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At least 703 records · Page 39

Identification of Inhibitors of the Association of ZAP-70 with the T Cell Receptor by High-Throughput Screen

ZAP-70 is a critical molecule in the transduction of T cell antigen receptor signaling and the activation of T cells. Upon activation of the T cell antigen receptor, ZAP-70 is recruited to the intracellular ζ-chains of the T cell receptor, where ZAP-70 is activated and colocalized with its substrates. Inhibitors of ZAP-70 could potentially function as treatments for autoimmune diseases or organ transplantation. In this work, we present the design, optimization, and implementation of a screen for inhibitors that would disrupt the interaction between ZAP-70 and the T cell antigen receptor. Finally, the screen is based on a fluorescence polarization assay for peptide binding to ZAP-70.

60 APPLIED LIFE SCIENCES↗

Rare key functional domain missense substitutions in MRE11A, RAD50 , and NBN contribute to breast cancer susceptibility: results from a Breast Cancer Family Registry case-control mutation-screening study

The MRE11A-RAD50-Nibrin (MRN) complex plays several critical roles related to repair of DNA double-strand breaks. Inherited mutations in the three components predispose to genetic instability disorders and the MRN genes have been implicated in breast cancer susceptibility, but the underlying data are not entirely convincing. Here, we address two related questions: (1) are some rare MRN variants intermediate-risk breast cancer susceptibility alleles, and if so (2) do the MRN genes follow a BRCA1/BRCA2 pattern wherein most susceptibility alleles are protein-truncating variants, or do they follow an ATM/CHEK2 pattern wherein half or more of the susceptibility alleles are missense substitutions? Using high-resolution melt curve analysis followed by Sanger sequencing, we mutation screened the coding exons and proximal splice junction regions of the MRN genes in 1,313 early-onset breast cancer cases and 1,123 population controls. Rare variants in the three genes were pooled using bioinformatics methods similar to those previously applied to ATM, BRCA1, BRCA2, and CHEK2, and then assessed by logistic regression. Results: Re-analysis of our ATM, BRCA1, and BRCA2 mutation screening data revealed that these genes do not harbor pathogenic alleles (other than modest-risk SNPs) with minor allele frequencies >0.1% in Caucasian Americans, African Americans, or East Asians. Limiting our MRN analyses to variants with allele frequencies of <0.1% and combining protein-truncating variants, likely spliceogenic variants, and key functional domain rare missense substitutions, we found significant evidence that the MRN genes are indeed intermediate-risk breast cancer susceptibility genes (odds ratio (OR) = 2.88, P = 0.0090). Key domain missense substitutions were more frequent than the truncating variants (24 versus 12 observations) and conferred a slightly higher OR (3.07 versus 2.61) with a lower P value (0.029 versus 0.14). These data establish that MRE11A, RAD50, and NBN are intermediate-risk breast cancer susceptibility genes. Like ATM and CHEK2, their spectrum of pathogenic variants includes a relatively high proportion of missense substitutions. However, the data neither establish whether variants in each of the three genes are best evaluated under the same analysis model nor achieve clinically actionable classification of individual variants observed in this study.

59 BASIC BIOLOGICAL SCIENCES↗

Synergistic effects of Al, Ga, and In doping on ZnO nanorod arrays grown via citrate-assisted hydrothermal technique for highly efficient and fast scintillator screens

To be used as efficient alpha particle scintillator in the fields of nuclear security, nuclear nonproliferation and high-energy physics, scintillator screens must have high light output and fast decay properties. While there has been a great deal of progress in scintillation efficiency, achieving fast decay time properties are still a challenge. In this work, the near band edge (NBE) UV luminescence and alpha particle induced scintillation properties of vertically aligned densely packed ZnO nanorods (NRs) doped with Al, Ga, and In have been thoroughly investigated. The high crystalline hexagonal wurtzite structure with a strong orientation through the c -axis plane (002) and aspect ratios in the range 13–22 have been observed for all ZnO NRs. Electron paramagnetic resonance (EPR) analysis exhibited paramagnetic signals at g ≈ 1.96 for all ZnO NRs. A cost effective green hydrothermal synthesis technique was employed to grow well-aligned NRs. Using citrate as an additive acting as a strong reducing agent in the solution during the crystal growth, defects on the surface are significantly suppressed, thereby enhancing the NBE UV emission. Significantly higher NBE UV emission was observed from the top surface of ZnO NRs in cathodoluminescence (CL) microscopy. Results show that citrate assisted donor doping of ZnO NRs not only reduces the defect emission and NBE self-absorption, but also induces fast decay time (~ 600–700 ps), which makes ZnO NRs a good candidate for fast alpha particle scintillator screens used in associated particle imaging for time and direction tagging of individual neutrons generated in D–T and D–D neutron generators.

36 MATERIALS SCIENCE↗

Development of a reverse genetics system for Sosuga virus allows rapid screening of antiviral compounds

Sosuga virus (SOSV) is a recently discovered zoonotic paramyxovirus isolated from a single human case in 2012; it has been ecologically and epidemiologically associated with transmission by the Egyptian rousette bat (Rousettus aegyptiacus). Bats have long been recognized as sources of novel zoonotic pathogens, including highly lethal paramyxoviruses like Nipah virus (NiV) and Hendra virus (HeV). The ability of SOSV to cause severe human disease supports the need for studies on SOSV pathogenesis to better understand the potential impact of this virus and to identify effective treatments. Here we describe a reverse genetics system for SOSV comprising a minigenome-based assay and a replication-competent infectious recombinant reporter SOSV that expresses the fluorescent protein ZsGreen1 in infected cells. First, we used the minigenome assay to rapidly screen for compounds inhibiting SOSV replication at biosafety level 2 (BSL-2). The antiviral activity of candidate compounds was then tested against authentic viral replication using the reporter SOSV at BSL-3. We identified several compounds with anti-SOSV activity, several of which also inhibit NiV and HeV. Alongside its utility in screening for potential SOSV therapeutics, the reverse genetics system described here is a powerful tool for analyzing mechanisms of SOSV pathogenesis, which will facilitate our understanding of how to combat the potential public health threats posed by emerging bat-borne paramyxoviruses.

59 BASIC BIOLOGICAL SCIENCES↗

BioSig3D: High Content Screening of Three-Dimensional Cell Culture Models

BioSig3D is a computational platform for high-content screening of three-dimensional (3D) cell culture models that are imaged in full 3D volume. It provides an end-to-end solution for designing high content screening assays, based on colony organization that is derived from segmentation of nuclei in each colony. BioSig3D also enables visualization of raw and processed 3D volumetric data for quality control, and integrates advanced bioinformatics analysis. The system consists of multiple computational and annotation modules that are coupled together with a strong use of controlled vocabularies to reduce ambiguities between different users. It is a web-based system that allows users to: design an experiment by defining experimental variables, upload a large set of volumetric images into the system, analyze and visualize the dataset, and either display computed indices as a heatmap, or phenotypic subtypes for heterogeneity analysis, or download computed indices for statistical analysis or integrative biology. BioSig3D has been used to profile baseline colony formations with two experiments: (i) morphogenesis of a panel of human mammary epithelial cell lines (HMEC), and (ii) heterogeneity in colony formation using an immortalized non-transformed cell line. These experiments reveal intrinsic growth properties of well-characterized cell lines that are routinely used for biological studies. BioSig3D is being released with seed datasets and video-based documentation.

Science & Technology - Other Topics↗

An effective “three-in-one” screening assay for testing drug and nanoparticle toxicity in human endothelial cells

Evaluating nanoparticle (NP) toxicity in human cell systems is a fundamental requirement for future NP biomedical applications. In this study, we have designed a screening assay for assessing different types of cell death induced by NPs in human umbilical vein endothelial cell (HUVEC) culture. This assay consists of WST-8, LDH and Hoechst 33342 staining, all performed in one well, which enables an evaluation of cell viability, necrosis and apoptosis, respectively, in the same cell sample. The 96-well format and automated processing of fluorescent images enhances the assay rapidity and reproducibility. After testing the assay functionality with agents that induced different types of cell death, we investigated the endothelial toxicity of superparamagnetic iron oxide nanoparticles (SPIONs, 8 nm), silica nanoparticles (SiNPs, 7–14 nm) and carboxylated multiwall carbon nanotubes (CNTCOOHs, 60 nm). Our results indicated that all the tested NP types induced decreases in cell viability after 24 hours at a concentration of 100 μg/ml. SPIONs caused the lowest toxicity in HUVECs. By contrast, SiNPs induced pronounced necrosis and apoptosis. A time course experiment showed the gradual toxic effect of all the tested NPs. CNTCOOHs inhibited tetrazolium derivatives at 100 μg/ml, causing false negative results from the WST-8 and LDH assay. In summary, our data demonstrate that the presented “three-in-one” screening assay is capable of evaluating NP toxicity effectively and reliably. Due to its simultaneous utilization of two different methods to assess cell viability, this assay is also capable of revealing, if NPs interfere with tetrazolium salts.

59 BASIC BIOLOGICAL SCIENCES↗

A miniaturized feedstocks-to-fuels pipeline for screening the efficiency of deconstruction and microbial conversion of lignocellulosic biomass

Sustainably grown biomass is a promising alternative to produce fuels and chemicals and reduce the dependency on fossil energy sources. However, the efficient conversion of lignocellulosic biomass into biofuels and bioproducts often requires extensive testing of components and reaction conditions used in the pretreatment, saccharification, and bioconversion steps. This restriction can result in a significant and unwieldy number of combinations of biomass types, solvents, microbial strains, and operational parameters that need to be characterized, turning these efforts into a daunting and time-consuming task. Here we developed a high-throughput feedstocks-to-fuels screening platform to address these challenges. The result is a miniaturized semi-automated platform that leverages the capabilities of a solid handling robot, a liquid handling robot, analytical instruments, and a centralized data repository, adapted to operate as an ionic-liquid-based biomass conversion pipeline. The pipeline was tested by using sorghum as feedstock, the biocompatible ionic liquid cholinium phosphate as pretreatment solvent, a “one-pot” process configuration that does not require ionic liquid removal after pretreatment, and an engineered strain of the yeast Rhodosporidium toruloides that produces the jet-fuel precursor bisabolene as a conversion microbe. By the simultaneous processing of 48 samples, we show that this configuration and reaction conditions result in sugar yields (~70%) and bisabolene titers (~1500 mg/L) that are comparable to the efficiencies observed at larger scales but require only a fraction of the time. We expect that this Feedstocks-to-Fuels pipeline will become an effective tool to screen thousands of bioenergy crop and feedstock samples and assist process optimization efforts and the development of predictive deconstruction approaches.

09 BIOMASS FUELS↗

Full Moment Tensor Solutions of U.S. Underground Nuclear Tests for Event Screening and Yield Estimation

Moment tensor (MT) solutions are proving increasingly valuable in explosion monitoring, especially now that they are more routinely calculated for the unconstrained, full (six component) MT. In this study, we have calculated MTs for U.S. underground nuclear tests conducted at the Nevada National Security Site using seismic recordings primarily from the Livermore Nevada Network. We are able to determine them for 130 nuclear explosions from 1970 to 1996 for a range of yields and under a variety of material conditions, which we have supplemented with 10 additional chemical explosions at the test site. The result is an extensive database of MTs that can be used to assess the performance of important monitoring tasks such as event identification and yield determination. We test the explosion event screening on the fundamental lune of the MT eigensphere and find MT screening to be a robust discriminant between earthquakes and explosions. We then explore the estimation of moment-derived yield, in which we find that material properties are the largest contributor to differences in the estimated moment-to-yield ratio. Additionally, further research conducted on this dataset can be used to develop, test, and improve various explosion monitoring methodologies.

58 GEOSCIENCES↗

Second Line of Defense Program, Secondary Screening Operations Field Data Collection

Secondary screening is a key component of the Second Line of Defense (SLD) Program’s capability to deter, detect, and interdict the illicit trafficking of radioactive materials. The primary detection system—the radiation portal monitor—makes alarm decisions based on gross counts of gamma and neutron radiation; it is able to provide little other information regarding what may have caused an elevated radiation level. Conducting a proper secondary screening with an instrument capable of identifying radioisotopes is the only way to determine what radioactive material is present—most importantly if it is a threat material. Only by knowing the radioactive material can a prudent decision be made with regard to disposition and response—and can the mistaken release of a threat be avoided.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Letter Report: LAW Simulant Development for Cast Stone Screening Test

More than 56 million gallons of radioactive and hazardous waste are stored in 177 underground storage tanks at the U.S. Department of Energy’s (DOE’s) Hanford Site in southeastern Washington State. The Hanford Tank Waste Treatment and Immobilization Plant (WTP) is being constructed to treat the wastes and immobilize them in a glass waste form. The WTP includes a pretreatment facility to separate the wastes into a small volume of high-level waste (HLW) containing most of the radioactivity and a larger volume of low-activity waste (LAW) containing most of the nonradioactive chemicals. The HLW will be converted to glass in the HLW vitrification facility for ultimate disposal at an offsite federal repository. At least a portion (~35%) of the LAW will be converted to glass in the LAW vitrification facility and will be disposed of onsite at the Integrated Disposal Facility (IDF). The pretreatment and HLW vitrification facilities will have the capacity to treat and immobilize the wastes destined for each facility. However, a second facility will be needed for the expected volume of additional LAW requiring immobilization. A cementitious waste form known as Cast Stone is being considered to provide the required additional LAW immobilization capacity. The Cast Stone waste form must be acceptable for disposal in the IDF. The Cast Stone waste form and immobilization process must be tested to demonstrate that the final Cast Stone waste form can comply with waste acceptance criteria for the IDF disposal facility and that the immobilization processes can be controlled to consistently provide an acceptable waste form product. Further, the waste form must be tested to provide the technical basis for understanding the long term performance of the waste form in the IDF disposal environment. These waste form performance data are needed to support risk assessment and performance assessment (PA) analyses of the long-term environmental impact of the waste disposal in the IDF. A testing program was developed in fiscal year (FY) 2012 describing in some detail the work needed to develop and qualify Cast Stone as a waste form for the solidification of Hanford LAW (Westsik et al. 2012). Included within Westsik et al. (2012) is a section on the near-term needs to address Tri-Party Agreement Milestone M-062-40ZZ. The objectives of the testing program to be conducted in FY 2013 and FY 2014 are to: • Determine an acceptable formulation for the LAW Cast Stone waste form. • Evaluate sources of dry materials for preparing the LAW Cast Stone. • Demonstrate the robustness of the Cast Stone waste form for a range of LAW compositions. • Demonstrate the robustness of the formulation for variability in the Cast Stone process. • Provide Cast Stone contaminant release data for PA and risk assessment evaluations. The first step in determining an acceptable formulation for the LAW Cast Stone waste form is to conduct screening tests to examine expected ranges in pretreated LAW composition, waste stream concentrations, dry-materials sources, and mix ratios of waste feed to dry blend. A statistically designed test matrix will be used to evaluate the effects of these key parameters on the properties of the Cast Stone as it is initially prepared and after curing. The second phase of testing will focus on selection of a baseline Cast Stone formulation for LAW and demonstrating that Cast Stone can meet expected waste form requirements for disposal in the IDF. It is expected that this testing will use the results of the screening tests to define a smaller suite of tests to refine the composition of the baseline Cast Stone formulation (e.g. waste concentration, water to dry mix ratio, waste loading).

Russell, Renee L.↗

Technology recommendations for pre-screening of IAEA swipe samples

Argonne and Oak Ridge National Laboratories have prepared an analysis of recommended, possible, and not recommended technologies for pre-screening and prioritizing IAEA swipes. The analytical techniques listed under the recommended technology list are the most promising techniques available to date. The recommended list is divided into two sections: Argonne’s recommended techniques and Oak Ridge’s recommended techniques. This list was divided based upon the expertise of staff in each subject area and/or the instrumentation available at each laboratory. The following section, titled Possible Techniques, is a list of analytical techniques that could be used for pre-screening and prioritizing swipes if additional instrumentation and effort were provided. These techniques are not necessarily top priority, but should not be discounted for future or expanded efforts. Lastly, a list of not recommended techniques is provided to outline the analytical methods and instrumentation that were investigated by each lab but deemed not suitable for this task. In addition to the recommendation list, a short procedure is provided outlining the steps followed for destructive analysis by the Network of Analytical Laboratories (NWAL) for determination of uranium concentrations, isotopic content of sample and swipe. Swipes generated for this project will be given to ORNL’s NWAL laboratory for analysis after analysis by other techniques at both laboratories.

98 NUCLEAR DISARMAMENT, SAFEGUARDS, AND PHYSICAL P↗

Can YAG screen accept LEReC bunch train?

LEReC RF diagnostic beamline is supposed to accept 250 us long pulse trains of 1.6 MeV – 2.6 MeV (kinetic energy) electrons. This beamline is equipped with YAG profile monitor. Since we are interested in observing only the last macro bunch in the train, one of the possibilities is to install a fast kicker and a dedicated dump upstream of the YAG screen (and related diagnostics equipment). This approach is expensive and challenging from engineering point of view. Another possibility is to send the whole pulse train to the YAG screen and to use a fast gated camera (such as Imperex B0610 with trigger jitter under 60ns) to observe the image from the last pulse only. In this paper we study the feasibility of the last approach.

43 PARTICLE ACCELERATORS↗

Large Scale Screening of Low Cost Ferritic Steel Designs For Advanced Ultra Supercritical Boiler Using First Principles Methods

Advanced Ultra Supercritical Boiler (AUSC) requires materials that can operate in corrosive environment at temperature and pressure as high as 760°C (or 1400°F) and 5000psi, respectively, while at the same time maintain good ductility at low temperature. We develop automated simulation software tools to enable fast large scale screening studies of candidate designs. While direct evaluation of creep rupture strength and ductility are currently not feasible, properties such as energy, elastic constants, surface energy, interface energy, and stack fault energy can be used to assess their relative ductility and creeping strength. We implemented software to automate the complex calculations to minimize human inputs in the tedious screening studies which involve model structures generation, settings for first principles calculations, results analysis and reporting. The software developed in the project and library of computed mechanical properties of phases found in ferritic steels, many are complex solid solutions estimated for the first time, will certainly help the development of low cost ferritic steel for AUSC.

36 MATERIALS SCIENCE↗

Optical Encoding Technology for Viral Screening Panels Final Report CRADA No TC02132.0

This was a collaborative effort between Lawrence Livermore National Security, LLC, Lawrence Livermore National Laboratory (LLNL) and Parallel Synthesis Technologies, Inc. (PSTI), to develop Optical Encoding Technology for Viral Screening Panels. The goal for this effort was to prepare a portable bead reader system that would enable the development of viral and bacterial screening panels which could be used for the detection of any desired set of bacteria or viruses in any location. The main objective was to determine if the combination of a bead-based, PCR suspension array technology, formulated from Parallume encoded beads and PSTI’s multiplex assay reader system (MARS), could provide advantages in terms of the number of simultaneously measured samples, portability, ruggedness, ease of use, accuracy, precision or cost as compared to the Luminexbased system developed at LLNL. The project underwent several no cost extensions however the overall goal of demonstrating the utility of this new system was achieved. As a result of the project a significant change to the type of bead PSTI used for the suspension system was implemented allowing better performance than the commercial Luminex system.

59 BASIC BIOLOGICAL SCIENCES↗

NDE Technology Development Program for Non-Visual Volumetric Inspection Technology: Technology Screening Report

The Pacific Northwest National Laboratory (PNNL) located in Richland, Washington, hosted and administered a Technology Screening test that allowed four different participants to demonstrate the NDE volumetric inspection technologies that were proposed in response to the WRPS EOI. This document provides a Technology Screening test report for the initial part of Phase I of a three-phase NDE Technology Development Program designed to identify and mature a system or set of non-visual volumetric NDE technologies for Hanford DST primary liner bottom inspection. Phase I of the program will baseline the performance of current or emerging non-visual volumetric NDE technologies for their ability to detect and characterize primary liner bottom flaws, and identify candidate technologies for adaptation and maturation for Phase II of the program.

42 ENGINEERING↗

Performance of BaFCl:Eu2+ scintillating composites for X-ray imaging screens

A cost-effective solution for improving X-ray imaging detectors is presented through the use of a translucent scintillating composite and pixelated screen structure. A large quantity of BaFCl:Eu 2+ polycrystalline scintillator powder was synthesized via flux growth and its optical properties characterized through photoluminescence (PL) and absolute quantum yield (QY). UV-curable composites containing the BaFCl:Eu 2+ material were fabricated with differing amounts of NOA1665 and NOA170 optical adhesives. It was determined that, when the two resins were combined, the translucency of the resulting composite could be enhanced. The BaFCl:Eu 2+ composite was also cast into a pixelated glass screen containing an array of 160 μm x 160 μm holes by two separate methods: composite paste application and powder-resin application. Based on visual inspection, using a composite paste to fill the apertures proved to be more effective in creating a uniform packing.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Washington State Highway Seismic Screening Tool (HSST)

The U.S. Department of Homeland Security’s Regional Resiliency Assessment Program (RRAP) undertook the Washington State Transportation Systems RRAP project in 2017 to assess the seismic vulnerabilities of that state’s transportation system to a Cascadia Subduction Zone (CSZ) earthquake. The specific goal of that project was to assess the vulnerability of, and prioritize, transportation infrastructure for moving goods and supplies from Incident Support Bases to Federal Staging Areas within Washington State following a CSZ earthquake. As part of this analysis, Argonne National Laboratory (Argonne), with input and support from the Washington State Department of Transportation (WSDOT) maintenance and pavements offices and state geotechnical engineer, developed the Highway Seismic Screening Tool (HSST) to inform a system-level analysis of highway recovery and reopening times. This report provides a full discussion of the HSST’s development, implementation, and results for the CSZ scenario earthquake. The primary outcome of the HSST is to determine the approximate reopening times of highway segments or bridges damaged by a CSZ magnitude 9.0 seismic event. This is done by calculating the permanent ground deformation (PGD) caused by soil liquefaction along state highways, which can cause significant damage or displacement of highway pavement structures. The term “reopening time” is different from “restoration time” and is intended to represent the approximate amount of time that would be needed to return highways to a minimum state of repair sufficient to support the limited movement of emergency and response vehicles. Additionally, the HSST is referred to as a “screening tool,” as it uses network- or system-level asset management information that WSDOT provided to assess seismic-related hazards and vulnerabilities of highways; it does not conduct an asset-level engineering analysis of seismic-related vulnerabilities. Accordingly, the results of the HSST analysis should only be used for system- or corridor-level planning and investment, and not for engineering or management decisions at the asset level. The HSST is also applied to assess the exposure of statewide rail systems to soil liquefaction and PGD hazards. However, little information is available about the sensitivity or vulnerability of railways and rail yards to such hazards, and therefore researchers were unable to determine approximate reopening times for the state’s rail system following a CSZ earthquake.

58 GEOSCIENCES↗