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At least 883 records · Page 49

Evaluation of a high-throughput method for processing sponge-stick samples to detect viable, non-spore-forming biothreat agents

After a bioterrorism incident, surface sampling is often used to determine the extent of contamination and exposure, guiding decontamination efforts and decisions for re-occupancy of affected sites. The sponge-stick (SS) is a preferred and commonly used device for sample collection to detect both spore-forming and non-spore-forming biothreat agents from non-porous surfaces. Here, in this study, a recently developed high-throughput method (HTM) for processing SS samples to detect viable Bacillus anthracis spores was adapted for detection of non-spore-forming biothreat agents, Yersinia pestis and Francisella tularensis. The scalable HTM was used to process up to 20 SS samples simultaneously, compared to the current stomacher-based method which processes one SS at a time. Comparisons of the HTM and the stomacher-based method were statistically indistinguishable for most experiments (P > 0.05) with HTM recoveries of 37–60 % for Y. pestis inoculated at 102–103 cells/SS and held 48 h at 4 °C to mimic sample transport/storage. The HTM was integrated with Rapid Viability-Polymerase Chain Reaction (RV-PCR) analysis to detect viable Y. pestis in the presence of particulate contamination (Arizona Test Dust, ATD). This approach detected Y. pestis inoculated at 20 cells/SS and ATD did not impact detection (P > 0.05). F. tularensis showed significantly lower recoveries between no-hold time and 48-h hold time (4 °C, P < 0.05) using the HTM, which further testing showed could be due to toxicity of the neutralizing buffer used for SS pre-wetting. With modifications, this method could enhance throughput capacity while maintaining similar recovery efficiencies to current methods for other non-spore-forming bacterial pathogens.

Biological and medical sciences↗

Sample Collection Metadata for Soil Cores from the East River Watershed, Colorado collected in 2017

This data package contains sample collection metadata for soil cores from the East River Watershed in Colorado used in biogeochemical analyses by the Watershed Function SFA. Soil cores were collected seasonally during autumn, winter, snowmelt, and spring at a predominately montane meadow, high altitude field site. Information provided for each soil core sample assigned an International Geo Sample Number (IGSN) includes: sample name, IGSN, material, description, collection method, core depth, location, field program, collectors, and date. The full list of 53 IGSN's is listed in the Methods section. There is also a complete core sample list and metadata for the set of baseline data.

54 ENVIRONMENTAL SCIENCES↗

Dispersion-enhanced sequential batch sampling for adaptive contour estimation

In computer simulation and optimal design, sequential batch sampling offers an appealing way to iteratively stipulate optimal sampling points based upon existing selections and efficiently construct surrogate modeling. Nonetheless, the issue of near duplicates poses tremendous quandary for sequential learning. It refers to the situation that selected critical points cluster together in each sampling batch, which are individually but not collectively informative towards the optimal design. Near duplicates severely diminish the computational efficiency as they barely contribute extra information towards update of the surrogate. To address this issue, we impose a dispersion criterion on concurrent selection of sampling points, which essentially forces a sparse distribution of critical points in each batch, and demonstrate the effectiveness of this approach in adaptive contour estimation. Specifically, we adopt Gaussian process surrogate to emulate the simulator, acquire variance reduction of the critical region from new sampling points as a dispersion criterion, and combine it with the modified expected improvement (EI) function for critical batch selection. The critical region here is the proximity of the contour of interest. This proposed approach is vindicated in numerical examples of a two-dimensional four-branch function, a four-dimensional function with a disjoint contour of interest and a time-delay dynamic system.

97 MATHEMATICS AND COMPUTING↗

Benchtop-compatible sample processing workflow for proteome profiling of < 100 mammalian cells

Extending proteomics to smaller samples can enable the mapping of protein expression across tissues with high spatial resolution and can reveal sub-group heterogeneity. However, despite the continually improving sensitivity of LC-MS instrumentation, in-depth profiling of samples containing low-nanogram amounts of protein has remained challenging due to analyte losses incurred during preparation and analysis. To address this, we recently developed nanodroplet processing in one pot for trace samples (nanoPOTS), a robotic/microfluidic platform that generates ready-to-analyze peptides from cellular material in ~200 nL droplets with greatly reduced sample losses. In combination with ultrasensitive LC-MS, nanoPOTS has enabled >3000 proteins to be confidently identified from as few as 10 cultured human cells and ~700 proteins from single cells. However, the nanoPOTS platform requires a highly skilled operator and a costly in-house-built robotic nanopipetting instrument. In this work, we sought to evaluate the extent to which the benefits of nanodroplet processing could be preserved when upscaling reagent dispensing volumes by a factor of 10 to those addressable by commercial micropipette. We characterized the resulting platform, termed microdroplet processing in one pot for trace samples (µPOTS), for the analysis of as few as ~25 cultured HeLa cells (4 ng total protein) or 50 µm square mouse liver tissue thin sections and found that ~1800 and ~1200 unique proteins were respectively identified with high reproducibility. As such, the reduced equipment requirements should facilitate broad dissemination of nanoproteomics workflows by obviating the need for a capital-intensive custom liquid handling system.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Cytological Sampling Versus Forceps Biopsy During Percutaneous Transhepatic Biliary Drainage and Analysis of Factors Predicting Success

Purpose: To assess the accuracy of cytological sampling and forceps biopsy in obstructing biliary lesions and to identify factors predictive of success. Methods: Consecutive patients (n = 119) with suspected malignant inoperable obstructive jaundice treated with percutaneous transhepatic biliary drainage during 7 years were included (60 male; mean age 72.5 years). All patients underwent forceps biopsy plus cytological sampling by washing the forceps device in cytological solution. Patient history, procedural and pathological records, and clinical follow-up were reviewed. Statistical analysis included chi-square test and multivariate regression analysis. Results: Histological diagnosis after forceps biopsy was more successful than cytology: Sensitivity was 78 versus 61%, and negative predictive value was 30 versus 19%. Cytology results were never positive when the forceps biopsy was negative. The cytological sample was negative and forceps sample positive in 2 cases of cholangiocarcinoma, 16 cases of pancreatic carcinoma, and 1 case of benign disease. Diagnostic accuracy was predicted by low bilirubin (p < 0.001), aspartate transaminase (p < 0.05), and white cell count (p {<=} 0.05). Conclusions: This technique is safe and effective and is recommended for histological diagnosis during PTBD in patients with inoperable malignant biliary strictures. Diagnostic yield is greater when bilirubin levels are low and there is no sepsis; histological diagnosis by way of forceps biopsy renders cytological sampling unnecessary.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Image Registration of Cone-Beam Computer Tomography and Preprocedural Computer Tomography Aids in Localization of Adrenal Veins and Decreasing Radiation Dose in Adrenal Vein Sampling

PurposeWe assessed whether image registration of cone-beam computed tomography (CT) (CBCT) and contrast-enhanced CT (CE-CT) images indicating the locations of the adrenal veins can aid in increasing the success rate of first-attempts adrenal vein sampling (AVS) and therefore decreasing patient radiation dose.Materials and Methods CBCT scans were acquired in the interventional suite (Philips Allura Xper FD20) and rigidly registered to the vertebra in previously acquired CE-CT. Adrenal vein locations were marked on the CT image and superimposed with live fluoroscopy and digital-subtraction angiography (DSA) to guide the AVS. Seventeen first attempts at AVS were performed with image registration and retrospectively compared with 15 first attempts without image registration performed earlier by the same 2 interventional radiologists. First-attempt AVS was considered successful when both adrenal vein samples showed representative cortisol levels. Sampling time, dose-area product (DAP), number of DSA runs, fluoroscopy time, and skin dose were recorded.ResultsWithout image registration, the first attempt at sampling was successful in 8 of 15 procedures indicating a success rate of 53.3 %. This increased to 76.5 % (13 of 17) by adding CBCT and CE-CT image registration to AVS procedures (p = 0.266). DAP values (p = 0.001) and DSA runs (p = 0.026) decreased significantly by adding image registration guidance. Sampling and fluoroscopy times and skin dose showed no significant changes.ConclusionGuidance based on registration of CBCT and previously acquired diagnostic CE-CT can aid in enhancing localization of the adrenal veins thereby increasing the success rate of first-attempt AVS with a significant decrease in the number of used DSA runs and, consequently, radiation dose required.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

A sensitivity analysis to predict the neutronics behavior of samples irradiated in the VTR rabbit system

We report a low-order neutronics model is developed to carry out hundreds of simulations efficiently and investigate the neutronics behavior of samples being irradiated in a test reactor setting under different geometrical constraints. The low-order model allowed for simulations that yield the expected neutronics behavior of any irradiated sample in any environment and allows for the calculation of highly accurate spatially averaged statistics and idealized spatial distributions in the neutron flux. Several benchmarks are performed to evaluate the performance and limitations of the low-order model revealing many important findings. The low-order model predicted the LHGR in the EBR-II driver fuel to within 2.34% by only simulating the fuel rod by itself, which served as a validation for the model. Sensitivity studies investigated 3% enriched UO 2 and U-10Zr being irradiated in the Versatile Test Reactor rabbit system. The analyses investigated a range of combinations of 15 radii and 5 heights for each sample in the rabbit system. Similar data sets are also provided for irradiations in the Advanced Test Reactor’s B-10 irradiation position, which is a thermal neutron spectrum environment. Generalized fits and fit coefficients are obtained for sample heating, reaction rate densities, and local multiplication rate characteristics, allowing the predictions of the neutronics behavior of the samples based on their geometrical constraints. The analyses and fits laid the groundwork for developing a user-end Multiphysics analysis framework to assist and accelerate irradiation experiment design and optimization.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Implicit sampling combined with reduced order modeling for the inversion of vadose zone hydrological data

Bayesian inverse modeling techniques are computationally expensive because many forward simulations are needed when sampling the posterior distribution of the parameters. Here, in this paper, we combine the implicit sampling method and generalized polynomial chaos expansion (gPCE) to significantly reduce the computational cost of performing Bayesian inverse modeling. There are three steps in this approach: (1) find the maximizer of the likelihood function using deterministic approaches; (2) construct a gPCE-based surrogate model using the results from a limited number of forward simulations; and (3) efficiently sample the posterior distribution of the parameters using implicit sampling method. The cost of constructing the gPCE-based surrogate model is further decreased by using sparse Bayesian learning to reduce the number of gPCE coefficients that have to be determined. We demonstrate the approach for a synthetic ponded infiltration experiment simulated with TOUGH2. The surrogate model is highly accurate with mean relative error that is < 0.035% in predicting saturation and < 0.25% in predicting the likelihood function. The posterior distribution of the parameters obtained using our proposed technique is nearly indistinguishable from the results obtained from either an implicit sampling method or a Markov chain Monte Carlo method utilizing the full mode.

54 ENVIRONMENTAL SCIENCES↗

Computing the partition function, ensemble averages, and density of states for lattice spin systems by sampling the mean

An algorithm to approximately calculate the partition function (and subsequently ensemble averages) and density of states of lattice spin systems through non-Monte-Carlo random sampling is developed. This algorithm (called the sampling-the-mean algorithm) can be applied to models where the up or down spins at lattice nodes interact to change the spin states of other lattice nodes, especially non-Ising-like models with long-range interactions such as the biological model considered here. Because it is based on the Central Limit Theorem of probability, the sampling-the-mean algorithm also gives estimates of the error in the partition function, ensemble averages, and density of states. Easily implemented parallelization strategies and error minimizing sampling strategies are discussed. The sampling-the-mean method works especially well for relatively small systems, systems with a density of energy states that contains sharp spikes or oscillations, or systems with little a priori knowledge of the density of states.

97 MATHEMATICS AND COMPUTING↗

Advantages and Disadvantages of using a Focused Ion Beam to Prepare TEM Samples From Irradiated U-10Mo Monolithic Nuclear Fuel

Transmission electron microscopy (TEM), scanning electron microscopy (SEM), and focused ion beam (FIB) milling were performed on an irradiated U-10Mo monolithic fuel to understand its irradiation microstructure. This is the first reported TEM work of irradiated fuel sample prepared using a FIB. Advantages and disadvantages of using the FIB to create TEM samples from this irradiated fuel will be presented along with some results from the work. Sample preparation techniques used to create SEM and FIB samples from the brittle irradiated monolithic sample will also be discussed.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

An efficient and cost-effective method for preparing transmission electron microscopy samples from powders

The preparation of transmission electron microcopy (TEM) samples from powders with particle sizes larger than ~100 nm poses a challenge. The existing methods are complicated and expensive, or have a low probability of success. Herein, we report a modified methodology for preparation of TEM samples from powders, which is efficient, cost-effective, and easy to perform. This method involves mixing powders with an epoxy on a piece of weighing paper, curing the powder–epoxy mixture to form a bulk material, grinding the bulk to obtain a thin foil, punching TEM discs from the foil, dimpling the discs, and ion milling the dimpled discs to electron transparency. Compared with the well established and robust grinding–dimpling–ion-milling method for TEM sample preparation for bulk materials, our modified approach for preparing TEM samples from powders only requires two additional simple steps. In this article, step-by-step procedures for our methodology are described in detail, and important strategies to ensure success are elucidated. Furthermore, our methodology has been applied successfully for preparing TEM samples with large thin areas and high quality for many different mechanically milled metallic powders.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Simple Sodium Dodecyl Sulfate-Assisted Sample Preparation Method for LC-MS-based Proteomic Applications

Sodium dodecyl sulfate (SDS) is one of the most popular laboratory reagents used for highly efficient biological sample extraction; however, SDS presents a significant challenge to LC-MS-based proteomic analyses due to its severe interference with reversed-phase LC separations and electrospray ionization interfaces. This study reports a simple SDS-assisted proteomic sample preparation method facilitated by a novel peptide-level SDS removal protocol. After SDS-assisted protein extraction and digestion, SDS was effectively (>99.9%) removed from peptides through ion substitution-mediated DS- precipitation with potassium chloride (KCl) followed by {approx}10 min centrifugation. Excellent peptide recovery (>95%) was observed for less than 20 {mu}g of peptides. Further experiments demonstrated the compatibility of this protocol with LC-MS/MS analyses. The resulting proteome coverage from this SDS-assisted protocol was comparable to or better than those obtained from other standard proteomic preparation methods in both mammalian tissues and bacterial samples. These results suggest that this SDS-assisted protocol is a practical, simple, and broadly applicable proteomic sample processing method, which can be particularly useful when dealing with samples difficult to solubilize by other methods.

59 BASIC BIOLOGICAL SCIENCES↗

High-Throughput Microfluidics Platform for Intracellular Delivery and Sampling of Biomolecules from Live Cells

Nondestructive cell membrane permeabilization systems enable the intracellular delivery of exogenous biomolecules for cell engineering tasks as well as the temporal sampling of cytosolic contents from live cells for the analysis of dynamic processes. Here, in this paper, we report a microwell array format live-cell analysis device (LCAD) that can perform localized-electroporation induced membrane permeabilization, for cellular delivery or sampling, and directly interfaces with surface-based biosensors for analyzing the extracted contents. We demonstrate the capabilities of the LCAD via an automated high-throughput workflow for multimodal analysis of live-cell dynamics, consisting of quantitative measurements of enzyme activity using self-assembled monolayers for MALDI mass spectrometry (SAMDI) and deep-learning enhanced imaging and analysis. By combining a fabrication protocol that enables robust assembly and operation of multilayer devices with embedded gold electrodes and an automated imaging workflow, we successfully deliver functional molecules (plasmid and siRNA) into live cells at multiple time-points and track their effect on gene expression and cell morphology temporally. Furthermore, we report sampling performance enhancements, achieving saturation levels of protein tyrosine phosphatase activity measured from as few as 60 cells, and demonstrate control over the amount of sampled contents by optimization of electroporation parameters using a lumped model. Lastly, we investigate the implications of cell morphology on electroporation-induced sampling of fluorescent molecules using a deep-learning enhanced image analysis workflow.

59 BASIC BIOLOGICAL SCIENCES↗

Design and Evaluation of a Tethered, Open Port Sampling Interface for Liquid Extraction-Mass Spectrometry Chemical Analysis

Presented in this work is a tethered, liquid-extraction-sampling interface designed for the mass spectrometric surface sampling/analysis of 3D objects. The tethered, open port sampling interface (TOPSI) incorporates a vacuum line between the sampling probe and ionization source, which enables the ability for an extended, tethered sample transfer line. Herein, several designs of the hand-held TOPSI are presented and evaluated on the basis of the analytical metrics of analyte transport time, peak width, and analyte sensitivity. The best analytical metrics were obtained with capillary flow resistances arranged in a particular order and the vacuum region set at 6.2 kPa. This TOPSI design incorporated a transfer capillary 1 m in length, while retaining a fast analyte transport time (12 s), short signal peak width (5 s baseline-to-baseline), and high analyte signal at 90% of that obtained with a regular open port sampling interface (OPSI). The hand-held TOPSI was demonstrated for the characterization of extracted small molecules and metabolites from the surface of mint and rosemary leaves.

47 OTHER INSTRUMENTATION↗

Improved explosive collection and detection with rationally assembled surface sampling materials

Sampling and detection of trace explosives is a key analytical process in modern transportation safety. In this work we have explored some of the fundamental analytical processes for collection and detection of trace level explosive on surfaces with the most widely utilized system, thermal desorption IMS. The performance of the standard muslin swipe material was compared with chemically modified fiberglass cloth. The fiberglass surface was modified to include phenyl functional groups. When compared to standard muslin, the phenyl functionalized fiberglass sampling material showed better analyte release from the sampling material as well as improved response and repeatability from multiple uses of the same swipe. The improved sample release of the functionalized fiberglass swipes resulted in a significant increase in sensitivity. Various physical and chemical properties were systematically explored to determine optimal performance. The results herein have relevance to improving the detection of other explosive compounds and potentially to a wide range of other chemical sampling and field detection challenges.

Explosive detection↗

Note: A sample holder design for sensitive magnetic measurements at high temperatures in a magnetic properties measurement system

A sample holder design for high temperature measurements in a commercial MPMS SQUID magnetometer from Quantum Design is presented. It fulfills the requirements for the simultaneous use of the oven and reciprocating sample option (RSO) options, thus allowing sensitive magnetic measurements up to 800 K. Alternating current susceptibility can also be measured, since the holder does not induce any phase shift relative to the ac driven field. It is easily fabricated by twisting Constantan Copyright-Sign wires into a braid nesting the sample inside. This design ensures that the sample be placed tightly into a tough holder with its orientation fixed, and prevents any sample displacement during the fast movements of the RSO transport, up to high temperatures.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Compact cold stage for micro-computerized tomography imaging of chilled or frozen samples

High resolution X-ray microCT (computerized tomography) can be used to image a variety of objects, including temperature-sensitive materials. In cases where the sample must be chilled or frozen to maintain sample integrity, either the microCT machine itself must be placed in a refrigerated chamber, or a relatively expensive commercial cold stage must be purchased. We describe here the design and construction of a low-cost custom cold stage suitable for use in a microCT imaging system. Our device uses a boron nitride sample holder, two-stage Peltier cooler, fan-cooled heat sink, and electronic controller to maintain sample temperatures as low as −25 °C ± 0.2 °C for the duration of a tomography acquisition. The design does not require modification to the microCT machine, and is easily installed and removed. Our custom cold stage represents a cost-effective solution for refrigerating CT samples for imaging, and is especially useful for shared equipment or machines unsuitable for cold room use.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

ANALYZING STAR CLUSTER POPULATIONS WITH STOCHASTIC MODELS: THE HUBBLE SPACE TELESCOPE/WIDE FIELD CAMERA 3 SAMPLE OF CLUSTERS IN M83

The majority of clusters in the universe have masses well below 10{sup 5} M{sub Sun }. Hence, their integrated fluxes and colors can be affected by the presence or absence of a few bright stars introduced by stochastic sampling of the stellar mass function. Specific methods are being developed to extend the analysis of cluster energy distributions into the low-mass regime. In this paper, we apply such a method to real observations of star clusters, in the nearby spiral galaxy M83. We reassess the ages and masses of a sample of 1242 clusters for which UBVIH{alpha} fluxes were obtained from observations with the Wide Field Camera 3 instrument on board the Hubble Space Telescope. Synthetic clusters with known properties are used to characterize the limitations of the method (valid range and resolution in age and mass, method artifacts). The ensemble of color predictions of the discrete cluster models are in good agreement with the distribution of observed colors. We emphasize the important role of the H{alpha} data in the assessment of the fraction of young objects, particularly in breaking the age-extinction degeneracy that hampers an analysis based on UBVI data only. We find the mass distribution of the cluster sample to follow a power law of index -2.1 {+-} 0.2, and the distribution of ages a power law of index -1.0 {+-} 0.2 for log (M/ M{sub Sun }) > 3.5, and ages between 10{sup 7} and 10{sup 9} yr. An extension of our main method, which makes full use of the probability distributions of age and mass obtained for the individual clusters of the sample, is explored. It produces similar power-law slopes and will deserve further investigation. Although the properties derived for individual clusters significantly differ from those obtained with traditional, non-stochastic models in about 30% of the objects, the first-order aspect of the age and mass distributions is similar to those obtained previously for this M83 sample in the range of overlap of the studies. We extend the power-law description to lower masses with better mass and age resolution and without most of the artifacts produced by the classical method.

79 ASTRONOMY AND ASTROPHYSICS↗