tensile strain. Fully printed and stretchable/wearable thin-film transistor arrays were also fabricated by employing the screen printed composite conductor as the source, drain, and gate, drop cast semiconducting carbon nanotubes as the channel, and a dielectric elastomer. The 10 × 6 thin-film transistor arrays had a fabrication yield of 91.7%, average mobility of 33.8 ± 3.7 cm 2 V -1 s -1 , ON/OFF ratio ~1000, and remained stable during 1,000 cycles of wearing on and peeling off a glass tube with 5 mm diameter.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗
DOE OSTI2022

Performance of borated scintillator screens for high-resolution neutron imaging

The most commonly used screens for neutron imaging consist of 6 LiF + ZnS. This type of screen yields the highest light output per detected neutron. For high resolution, gadolinium oxysulfide (GOS, Gadox) screens are employed, which have a much higher detection efficiency, but a light output so much lower than LiF + ZnS that measurements are often limited by photon statistics. Historically, screens using boron as a neutron-sensitive material have not been very successful. However, a new preparation method was introduced recently that produces light output higher than Gadox with detection efficiency greater than LiF + ZnS. Measurements of these new borated screens were performed at the NeXT facility at ILL, Grenoble, in comparison to a high resolution Gadox screen.

10B↗
DOE OSTI2014

Compensatory mitigation and screening rules in environmental impact assessment

Concerns about the effectiveness of environmental impact assessment (EIA) have prompted proposals to improve its performance by limiting the discretion of decision-makers in screening. To investigate whether such proposals are likely to generate the desired results, we conducted an evaluation of the screening process under the Australian government's EIA regime from its introduction on 16 July 2000 to 30 June 2013 (study period). Almost 1 in 5 ‘particular manner’ decisions—a type of screening decision under the regime—were found to be unlawful. The extent of non-compliance is explained on the basis of convenience. The department was required to assess a large number of projects under tight timeframes and with limited resources, while being pressured by proponents to allow their projects to bypass EIA. These pressures resulted in the development of an informal custom whereby the formal compensatory mitigation restrictions were frequently ignored. The results highlight the relative significance of formal and informal institutions in EIA. Formal EIA rules typically provide a mere outline of the process. The informal institutions adopted by administrators often have a greater influence on how the process operates and what it achieves. - Highlights: • Concerns about the effectiveness of environmental impact assessment (EIA) have prompted proposals to improve its performance by limiting the discretion of decision-makers in screening. • To investigate whether such proposals are likely to generate the desired results, we conducted an evaluation of the Australian government's screening process, looking at the extent of compliance with a formal prohibition on the consideration of compensatory mitigation. • Almost 1 in 5 ‘particular manner’ decisions – a type of screening decision under the regime – were found to be unlawful (with a 95% confidence interval of between 1:4 and 1:7) because of a failure to abide by the compensatory mitigation restrictions. For urban development actions, the ratio between lawful and unlawful particular manner decisions was worse than 1:1. • The extent of non-compliance is explained on the basis of convenience. Budget and interest group pressures resulted in the development of an informal custom whereby the formal compensatory mitigation restrictions were ignored. • The results highlight the relative significance of formal and informal institutions in EIA. Formal EIA rules typically provide a mere outline of the process. The informal institutions adopted by administrators often have a greater influence on how the process operates and what it achieves.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗
DOE OSTI2017

Land take and the effectiveness of project screening in Environmental Impact Assessment: Findings from an empirical study

Land take is emerging as a global environmental concern, and is particularly critical in intensively developed and land-scarce regions. This paper seeks to understand the effectiveness of the screening stage of Environmental Impact Assessment (EIA) in addressing land take. Screening is the stage where a decision is made as to whether an EIA is required for a project. In many jurisdictions, screening results in three pathways: full EIA directly, preliminary EIA only, or preliminary EIA followed by full EIA. We compared the land take of 217 projects triggering the different pathways in a study region in Italy over a 15-year time interval. Land take was quantified by overlaying the footprint of the projects with a land cover map. The results show that while more attention was given to projects with larger land take impacts overall, the cumulative land take from smaller projects not triggering full EIA was considerable (40% of overall land take). The case-by-case examination conducted through the preliminary EIA was found to work better for some project types (ski areas and small urban development), than for others (quarries). Our findings lead us to advocate improvements in current screening procedures to ensure that the land take impacts are quantified and made explicit in preliminary EIA reports. Our evidence-based approach to determining land take in EIA provides a compelling basis for understanding ways to improve EIA policies, guidance and practice. - Highlights: •We quantified the land take of 217 projects triggering different EIA screening pathways. •Projects that “fly under the radar” of full EIA caused 40% of the overall land take. •The case-by-case examination works better for some project types than others. •We recommend improvements in screening procedures to ensure that land take impacts are quantified.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2019

High-throughput screening and chemotype-enrichment analysis of ToxCast phase II chemicals evaluated for human sodium-iodide symporter (NIS) inhibition

In support of the Endocrine Disruptor Screening Program (EDSP), the U.S.EPA's Office of Research and Development (ORD) is developing high-throughput screening (HTS) approaches to identify chemicals that alter target sites in the thyroid hormone (TH) pathway. The sodium iodide symporter (NIS) is a transmembrane glycoprotein that mediates iodide uptake into the thyroid as the initial step of TH biosynthesis. Previously, we screened 293 ToxCast chemicals (ph1v2) using a HEK293T cell line expressing human NIS in parallel radioactive iodide uptake (RAIU) and cell viability assays to identify potential environmental NIS inhibitors. Here, we expanded NIS inhibitor screening for a set of 768 ToxCast Phase II (ph2) chemicals, and applied a novel computational toxicology approach based on the ToxPrint chemotype to identify chemical substructures associated with NIS inhibition. Following single-concentration screening (at 1 × 10 -4 M with a 20% inhibition cutoff), 235 samples (228 chemicals) were further tested in multipleconcentration (1 × 10 -9 - 1 × 10 -4 M) format in both RAIU and cell viability assays. The 167 chemicals that exhibited significant RAIU inhibition were then prioritized using combined RAIU and cell viability responses that were normalized relative to the known NIS inhibitor sodium perchlorate. Some of the highest ranked chemicals, such as PFOS, tributyltin chloride, and triclocarban, have been previously reported to be thyroid disruptors. In addition, several novel chemicals were identified as potent NIS inhibitors. The present results were combined with the previous ph1v2 screening results to produce two sets of binary hit-calls for 1028 unique chemicals, consisting of 273 positives exhibiting significant RAIU inhibition, and 63 positives following application of a cell viability filter. A ToxPrint chemotype-enrichment analysis identified > 20 distinct chemical substructural features, represented in > 60% of the active chemicals, as significantly enriched in each NIS inhibition hit-call space. A shared set of 9 chemotypes enriched in both hit-call sets indicates stable chemotype signals (insensitive to cytotoxicity filters) that can help guide structure-activity relationship (SAR) investigations and inform future research.

54 ENVIRONMENTAL SCIENCES↗
DOE OSTI2017

Berkeley Screen: a set of 96 solutions for general macromolecular crystallization

Using statistical analysis of the Biological Macromolecular Crystallization Database, combined with previous knowledge about crystallization reagents, a crystallization screen called the Berkeley Screen has been created. Correlating crystallization conditions and high-resolution protein structures, it is possible to better understand the influence that a particular solution has on protein crystal formation. Ions and small molecules such as buffers and precipitants used in crystallization experiments were identified in electron density maps, highlighting the role of these chemicals in protein crystal packing. The Berkeley Screen has been extensively used to crystallize target proteins from the Joint BioEnergy Institute and the Collaborative Crystallography program at the Berkeley Center for Structural Biology, contributing to several Protein Data Bank entries and related publications. The Berkeley Screen provides the crystallographic community with an efficient set of solutions for general macromolecular crystallization trials, offering a valuable alternative to the existing commercially available screens. The Berkeley Screen provides an efficient set of solutions for general macromolecular crystallization trials.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2014

Protein crystallization with microseed matrix screening: application to human germline antibody Fabs

The power of microseed matrix screening is demonstrated in the crystallization of a panel of antibody Fab fragments. The crystallization of 16 human antibody Fab fragments constructed from all pairs of four different heavy chains and four different light chains was enabled by employing microseed matrix screening (MMS). In initial screening, diffraction-quality crystals were obtained for only three Fabs, while many Fabs produced hits that required optimization. Application of MMS, using the initial screens and/or refinement screens, resulted in diffraction-quality crystals of these Fabs. Five Fabs that failed to give hits in the initial screen were crystallized by cross-seeding MMS followed by MMS optimization. The crystallization protocols and strategies that resulted in structure determination of all 16 Fabs are presented. These results illustrate the power of MMS and provide a basis for developing future strategies for macromolecular crystallization.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗
DOE OSTI2017

Screening in a multicomponent plasma by the example of a wet air plasma

A general theory of screening of a dust-particle charge in a multicomponent plasma is developed based on the asymptotic screening theory. In the asymptotic theory, the balance equations for the number of charged plasma particles in the diffusion-drift approximation supplemented with point sinks of plasma particles on a dust particle and the Poisson equation for the self-consistent electric field potential are solved by the linearization method using the three-dimensional Fourier transform. The coefficients of a secular or a characteristic polynomial, whose zeroes determine screening constants in a multicomponent plasma, were calculated by the Leverrier–Faddeev method, allowing one to find these coefficients for any number of initial balance equations for the number of charged plasma particles. By the example of a wet air plasma produced by an external gas-ionization source, the procedure is considered for constructing the truncated system of balance equations for main plasma ions and screening constants are determined. The influence of the number of main ions on screening constants is considered and the physical meaning of all screening constants is found.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗
DOE OSTI2020

Design and feasibility of a novel program of cervical screening in Nigeria: self-sampled HPV testing paired with visual triage

Background: Accelerated global control of cervical cancer would require primary prevention with human papillomavirus (HPV) vaccination in addition to novel screening program strategies that are simple, inexpensive, and effective. We present the feasibility and outcome of a community-based HPV self-sampled screening program. Methods: In Ile Ife, Nigeria, 9406 women aged 30–49 years collected vaginal self-samples, which were tested for HPV in the local study laboratory using Hybrid Capture-2 (HC2) (Qiagen). HPV-positive women were referred to the colposcopy clinic. Gynecologist colposcopic impression dictated immediate management; biopsies were taken when definite acetowhitening was present to produce a histopathologic reference standard of precancer (and to determine final clinical management). Retrospective linkage to the medical records identified 442 of 9406 women living with HIV (WLWH). Results: With self-sampling, it was possible to screen more than 100 women per day per clinic. Following an audio-visual presentation and in-person instructions, overall acceptability of self-sampling was very high (81.2% women preferring self-sampling over clinician collection). HPV positivity was found in 17.3% of women. Intensive follow-up contributed to 85.9% attendance at the colposcopy clinic. Of those referred, 8.2% were initially treated with thermal ablation and 5.6% with large loop excision of transformation zone (LLETZ). Full visibility of the squamocolumnar junction, necessary for optimal visual triage and ablation, declined from 68.5% at age 30 to 35.4% at age 49. CIN2+ and CIN3+ (CIN- Cervical intraepithelial neoplasia), including five cancers, were identified by histology in 5.9 and 3.2% of the HPV-positive women, respectively (0.9 and 0.5% of the total screening population), leading to additional treatment as indicated. The prevalences of HPV infection and CIN2+ were substantially higher (40.5 and 2.5%, respectively) among WLWH. Colposcopic impression led to over- and under-treatment compared to the histopathology reference standard. Conclusion: A cervical cancer screening program using self-sampled HPV testing, with colposcopic immediate management of women positive for HPV, proved feasible in Nigeria. Based on the collected specimens and images, we are now evaluating the use of a combination of partial HPV typing and automated visual evaluation (AVE) of cervical images to improve the accuracy of the screening program.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Fast Neutron Scintillator Screens for Neutron Imaging Using a Layered Polymer-Phosphor Architecture

Fast neutrons enable a nondestructive examination of dense, large, and highly attenuating samples due to their lower interaction probability compared to thermal neutrons. However, this also creates a challenge in fast neutron imaging, as the thicker sensors necessary to detect fast neutrons degrade an image’s spatial resolution due to scattering within the sensor and the indeterminate depth of interaction in the sensor. This work explores the advantages of a fast neutron imaging screen with a layered polymer-phosphor screen approach as opposed to a mixed polymer-phosphor screen typically used in fast neutron imaging. Proton recoil is the primary conversion mechanism for fast neutron imaging. Simulations showed that the recoil proton range of typical fast neutrons is approximately 200 µm, however, tests at Idaho National Laboratory revealed that the light output of these screens increased at much greater polymer thicknesses. The NECTAR fast neutron beamline at FRM II was used to test the imaging performance of layered fast neutron imaging screens. Distinguishing between the fast-neutron and γ-ray signals is a major challenge in fast neutron imaging because all fast neutron sources also produce γ-rays. A relative comparison between a control plate and the fast neutron screen was made to distinguish between a γ-ray and fast neutron signals. MCNP modeling quantified the γ-ray and fast neutron contributions to the images measured at NECTAR, which were approximately a 75% γ-ray image.

73 NUCLEAR PHYSICS AND RADIATION PHYSICS↗
DOE OSTI2022

Improved high-throughput screening technique to rapidly isolate Chlamydomonas transformants expressing recombinant proteins

Abstract The single-celled eukaryotic green alga Chlamydomonas reinhardtii has long been a model system for developing genetic tools for algae, and is also considered a potential platform for the production of high-value recombinant proteins. Identifying transformants with high levels of recombinant protein expression has been a challenge in this organism, as random integration of transgenes into the nuclear genome leads to low frequency of cell lines with high gene expression. Here, we describe the design of an optimized vector for the expression of recombinant proteins in Chlamydomonas , that when transformed and screened using a dual antibiotic selection, followed by screening using fluorescence activated cell sorting (FACS), permits rapid identification and isolation of microalgal transformants with high expression of a recombinant protein. This process greatly reduces the time required for the screening process, and can produce large populations of recombinant algae transformants with between 60 and 100% of cells producing the recombinant protein of interest, in as little as 3 weeks, that can then be used for whole population sequencing or individual clone analysis. Utilizing this new vector and high-throughput screening (HTS) process resulted in an order of magnitude improvement over existing methods, which normally produced under 1% of algae transformants expressing the protein of interest. This process can be applied to other algal strains and recombinant proteins to enhance screening efficiency, thereby speeding up the discovery and development of algal-derived recombinant protein products. Key points • A protein expression vector using double-antibiotic resistance genes was designed • Double antibiotic selection causes fewer colonies with more positive for phenotype • Coupling the new vector with FACS improves microalgal screening efficiency > 60%

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2019

Comparison of extracolonic findings and clinical outcomes in a screening and diagnostic CT colonography population

Purpose: To compare the distribution of extracolonic findings and clinical outcomes between screening and diagnostic CT colonography (CTC) populations. Methods: 388 consecutive patients (369 men, 19 women; mean ± SD age 67.8 ± 10 years) who underwent first-time CTC (4/2011–4/2017) at a Veteran’s Affairs Medical Center were divided into screening (asymptomatic) or diagnostic (symptomatic) cohorts based on CTC indication. CTC reporting and data system E-scores for extracolonic findings were retrospectively assigned based on prospective CTC radiologic reports. Multinomial logistic regression was used to examine the association between E-scores and CTC indication. Electronic medical records of all patients with E3 or E4 scores were reviewed (median follow-up 2.8 years) to determine clinical outcomes. Results: 68% (262/388) underwent screening and 32% (126/388) diagnostic CTC. 7.2% (28/388) had extracolonic findings considered potentially significant (E4), 4.4% (17/388) had indeterminate but likely unimportant findings (E3), and 88.4% (347/388) had normal or unimportant findings (E1 or E2). E-scores were not significantly different between screening and diagnostic CTC when adjusted for age, gender, and prior imaging (p = 0.44). 4.6% (12/262) of patients with E3/E4 findings in the screening cohort demonstrated clinically significant outcomes, compared with 4.0% (5/126) in the diagnostic cohort, including a total of three extracolonic malignancies (0.8%) and three abdominal aortic aneurysms (0.8%). 4.6% (18/388) underwent follow-up imaging studies to confirm a benign outcome after detection of a category E3/E4 finding. Conclusions: The distribution of extracolonic findings and clinical outcomes were not statistically significantly different between screening and diagnostic CTC populations.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2024

Aviation security screening optimizer for risk and throughput (ASSORT)

The increasing number of air travelers each year presents a challenge as many airports are near their capacity in terms of resources and space for passenger screening. Fortunately, advancements in technologies like next-generation millimeter wave scanning offer solutions to ease this strain. The focus remains on managing risk while enhancing the passenger experience for the traveling public. The risk model presented in this paper known as the Aviation Security Screening Optimizer for Risk and Throughput (ASSORT) is designed to assess risk-based approaches for passenger screening and checkpoint operations. Additionally, ASSORT is exploring various traveler categories — general, trusted, and trusted-plus — along with different checkpoint screening Concept of Operations tailored to each traveler type. For instance, travelers with a higher trust level may experience fewer screening technologies, resulting in quicker processing times at the checkpoint. The output of ASSORT provides a risk score for predefined threat scenarios, as well as the overall risk to the checkpoint, aircraft, and airport by traveler type. In conclusion, benefits of using this tool include assessing the trade-offs between the overall risk associated with checkpoints and the throughput rate of passengers screened. We show for example the impact that different passenger volumes at the checkpoint can have on risk.

99 GENERAL AND MISCELLANEOUS↗
DOE OSTI2014

3D high-content screening for the identification of compounds that target cells in dormant tumor spheroid regions

Cancer cells in poorly vascularized tumor regions need to adapt to an unfavorable metabolic microenvironment. As distance from supplying blood vessels increases, oxygen and nutrient concentrations decrease and cancer cells react by stopping cell cycle progression and becoming dormant. As cytostatic drugs mainly target proliferating cells, cancer cell dormancy is considered as a major resistance mechanism to this class of anti-cancer drugs. Therefore, substances that target cancer cells in poorly vascularized tumor regions have the potential to enhance cytostatic-based chemotherapy of solid tumors. With three-dimensional growth conditions, multicellular tumor spheroids (MCTS) reproduce several parameters of the tumor microenvironment, including oxygen and nutrient gradients as well as the development of dormant tumor regions. We here report the setup of a 3D cell culture compatible high-content screening system and the identification of nine substances from two commercially available drug libraries that specifically target cells in inner MCTS core regions, while cells in outer MCTS regions or in 2D cell culture remain unaffected. We elucidated the mode of action of the identified compounds as inhibitors of the respiratory chain and show that induction of cell death in inner MCTS core regions critically depends on extracellular glucose concentrations. Finally, combinational treatment with cytostatics showed increased induction of cell death in MCTS. The data presented here shows for the first time a high-content based screening setup on 3D tumor spheroids for the identification of substances that specifically induce cell death in inner tumor spheroid core regions. This validates the approach to use 3D cell culture screening systems to identify substances that would not be detectable by 2D based screening in otherwise similar culture conditions. - Highlights: • Establishment of a novel method for 3D cell culture based high-content screening. • First reported high-content screen for the identification of compounds that target cells in dormant tumor spheroid regions. • Metabolite profiling by NMR spectroscopy on 3D tumor spheroids. • Identification of respiratory chain inhibitors to specifically induce cell death in inner tumor spheroid regions. • Respiratory chain inhibitors enhance cytostatic based therapy in vitro.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2019

Energy-Screened Many-Body Expansion: A Practical Yet Accurate Fragmentation Method for Quantum Chemistry

We introduce an implementation of the truncated many-body expansion, MBE(n), in which the n-body corrections are screened using the effective fragment potential force field, and only those that exceed a specified energy threshold are computed at a quantum-mechanical level of theory. This energy-screened MBE(n) approach is tested at the n = 3 level for a sequence of water clusters, (H2O) N=6–34 . A threshold of 0.25 kJ/mol eliminates more than 80% of the subsystem electronic structure calculations and is even more efficacious in that respect than is distance-based screening. Even so, the energy-screened MBE(3) method is faithful to a full-system quantum chemistry calculation to within 1–2 kJ/mol/monomer, even in good quality basis sets such as aug-cc-pVTZ. These errors can be reduced by means of a two-layer approach that involves a Hartree–Fock calculation for the entire cluster. Such a correction proves to be necessary in order to obtain accurate relative energies for conformational isomers of (H2O)20, but the cost of a full-system Hartree–Fock calculation remains smaller than the cost of three-body subsystem calculations at correlated levels of theory. At the level of second-order Møller–Plesset perturbation theory (MP2), a screened MBE(3) calculation plus a full-system Hartree–Fock calculation is less expensive than a full-system MP2 calculation starting at N = 12 water molecules. This is true even if all MBE(3) subsystem calculations are performed on a single 40-core compute node, i.e., without significant parallelization. Energy-screened MBE(n) thus provides a fragment-based method that is accurate, stable in large basis sets, and low in cost, even when the latter is measured in aggregate computer time.

74 ATOMIC AND MOLECULAR PHYSICS↗
DOE OSTI2024

Pivotal Roles of Triple Screening-Topological, Electrostatic, and Hydrodynamic-On Dynamics in Semidilute Polyelectrolyte Solutions

For semidilute polyelectrolyte solutions, it is generally assumed that topological, electrostatic, and hydrodynamic interactions are screened (called triple screening). Despite a large body of research focused on polyelectrolyte solutions, the concept of triple screening has never been rigorously verified. In this work, we test the concept by probing concentration fluctuations in aqueous solutions containing a well-studied polyelectrolyte, sodium poly(styrenesulfonate) (NaPSS) with neutron scattering, theory, and molecular dynamics simulations. Neutron spin–echo (NSE) and small-angle neutron scattering (SANS) data from semidilute solutions of NaPSS are presented at different polymer and salt (NaCl) concentrations. A combined theory for structure (J. Chem. Phys. 105, 5183 (1996)) and dynamics (J. Chem. Phys. 107, 2619 (1997)), which captures effects of hydrodynamic, topological, and electrostatic screening, is used to interpret the experimental results. Here, the theory quantitatively predicts the decay rate obtained from the NSE measurements while capturing the shape and concentration dependencies of the polyelectrolyte peak observed in the SANS spectra. Detailed comparisons of the theory and the experiments reveal that the wavevector-dependent decay rate of concentration fluctuations in semidilute solutions of polyelectrolytes is dictated by the screening of hydrodynamic, topological, and electrostatic interactions. This conclusion is corroborated by coarse-grained molecular dynamics simulations, executed without any hydrodynamic interactions, which fail to capture the correct wavevector dependence of the decay rate. These results highlight that the theories based on the concept of triple screening provide a quantitative framework for predicting a relation between the structure and dynamics of polyelectrolyte solutions.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2021

Click-Chemistry-Based Free Azide versus Azido Sugar Detection Enables Rapid In Vivo Screening of Glycosynthase Activity

Engineering of carbohydrate-active enzymes such as glycosynthases to enable chemoenzymatic synthesis of bespoke oligosaccharides has been limited by the lack of suitable ultrahigh-throughput screening methods capable of robustly detecting either starting substrates or end-products of the glycosidic bond formation reaction. Currently, there are limited screening methods available for rapid and highly sensitive single-cell-based screening of glycosynthase enzymes employing azido sugars as activated donor glycosyl substrates. Here, we report a fluorescence-based approach employing click-chemistry for the selective detection of glycosyl azides as substrates versus free inorganic azides as reaction products that facilitated an ultrahigh-throughput in vivo single-cell-based assay of glycosynthase activity. This assay was developed based on the distinct differences observed in relative fluorescence intensity of the triazole-containing fluorophore product formed during the click-chemistry reaction of organic glycosyl azides versus inorganic azides. This discovery formed the basis for proof of concept validation of a directed evolution methodology for screening and sorting glycosynthase mutants capable of synthesis of targeted fucosylated oligosaccharides. Our screening approach facilitated fluorescence-activated cell sorting of an error-prone polymerase chain reaction-based mutant library of fucosynthases expressed in Escherichia coli to identify several novel mutants that showed increased activity for ß-fucosyl azide-activated donor sugars toward desired acceptor sugars (e.g., pNP-xylose and lactose). Finally, we discuss avenues for improving this proof of concept in vivo assay method to identify better glycosynthase mutants and further demonstrate the broader applicability of this screening methodology for synthesis of bespoke glycans.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗